Combinations for the treatment and prevention of cancer

Abstract

The present disclosure provides a method for treating or preventing cancer, comprising administering an antigen-binding molecule that binds to HER3 and an antigen-binding molecule that binds to EGFR. Also provided are pharmaceutical combinations containing the molecules, and therapeutic and prophylactic methods using the pharmaceutical combinations.

Description

[Technical Field]

[0001] This application claims priority from US63 / 430942, filed December 7, 2022, the contents and elements of which are incorporated herein by reference for all purposes.

[0002] The present disclosure relates particularly to the medical treatment and prevention of cancer. [Background technology]

[0003] Forster et al., Eur J Cancer. (2019) 123:36-47, discloses the treatment of recurrent or metastatic squamous cell carcinoma of the head and neck (SCCHN) using a combination of the anti-HER3 antibody patritumab and the anti-EGFR antibody cetuximab. Patritumab (also known as U-1287 and AMG-888) has been shown to inhibit HER3-mediated signaling by blocking heregulin (HRG) binding to HER3 (see, e.g., Shimizu et al., Cancer Chemother Pharmacol. (2017) 79(3):489-495).

[0004] Cleary et al., Investigational New Drugs (2017) 35:68-78, discloses the administration of a combination of the anti-HER3 antibodies seribantumab and cetuximab for the treatment of EGFR-dependent cancers. Similar to patritumab, seribantumab (also known as MM-121) has been shown to inhibit HER3-mediated signaling by blocking the binding of heregulin (HRG) to HER3 (Schoeberl et al., Sci. Signal. (2009) 2(77):ra31).

[0005] Meulendijks et al., Clin Cancer Res. (2017) 23(18):5406-5415, discloses the administration of a combination of the anti-HER3 antibodies lumuletuzumab and cetuximab for the treatment of advanced HER3-positive cancer. Only minimal clinical activity was observed. Lumuletuzumab (also known as RG7116 and RO-5479599) recognizes an epitope in subdomain I of the HER3 extracellular domain (see, e.g., Mirschberger et al., Cancer Research (2013) 73(16) 5183-5194).

[0006] Bauman et al., Cancers (Basel) (2022) 14(10):2355, reported the results of a phase II trial of the anti-HER3 antibody CDX-3379 in combination with cetuximab for the treatment of recurrent / metastatic, HPV-negative, cetuximab-resistant head and neck squamous cell carcinoma (HNSCC). Objective response rates were modest. CDX-3379 (also known as KTN3379) binds to HER3 through interactions with amino acid residues in subdomain III (corresponding to the following positions in SEQ ID NO: 1: Gly476, Pro477, Arg481, Gly452, Arg475, Ser450, Gly420, Ala451, Gly419, Arg421, Thr394, Leu423, Arg426, Gly427, Lys356, Leu358, Leu358, Lys356, Ala330, Lys329, and Gly337) and Met310, Glu311, and Pro328 in subdomain II (see Lee et al., Proc Natl Acad Sci USA. 2015 Oct 27;112(43):13225).

[0007] Garner et al., Cancer Res. (2013) 73(19):6024-6035, discloses the treatment of HNSCC using a combination of the anti-HER3 antibody LJM-716 and cetuximab. LJM-716 binds to an epitope on subdomains II and IV of the HER3 extracellular domain, locking HER3 in an inactive conformation (Garner et al., Cancer Res (2013) 73:6024-6035).

[0008] Papadopoulos et al., Journal of Clinical Oncology (2014) 32(15_suppl): 2516-2516, discloses the administration of a combination of the anti-HER3 antibody REGN1400 and cetuximab for the treatment of advanced non-small cell lung cancer (NSCLC), colorectal cancer (CRC), or SCCHN. REGN1400 also inhibits the binding of ligands to HER3 (see Zhang et al., Mol Cancer Ther (2014) 13: 1345-1355).

[0009] Kim et al., Annals of Oncology (2020) 31(suppl_4):S599-S628, disclose the treatment of recurrent or metastatic HNSCC using a combination of the anti-HER3 antibody ISU104 and cetuximab. ISU104 (also known as varsetamab) binds primarily to domain III of HER3, interacts weakly with domain I, and exhibits dose-dependent inhibition of HRG binding to HER3 (see Kim et al., Cancer Research (2018) 78(13 Supplement):830-830). Summary of the Invention [Means for solving the problem]

[0010] In a first aspect, the present disclosure provides an antigen-binding molecule that binds to HER3 for use in a method for treating or preventing cancer, the method comprising administering an antigen-binding molecule that binds to EGFR, wherein the antigen-binding molecule that binds to HER3 binds to a region of HER3 set forth in SEQ ID NO: 77. In some embodiments, the method for treating or preventing cancer further comprises administering docetaxel.

[0011] The present disclosure also provides use of an antigen-binding molecule that binds to HER3 in the manufacture of a medicament for use in a method for treating or preventing cancer, the method comprising administering an antigen-binding molecule that binds to EGFR, wherein the antigen-binding molecule that binds to HER3 binds to a region of HER3 set forth in SEQ ID NO: 77. In some embodiments, the method for treating or preventing cancer further comprises administering docetaxel.

[0012] The present disclosure also provides a method for treating or preventing cancer, the method comprising administering to a subject therapeutically or prophylactically effective amounts of (i) an antigen-binding molecule that binds to HER3 and (ii) an antigen-binding molecule that binds to EGFR, wherein the antigen-binding molecule that binds to HER3 binds to a region of HER3 set forth in SEQ ID NO: 77. In some embodiments, the method for treating or preventing cancer further comprises administering docetaxel.

[0013] The present disclosure also provides a pharmaceutical combination comprising an antigen-binding molecule that binds to HER3 and an antigen-binding molecule that binds to EGFR; wherein the antigen-binding molecule that binds to HER3 binds to a region of HER3 set forth in SEQ ID NO: 77.

[0014] The present disclosure also provides a pharmaceutical combination according to the present disclosure for use in a method for treating or preventing cancer. In some embodiments, the method for treating or preventing cancer further comprises administering docetaxel.

[0015] The present disclosure also provides the use of a pharmaceutical combination according to the present disclosure in the manufacture of a medicament for treating or preventing cancer. In some embodiments, the method of treating or preventing cancer further comprises administering docetaxel.

[0016] The present disclosure also provides a method of treating or preventing cancer, comprising administering to a subject a therapeutically or prophylactically effective amount of a pharmaceutical combination according to the present disclosure. In some embodiments, the method of treating or preventing cancer further comprises administering docetaxel.

[0017] In some embodiments, the antigen binding molecule that binds to HER3 is (i) The following CDR: HC-CDR1 having the amino acid sequence of SEQ ID NO: 40 HC-CDR2 having the amino acid sequence of SEQ ID NO: 43 HC-CDR3 having the amino acid sequence of SEQ ID NO: 48 a heavy chain variable (VH) region incorporating (ii) the following CDR: LC-CDR1 having the amino acid sequence of SEQ ID NO: 66 LC-CDR2 having the amino acid sequence of SEQ ID NO: 69 LC-CDR3 having the amino acid sequence of SEQ ID NO: 74 A light chain variable (VL) region incorporating Includes.

[0018] In some embodiments, the antigen binding molecule that binds to HER3 is (i) The following CDR: HC-CDR1 having the amino acid sequence of SEQ ID NO: 38 HC-CDR2 having the amino acid sequence of SEQ ID NO: 42 HC-CDR3 having the amino acid sequence of SEQ ID NO: 45 and a VH region incorporating (ii) the following CDR: LC-CDR1 having the amino acid sequence of SEQ ID NO: 63 LC-CDR2 having the amino acid sequence of SEQ ID NO: 67 LC-CDR3 having the amino acid sequence of SEQ ID NO: 70 The VL region incorporating Includes.

[0019] In some embodiments, the antigen binding molecule that binds to HER3 is a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 33; and A VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 58. Includes.

[0020] In some embodiments, the antigen binding molecule that binds to HER3 is A polypeptide comprising or consisting of an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 75, and A polypeptide comprising or consisting of an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 76. Includes.

[0021] In some embodiments, the antigen binding molecule that binds to EGFR is (i) The following CDR: HC-CDR1 having the amino acid sequence of SEQ ID NO: 92 HC-CDR2 having the amino acid sequence of SEQ ID NO: 93 HC-CDR3 having the amino acid sequence of SEQ ID NO: 94 a heavy chain variable (VH) region incorporating (ii) the following CDR: LC-CDR1 having the amino acid sequence of SEQ ID NO: 96 LC-CDR2 having the amino acid sequence of SEQ ID NO: 97 LC-CDR3 having the amino acid sequence of SEQ ID NO: 98 A light chain variable (VL) region incorporating Includes.

[0022] In some embodiments, the antigen binding molecule that binds to EGFR is a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 91; and A VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 95. Includes.

[0023] In some embodiments, the antigen binding molecule that binds to EGFR is A polypeptide comprising or consisting of an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 99; and A polypeptide comprising or consisting of an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 100. Includes.

[0024] In some embodiments, the cancer is selected from the group consisting of cancers comprising cells that express / overexpress an EGFR family member, cancers comprising cells that express / overexpress HER3, cancers comprising cells that express / overexpress EGFR, cancers comprising cells that express / overexpress HER3 and EGFR, cancers comprising cells with a mutation that results in increased expression of a ligand for HER3, cancers comprising cells with a mutation that results in increased expression of a ligand for EGFR, cancers comprising cells with an NRG gene fusion, solid tumors, hematologic cancers, squamous cell carcinoma, EGFR-amplified squamous cell carcinoma, breast cancer, breast cancer, invasive breast cancer, ductal carcinoma, metastatic breast cancer, triple-negative breast cancer, HER2-positive breast cancer, gastric cancer, gastric adenocarcinoma, gastrointestinal adenocarcinoma, colorectal cancer, metastatic colorectal cancer, colon cancer, colorectal cancer, colorectal adenocarcinoma, colon adenocarcinoma, head and neck cancer, head and neck squamous cell carcinoma, lung Cancer, non-small cell lung cancer, lung adenocarcinoma, invasive mucinous lung adenocarcinoma, lung squamous cell carcinoma, ovarian cancer, ovarian cancer, serous ovarian adenocarcinoma, serous ovarian cystadenocarcinoma, fallopian tube cancer, kidney cancer, renal cell carcinoma, renal clear cell carcinoma, renal cell adenocarcinoma, papillary renal cell carcinoma, pancreatic cancer, pancreatic adenocarcinoma, pancreatic ductal adenocarcinoma, cervical cancer, cervical squamous cell carcinoma, skin cancer, melanoma, oral cancer, oropharyngeal cancer, esophageal cancer, esophageal squamous cell carcinoma, esophageal adenocarcinoma, liver cancer, hepatocellular carcinoma, bile duct cancer, gallbladder cancer, uterine cancer , endometrial cancer, endometrial carcinoma, uterine carcinosarcoma, thyroid cancer, thyroid cancer, thyroid carcinoma, pheochromocytoma, paraganglioma, bladder cancer, bladder urothelial carcinoma, prostate cancer, prostate adenocarcinoma, retinoblastoma, sarcoma, soft tissue sarcoma, peritoneal cancer, thymoma, neuroendocrine tumor, nasopharyngeal neuroendocrine tumor, squamous cell carcinoma of the skin, astrocytoma, low-grade astrocytoma, high-grade astrocytoma, anaplastic astrocytoma, and glioblastoma multiforme.

[0025] In some embodiments, the cancer is selected from cancer comprising cells that express / overexpress HER3, cancer comprising cells that express / overexpress EGFR, cancer comprising cells that express / overexpress HER3 and EGFR, squamous cell carcinoma, EGFR-amplified squamous cell carcinoma, esophageal cancer, esophageal squamous cell carcinoma, head and neck cancer, head and neck squamous cell carcinoma, colorectal cancer, metastatic colorectal cancer, colon adenocarcinoma, pancreatic cancer, pancreatic adenocarcinoma, pancreatic ductal adenocarcinoma, lung cancer, and lung squamous cell carcinoma. DETAILED DESCRIPTION OF THE INVENTION

[0026] The present disclosure relates to a combination treatment for cancer, comprising an antigen-binding molecule that binds to HER3 and an antigen-binding molecule that binds to EGFR. In a preferred embodiment, the antigen-binding molecule that binds to HER3 is 10D1F, and the antigen-binding molecule that binds to EGFR is cetuximab.

[0027] 10D1F binds to an epitope on HER3 that is different from the epitope of anti-HER3 antibodies used in combination treatments disclosed in the prior art, including anti-HER3 antibodies and cetuximab. More specifically, 10D1F binds to the dimerization interface of HER3, whereas other anti-HER3 antibodies bind to various sites in the HER3 extracellular domain (e.g., the ligand-binding domain, domain II / IV). 10D1F inhibits the dimerization of HER3 with its receptor partner, blocking both ligand-dependent and ligand-independent activation of HER3, resulting in a more complete shutdown of HER3-mediated signaling.

[0028] In the experimental examples of the present application, the inventors demonstrate that the combination of 10D1F and cetuximab provides unexpectedly advantageous properties over combination treatments comprising an anti-HER3 antibody and cetuximab disclosed in the prior art, and the combination of 10D1F and cetuximab provides unexpectedly advantageous effects as an intervention for the treatment / prevention of a variety of different cancers compared to either component of the combination used as a monotherapy.

[0029] HER3 HER3 (also known as, for example, ERBB3, LCCS2, MDA-BF-1) is a protein identified by UniProt P21860.

[0030] The structure and function of HER3 are described, for example, in Cho and Leahy Science (2002) 297(5585):1330-1333, Singer et al., Journal of Biological Chemistry (2001) 276, 44266-44274, Roskoski et al., Pharmacol. Res. (2014) 79:34-74, Bazley and Gullick Endocrine-Related Cancer (2005) S17-S27, and Mujoo et al., Oncotarget (2014) 5(21):10222-10236, each of which is incorporated herein by reference in its entirety. HER3 is a single-pass transmembrane ErbB receptor tyrosine kinase with an N-terminal extracellular region (SEQ ID NO: 9) containing two leucine-rich subdomains (domains I and III, shown in SEQ ID NOs: 15 and 17, respectively) and two cysteine-rich subdomains (domains II and IV, shown in SEQ ID NOs: 16 and 18, respectively). Domain II contains a β-hairpin dimerization loop (SEQ ID NO: 19) that is involved in intermolecular interactions with other HER receptor molecules. The extracellular region is connected to the cytoplasmic region (SEQ ID NO: 11) via a transmembrane region (SEQ ID NO: 10). The cytoplasmic region contains a membrane-proximal segment (SEQ ID NO: 12), a protein kinase domain (SEQ ID NO: 13), and a C-terminal segment (SEQ ID NO: 14).

[0031] As used herein, "HER3" refers to HER3 from any species, and includes isoforms, fragments, variants (including mutants), or homologs of HER3 from any species.

[0032] As used herein, a "fragment," "variant," or "homologue" of a protein may optionally be characterized as having at least 60%, preferably 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% amino acid sequence identity to the amino acid sequence of a reference protein (e.g., a reference isoform). In some embodiments, fragments, variants, isoforms, and homologues of a reference protein may be characterized by their ability to perform a function performed by the reference protein.

[0033] A "fragment" generally refers to a portion of a reference protein. A "variant" generally refers to a protein having an amino acid sequence that contains one or more amino acid substitutions, insertions, deletions, or other modifications compared to the amino acid sequence of the reference protein, but retains a significant degree of sequence identity (e.g., at least 60%) to the amino acid sequence of the reference protein. An "isoform" generally refers to a variant of a reference protein that is expressed by the same species as the reference protein (e.g., human HER3 isoforms 1-5 are all isoforms of each other). A "homologue" generally refers to a variant of a reference protein that is produced by a different species compared to the species of the reference protein. For example, human HER3 isoform 1 (P21860-1, v1; SEQ ID NO: 1) and rhesus monkey HER3 (UniProt: F7HEH3-1, v2; SEQ ID NO: 20) are homologs of each other. Homologues include orthologs.

[0034] A "fragment" of a reference protein may be of any length (by number of amino acids), but may optionally be at least 20% of the length of the reference protein (i.e., the protein from which the fragment is derived), and may have a maximum length of one of 50%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the length of the reference protein.

[0035] A fragment of HER3 may have a minimum length of one of 10, 20, 30, 40, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800, 900, 1000, 1100, 1200 amino acids and a maximum length of one of 20, 30, 40, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800, 900, 1000, 1100, 1200, or 1300 amino acids.

[0036] In some embodiments, the HER3 is mammalian-derived HER3 (e.g., primate (rhesus monkey, cynomolgus monkey, non-human primate, or human) and / or rodent (e.g., rat or mouse) HER3). HER3 isoforms, fragments, variants, or homologs may optionally be characterized as having at least 70%, preferably 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% amino acid sequence identity to the amino acid sequence of an immature or mature HER3 isoform from a given species, e.g., human.

[0037] The isoform, fragment, variant, or homologue may optionally be a functional isoform, fragment, variant, or homologue that has the functional property / activity of a reference HER3 (e.g., human HER3 isoform 1), e.g., as determined by analysis with an assay appropriate for the functional property / activity. For example, an isoform, fragment, variant, or homologue of HER3 may exhibit association with one or more of HER2, NRG1 (types I, II, III, IV, V, or VI), or NRG2 (α or β).

[0038] In some embodiments, HER3 comprises or consists of an amino acid sequence having at least 70%, preferably 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% amino acid sequence identity to one of SEQ ID NOs: 1-8.

[0039] In some embodiments, the fragment of HER3 comprises or consists of an amino acid sequence having at least 70%, preferably 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% amino acid sequence identity to one of SEQ ID NOs: 9-19, e.g., one of SEQ ID NOs: 9, 16, or 19.

[0040] Signal transduction through HER3 involves receptor heteromultimerization (i.e., with other ErB receptors, e.g., HER2, EGFR) and the resulting autophosphorylation of tyrosine residues in the cytoplasmic domain by the protein kinase domain. HER3 lacks kinase activity and does not form stable homodimers. Therefore, HER3 must be transphosphorylated by binding to a kinase-active heterodimeric partner (e.g., EGFR or HER2) to initiate signal transduction (Berger MB et al., FEBS Lett 2004;569:332-6; Kim HH et al., Biochem J 1998;334:189-95).

[0041] Multimerization (e.g., dimerization) of HER receptor family members is required to activate cell growth signaling pathways, and HER3 can dimerize with other HER family members in both ligand-dependent and ligand-independent manners. The extracellular domain (ECD) of HER3 exists in a reversible equilibrium between a "closed" inactive conformation and an "open" active conformation, exposing dimerization arms within domain II and allowing dimerization along the dimerization interface of domain II, particularly through the cysteine-rich CR1 region (Carraway, KL et al., Nature, 1997. 387(6632):512-6; Riese, DJ et al., Mol Cell Biol, 1995. 15(10):5770-6; Harari, D. et al., Oncogene, 1999. 18(17):2681-9; Zhang, D. et al., Proc Natl Acad Sci USA, 1997. 94(18):9562-7; Meyer et al., Nature, 1995. 378(6555):386-90; Jura, N. et al., Proc Natl Acad Sci USA, 2009.106(51):21608-13; Fornaro, L. et al., Nat Rev Gastroenterol Hepatol, 2011.8(7):369-83; Mota et al., Oncotarget (2015) 5:89284-306). HER3 is "activated" when the equilibrium shifts in favor of the open conformation, increasing the likelihood of forming an active heterodimer. The traditional model of activation is ligand-dependent; the equilibrium shifts when HER3 in the open conformation is stabilized by binding to its ligand, such as neuregulin (NRG), e.g., NRG1 (also known as heregulin, HRG) or NRG2. In addition, the presence of either dimerization partner at sufficient concentrations transiently binds and stabilizes HER3 in the open conformation, thus shifting the equilibrium in favor of the open conformation.This is known as ligand-independent activation (Jura, N. et al., Proc Natl Acad Sci USA, 2009.106(51):21608-13; Fornaro, L. et al., Nat Rev Gastroenterol Hepatol, 2011.8(7):369-83; Mota et al., Oncotarget (2015) 5:89284-306).

[0042] As used herein, "HER3-mediated signal transduction" refers to signal transduction mediated by HER3 and / or a multimeric ErbB family member receptor complex containing HER3. "Signal transduction" refers to signal transduction and other cellular processes that control cellular activity. HER3-mediated signal transduction can be mediated by a HER3 receptor-containing complex, for example, a heteromultimeric complex containing HER3 and other HER receptors (e.g., HER2, EGFR). HER3-mediated signal transduction can be ligand-dependent, for example, caused by the binding of NRG (e.g., NRG1, NRG2), or can be ligand-independent.

[0043] HER3-mediated signal transduction occurs intracellularly via the MAPK / ERK and PI3K / AKT / mTOR pathways to promote cell survival and proliferation. HER3-mediated signal transduction is described, for example, in Gala and Chandarlapaty, Clin Cancer Res. (2014) 20 (6): 1410-1416; Mishra et al., Oncol Rev. (2018) 12 (1): 355; Baselga et al., Nat Rev Cancer (2009) 9: 463-75; Yarden et al., Nat Rev Mol Cell Biol (2001) 2: 35052073; Mota et al., Oncotarget (2015) 5: 89284-306; and Haikala and Janne, Clin.Cancer Res. (2021) 27: 3528-39, all of which are incorporated herein by reference in their entirety.

[0044] Phosphorylated tyrosine residues in the protein kinase domain of the HER3-containing receptor complex recruit the adaptor / effector protein GRB2 through interaction with its SH2 domain. Upon ligand stimulation, the activated receptor (EGFR / HER2) undergoes autophosphorylation, providing the phosphotyrosine residue for recruitment of GRB2. GRB2 binds to the guanine nucleotide exchange factor SOS through its SH3 domain. Activated SOS in the GRB2-SOS complex promotes the removal of GDP from Ras family GTPases, such as H-Ras, N-Ras, and K-Ras, thereby activating them. Activated Ras GTPases then activate RAF kinases, such as A-Raf, B-Raf, and C-Raf. RAF kinases then phosphorylate and activate MEK1 and MEK2, which then phosphorylate and activate MAPKs (also known as ERKs). Activated MAPKs can directly regulate the activity of transcription factors such as c-Myc. Activated MAPK also upregulates mRNA translation through the phosphorylation of RSK and the consequent phosphorylation and activation of the 40S ribosomal protein S6. Activated MAPK also phosphorylates and activates MNK, which then phosphorylates and activates the transcription factor CREB.

[0045] Phosphorylated tyrosine residues in the protein kinase domain of HER3 also recruit the p85 subunit of PI3K via its SH2 domain. Association of p85 triggers allosteric activation of the lipid kinase p100α subunit of PI3K. Activated PI3K converts PIP2 to PIP3, which recruits AKT, which is phosphorylated and activated by mTORC2 and PDK1. Phosphorylated AKT has many activities, including activating CREB and mTOR. PTEN antagonizes signaling through the PI3K / AKT / mTOR pathway by dephosphorylating PIP3 to PIP2, and PP2A inhibits the PI3K / AKT / mTOR pathway by dephosphorylating AKT.

[0046] Oncogenic Src homology domain 2 protein tyrosine phosphatase 2 (SHP2) promotes tumor progression and functions as a pivotal hub connecting multiple oncogenic signaling pathways, including PI3K / AKT and Ras / Raf / MAPK (Dong et al., Front. Cell Dev. Biol., 11 March 2021). GAB2 binds to GRB2 and becomes phosphorylated on multiple tyrosine residues, making it available to bind to the SH2 domains of SHP2 and p85 (Adams et al., Mol Cancer Res. 2012 Oct;10(10):1265-70; (Liu et al., Proc. Natl. Acad. Sci. USA (2016) 113, 984-989). The interaction induces conformational changes that relieve autoinhibition of the SHP2 catalytic site (Neel et al., Trends Biochem Sci. 2003 Jun;28(6):284-93) and p85 inhibition of the p110 catalytic subunit of PI3K (Cuevas et al., J Biol Chem. 2001 Jul 20;276(29):2745-6). SHP2 also directly dephosphorylates RAS (Bunda et al., Nat Commun. 2015 Nov). 30;6:8859), RASGAP (RAS GTPase-activating protein) (Neel et al., Trends Biochem Sci. 2003 Jun;28(6):284-93), and SPRY (Hanafusa et al., J Biol Chem. 2004 May 28;279(22):22992-5). Overexpression of SHP2 has been shown to enhance tumor invasion by activating the PI3K / AKT axis (Hu et al., Onco Targets Ther. (2017) 10, 3881-3891), while knockdown of SHP2 inhibits cell migration, suggesting that the tumor-promoting effects of SHP2 are partly related to AKT signaling (Cao et al., Pathol. Res. Pract. (2019) 215:152-21).

[0047] STAT3 and 5 proteins are transcription factors that enhance the expression of p85α, p110α, and AKT1, thereby increasing signaling through the PI3K / AKT signaling cascade (Radler et al., Mol Cell Endocrinol. 2017 August 15;451:31-39). Upon activation by JAK2, phosphorylated STAT5 binds to the SH2 domain of the p85α regulatory subunit of PI3K in a PRL signaling-dependent manner, suggesting that STAT5 may also be directly involved in signaling through the PI3K complex. Another kinase that phosphorylates EGFR is the cytokine-regulated tyrosine kinase Jak2, thus enabling MAPK activation even by kinase-deficient mutants of EGFR (Mishra et al., Oncol Rev. (2018) 12(1):355; Baselga et al., Nat Rev Cancer (2009) 9:463-75). Collective observations in genetic models that overexpress or delete active STAT5 and AKT or express mutant PTEN support the idea that STAT5 function as a survival factor during normal mammary gland development and as an oncogene during breast cancer development is mediated by the PI3K / AKT pathway (Radler et al., Mol Cell Endocrinol. 2017 August 15;451:31-39).

[0048] EGFR EGFR (also known as, for example, ERBB1, HER1) is a protein identified by UniProt P00533.

[0049] The structure and function of EGFR are described, for example, in Sabbah et al., Curr Top Med Chem. (2020) 20(10):815-834, and Sigismund et al., Mol Oncol. (2018) 12(1):3-20, both of which are incorporated herein by reference in their entireties. EGFR is a single-pass transmembrane ErbB receptor tyrosine kinase with an N-terminal extracellular domain (SEQ ID NO: 88) connected to a cytoplasmic domain (SEQ ID NO: 90) via a transmembrane domain (SEQ ID NO: 89).

[0050] As used herein, "EGFR" refers to EGFR from any species, and includes isoforms, fragments, variants (including mutants), or homologs of EGFR from any species.

[0051] A fragment of EGFR may have a minimum length of one of 10, 20, 30, 40, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800, 900, 1000, 1100, 1200 amino acids and a maximum length of one of 20, 30, 40, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800, 900, 1000, 1100, or 1200 amino acids.

[0052] In some embodiments, the EGFR is a mammalian EGFR (e.g., a primate (rhesus monkey, cynomolgus monkey, non-human primate, or human) and / or rodent (e.g., rat or mouse) EGFR). EGFR isoforms, fragments, variants, or homologs may optionally be characterized as having at least 70%, preferably 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% amino acid sequence identity to the amino acid sequence of an immature or mature EGFR isoform from a given species, e.g., human.

[0053] The isoform, fragment, variant, or homologue may optionally be a functional isoform, fragment, variant, or homologue that has a functional property / activity of a reference EGFR (e.g., human EGFR isoform 1), e.g., as determined by analysis with an assay appropriate for the functional property / activity. For example, an isoform, fragment, variant, or homologue of EGFR may exhibit association with one or more of HER3, HER2, EGF, TGFα, and amphiregulin.

[0054] In some embodiments, the EGFR comprises or consists of an amino acid sequence having at least 70%, preferably 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% amino acid sequence identity to one of SEQ ID NOs: 80-87.

[0055] In some embodiments, a fragment of EGFR comprises or consists of an amino acid sequence having at least 70%, preferably 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% amino acid sequence identity to one of SEQ ID NOs: 88, 89, or 90.

[0056] EGFR-mediated signaling is described, for example, in Sigismund et al., Mol Oncol. (2018) 12(1):3-20, and Kovacs et al., Annu Rev Biochem. (2015) 84:739-764, both of which are incorporated herein in their entireties. Canonical EGFR signaling is important for various cellular functions, including survival, proliferation, differentiation, and motility. In the absence of ligand, EGFR resides primarily in an autoinhibited, dimerization-incompetent state at the plasma membrane. Ligand binding induces receptor dimerization and aggregation of the catalytic domain, leading to transautophosphorylation of key tyrosine residues in the cytoplasmic domain and triggering an intracellular signaling cascade. There are seven known EGFR ligands, each of which differs in the type and intensity of downstream signaling they trigger. EGFR can also heterodimerize with HER2, HER3, and HER4. Signaling from heterodimers is predicted to be more oncogenic than signaling from EGFR homodimers.

[0057] Activation of EGFR triggers multiple intracellular signaling pathways, including the Ras / Raf / MAPK pathway, the PI3K / AKT pathway, and the phospholipase C (PLC) / protein kinase C (PKC) signaling cascade.

[0058] antigen binding molecule The present disclosure relates to therapeutic and prophylactic uses of antigen-binding molecules that bind to HER3 and antigen-binding molecules that bind to EGFR.

[0059] An "antigen-binding molecule" refers to a molecule that binds to a given target antigen. Antigen-binding molecules include antibodies (i.e., immunoglobulins (Ig)) and antigen-binding fragments thereof. As used herein, "antibody" includes monoclonal antibodies, polyclonal antibodies, monospecific and multispecific (e.g., bispecific, trispecific, etc.) antibodies, as well as antigen-binding molecules derived from antibodies such as scFv, scFab, diabodies, triabodies, scFv-Fc, minibodies, and single-domain antibodies (e.g., VhH, etc.). Antigen-binding fragments of antibodies include, for example, Fv, Fab, F(ab')2, and F(ab')2 fragments. In some embodiments, the antigen-binding molecule may be an antibody or an antigen-binding fragment thereof.

[0060] Antigen-binding molecules according to the present disclosure also include antibody-derived molecules, e.g., molecules comprising an antigen-binding region / domain derived from an antibody. Antibody-derived antigen-binding molecules may comprise an antigen-binding region / domain that comprises or consists of the antigen-binding region of an antibody (e.g., an antigen-binding fragment of an antibody). In some embodiments, the antigen-binding region / domain of an antibody-derived antigen-binding molecule may be or comprise the Fv (e.g., provided as an scFv) or Fab region of an antibody, or the entire antibody. For example, antigen-binding molecules according to the present disclosure include antibody-drug conjugates (ADCs) comprising a (cytotoxic) drug moiety (e.g., as described herein below). Antigen-binding molecules according to the present disclosure also include multispecific antigen-binding molecules such as immune cell engager molecules that contain domains for recruiting (effector) immune cells, including BiTE, BiKE, and TriKE (reviewed, e.g., in Goebeler and Bargou, Nat. Rev. Clin. Oncol. (2020) 17:418-434 and Ellerman, Methods (2019) 154:102-117, both of which are incorporated by reference in their entirety). Antigen-binding molecules according to the present disclosure also include chimeric antigen receptors (CARs), which are recombinant receptors that provide both antigen-binding and T-cell activation functions (the structure, function, and operation of CARs are reviewed, for example, in Dotti et al., Immunol Rev (2014) 257(1), and Jayaraman et al., EBioMedicine (2020) 58:102931, both of which are incorporated herein by reference in their entireties).

[0061] The antigen-binding molecules of the present disclosure comprise one or more moieties capable of binding to a target antigen. In some embodiments, the moiety capable of binding to the target antigen comprises an antibody heavy chain variable region (VH) and an antibody light chain variable region (VL) of an antibody capable of specifically binding to the target antigen. In some embodiments, the moiety capable of binding to the target antigen comprises or consists of an aptamer, e.g., a nucleic acid aptamer (reviewed, e.g., in Zhou and Rossi Nat Rev Drug Discov. 2017 16(3):181-202), capable of binding to the target antigen. In some embodiments, the moiety capable of binding to a target antigen comprises or consists of an antigen-binding peptide / polypeptide, such as a peptide aptamer, thioredoxin, monobody, anticalin, Kunitz domain, avimer, knottin, fynomer, atrimer, DARPin, affibody, nanobody (i.e., single domain antibody (sdAb)), affilin, armadillo repeat protein (ArmRP), OBody, or fibronectin (e.g., as reviewed in Reverdatto et al., Curr Top Med Chem., 2015;15(12):1082-1101, which are incorporated by reference in their entireties (see also, e.g., Boersma et al., J Biol Chem (2011) 286:41273-85 and Emanuel et al., Mabs (20113:38-48)).

[0062] As used herein, "peptide" refers to a chain of two or more amino acid monomers linked by peptide bonds. Peptides typically have a length in the region of about 2 to 50 amino acids. A "polypeptide" is a polymeric chain of two or more peptides. Polypeptides are typically greater than about 50 amino acids in length.

[0063] The antigen-binding molecules of the present disclosure generally comprise an antigen-binding domain comprising VH and VL of an antibody capable of specifically binding to a target antigen. Herein, the antigen-binding domain formed by VH and VL may also be referred to as an Fv region.

[0064] An antigen-binding molecule may be or comprise an antigen-binding polypeptide or an antigen-binding polypeptide complex. An antigen-binding molecule may comprise more than one polypeptide that together form an antigen-binding domain. The polypeptides may be covalently or non-covalently associated. In some embodiments, the polypeptide forms part of a larger polypeptide that comprises the polypeptide (e.g., in the case of an scFv comprising a VH and a VL, or in the case of an scFab comprising a VH-CH1 and a VL-CL).

[0065] An antigen-binding molecule may refer to a non-covalent or covalent complex of IgG-like antigen-binding molecules that includes more than one polypeptide (e.g., two, three, four, six, or eight polypeptides), e.g., two heavy chain polypeptides and two light chain polypeptides.

[0066] The antigen-binding molecules of the present disclosure can be designed and prepared using the sequence of a monoclonal antibody (mAb) that can bind to a given target antigen (e.g., HER3 or EGFR). Antigen-binding regions of antibodies, such as single-chain variable fragments (scFv), Fab, and F(ab')2 fragments, can also be used / derived. An "antigen-binding region" is any fragment of an antibody that binds to the target for which the given antibody is specific.

[0067] Antibodies generally contain six complementarity-determining regions, or CDRs: three in the heavy chain variable (VH) region: HC-CDR1, HC-CDR2, and HC-CDR3, and three in the light chain variable (VL) region: LC-CDR1, LC-CDR2, and LC-CDR3. Together, the six CDRs define the antibody paratope, which is the portion of the antibody that binds to the target antigen.

[0068] The VH and VL regions comprise, on either side of each CDR, framework regions (FRs) that provide a scaffold for the CDRs. From N- to C-terminus, the VH region comprises the following structure: N-terminus-[HC-FR1]-[HC-CDR1]-[HC-FR2]-[HC-CDR2]-[HC-FR3]-[HC-CDR3]-[HC-FR4]-C-terminus, and the VL region comprises the following structure: N-terminus-[LC-FR1]-[LC-CDR1]-[LC-FR2]-[LC-CDR2]-[LC-FR3]-[LC-CDR3]-[LC-FR4]-C-terminus.

[0069] There are several different conventions for defining the CDRs and FRs of antibodies, such as those described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD (1991), Chothia et al., J. Mol. Biol. 196:901-917 (1987), and VBASE2, described in Retter et al., Nucl. Acids Res. (2005) 33(suppl 1):D671-D674. The CDRs and FRs of the VH and VL regions of the antibody clones described herein were defined according to the international IMGT (ImMunoGeneTics) information system (LeFranc et al., Nucleic Acids Res. (2015) 43 (Database issue): D413-22), using the IMGT V-domain numbering convention described in Lefranc et al., Dev. Comp. Immunol. (2003) 27: 55-77. In a preferred embodiment, the CDRs and FRs of the antigen-binding molecules referred to herein are defined according to the IMGT information system.

[0070] The VH and VL regions of the antigen-binding region of an antibody together constitute an Fv region. In some embodiments, an antigen-binding molecule according to the present disclosure comprises or consists of an Fv region that binds to HER3. In some embodiments, an antigen-binding molecule according to the present disclosure comprises or consists of an Fv region that binds to EGFR. In some embodiments, the VH and VL regions of the Fv are provided as a single polypeptide joined by a linker sequence, i.e., a single-chain Fv (scFv).

[0071] The VL and light chain constant (CL) regions, and the VH region and heavy chain constant 1 (CH1) region of the antigen-binding region of an antibody together constitute a Fab region. In some embodiments, the antigen-binding molecule comprises a Fab region comprising a VH, CH1, VL, and CL (e.g., CK or Cλ). In some embodiments, the Fab region comprises a polypeptide comprising a VH and CH1 (e.g., a VH-CH1 fusion polypeptide) and a polypeptide comprising a VL and CL (e.g., a VL-CL fusion polypeptide). In some embodiments, the Fab region comprises a polypeptide comprising a VH and CL (e.g., a VH-CL fusion polypeptide) and a polypeptide comprising a VL and CH (e.g., a VL-CH1 fusion polypeptide); i.e., in some embodiments, the Fab region is a CrossFab region. In some embodiments, the VH, CH1, VL, and CL regions of a Fab or CrossFab are provided as a single polypeptide joined by a linker region, i.e., as a single-chain Fab (scFab) or single-chain CrossFab (scCrossFab).

[0072] In some embodiments, the antigen-binding molecules described herein comprise or consist of a whole antibody that binds to HER3. In some embodiments, the antigen-binding molecules described herein comprise or consist of a whole antibody that binds to EGFR. As used herein, "whole antibody" refers to an antibody having a structure substantially similar to that of an immunoglobulin (Ig). Different types of immunoglobulins and their structures are described, for example, in Schroeder and Cavacini, J Allergy Clin Immunol. (2010) 125(202):S41-S52, which is incorporated herein by reference in its entirety.

[0073] G-type immunoglobulins (i.e., IgG) are glycoproteins of approximately 150 kDa that contain two heavy chains and two light chains. From the N-terminus to the C-terminus, the heavy chain contains a VH followed by a heavy chain constant region containing three constant domains (CH1, CH2, and CH3); similarly, the light chain contains a VL followed by a CL. Depending on the heavy chain, immunoglobulins can be classified as IgG (e.g., IgG1, IgG2, IgG3, IgG4), IgA (e.g., IgA1, IgA2), IgD, IgE, or IgM. The light chain can be kappa (κ) or lambda (λ).

[0074] In some embodiments, the antigen-binding molecule comprises or consists of an IgG (e.g., IgG1, IgG2, IgG3, IgG4), IgA (e.g., IgA1, IgA2), IgD, IgE, or IgM that binds to HER3. In some embodiments, the antigen-binding molecule comprises or consists of an IgG (e.g., IgG1, IgG2, IgG3, IgG4), IgA (e.g., IgA1, IgA2), IgD, IgE, or IgM that binds to EGFR.

[0075] In some embodiments described herein, one or more amino acids of an amino acid sequence referred to herein (e.g., an amino acid sequence of an antigen-binding molecule, e.g., an amino acid sequence of a CDR or VH / VL region) are substituted with another amino acid. Substitution includes replacing an amino acid residue with a non-identical "replacement" amino acid residue. The replacement amino acid residue for a substitution according to the present disclosure may be a naturally occurring amino acid residue (i.e., encoded by the genetic code) that is non-identical to the amino acid residue at the relevant position of the equivalent unsubstituted amino acid sequence selected from alanine (Ala), arginine (Arg), asparagine (Asn), aspartic acid (Asp), cysteine ​​(Cys), glutamine (Gln), glutamic acid (Glu), glycine (Gly), histidine (His), isoleucine (Ile), leucine (Leu), lysine (Lys), methionine (Met), phenylalanine (Phe), proline (Pro), serine (Ser), threonine (Thr), tryptophan (Trp), tyrosine (Tyr), and valine (Val). In some embodiments, the replacement amino acid may be a non-naturally occurring amino acid residue, i.e., an amino acid residue other than those listed above. Examples of non-naturally occurring amino acid residues include norleucine, ornithine, norvaline, homoserine, aib, and other amino acid residue analogs such as those described in Ellman et al., Meth. Enzym. 202 (1991) 301-336.

[0076] In some embodiments, substitutions can be biochemically conservative. In some embodiments, if the amino acid to be substituted is provided in one of lines 1-5 of the table below, the replacement amino acid for the substitution is another non-identical amino acid provided in the same line:

[0077] [Table 1] By way of example, in some embodiments where the substitution is a Met residue, the replacement amino acid may be selected from Ala, Val, Leu, Ile, Trp, Tyr, Phe, and norleucine.

[0078] In some embodiments, the replacement amino acid in a substitution may have the same side chain polarity as the amino acid residue it replaces. In some embodiments, the replacement amino acid in a substitution may have the same side chain charge (at pH 7.4) as the amino acid residue it replaces:

[0079] [Table 2] That is, in some embodiments, a nonpolar amino acid is substituted with another, non-identical nonpolar amino acid; in some embodiments, a polar amino acid is substituted with another, non-identical polar amino acid; in some embodiments, an acidic polar amino acid is substituted with another, non-identical acidic polar amino acid; in some embodiments, a basic polar amino acid is substituted with another, non-identical basic polar amino acid; in some embodiments, a neutral amino acid is substituted with another, non-identical neutral amino acid; in some embodiments, a positive amino acid is substituted with another, non-identical positive amino acid; in some embodiments, a negative amino acid is substituted with another, non-identical negative amino acid.

[0080] In some embodiments, substitutions may be functionally conservative, i.e., in some embodiments, the substitution may not affect (or may not substantially affect) one or more functional properties (e.g., target binding) of an antigen-binding molecule comprising the substitution, compared to a comparable unsubstituted molecule.

[0081] Antigen-binding molecules that bind to HER3 The present disclosure provides antigen-binding molecules that bind to HER3. In some embodiments, the antigen-binding molecule comprises the CDRs of an antigen-binding molecule capable of binding to HER3. In some embodiments, the antigen-binding molecule comprises the FRs of an antigen-binding molecule capable of binding to HER3. In some embodiments, the antigen-binding molecule comprises the CDRs and FRs of an antigen-binding molecule capable of binding to HER3. That is, in some embodiments, the antigen-binding molecule comprises the VH region and VL region of an antigen-binding molecule capable of binding to HER3.

[0082] In some embodiments, an antigen-binding molecule capable of binding to HER3 according to the present disclosure is any of the embodiments of the antigen-binding molecules described in WO2019 / 185878A1 (which is incorporated herein by reference in its entirety), 10D1F (e.g., as described in WO2019 / 185878A1), seribantumab (also known as MM-121 and described, e.g., in Schoeberl et al., Sci. Signal. (2009) 2(77):ra31; DrugBank Acc. No. DB11857), elgemtumab (also known as LJM-716 and described, e.g., in Garner et al., Cancer Res (2013) 73:6024-6035; DrugBank Acc. No. DB15430), patritumab (also known as U-1287 and AMG-888 and described, e.g., in Shimizu et al., Cancer Chemother. Pharmacol. (2017) 79(3):489-495; DrugBank Acc. No. DB12090), GSK2849330 (described, for example, in Clarke et al., Eur J Cancer. (2014) 50:98-9), lumletuzumab (also known as RG7116 and RO-5479599, and described, for example, in Mirschberger et al., Cancer Research (2013) 73(16) 5183-5194; DrugBank Acc. No. DB12683), CDX-3379 (also known as KTN3379, and described, for example, in Lee et al., Proc Natl Acad Sci USA. 2015 Oct 27;112(43):13225), AV-203 (also known as CAN-017, and described, for example, in Meetze et al., Eur J Cancer 2012;48:126), balacetamab (also known as ISU104 and described, for example, in Kim et al., Cancer Res (2018) 78(13 Suppl):Abstract #830), TK-A3, TK-A4 (described, for example, in Malm et al., MAbs (2016) 8:1195-209), MP-EV20 (described, for example, in Sala et al., Transl. Oncol.(2013) 6:676-84), 1A5-3D4 (described, for example, in Wang et al., Cancer Lett (2016) 380:20-30), 9F7-F11, 16D3-C1 (described, for example, in Lazrek et al., Neoplasia (2013) 15:335-47), NG33, A5, F4 (described, for example, in Gaborit et al., PNAS USA (2015) 112:839-44), huHER3-8 (described, for example, in Kugel et al., Cancer Res. (2014) 74:4122-32), REGN1400 (described, for example, in Zhang et al., Mol Cancer Ther (2014) 13:1345-1355), and xenoctuzumab (also known as MCLA-128, described, for example, in de Vries Schultink et al., Clin Pharmacokinet. (2020) 59:875-884; DrugBank Acc. No. DB15559). In some embodiments, the antigen-binding molecule is 10D1F.

[0083] In some embodiments, the antigen-binding molecule binds to the extracellular region of HER3 (e.g., the region set forth in SEQ ID NO: 9). In some embodiments, the antigen-binding molecule binds to subdomain II of the extracellular region of HER3 (e.g., the region set forth in SEQ ID NO: 16).

[0084] In some embodiments, the antigen-binding molecule binds to a region of HER3 set forth in SEQ ID NO: 77. In some embodiments, the antigen-binding molecule contacts one or more amino acid residues in the region of HER3 set forth in SEQ ID NO: 77. In some embodiments, the antigen-binding molecule binds to a region of HER3 set forth in SEQ ID NOs: 78 and 79. In some embodiments, the antigen-binding molecule contacts one or more amino acid residues in the region of HER3 set forth in SEQ ID NO: 78 and 79. In some embodiments, the antigen-binding molecule binds to a region of HER3 set forth in SEQ ID NO: 78. In some embodiments, the antigen-binding molecule contacts one or more amino acid residues in the region of HER3 set forth in SEQ ID NO: 78. In some embodiments, the antigen-binding molecule binds to a region of HER3 set forth in SEQ ID NO: 79. In some embodiments, the antigen-binding molecule contacts one or more amino acid residues in the region of HER3 set forth in SEQ ID NO: 79.

[0085] In some embodiments, the antigen-binding molecule does not bind to the region of HER3 corresponding to positions 260 to 279 of SEQ ID NO: 1. In some embodiments, the antigen-binding molecule does not contact amino acid residues in the region of HER3 corresponding to positions 260 to 279 of SEQ ID NO: 1.

[0086] The region of the peptide / polypeptide that the antibody binds can be determined by those skilled in the art using various methods well known in the art, including X-ray cocrystallography of antibody-antigen complex, peptide scanning, mutagenesis mapping, hydrogen-deuterium exchange analysis by mass spectrometry, phage display, competitive ELISA, and proteolysis-based "protection" method. Such methods are described, for example, in Gershoni et al., BioDrugs, 2007, 21(3):145-156, which is incorporated herein by reference in its entirety.

[0087] In some embodiments, the antigen-binding molecule can bind to the same region of HER3, or overlapping regions of HER3, as bound by an antibody comprising the VH and VL sequences of one of antibody clones 10D1_c89, 10D1, 10D1_c75, 10D1_c76, 10D1_c77, 10D1_c78v1, 10D1_c78v2, 10D1_11B, 10D1_c85v1, 10D1_c85v2, 10D1_c85o1, 10D1_c85o2, 10D1_c87, 10D1_c90, 10D1_c91, 10D1_c92, and 10D1_c93 described herein. In some embodiments, the antigen-binding molecule can bind to the same region of HER3, or overlapping regions of HER3, as bound by an antibody comprising the VH and VL sequences of antibody clone 10D1_c89.

[0088] In some embodiments, the antigen-binding molecule is capable of binding to a polypeptide comprising or consisting of the amino acid sequence of one of SEQ ID NOs: 1, 3, 4, 6, or 8. In some embodiments, the antigen-binding molecule is capable of binding to a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 9. In some embodiments, the antigen-binding molecule is capable of binding to a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 16. In some embodiments, the antigen-binding molecule is capable of binding to a peptide / polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 77. In some embodiments, the antigen-binding molecule is capable of binding to a peptide / polypeptide comprising or consisting of the amino acid sequence of SEQ ID NOs: 78 and 79. In some embodiments, the antigen-binding molecule is capable of binding to a peptide / polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 78. In some embodiments, the antigen-binding molecule is capable of binding to a peptide / polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 79.

[0089] In some embodiments, the antigen-binding molecule is unable to bind to a peptide consisting of the amino acid sequence corresponding to positions 260 to 279 of SEQ ID NO:1. The ability of an antigen-binding molecule to bind to a given peptide / polypeptide can be analyzed by methods well known to those skilled in the art, including ELISA, immunoblot (e.g., Western blot), immunoprecipitation, surface plasmon resonance (SPR; see, e.g., Hearty et al., Methods Mol Biol (2012) 907:411-442), or biolayer interferometry (see, e.g., Lad et al., (2015) J Biomol Screen 20(4):498-507).

[0090] In embodiments in which the antigen-binding molecule can bind to a peptide / polypeptide comprising a reference amino acid sequence, the peptide / polypeptide may comprise one or more additional amino acids at one or both termini of the reference amino acid sequence. In some embodiments, the peptide / polypeptide comprises, for example, 1 to 5, 1 to 10, 1 to 20, 1 to 30, 1 to 40, 1 to 50, 5 to 10, 5 to 20, 5 to 30, 5 to 40, 5 to 50, 10 to 20, 10 to 30, 10 to 40, 10 to 50, 20 to 30, 20 to 40, or 20 to 50 additional amino acids at one or both termini of the reference amino acid sequence.

[0091] In some embodiments, the additional amino acids provided at one or both ends (ie, the N-terminus and C-terminus) of the reference sequence correspond to positions at the ends of the reference sequence in the context of the amino acid sequence of HER3.

[0092] In some embodiments, the antigen-binding molecule is capable of binding to a peptide / polypeptide bound by an antibody comprising the VH and VL sequences of one of the antibody clones 10D1_c89, 10D1, 10D1_c75, 10D1_c76, 10D1_c77, 10D1_c78v1, 10D1_c78v2, 10D1_11B, 10D1_c85v1, 10D1_c85v2, 10D1_c85o1, 10D1_c85o2, 10D1_c87, 10D1_c90, 10D1_c91, 10D1_c92, and 10D1_c93 described herein. In some embodiments, the antigen-binding molecule is capable of binding to a peptide / polypeptide bound by an antibody comprising the VH and VL sequences of antibody clone 10D1_c89.

[0093] In some embodiments, the antigen-binding molecule comprises the CDRs of a HER3-binding antibody clone selected from 10D1_c89, 10D1, 10D1_c75, 10D1_c76, 10D1_c77, 10D1_c78v1, 10D1_c78v2, 10D1_11B, 10D1_c85v1, 10D1_c85v2, 10D1_c85o1, 10D1_c85o2, 10D1_c87, 10D1_c90, 10D1_c91, 10D1_c92, and 10D1_c93, or comprises the VH and VL thereof.

[0094] In some embodiments, the antigen-binding molecule is (1) The following CDR: HC-CDR1 having the amino acid sequence of SEQ ID NO: 40 HC-CDR2 having the amino acid sequence of SEQ ID NO: 43 HC-CDR3 having the amino acid sequence of SEQ ID NO: 48; or variants thereof in which one, two, or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are replaced with another amino acid. and a VH region incorporating The following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 66 LC-CDR2 having the amino acid sequence of SEQ ID NO: 69 LC-CDR3 having the amino acid sequence of SEQ ID NO: 74; or variants thereof in which one, two or three amino acids in one or more of LC-CDR1, LC-CDR2 or LC-CDR3 are replaced with another amino acid. The VL region incorporating (2) The following CDR: HC-CDR1 having the amino acid sequence of SEQ ID NO: 38 HC-CDR2 having the amino acid sequence of SEQ ID NO: 41 HC-CDR3 having the amino acid sequence of SEQ ID NO: 44; or variants thereof in which one, two, or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are replaced with another amino acid. and a VH region incorporating The following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 63 LC-CDR2 having the amino acid sequence of SEQ ID NO: 67 LC-CDR3 having the amino acid sequence of SEQ ID NO: 70; or variants thereof in which one, two or three amino acids in one or more of LC-CDR1, LC-CDR2 or LC-CDR3 are replaced with another amino acid. The VL region incorporating (3) The following CDR: HC-CDR1 having the amino acid sequence of SEQ ID NO: 38 HC-CDR2 having the amino acid sequence of SEQ ID NO: 41 HC-CDR3 having the amino acid sequence of SEQ ID NO: 44; or variants thereof in which one, two, or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are replaced with another amino acid. and a VH region incorporating The following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 64 LC-CDR2 having the amino acid sequence of SEQ ID NO: 67 LC-CDR3 having the amino acid sequence of SEQ ID NO: 70; or variants thereof in which one, two or three amino acids in one or more of LC-CDR1, LC-CDR2 or LC-CDR3 are replaced with another amino acid. The VL region incorporating (4) The following CDR: HC-CDR1 having the amino acid sequence of SEQ ID NO: 38 HC-CDR2 having the amino acid sequence of SEQ ID NO: 41 HC-CDR3 having the amino acid sequence of SEQ ID NO: 44; or variants thereof in which one, two, or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are replaced with another amino acid. and a VH region incorporating The following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 65 LC-CDR2 having the amino acid sequence of SEQ ID NO: 67 LC-CDR3 having the amino acid sequence of SEQ ID NO: 71; or variants thereof in which one, two or three amino acids in one or more of LC-CDR1, LC-CDR2 or LC-CDR3 are replaced with another amino acid. The VL region incorporating (5) The following CDR: HC-CDR1 having the amino acid sequence of SEQ ID NO: 38 HC-CDR2 having the amino acid sequence of SEQ ID NO: 42 HC-CDR3 having the amino acid sequence of SEQ ID NO: 45; or variants thereof in which one, two, or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are replaced with another amino acid. and a VH region incorporating The following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 63 LC-CDR2 having the amino acid sequence of SEQ ID NO: 67 LC-CDR3 having the amino acid sequence of SEQ ID NO: 70; or variants thereof in which one, two or three amino acids in one or more of LC-CDR1, LC-CDR2 or LC-CDR3 are replaced with another amino acid. The VL region incorporating (6) The following CDR: HC-CDR1 having the amino acid sequence of SEQ ID NO: 39 HC-CDR2 having the amino acid sequence of SEQ ID NO: 42 HC-CDR3 having the amino acid sequence of SEQ ID NO: 45; or variants thereof in which one, two, or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are replaced with another amino acid. and a VH region incorporating The following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 63 LC-CDR2 having the amino acid sequence of SEQ ID NO: 67 LC-CDR3 having the amino acid sequence of SEQ ID NO: 70; or variants thereof in which one, two or three amino acids in one or more of LC-CDR1, LC-CDR2 or LC-CDR3 are replaced with another amino acid. The VL region incorporating (7) The following CDR: HC-CDR1 having the amino acid sequence of SEQ ID NO: 38 HC-CDR2 having the amino acid sequence of SEQ ID NO: 42 HC-CDR3 having the amino acid sequence of SEQ ID NO: 44; or variants thereof in which one, two, or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are replaced with another amino acid. and a VH region incorporating The following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 63 LC-CDR2 having the amino acid sequence of SEQ ID NO: 68 LC-CDR3 having the amino acid sequence of SEQ ID NO: 70; or variants thereof in which one, two or three amino acids in one or more of LC-CDR1, LC-CDR2 or LC-CDR3 are replaced with another amino acid. The VL region incorporating (8) The following CDR: HC-CDR1 having the amino acid sequence of SEQ ID NO: 38 HC-CDR2 having the amino acid sequence of SEQ ID NO: 42 HC-CDR3 having the amino acid sequence of SEQ ID NO: 46; or variants thereof in which one, two, or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are replaced with another amino acid. and a VH region incorporating The following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 63 LC-CDR2 having the amino acid sequence of SEQ ID NO: 68 LC-CDR3 having the amino acid sequence of SEQ ID NO: 70; or variants thereof in which one, two or three amino acids in one or more of LC-CDR1, LC-CDR2 or LC-CDR3 are replaced with another amino acid. The VL region incorporating (9) The following CDR: HC-CDR1 having the amino acid sequence of SEQ ID NO: 38 HC-CDR2 having the amino acid sequence of SEQ ID NO: 42 HC-CDR3 having the amino acid sequence of SEQ ID NO: 47; or variants thereof in which one, two, or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are replaced with another amino acid. and a VH region incorporating The following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 63 LC-CDR2 having the amino acid sequence of SEQ ID NO: 68 LC-CDR3 having the amino acid sequence of SEQ ID NO: 70; or variants thereof in which one, two or three amino acids in one or more of LC-CDR1, LC-CDR2 or LC-CDR3 are replaced with another amino acid. The VL region incorporating (10) The following CDR: HC-CDR1 having the amino acid sequence of SEQ ID NO: 38 HC-CDR2 having the amino acid sequence of SEQ ID NO: 42 HC-CDR3 having the amino acid sequence of SEQ ID NO: 45; or variants thereof in which one, two, or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are replaced with another amino acid. and a VH region incorporating The following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 63 LC-CDR2 having the amino acid sequence of SEQ ID NO: 67 LC-CDR3 having the amino acid sequence of SEQ ID NO: 72; or variants thereof in which one, two or three amino acids in one or more of LC-CDR1, LC-CDR2 or LC-CDR3 are replaced with another amino acid. The VL region incorporating (11) The following CDR: HC-CDR1 having the amino acid sequence of SEQ ID NO: 38 HC-CDR2 having the amino acid sequence of SEQ ID NO: 41 HC-CDR3 having the amino acid sequence of SEQ ID NO: 44; or variants thereof in which one, two, or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are replaced with another amino acid. and a VH region incorporating The following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 63 LC-CDR2 having the amino acid sequence of SEQ ID NO: 67 LC-CDR3 having the amino acid sequence of SEQ ID NO: 73; or variants thereof in which one, two or three amino acids in one or more of LC-CDR1, LC-CDR2 or LC-CDR3 are replaced with another amino acid. The VL region incorporating Includes.

[0095] In some embodiments, the antigen-binding molecule comprises: (12) A VH region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, to the amino acid sequence of SEQ ID NO: 21; and a VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, to the amino acid sequence of SEQ ID NO: 49.

[0096] (13) A VH region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, to the amino acid sequence of SEQ ID NO: 22; and a VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, to the amino acid sequence of SEQ ID NO: 50.

[0097] (14) A VH region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, to the amino acid sequence of SEQ ID NO: 23; and a VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, to the amino acid sequence of SEQ ID NO: 51.

[0098] (15) A VH region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, to the amino acid sequence of SEQ ID NO: 24; and a VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, to the amino acid sequence of SEQ ID NO: 52.

[0099] (16) A VH region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, to the amino acid sequence of SEQ ID NO: 25; and a VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, to the amino acid sequence of SEQ ID NO: 53.

[0100] (17) A VH region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, to the amino acid sequence of SEQ ID NO: 26; and a VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, to the amino acid sequence of SEQ ID NO: 53.

[0101] (18) A VH region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, to the amino acid sequence of SEQ ID NO: 27; and a VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, to the amino acid sequence of SEQ ID NO: 53.

[0102] (19) A VH region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, to the amino acid sequence of SEQ ID NO: 28; and a VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, to the amino acid sequence of SEQ ID NO: 54.

[0103] (20) A VH region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, to the amino acid sequence of SEQ ID NO: 29; and a VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, to the amino acid sequence of SEQ ID NO: 54.

[0104] (21) A VH region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, to the amino acid sequence of SEQ ID NO: 30; and a VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, to the amino acid sequence of SEQ ID NO: 55.

[0105] (22) A VH region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, to the amino acid sequence of SEQ ID NO: 31; and a VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, to the amino acid sequence of SEQ ID NO: 56.

[0106] (23) A VH region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, to the amino acid sequence of SEQ ID NO: 32; and a VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, to the amino acid sequence of SEQ ID NO: 57.

[0107] (24) A VH region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, to the amino acid sequence of SEQ ID NO: 33; and a VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, to the amino acid sequence of SEQ ID NO: 58.

[0108] (25) A VH region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, to the amino acid sequence of SEQ ID NO: 34; and a VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, to the amino acid sequence of SEQ ID NO: 59.

[0109] (26) A VH region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, to the amino acid sequence of SEQ ID NO: 35; and a VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, to the amino acid sequence of SEQ ID NO: 60.

[0110] (27) A VH region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, to the amino acid sequence of SEQ ID NO: 36; and a VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, to the amino acid sequence of SEQ ID NO: 61.

[0111] (28) A VH region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, to the amino acid sequence of SEQ ID NO: 37; and a VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, to the amino acid sequence of SEQ ID NO: 62.

[0112] In some embodiments, the antigen-binding molecule is (29)(i) one or more (e.g., two) polypeptides comprising or consisting of an amino acid sequence having at least 70%, preferably 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% amino acid sequence identity to the amino acid sequence of SEQ ID NO: 75; and (ii) one or more (e.g., two) polypeptides comprising or consisting of an amino acid sequence having at least 70%, preferably 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% amino acid sequence identity to the amino acid sequence of SEQ ID NO: 76. It comprises or consists of:

[0113] Antigen-binding molecules that bind to EGFR The present disclosure provides antigen-binding molecules that bind to EGFR. In some embodiments, the antigen-binding molecule comprises the CDRs of an antigen-binding molecule capable of binding to EGFR. In some embodiments, the antigen-binding molecule comprises the FRs of an antigen-binding molecule capable of binding to EGFR. In some embodiments, the antigen-binding molecule comprises the CDRs and FRs of an antigen-binding molecule capable of binding to EGFR. That is, in some embodiments, the antigen-binding molecule comprises the VH region and VL region of an antigen-binding molecule capable of binding to EGFR.

[0114] In some embodiments, an antigen-binding molecule capable of binding to EGFR according to the present disclosure is any of the embodiments of the antigen-binding molecules described in US 6,217,866 B1 (which is incorporated herein by reference in its entirety), cetuximab (e.g., as described in US 6,217,866 B1 and Wong et al., Clin Ther. (2005) 27(6):684-694; DrugBank Acc. No. DB00002), panitumumab (e.g., as described in Foon et al., Int J Radiat Oncol Biol Phys. (2004) 58(3):984-990; DrugBank Acc. No. DB01269), zalutumumab (e.g., as described in Bastholt et al., Radiother Oncol. (2007) 85(1):24-28; DrugBank Acc. No. DB12202), necitumumab (e.g., as described in Kuenen et al., Clin Cancer Res. (2010) 16(6):1915-1923; DrugBank Acc. No. DB09559), nimotuzumab (e.g., as described in Ramakrishnan et al., mAbs (2009) 1(1) 41-48; DrugBank Acc. No. DB06192), durigotuzumab (e.g., as described in Fayette et al., Front Oncol. (2016) 6:232; DrugBank Acc. No. DB12142), and matuzumab (DrugBank Acc. No. DB05101). In some embodiments, the antigen-binding molecule is cetuximab.

[0115] In some embodiments, the antigen-binding molecule can bind to the same region of EGFR, or overlapping regions of EGFR, as bound by an antibody comprising the VH and VL sequences of cetuximab.

[0116] In some embodiments, the antigen-binding molecule comprises the CDRs of cetuximab or comprises the VH and VL of cetuximab. In some embodiments, the antigen-binding molecule is (30) The following CDR: HC-CDR1 having the amino acid sequence of SEQ ID NO: 92 HC-CDR2 having the amino acid sequence of SEQ ID NO: 93 HC-CDR3 having the amino acid sequence of SEQ ID NO: 94 VH region incorporating or a variant thereof in which one, two, or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are replaced with another amino acid; and The following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 96 LC-CDR2 having the amino acid sequence of SEQ ID NO: 97 LC-CDR3 having the amino acid sequence of SEQ ID NO: 98 The VL region incorporating or a variant thereof in which one, two, or three amino acids in one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are replaced with another amino acid. Includes.

[0117] In some embodiments, the antigen-binding molecule is (31) A VH region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 91; and a VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 95; Includes.

[0118] In some embodiments, the antigen-binding molecule is (32)(i) one or more (e.g., two) polypeptides comprising or consisting of an amino acid sequence having at least 70%, preferably 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% amino acid sequence identity to the amino acid sequence of SEQ ID NO: 99; and (ii) one or more (e.g., two) polypeptides comprising or consisting of an amino acid sequence having at least 70%, preferably 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% amino acid sequence identity to the amino acid sequence of SEQ ID NO: 100. It comprises or consists of:

[0119] Fc area In some embodiments, the antigen-binding molecules of the present disclosure comprise an Fc region. The Fc region is composed of CH2 and CH3 regions from one polypeptide and CH2 and CH3 regions from another polypeptide. The CH2 and CH3 regions from the two polypeptides together form the Fc region.

[0120] Fc-mediated functions include binding to Fc receptors, antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cell-mediated phagocytosis (ADCP), complement-dependent cytotoxicity (CDC), formation of the membrane attack complex (MAC), cell degranulation, cytokine and / or chemokine production, and antigen processing and presentation.

[0121] Pharmaceutical Combinations and Compositions The present disclosure provides a combination comprising (i) an antigen-binding molecule that binds to HER3, and (ii) an antigen-binding molecule that binds to EGFR. The present disclosure also provides a composition comprising (i) an antigen-binding molecule that binds to HER3, and (ii) an antigen-binding molecule that binds to EGFR.

[0122] In some aspects and embodiments, the combination further comprises (iii) a chemotherapeutic agent. In some aspects and embodiments, the composition further comprises (iii) a chemotherapeutic agent. Preferably, the chemotherapeutic agent is a microtubule targeting agent, such as a taxane. More preferably, the chemotherapeutic agent is a taxane, such as paclitaxel, docetaxel, or cabazitaxel. Most preferably, the chemotherapeutic agent is docetaxel.

[0123] It will be understood that the antigen-binding molecule that binds to HER3 may be an antigen-binding molecule that binds to HER3 according to any embodiment described herein, and similarly, the antigen-binding molecule that binds to EGFR may be an antigen-binding molecule that binds to EGFR according to any embodiment described herein. In preferred embodiments of the combinations and compositions of the previous paragraphs, the antigen-binding molecule that binds to HER3 is selected from one of (1) to (29) above, and the antigen-binding molecule that binds to EGFR is selected from any one of (30) to (32) above.

[0124] In some aspects and embodiments, the combination is a pharmaceutical combination.As used herein, " pharmaceutical combination " refers to the product that contains multiple (typically two or three in this specification) different active (i.e., therapeutic / prophylactic) agents, and they are intended to be used in combination.The drugs of pharmaceutical combination can be formulated together or separately, but are typically packaged together, and are typically packaged with a package insert that describes the instructions for using the drugs in combination.

[0125] In some embodiments, the agents of the pharmaceutical combination are contained in a single composition, e.g., a pharmaceutical composition containing both / all agents. In some embodiments, the agents of the pharmaceutical combination are contained in separate compositions; for example, the pharmaceutical combination may be provided as (i) a pharmaceutical composition containing an antigen-binding molecule that binds to HER3, and (ii) a pharmaceutical composition containing an antigen-binding molecule that binds to EGFR. In another example, the pharmaceutical combination may be provided as (i) a pharmaceutical composition containing an antigen-binding molecule that binds to HER3, (ii) a pharmaceutical composition containing an antigen-binding molecule that binds EGFR, and (iii) a chemotherapeutic agent (e.g., docetaxel).

[0126] The present disclosure also provides compositions (e.g., pharmaceutical compositions and medicaments) comprising the agents described herein (i.e., antigen-binding molecules that bind to HER3, antigen-binding molecules that bind to EGFR). Such compositions may include related articles in formulations suitable for clinical use.

[0127] The compositions of the present disclosure may be formulated in a pharmaceutical composition comprising one or more pharmaceutically acceptable carriers (e.g., liposomes, micelles, microspheres, nanoparticles), diluents / excipients (e.g., starch, cellulose, cellulose derivatives, polyols, dextrose, maltodextrin, magnesium stearate), adjuvants, fillers, buffering agents, preservatives (e.g., vitamin A, vitamin E, vitamin C, retinyl palmitate, selenium, cysteine, methionine, citric acid, sodium citrate, methylparaben, propylparaben), antioxidants, or the like. The composition may contain an inhibitor (e.g., vitamin A, vitamin E, vitamin C, retinyl palmitate, selenium), a lubricant (e.g., magnesium stearate, talc, silica, stearic acid, vegetable stearin), a binder (e.g., sucrose, lactose, starch, cellulose, gelatin, polyethylene glycol (PEG), polyvinylpyrrolidone (PVP), xylitol, sorbitol, mannitol), a stabilizer, a solubilizer, a surfactant (e.g., a wetting agent), a masking agent, or a colorant (such as titanium oxide).

[0128] As used herein, the term "pharmaceutically acceptable" refers to compounds, ingredients, materials, compositions, dosage forms, etc., that are suitable, within the scope of sound medical judgment, for use in contact with the tissues of a subject (e.g., a human subject) without undue toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio. Each carrier, diluent, excipient, adjuvant, filler, buffer, preservative, antioxidant, lubricant, binder, stabilizer, solubilizer, surfactant, masking agent, colorant, flavoring, or sweetener of a composition in accordance with the present disclosure must also be "acceptable" in the sense of being compatible with the other ingredients of the formulation. Suitable carriers, diluents, excipients, adjuvants, fillers, buffers, preservatives, antioxidants, lubricants, binders, stabilizers, solubilizers, surfactants, masking agents, colorants, flavorants, or sweeteners can be found in standard pharmaceutical texts, such as Remington's "The Science and Practice of Pharmacy" (Ed. A. Adejare), 23rd Edition (2020), Academic Press.

[0129] Pharmaceutical compositions / medicaments according to the present disclosure can be formulated for administration to a subject, e.g., via a route of administration appropriate to the nature of the composition / medicament and the disease / condition to be treated / prevented. In some embodiments, pharmaceutical compositions / medicaments can be formulated for parenteral, systemic, topical, intracavitary, intravascular, intravenous, intraarterial, intramuscular, intrathecal, intraocular, intraconjunctival, intratumoral, subcutaneous, intradermal, oral, or transdermal administration. In some embodiments, pharmaceutical compositions / medicaments can be formulated for administration by injection or infusion, or by ingestion.

[0130] Medicaments and pharmaceutical compositions can be formulated for administration to a blood vessel, or a tissue / organ of interest (e.g., a tissue / organ affected by a condition, affected by a disease / condition; e.g., a tissue / organ where symptoms of a disease / condition are manifested), or a tumor.

[0131] The pharmaceutical composition / medicament may contain an agent (i.e., an antigen-binding molecule that binds to HER3 and / or an antigen-binding molecule that binds to EGFR) in a sterile or isotonic medium. The pharmaceutical composition / medicament may be provided in a fluid, including a gel form. Fluid formulations may be formulated for administration by injection or infusion (e.g., via a cannula) into a blood vessel or selected region or tumor of the human or animal body. The pharmaceutical composition / medicament may be provided in a solid form, for example, a lyophilized form.

[0132] Functional properties The combinations and compositions described herein can be characterized by certain functional properties. In some embodiments, the combinations / compositions described herein can have one or more of the following properties: Increased killing of cells expressing HER3 and / or EGFR; Increases ADCC of cells expressing HER3 and / or EGFR; inhibiting tumor growth and / or reducing tumor size / volume (e.g., in HER3- and / or EGFR-expressing cancers); Prolonging survival in subjects with cancer (e.g., cancers that express HER3 and / or EGFR); inhibits tumor growth and / or reduces tumor size / volume (e.g., of HER3- and / or EGFR-expressing cancers) to a degree that is greater than the tumor growth inhibition / reduction in tumor size / volume observed when the components of the combination / composition are used alone; prolonging the survival of subjects with cancer (e.g., cancers that express HER3 and / or EGFR) to an extent greater than the survival observed when the components of the combination / composition are used alone; synergistically inhibit tumor growth and / or synergistically reduce tumor size / volume (e.g., of HER3- and / or EGFR-expressing cancers) compared to the tumor growth inhibition / reduction in tumor size / volume observed when the components of the combination / composition are used alone; and / or The combination / composition synergistically extends the survival of subjects with cancer (e.g., cancers that express HER3 and / or EGFR) compared to the extension of survival observed when the components of the combination / composition are used alone.

[0133] It will be understood that a given combination / composition may exhibit more than one of the properties listed in the previous paragraph. A given combination / composition may be evaluated for the properties listed in the previous paragraph using an appropriate assay. For example, the assay may be, for example, an in vitro assay, optionally a cell-based assay or a cell-free assay. In some embodiments, the assay may be, for example, an in vivo assay, i.e., performed in an animal other than a human. In some embodiments, the assay may be, for example, an ex vivo assay, i.e., performed using cells / tissues / organs obtained from a subject.

[0134] If the assay is a cell-based assay, the assay may include treating cells with the combination / composition to determine whether the combination / composition exhibits one or more of the listed properties. The assay may use a biological species labeled with a detectable entity to facilitate detection. The assay may include treating cells separately with a range of amounts / concentrations (e.g., a dilution series) of a given combination / composition and then evaluating the listed properties.

[0135] Analysis of the results of such assays may include determining the concentration at which 50% of the maximal level of the relevant activity is achieved. The concentration of a given drug at which 50% of the maximal level of the relevant activity is achieved is sometimes referred to as the "half-maximal effective concentration" of the drug for the relevant activity, which is known as the "EC 50 Depending on the characteristics, EC 50 is the "half maximal inhibitory concentration" or "IC 50 ", which is the concentration of drug at which 50% of the maximal level of inhibition of a given property is observed.

[0136] In some embodiments, combinations / compositions according to the present disclosure enhance (ie, upregulate, enhance) cell killing of cells containing / expressing HER3 and / or EGFR. In some embodiments, the combinations / compositions according to the present disclosure may inhibit the growth or reduce metastasis of cancers comprising cells comprising / expressing HER3 and / or EGFR. In some embodiments, the combinations / compositions may enhance (i.e., upregulate, enhance) cell killing of cells comprising / expressing HER3 and / or EGFR. In some embodiments, the combinations / compositions may inhibit the growth of cells of cancers comprising cells comprising / expressing HER3 and / or EGFR or inhibit tumor growth. In some embodiments, the combinations / compositions may inhibit metastasis of cancers / tumors comprising cells comprising / expressing HER3 and / or EGFR.

[0137] Cell killing can be determined, for example, using any of the methods outlined in Zaritskaya et al., Expert Rev Vaccines (2011), 9(6):601-616, which is incorporated herein by reference in its entirety. Examples of in vitro cytotoxicity / cell killing assays include: 51These assays include release assays such as Cr release assay, lactate dehydrogenase (LDH) release assay, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) release assay, and calcein-acetoxymethyl (calcein-AM) release assay. These assays measure cell killing based on the detection of factors released from lysed cells. Cell killing of a given test cell type by a given effector immune cell type can be analyzed, for example, by co-culturing the test cells with the effector immune cells and measuring the number / percentage of live / dead (e.g., lysed) test cells after an appropriate period of time. Other suitable assays include the xCELLigence real-time cytolysis in vitro potency assay described in Cerignoli et al., PLoS One. (2018) 13(3):e0193498 (incorporated herein by reference in its entirety).

[0138] In some embodiments, the combinations / compositions according to the present disclosure can reduce the number / proportion of cells that express HER3 and / or EGFR, hi some embodiments, the combinations / compositions according to the present disclosure can deplete / enhance the depletion of such cells.

[0139] The antigen-binding molecules constituting the combinations / compositions according to the present disclosure may contain one or more moieties for enhancing the reduction in the number / proportion of cells expressing HER3 and / or EGFR. For example, the antigen-binding molecules may contain, for example, an Fc region and / or a drug moiety.

[0140] The Fc region interacts with Fc receptors and other molecules of the immune system to produce functional effects. IgG Fc-mediated effector functions are outlined, for example, in Jefferis et al., Immunol Rev 1998 163:59-76 (incorporated herein by reference in its entirety), and include the Fc-mediated recruitment and activation of immune cells (e.g., macrophages, dendritic cells, neutrophils, basophils, eosinophils, platelets, mast cells, NK cells, and T cells) through the interaction of the Fc region with Fc receptors expressed by immune cells, the recruitment of complement pathway components through the binding of the Fc region to the complement protein C1q, and the resulting activation of the complement cascade. Fc-mediated functions include binding to Fc receptors, antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cell-mediated phagocytosis (ADCP), complement-dependent cytotoxicity (CDC), formation of the membrane attack complex (MAC), cell degranulation, cytokine and / or chemokine production, and antigen processing and presentation.

[0141] In some embodiments, the antigen-binding molecules of the combinations / compositions according to the present disclosure comprise an Fc region that is capable of enhancing / inducing one or more of ADCC, ADCP, CDC against cells that express HER3 and / or EGFR (e.g., cells that express HER3 and / or EGFR on their cell surface), and / or that is capable of enhancing MAC formation or cell degranulation on cells that express HER3 and / or EGFR (e.g., cells that express HER3 and / or EGFR on their cell surface).

[0142] In some embodiments, the antigen-binding molecules of the combinations / compositions according to the present disclosure are capable of enhancing / inducing ADCC against cells expressing HER3 and / or EGFR. The ability of a given antigen-binding molecule to induce ADCC of a given target cell type, and the extent to which it does so, can be determined, for example, according to the methods described in Yamashita et al., Scientific Reports (2016) 6:19772 (incorporated herein by reference in its entirety), or, for example, according to the methods described in Jedema et al., Blood (2004) 103:2677-82 (incorporated herein by reference in its entirety). 51 The ability and extent of a given antigen-binding molecule to induce ADCP can be analyzed by Cr release assay. The ability and extent of a given antigen-binding molecule to induce CDC can be analyzed, for example, according to the method described in Kamen et al., J Immunol (2017) 198 (1 Supplement) 157.17 (incorporated herein by reference in its entirety). The ability and extent of a given antigen-binding molecule to induce CDC can be analyzed, for example, using a C1q binding assay, as described in Schlothauer et al., Protein Engineering, Design and Selection (2016), 29 (10): 457-466 (incorporated herein by reference in its entirety).

[0143] In some embodiments, the combination / composition of the present disclosure exhibits anti-cancer activity. In some embodiments, the combination / composition increases the killing of cancer cells. In some embodiments, the combination / composition causes a reduction in the number of cancer cells in vivo, for example, compared to an appropriate control condition. The cancer may be a cancer described herein, for example, a cancer that expresses / overexpresses HER3 and / or EGFR.

[0144] In some embodiments, the combinations / compositions according to the present disclosure reduce / inhibit cancer and / or cancer tumor growth. In some embodiments, the combinations / compositions reduce tissue invasion by cancer cells. In some embodiments, the combinations / compositions reduce cancer metastasis. In some embodiments, the combinations / compositions exhibit anti-cancer activity. In some embodiments, the combinations / compositions reduce cancer cell growth / proliferation. In some embodiments, the combinations / compositions reduce cancer cell survival. In some embodiments, the combinations / compositions increase cancer cell killing. In some embodiments, the combinations / compositions of the present disclosure cause a reduction in the number of cancer cells, for example, in vivo. The cancer may be a cancer comprising cells expressing HER3 and / or EGFR.

[0145] The combinations / compositions of the present disclosure can be analyzed for the properties described in the previous paragraphs in suitable assays. Such assays include, for example, in vivo models. By way of illustration, Example 2 herein describes the evaluation of tumor growth inhibition by the combination of the HER3-binding molecule HMBD-001 IgG1 and the EGFR-binding molecule cetuximab in models derived from human cancer cells of various different cancers.

[0146] In some embodiments, administration of a combination / composition according to the present disclosure may result in one or more of: inhibiting cancer development / progression, delaying / preventing cancer onset, reducing / delaying / preventing tumor growth, reducing / delaying / preventing tissue invasion, reducing / delaying / preventing metastasis, reducing the severity of one or more symptoms of cancer, reducing the number of cancer cells, reducing the cancer burden, reducing the tumor size / volume, and / or increasing the survival (e.g., progression-free survival or overall survival) of a subject with cancer, e.g., as determined in an appropriate model.

[0147] It will be understood that the properties listed in the previous paragraphs will be evaluated after a period of time sufficient to observe the effects associated with treatment with the combination / composition. Tumor growth can be monitored by examining tumor volume over time, for example, as described in Example 2 herein. Tumor growth can be measured by measuring tumor volume (e.g., mm) over time. 3 It can be evaluated by measuring the

[0148] In some embodiments, the combinations / compositions of the disclosure may reduce tumor size / volume in a given assay (e.g., mean tumor size / volume of a treatment group in an in vivo model, e.g., a cancer expressing HER3 and / or EGFR) by less than 1-fold, e.g., <0.99-fold, <0.95-fold, <0.9-fold, <0.85-fold, <0.8-fold, <0.75-fold, <0.7-fold, <0.65-fold, <0.6-fold, <0.55-fold, <0.5-fold, <0.45-fold, <0.4-fold, <0.35-fold, <0.3-fold, <0.25-fold, <0.2-fold, <0.15-fold, <0.1-fold, <0.05-fold, or <0.01-fold, the tumor size / volume observed over the same period of time in the absence of treatment with the combination / composition (or after treatment with a suitable control composition known to have no effect on tumor growth). In some embodiments, assessment of tumor size / volume for such comparison purposes is performed in the relevant model after more than 5 days, for example, ≧10 days, ≧15 days, ≧20 days, ≧25 days, ≧30 days, ≧35 days, ≧40 days, ≧35 days, ≧50 days, ≧55 days, ≧60 days, ≧65 days, ≧70 days, ≧75 days, ≧80 days, ≧85 days, ≧90 days, ≧95 days, or ≧100 days after administration of the first dose of the combination / composition.

[0149] In some embodiments, the combinations / compositions of the present disclosure achieve a level of tumor growth inhibition (e.g., expressed as % tumor growth inhibition, e.g., calculated relative to tumor growth observed with treatment with an isotype-matched control antibody) in a given assay that is greater than 1-fold, e.g., ≥ 1.01-fold, ≥ 1.02-fold, ≥ 1.03-fold, ≥ 1.04-fold, ≥ 1.05-fold, ≥ 1.1-fold, ≥ 1.2-fold, ≥ 1.3-fold, ≥ 1.4-fold, ≥ 1.5-fold, ≥ 1.6-fold, ≥ 1.7-fold, ≥ 1.8-fold, ≥ 1.9-fold, ≥ 2-fold, ≥ 3-fold, ≥ 4-fold, ≥ 5-fold, ≥ 6-fold, ≥ 7-fold, ≥ 8-fold, ≥ 9-fold, or ≥ 10-fold the level of tumor growth inhibition observed at the same time point in the absence of treatment with the combination / composition (or following treatment with a suitable control composition known to have no effect on tumor growth). In some embodiments, assessment of tumor growth inhibition for such comparative purposes is performed in the relevant model after more than 5 days, e.g., ≧10 days, ≧15 days, ≧20 days, ≧25 days, ≧30 days, ≧35 days, ≧40 days, ≧35 days, ≧50 days, ≧55 days, ≧60 days, ≧65 days, ≧70 days, ≧75 days, ≧80 days, ≧85 days, ≧90 days, ≧95 days, or ≧100 days, after administration of the first dose of the combination / composition.

[0150] In some embodiments, the combinations / compositions of the present disclosure can increase the median survival of subjects with cancer (e.g., in an in vivo model, e.g., a cancer expressing HER3 and / or EGFR) in a given assay by more than 1-fold, e.g., ≧1.01-fold, ≧1.02-fold, ≧1.03-fold, ≧1.04-fold, ≧1.05-fold, ≧1.1-fold, ≧1.2-fold, ≧1.3-fold, ≧1.4-fold, ≧1.5-fold, ≧1.6-fold, ≧1.7-fold, ≧1.8-fold, ≧1.9-fold, ≧2-fold, ≧3-fold, ≧4-fold, ≧5-fold, ≧6-fold, ≧7-fold, ≧8-fold, ≧9-fold, or ≧10-fold, the median survival observed in the absence of treatment with the combination / composition (or after treatment with a suitable control composition known to have no effect on survival of subjects with cancer). Median survival can be expressed in days from the start of the experiment for subjects in the relevant treatment group.

[0151] In some embodiments, the combinations / compositions of the present disclosure reduce tumor growth, delay tumor growth, prevent tumor growth, reduce the severity of a symptom of cancer, reduce the number of cancer cells, reduce the cancer burden, reduce tumor size / volume, and / or prolong the survival of a subject with cancer to a greater extent than the components of the combination / composition used alone. In some embodiments, the combinations / compositions reduce tumor growth, delay tumor growth, prevent tumor growth, reduce the severity of one or more symptoms of cancer, reduce the number of cancer cells, reduce the cancer burden, reduce tumor size / volume, and / or prolong the survival of a subject with cancer to a greater extent than the HER3-binding molecule component of the combination / composition used as a monotherapy and / or to a greater extent than the EGFR-binding molecule component of the combination / composition used as a monotherapy.

[0152] In some embodiments, the combinations / compositions of the present disclosure inhibit tumor growth and / or reduce tumor size / volume to a degree that is greater than the tumor growth inhibition / reduction in tumor size / volume observed when the components of the combination / composition are used alone. In some embodiments, the combinations / compositions exhibit improved tumor growth inhibition and / or improved reduction in tumor size / volume compared to the levels observed when the HER3-binding molecule component of the combination / composition is used as a monotherapy and / or compared to the levels observed when the EGFR-binding molecule component of the combination / composition is used as a monotherapy.

[0153] In some embodiments, the combinations / compositions of the present disclosure extend the survival of a subject with cancer to a degree greater than the extension of survival observed when the components of the combination / composition are used alone. In some embodiments, the combinations / compositions extend the survival of a subject with cancer compared to the extension of survival observed when the HER3-binding molecule components of the combination / composition are used as monotherapy and / or compared to the extension of survival observed when the EGFR-binding molecule components of the combination / composition are used as monotherapy.

[0154] For the purpose of such comparison, the monotherapy preferably uses the same dose of the relevant drug as that used in the combination therapy. For example, in the experiment described in Example 2 herein, HMBD-001 IgG1 (anti-HER3 antibody) is administered at 20 mg / kg body weight per dose as a monotherapy, and in combination therapy with cetuximab, HMBD-001 IgG1 is also administered at 20 mg / kg body weight per dose. Similarly, cetuximab (anti-EGFR antibody) is administered at 10 mg / kg body weight per dose as a monotherapy, and in combination therapy with HMBD-001 IgG1, cetuximab is also administered at 10 mg / kg body weight per dose.

[0155] In some embodiments, the combinations / compositions of the disclosure can reduce tumor size / volume (e.g., mean tumor size / volume for a treatment group, e.g., in an in vivo model of a cancer that expresses HER3 and / or EGFR) to less than 1-fold, e.g., <0.99-fold, <0.95-fold, <0.9-fold, <0.85-fold, <0.8-fold, <0.75-fold, <0.7-fold, <0.65-fold, <0.6-fold, <0.55-fold, <0.5-fold, <0.45-fold, <0.4-fold, <0.35-fold, <0.3-fold, <0.25-fold, <0.2-fold, <0.15-fold, <0.1-fold, <0.05-fold, or <0.01-fold, the size / volume observed at the same time point after treatment of the subject with the same amount of one of the components of the combination / composition (i.e., the antigen-binding molecule that binds to HER3 or the antigen-binding molecule that binds EGFR) as a monotherapy. In some embodiments, assessment of tumor size / volume for such comparison purposes is performed in the relevant model more than 5 days, e.g., ≧10 days, ≧15 days, ≧20 days, ≧25 days, ≧30 days, ≧35 days, ≧40 days, ≧35 days, ≧50 days, ≧55 days, ≧60 days, ≧65 days, ≧70 days, ≧75 days, ≧80 days, ≧85 days, ≧90 days, ≧95 days, or ≧100 days after administration of the first dose of the combination / composition.

[0156] In some embodiments, the combinations / compositions of the present disclosure achieve a level of tumor growth inhibition (e.g., expressed as % tumor growth inhibition, e.g., calculated relative to the tumor growth observed with treatment with an isotype-matched control antibody) that is greater than 1-fold, e.g., one of: ≥1.01-fold, ≥1.02-fold, ≥1.03-fold, ≥1.04-fold, ≥1.05-fold, ≥1.1-fold, ≥1.2-fold, ≥1.3-fold, ≥1.4-fold, ≥1.5-fold, ≥1.6-fold, ≥1.7-fold, ≥1.8-fold, ≥1.9-fold, ≥2-fold, ≥3-fold, ≥4-fold, ≥5-fold, ≥6-fold, ≥7-fold, ≥8-fold, ≥9-fold, or ≥10-fold the level of tumor growth inhibition observed following treatment of a subject with the same amount of one of the components of the combination / composition (i.e., the antigen-binding molecule that binds HER3 or the antigen-binding molecule that binds EGFR) as a monotherapy. In some embodiments, assessment of tumor growth inhibition for purposes of such comparison is performed in the relevant model more than 5 days after administration of the first dose of the combination / composition, e.g., one of: ≧10 days, ≧15 days, ≧20 days, ≧25 days, ≧30 days, ≧35 days, ≧40 days, ≧35 days, ≧50 days, ≧55 days, ≧60 days, ≧65 days, ≧70 days, ≧75 days, ≧80 days, ≧85 days, ≧90 days, ≧95 days, or ≧100 days.

[0157] In some embodiments, the combinations / compositions of the present disclosure can extend the survival of a subject with cancer (e.g., the median survival of a subject with cancer, e.g., as determined in an in vivo model, e.g., of a cancer that expresses HER3 and / or EGFR) by more than 1-fold, e.g., ≧1.01-fold, ≧1.02-fold, ≧1.03-fold, ≧1.04-fold, ≧1.05-fold, ≧1.1-fold, ≧1.2-fold, ≧1.3-fold, ≧1.4-fold, ≧1.5-fold, ≧1.6-fold, ≧1.7-fold, ≧1.8-fold, ≧1.9-fold, ≧2-fold, ≧3-fold, ≧4-fold, ≧5-fold, ≧6-fold, ≧7-fold, ≧8-fold, ≧9-fold, or ≧10-fold, the survival observed after treating the subject with the same amount of one of the components of the combination / composition (i.e., the antigen-binding molecule that binds HER3 or the antigen-binding molecule that binds EGFR) as a monotherapy.

[0158] In the previous three paragraphs, "the same amount" refers to the amount of the relevant drug used in the combination / composition. For example, if a subject receiving the combination is administered 20 mg / kg of an antigen-binding molecule that binds to HER3 and 10 mg / kg of an antigen-binding molecule that binds to EGFR, monotherapy with the "same amount" of the antigen-binding molecule that binds to HER3 is monotherapy with 20 mg / kg of the antigen-binding molecule that binds to HER3. Similarly, monotherapy with the "same amount" of the antigen-binding molecule that binds to EGFR is monotherapy with 10 mg / kg of the antigen-binding molecule that binds to EGFR.

[0159] In some embodiments, the combinations / compositions of the present disclosure achieve a synergistic therapeutic and / or prophylactic effect, i.e., in some embodiments, the combinations / compositions achieve a treatment effect that is synergistic (i.e., super-additive) compared to that observed when the HER3-binding molecule components of the combination / composition are used as monotherapy and / or compared to that observed when the EGFR-binding molecule components of the combination / composition are used as monotherapy.

[0160] As used herein, a "synergistic" or "superadditive" level of associated effect (e.g., tumor growth inhibition, reduction in tumor size / volume, extended survival) for a given combination / composition refers to a level of effect that is greater than the sum of the effects observed for the individual components of the combination / composition.

[0161] Quantitative methods for assessing synergy are described, for example, in Tallarida, Genes Cancer. (2011) 2(11):1003-1008 and Chou, Cancer Res (2010) 70:440-446, both of which are incorporated herein by reference in their entireties. Additive, synergistic, and antagonistic effects can be evaluated in experiments in which different dose ranges of the combination / composition and its individual components are evaluated for relative effects. Dose-response curves can be plotted and evaluated to determine whether the combination / composition achieves a synergistic level of relative effect relative to the individual components of the combination / composition used alone (i.e., as monotherapy). In some embodiments, synergistic effects can be assessed using combination index (CI) values ​​calculated using the Chou-Talalay method described in Chou, Cancer Res (2010) 70:440-446. According to the Chou-Talalay method, for a given combination, CI=1 indicates an additive effect, CI<1 indicates a synergistic effect, and CI>1 indicates an antagonistic effect.

[0162] In some embodiments, the combinations / compositions of the present disclosure achieve a synergistic (i.e., super-additive) reduction in tumor growth, a delay in tumor growth, prevention of tumor growth, a reduction in the severity of one or more symptoms of cancer, a reduction in the number of cancer cells, a reduction in cancer burden, a reduction in tumor size / volume, and / or an increase in the survival of a subject with cancer compared to that observed when either component of the combination / composition is used alone. In some embodiments, the combinations / compositions achieve a synergistic (i.e., super-additive) reduction in tumor growth, a delay in tumor growth, prevention of tumor growth, a reduction in the severity of one or more symptoms of cancer, a reduction in the number of cancer cells, a reduction in cancer burden, a reduction in tumor size / volume, and / or an increase in the survival of a subject with cancer compared to that observed when the HER3-binding molecule component of the combination / composition is used as a monotherapy and / or compared to that observed when the EGFR-binding molecule component of the combination / composition is used as a monotherapy.

[0163] Therapeutic and prophylactic uses The present disclosure provides methods and articles (eg, agents, combinations, and compositions of the present disclosure) for the treatment and / or prevention of diseases, such as cancer.

[0164] Thus, the present disclosure provides an antigen-binding molecule that binds to HER3 for use in a method for treating or preventing cancer (e.g., a cancer described herein), wherein the method further comprises the step of administering an antigen-binding molecule that binds EGFR. Also provided is an antigen-binding molecule that binds EGFR for use in a method for treating or preventing cancer (e.g., a cancer described herein), wherein the method further comprises the step of administering an antigen-binding molecule that binds HER3.

[0165] Also provided is the use of an antigen-binding molecule that binds to HER3 in the manufacture of a medicament for use in a method of treating or preventing cancer (e.g., a cancer described herein), wherein the method further comprises the step of administering an antigen-binding molecule that binds EGFR.Also provided is the use of an antigen-binding molecule that binds EGFR in the manufacture of a medicament for use in a method of treating or preventing cancer (e.g., a cancer described herein), wherein the method further comprises the step of administering an antigen-binding molecule that binds HER3.

[0166] Further provided is a method for treating or preventing cancer (e.g., a cancer described herein), comprising administering to a subject in need thereof a therapeutically or prophylactically effective amount of (i) an antigen-binding molecule that binds to HER3, and (ii) an antigen-binding molecule that binds to EGFR.

[0167] The present disclosure also provides (i) an antigen-binding molecule that binds HER3 and (ii) an antigen-binding molecule that binds EGFR for use in a method of treating or preventing cancer (e.g., a cancer described herein) in a subject. Also provided is the use of (i) an antigen-binding molecule that binds HER3 and (ii) an antigen-binding molecule that binds EGFR in the manufacture of a medicament for use in treating or preventing cancer (e.g., a cancer described herein) in a subject. Also provided is a method of treating or preventing cancer (e.g., a cancer described herein) in a subject, comprising administering to the subject therapeutically or prophylactically effective amounts of (i) an antigen-binding molecule that binds HER3 and (ii) an antigen-binding molecule that binds EGFR.

[0168] In embodiments according to the aspects of the preceding paragraph, the provision of (i) and (ii) may be as a combination therapy. In some embodiments, (i) and (ii) may be provided simultaneously or sequentially.

[0169] The present disclosure is directed to methods and articles (eg, agents, combinations, and compositions of the present disclosure) for the treatment and / or prevention of cancer. As used herein, "cancer" may be or include any unwanted cell proliferation (or any disease that manifests itself due to unwanted cell proliferation), neoplasm, or tumor. Cancer may be benign or malignant. Cancer may be primary or secondary (e.g., metastatic). A neoplasm or tumor may be any abnormal growth or proliferation of cells and may be located in any tissue. The cancer can be, for example, a cancer of tissue / cells derived from the adrenal gland, adrenal medulla, anus, appendix, bladder, blood, bone, bone marrow, brain, breast, cecum, central nervous system (including or excluding the brain), cerebellum, cervix, colon, duodenum, endometrium, epithelial cells (e.g., renal epithelium), gallbladder, esophagus, glial cells, heart, ileum, jejunum, kidney, lacrimal gland, larynx, liver, lung, lymph, lymph node, lymphoblast, jaw, mediastinum, mesentery, myometrium, nasopharynx, omentum, oral cavity, ovary, pancreas, parotid gland, peripheral nervous system, peritoneum, pleura, prostate, salivary gland, sigmoid colon, skin, small intestine, soft tissue, spleen, stomach, testis, thymus, thyroid, tongue, tonsils, trachea, uterus, vulva, white blood cells.

[0170] The tumor to be treated may be a nervous system tumor or a non-nervous system tumor.Nervous system tumors may originate from either the central nervous system or the peripheral nervous system, and may be, for example, glioma, medulloblastoma, meningioma, neurofibroma, ependymoma, schwannoma, neurofibrosarcoma, astrocytoma, and oligodendroglioma.Non-nervous system cancers / tumors may originate from any other non-nervous system tissue; examples include melanoma, mesothelioma, lymphoma, myeloma, leukemia, non-Hodgkin's lymphoma (NHL), Hodgkin's lymphoma, chronic myeloid leukemia (CML), acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), cutaneous T-cell lymphoma (CTCL), chronic lymphocytic leukemia (CLL), liver cancer, epidermoid carcinoma, prostate cancer, breast cancer, lung cancer, colon cancer, ovarian cancer, pancreatic cancer, thymic carcinoma, NSCLC, blood cancer, and sarcoma.

[0171] In some embodiments, the cancer to be treated / prevented comprises cells that express an EGFR family member (e.g., HER3, EGFR, HER2, or HER4) and / or cells that express a ligand for an EGFR family member. In some embodiments, the cancer to be treated / prevented is a cancer that is positive for an EGFR family member. In some embodiments, the cancer comprises cells that overexpress an EGFR family member and / or a ligand for an EGFR family member. Overexpression can be determined by detecting an expression level that is greater than the expression level by comparable non-cancer cells / non-tumor tissues.

[0172] Expression can be determined by any suitable means.Expression can be gene expression or protein expression.Gene expression can be determined, for example, by quantitative real-time PCR (qRT-PCR), for example, by detecting the mRNA encoding HER3.For example, protein expression can be determined by antibody-based methods, for example, Western blot, immunohistochemistry, immunocytochemistry, flow cytometry or ELISA.

[0173] In some embodiments, the cancer is a cancer in which HER3 and / or EGFR are pathologically involved. That is, in some embodiments, the cancer is a cancer caused or exacerbated by the expression of HER3 and / or EGFR, a cancer in which the expression of HER3 and / or EGFR is a risk factor, and / or a cancer in which the expression of HER3 and / or EGFR is positively associated with the onset, development, progression, severity, or metastasis of cancer. The cancer can be characterized by the expression of HER3 and / or EGFR, for example, the cancer can contain cells (e.g., cells of tumor tissue) that express HER3 and / or EGFR. Such cancers can be referred to as HER3 and / or EGFR positive. A cancer that is "positive" for HER3 and / or EGFR can be a cancer that contains cells that express HER3 and / or EGFR (e.g., on the cell surface). A cancer that is "positive" for HER3 and / or EGFR can overexpress HER3 and / or EGFR.

[0174] In some embodiments, the cancers being treated / prevented include cells harboring a genetic variant (e.g., a mutation) that causes increased HER3 and / or EGFR (gene and / or protein) expression and / or activity compared to comparable cells harboring a reference allele that does not contain the genetic variant (e.g., a non-mutated, or "wild-type," allele). The genetic variant may be or may include an insertion, deletion, substitution, or larger translocation / rearrangement of a nucleotide sequence relative to the reference allele.

[0175] Mutations that "result in" increased expression of HER3 and / or EGFR may be known, predicted, or associated with increased gene / protein expression of HER3 and / or EGFR. Mutations that "result in" increased activity of HER3 and / or EGFR may be known, predicted, or associated with increased HER3-mediated signaling and / or EGFR-mediated signaling. Mutations that result in increased expression and / or activity of HER3 and / or EGFR may be referred to as "activating" mutations.

[0176] Mutations that cause increased expression of HER3 and / or EGFR may result in expression of a HER3 and / or EGFR gene or protein that is not expressed by, and / or encoded by, a comparable cell that does not carry the mutation. That is, HER3 and / or EGFR may be neoantigens that arise as a result of the mutation, and thus "increased expression" may be absent.

[0177] A mutation that causes increased expression of HER3 and / or EGFR can result in increased expression of the HER3 and / or EGFR gene or protein expressed by and / or encoded by its genomic nucleic acid in a comparable cell that does not contain the mutation. Illustratively, a cell can contain a mutation that results in increased transcription levels of a nucleic acid encoding HER3 and / or EGFR compared to the transcription levels of a nucleic acid encoding HER3 and / or EGFR by a comparable cell that does not contain the mutation.

[0178] In some embodiments, mutations that cause increased expression of HER3 and / or EGFR may cause increased gene expression of HER3 and / or EGFR compared to comparable cells that do not contain the mutation. In some embodiments, mutations that cause increased expression of HER3 and / or EGFR may cause increased protein expression of HER3 and / or EGFR compared to comparable cells that do not contain the mutation.

[0179] In some embodiments, a mutation that causes increased expression of HER3 and / or EGFR may cause increased levels of HER3 and / or EGFR on or at the cell surface of cells that contain the mutation compared to comparable cells that do not contain the mutation.

[0180] Cells that have increased expression of HER3 and / or EGFR (e.g., as a result of a mutation) compared to the expression levels of HER3 and / or EGFR by reference cells may be described as having "overexpression" of HER3 and / or EGFR, or "upregulated expression" of HER3 and / or EGFR. For example, a cancer containing cells that harbor a mutation that results in increased expression of HER3 and / or EGFR compared to comparable cells lacking the mutation may be described as a cancer containing cells exhibiting overexpression / upregulated expression of HER3 and / or EGFR. In some embodiments, the reference cells lacking the mutation may be non-cancerous cells (e.g., of a comparable cell type) or cancer cells (e.g., of a comparable cancer type).

[0181] Mutations that cause increased activity of HER3 and / or EGFR can result in increased HER3-mediated signaling and / or EGFR-mediated signaling compared to the level of HER3-mediated signaling and / or EGFR-mediated signaling by comparable cells that do not contain the mutation.

[0182] In some embodiments, cancers treated / prevented in accordance with the present disclosure may be characterized, for example, by increased expression and / or activity (i.e., gene and / or protein expression) of HER3 and / or EGFR in organs / tissues / subjects affected by the disease / condition compared to normal organs / tissues / subjects (i.e., in the absence of the disease / condition). In some embodiments, cancer cells and / or tumors treated / prevented may be characterized, for example, by increased expression and / or activity of HER3 and / or EGFR compared to the levels of expression and / or activity observed in comparable non-cancerous cells / non-tumor tissues.

[0183] Cancers that overexpress HER3 may do so as a result of amplification of the HER3 gene, and similarly, cancers that overexpress EGFR may do so as a result of amplification of the EGFR gene.

[0184] In some embodiments, the cancer treated or prevented according to the present disclosure is a HER3-amplified cancer. In some embodiments, the cancer is an EGFR-amplified cancer. In some embodiments, the cancer is a cancer comprising HER3 and EGFR amplification. In some embodiments, the cancer treated or prevented according to the present disclosure is a TP63-amplified cancer.

[0185] HER3, EGFR, and / or TP63 amplification can be identified using techniques well known in the art, such as in situ hybridization. For example, HER3 amplification can be assessed by fluorescence in situ hybridization, as described, for example, in Chung et al., J Gynecol Oncol. (2019) 30(5):e75. HER3-amplified cancers can include a 12q13.2 to chromosome 12 centromere ratio of 2 or greater, as determined by ISH. EGFR amplification can similarly be assessed by in situ hybridization, as described, for example, in French et al., Neuro-Oncology (2019) 21(10):1263-1272. EGFR-amplified cancers can include a 7p11.2-7p12 to chromosome 7 centromere ratio of 2 or greater, as determined by ISH. For example, TP63 amplification can be assessed by fluorescence in situ hybridization, e.g., as described in Massion et al., Cancer Res. (2003) 63(21):7113-21. TP63-amplified cancers can include a 3q26-3qter to chromosome 3 centromere ratio of 2 or greater, as determined by ISH.

[0186] EGFR and its association with and role in cancer are reviewed, for example, in Uribe et al., Cancers (Basel) (2021) 13(11):2748, Sigismund et al., Mol Oncol. (2018) 12(1):3-20, and da Silva Santos et al., Int J Pharm. (2021) 592:120082, which are incorporated by reference in their entireties. da Silva Santos et al., Int J Pharm. (2021) 592:120082 describes EGFR-targeted interventions for the treatment of cancer, including monoclonal anti-EGFR antibody therapy.

[0187] HER3 and its association with and role in cancer are reviewed, for example, in Mishra et al., Oncol Rev. (2018) 12(1):355, Karachaliou et al., BioDrugs. (2017) 31(1):63-73, and Zhang et al., Acta Biochimica et al., Biophysica Sinica (2016) 48(1):39-48, all of which are incorporated herein by reference in their entireties. Mishra et al., Oncol Rev. (2018) 12(1):355 also describes HER3-targeted interventions for the treatment of cancer, including monoclonal anti-HER3 antibody therapy.

[0188] In some embodiments, the cancer to be treated / prevented comprises cells that express a ligand for HER3 (e.g., NRG1 and / or NRG2). In some embodiments, the cancer to be treated / prevented comprises cells that express a higher expression level of NRG1 and / or NRG2 than the expression level by comparable non-cancer cells / non-tumor tissues. The cancer may be described as comprising cells that overexpress NRG1 and / or NRG2.

[0189] The HER3-binding antigen-binding molecules described herein bind to HER3 with extremely high affinity both when HER3 is bound by NRG (i.e., when HER3 is provided in an "open" conformation) and when HER3 is not bound by NRG (i.e., when HER3 is provided in a "closed" conformation). Thus, they are particularly useful for treating / preventing cancers characterized by expression / overexpression of HER3 ligands, for example, cancers / tumors containing cells that express / overexpress a ligand for HER3.

[0190] In some embodiments, the cancer being treated / prevented comprises cells harboring a genetic variant (e.g., a mutation) that causes increased (gene and / or protein) expression of a ligand for HER3 compared to comparable cells harboring a reference allele that does not contain the genetic variant (e.g., a non-mutated or "wild-type" allele). The genetic variant may be or may include an insertion, deletion, substitution, or larger translocation / rearrangement of a nucleotide sequence relative to the reference allele.

[0191] A mutation that "results in" increased expression of a ligand for HER3 may be known, predicted, or associated with increased gene / protein expression of the ligand for HER3. A mutation that results in increased expression of a ligand for HER3 may be referred to as an "activating" mutation.

[0192] A mutation that causes increased expression of a ligand for HER3 can result in gene or protein expression of a ligand for HER3 that is not expressed by a comparable cell that does not carry the mutation and / or is not encoded by its genomic nucleic acid. That is, the ligand for HER3 may be a neoantigen that arises as a result of the mutation, and thus "increased expression" may be absent. Illustratively, cells containing a CD74-NRG1 gene fusion exhibit increased expression of the CD74-NRG1 fusion polypeptide encoded by the gene fusion compared to cells lacking the CD74-NRG1 gene fusion.

[0193] A mutation that causes increased expression of a ligand for HER3 can result in increased gene or protein expression of the ligand for HER3 expressed by and / or encoded by its genomic nucleic acid in a comparable cell that does not contain the mutation. Illustratively, a cell can contain a mutation that results in increased transcription levels of a nucleic acid encoding NRG1 compared to the transcription levels of the nucleic acid encoding NRG1 by a comparable cell that does not contain the mutation.

[0194] In some embodiments, a mutation that causes increased expression of a ligand for HER3 may cause increased gene expression of the ligand for HER3 compared to a comparable cell that does not contain the mutation. In some embodiments, a mutation that causes increased expression of a ligand for HER3 may cause increased protein expression of the ligand for HER3 compared to a comparable cell that does not contain the mutation.

[0195] In some embodiments, a mutation that causes increased expression of a ligand for HER3 may cause increased levels of a ligand for HER3 on or at the cell surface of cells containing the mutation compared to comparable cells that do not contain the mutation. In some embodiments, a mutation that causes increased expression of a ligand for HER3 may cause increased levels of secretion of a ligand for HER3 from cells containing the mutation compared to comparable cells that do not contain the mutation.

[0196] Cells that have increased expression of a ligand for HER3 (e.g., as a result of a mutation) compared to the expression level of the ligand for HER3 by a reference cell may be described as "overexpressing" the ligand for HER3 or having "upregulated expression" of the ligand for HER3. For example, a cancer containing cells that harbor a mutation that results in increased expression of a ligand for HER3 compared to comparable cells that lack the mutation may be described as a cancer containing cells that exhibit overexpression / upregulated expression of the ligand for HER3. In some embodiments, the reference cells that lack the mutation may be non-cancerous cells (e.g., of a comparable cell type) or cancer cells (e.g., of a comparable cancer type).

[0197] As used herein, a "ligand for HER3" is generally intended to refer to a molecule that can bind to HER3 via the ligand-binding region of HER3 formed by domains I and III of HER3. In some embodiments, a ligand for HER3 binds to HER3 through interaction with domains I and / or III of HER3. Exemplary ligands for HER3 include neuregulins such as NRG1 and NRG2, which bind to HER3 via interaction between their EGF-like domains and the ligand-binding region of HER3.

[0198] The HER3 ligand preferably binds to and induces signal transduction through the HER3 receptor and / or a receptor complex containing HER3. As will be apparent from the present disclosure, the receptor complex containing HER3 may further include an interaction partner for HER3 described herein, such as HER3, HER2, EGFR, HER4, HGFR, IGF1R, and / or cMet.

[0199] In some embodiments, the ligand for HER3 can bind to a HER3 receptor / receptor complex expressed by a cell other than a cell with increased expression of a HER3 ligand. For example, in some embodiments, the ligand for HER3 can bind to a cancer cell that expresses HER3.

[0200] In some embodiments, the ligand for HER3 is capable of binding to a HER3 receptor / receptor complex expressed by cells that have increased expression of a HER3 ligand. In some embodiments, the cancer being treated / prevented comprises (i) cells that express HER3 and (ii) cells that express a ligand for HER3 (e.g., cells that have increased expression of a ligand for HER3 as a result of, e.g., a mutation that results in increased expression of a ligand for HER3).

[0201] In some embodiments, the cancer being treated / prevented comprises cells that (i) express HER3 and (ii) also express a ligand for HER3 (e.g., increased expression of the ligand for HER3 as a result of a mutation that results in increased expression of the ligand for HER3).

[0202] In some embodiments, the ligand for HER3 comprises or consists of the amino acid sequence of a HER3-binding region of a ligand for HER3, or an amino acid sequence derived from the HER3-binding region of a ligand for HER3. The amino acid sequence derived from the HER3-binding region of a ligand for HER3 may comprise at least 60% (e.g., 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) amino acid sequence identity to the amino acid sequence from which it is derived.

[0203] In some embodiments, the ligand for HER3 comprises an EGF-like domain capable of binding to HER3, or a HER3-binding fragment thereof. In some embodiments, the HER3-binding EGF-like domain / fragment is or is derived from an EGF family member (e.g., heparin-binding EGF-like growth factor (HB-EGF), transforming growth factor-α (TGF-α), amphiregulin (AR), epiregulin (EPR), epigen, betacellulin (BTC), NRG1, NRG2, NRG3, or NRG4).

[0204] Exemplary ligands for HER3 include neuregulins (NRGs). Neuregulins include NRG1 (including its alpha, alpha2b, and alpha3 isoforms), NRG2, NRG3, and NRG4. In some embodiments, NRGs are selected from NRG1, NRG2, NRG3, and NRG4. In some embodiments, NRGs are selected from NRG1 and NRG2.

[0205] The EGF-like domain of human NRG1, which binds to HER3, is formed by positions 178-222 of UniProt:Q02297-1. The EGF-like domain of human NRG2 is formed by positions 341-382 of UniProt:O14511-1. The EGF-like domain of human NRG3 is formed by positions 286-329 of UniProt:B9EGV5-1. The EGF-like domain of human NRG4 is formed by positions 5-46 of UniProt:Q8WWG1-1. In some embodiments, the EGF-like domain / fragment comprises or consists of an amino acid sequence having at least 60% (e.g., 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) amino acid sequence identity to the EGF-like domain of an NRG (NRG1, NRG2, NRG3, or NRG4).

[0206] In some embodiments, the ligand for HER3 is not an EGFR family protein (e.g., HER3, HER2, EGFR, HER4, HGFR, IGF1R, cMet).

[0207] In some embodiments, the mutation that results in increased expression of a ligand for HER3 is an NRG gene fusion. In some embodiments, the ligand for HER3 is the product of (i.e., the polypeptide encoded by) an NRG gene fusion. In some embodiments, the cancer comprises cells having an NRG gene fusion. As used herein, "NRG gene fusion" refers to a genetic variant that encodes a polypeptide that includes (i) the amino acid sequence of an NRG protein (e.g., NRG1, NRG2, NRG3, or NRG4; e.g., NRG1 or NRG2), and (ii) the amino acid sequence of a protein other than an NRG protein.

[0208] It will be understood that the NRG gene fusion preferably encodes a HER3 ligand as described herein. In some embodiments, the NRG gene fusion encodes a polypeptide comprising the HER3-binding region of the NRG protein. In some embodiments, the NRG gene fusion encodes the EGF-like domain of the NRG protein, or a polypeptide that can bind to HER3 and comprises an amino acid sequence having at least 60% (e.g., 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) amino acid sequence identity to the EGF-like domain of the NRG protein.

[0209] In some embodiments, the NRG gene fusion encodes a fusion polypeptide that includes a transmembrane domain. In some embodiments, the NRG gene fusion encodes a fusion polypeptide that includes a transmembrane domain of a protein other than an NRG protein.

[0210] In some embodiments, the NRG gene fusion is an NRG1 gene fusion. In some embodiments, the NRG1 gene fusion encodes an EGF-like domain of NRG1 or a polypeptide that can bind to HER3 and includes an amino acid sequence having at least 60% (e.g., 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) amino acid sequence identity to the EGF-like domain of NRG1.

[0211] NRG1 gene fusions are described, for example, in WO2021 / 048274A1, WO2018 / 182422A1, WO2019 / 051155A1, Dhanasekaran et al., Nat Commun. (2014) 5:5893, Drilon et al., Cancer Discov. (2018) 8(6):686-695, Nagasaka et al., Journal of Thoracic Oncology (2019) 14(8):1354-1359, and Jonna et al., Clin Cancer Res. (2019) 25(16):4966-4972, all of which are incorporated by reference in their entireties. The diversity of NRG1 gene fusions may be due to NRG1 being located on chromosome 8, which is particularly susceptible to genomic translocation events (Adelaide et al., Genes Chromosomes Cancer. (2003) 37(4):333-45).

[0212] In some embodiments, the NRG1 gene fusion is selected from the group consisting of CLU-NRG1, CD74-NRG1, DOC4-NRG1, SLC3A2-NRG1, RBPMS-NRG1, WRN-NRG1, SDC4-NRG1, RAB2IL1-NRG1, VAMP2-NRG1, KIF13B-NRG1, THAP7-NRG1, SMAD4-NRG1, MDK-NRG1, TNC-NRG1, DIP2B-NRG1, MRPL13 In some embodiments, the NRG1 gene fusion is selected from the group consisting of CLU-NRG1, PARP8-NRG1, ROCK1-NRG1, DPYSL2-NRG1, ATP1B1-NRG1, CDH6-NRG1, APP-NRG1, AKAP13-NRG1, THBS1-NRG1, FOXA1-NRG1, PDE7A-NRG1, RAB3IL1-NRG1, CDK1-NRG1, BMPRIB-NRG1, TNFRSF10B-NRG1, and MCPH1-NRG1. In some embodiments, the NRG1 gene fusion is CLU-NRG1.

[0213] CD74-NRG1 gene fusions are described, for example, in Fernandez-Cuesta et al., Cancer Discov. (2014) 4:415-22, and Nakaoku et al., Clin Cancer Res (2014) 20:3087-93. DOC4-NRG1 gene fusions are described, for example, in Liu et al., Oncogene. (1999) 18(50):7110-4, and Wang et al., Oncogene. (1999) 18(41):5718-21. SLC3A2-NRG1 gene fusions are described, for example, in Nakaoku et al., Clin Cancer Res (2014) 20:3087-93, Shin et al., Oncotarget (2016) 7:69450-65, and Shin et al., Mol Cancer Ther. (2018) 17(9):2024-2033. RBPMS-NRG1, WRN-NRG1, RAB2IL1-NRG1, and SDC4-NRG1 gene fusions are described, for example, in Dhanasekaran et al., Nat Commun. (2014) 5:5893. VAMP2-NRG1 gene fusions are described, for example, in Jung et al., J Thorac Oncol. (2015) 10(7):1107-11, and Shim et al., J Thorac Oncol. (2015) 10(8):1156-62. KIF13B-NRG1 gene fusions are described, for example, in Xia et al., Int J Surg Pathol. (2017) 25(3):238-240. SMAD4-NRG1, AKAP13-NRG1, THBS1-NRG1, FOXA1-NRG1, PDE7A-NRG1, RAB3IL1-NRG1, and THAP7-NRG1 gene fusions are described, for example, in Drilon et al., Cancer Discov. (2018) 8(6):686-695. MDK-NRG1, TNC-NRG1, DIP2B-NRG1, MRPL13-NRG1, PARP8-NRG1, ROCK1-NRG1, and DPYSL2-NRG1 gene fusions are described, for example, in Jonna et al., Clin Cancer Res. (2019) 25(16):4966-4972.ATP1B1-NRG1 gene fusions are described, for example, in Drilon et al., Cancer Discov. (2018) 8(6):686-695, and Jones et al., Annals of Oncology (2017) 28:3092-3097. CLU-NRG1 gene fusions are described, for example, in Drilon et al., Cancer Discov. (2018) 8(6):686-695, and Nagasaka et al., Journal of Thoracic Oncology (2019) 14(8):1354-1359.

[0214] In some embodiments, the NRG gene fusion is an NRG2 gene fusion. In some embodiments, the NRG2 gene fusion encodes an EGF-like domain of NRG2 or a polypeptide that can bind to HER3 and includes an amino acid sequence having at least 60% (e.g., 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) amino acid sequence identity to the EGF-like domain of NRG2.

[0215] NRG2 gene fusions include, for example, SLC12A2-NRG2, as described in WO2021 / 048274A1 and WO2015 / 093557A1, and ZNF208-NRG2, as described in Dupain et al., Mol Ther. (2019) 27(1):200-218.

[0216] The cancer comprising cells with a mutation that results in increased expression of a ligand for HER3 (e.g., comprising cells with an NRG gene fusion, e.g., an NRG1 gene fusion or an NRG2 gene fusion) can be any cancer described herein. In some embodiments, such a cancer can be a cancer of tissue / cells derived from the lung, breast, head, neck, kidney, ovary, pancreas, prostate, uterus, gallbladder, colon, rectum, bladder, soft tissue, or nasopharynx.

[0217] In some embodiments, the cancer comprising cells with a mutation that results in increased expression of a ligand for HER3 (e.g., comprising cells with an NRG gene fusion, e.g., an NRG1 gene fusion, or an NRG2 gene fusion) is selected from lung cancer, non-small cell lung cancer, lung adenocarcinoma, invasive mucinous lung adenocarcinoma, lung squamous cell carcinoma, breast cancer, breast cancer, invasive breast carcinoma, head and neck cancer, head and neck squamous cell carcinoma, renal cancer, renal clear cell carcinoma, ovarian cancer, serous ovarian cystadenocarcinoma, pancreatic cancer, pancreatic adenocarcinoma, pancreatic ductal adenocarcinoma, prostate cancer, prostate adenocarcinoma, endometrial cancer, uterine carcinosarcoma, gallbladder cancer, bile duct carcinoma, colorectal cancer, metastatic colorectal cancer, bladder cancer, urothelial bladder cancer, sarcoma, soft tissue sarcoma, neuroendocrine tumor, and nasopharyngeal neuroendocrine tumor.

[0218] In some embodiments, the cancer being treated / prevented is a lung cancer (e.g., non-small cell lung cancer, lung adenocarcinoma, invasive mucinous lung adenocarcinoma, or lung squamous cell carcinoma) comprising cells with NRG1 gene fusions.

[0219] It will be understood that in embodiments herein, a cancer comprising cells with particular characteristics may be or may include a tumor comprising cells with those characteristics. As is common in the art, a cancer / tumor containing cells with particular characteristics may be referred to herein simply as a cancer / tumor having those characteristics. By way of example, a cancer / tumor containing cells with an NRG1 gene fusion may be referred to simply as an "NRG1 gene fusion-containing cancer / tumor" or an "NRG1 gene fusion cancer / tumor."

[0220] In some embodiments, the cancer to be treated / prevented contains a mutation that confers resistance to treatment with a BRAF inhibitor. In some embodiments, the mutation is a mutation in BRAF V600. In some embodiments, the mutation is BRAF V600E or V600K. The cancer may be thyroid cancer or colon cancer, for example, RAS wild-type colorectal cancer. In some embodiments, the cancer to be treated / prevented contains a mutation that confers resistance to treatment with a BRAF inhibitor (e.g., a mutation in BRAF V600), and the treatment includes administration of vemurafenib or darafenib.

[0221] In squamous cell carcinoma (SCC), the PI3K / AKT signaling pathway is commonly altered by gene amplification and / or mutation. The frequently amplified 3q26 / 28 chromosomal region, where PIK3CA is located, also contains the TP63 and SOX2 lineage genes. TP63 is a member of the TP53 gene family and is expressed in the skin, esophagus, pulmonary airways, and the basal compartment of the larynx during development and homeostasis. TP63 is used as a diagnostic marker for squamous and adenocarcinoma types of lung and esophageal cancer. Preclinical data suggest that TP63 regulates NRG1 expression in SCC, and the HER3 signaling pathway is active in TP63-amplified squamous cell carcinoma. Furthermore, the association between high NRG1 levels and response rates to anti-HER3 antibodies and EGFR overexpression in a subset of squamous cell carcinomas supports the rationale for combining 10D1F with cetuximab in EGFR-amplified squamous cell carcinoma.

[0222] In some embodiments, the cancer to be treated / prevented is squamous cell carcinoma, i.e., squamous cell carcinoma. In some embodiments, the cancer is advanced or metastatic squamous cell carcinoma. In some embodiments, the squamous cell carcinoma is selected from EGFR-amplified squamous cell carcinoma, head and neck squamous cell carcinoma (HNSCC), lung squamous cell carcinoma (LUSC), and esophageal squamous cell carcinoma (ESCC), cervical squamous cell carcinoma, cutaneous squamous cell carcinoma (cSCC), thyroid squamous cell carcinoma (SCTC), vaginal squamous cell carcinoma (SCCV), prostate squamous cell carcinoma, and penile squamous cell carcinoma. In some embodiments, the squamous cell carcinoma is selected from EGFR-amplified squamous cell carcinoma, head and neck squamous cell carcinoma (HNSCC), lung squamous cell carcinoma (LUSC), and esophageal squamous cell carcinoma (ESCC).

[0223] In some embodiments, the cancer being treated / prevented is an NRG1 and HER3 expressing / overexpressing cancer. In some embodiments, the cancer is an NRG1 and HER3 expressing / overexpressing squamous cell carcinoma. In some embodiments, the cancer is an NRG1 and HER3 expressing / overexpressing head and neck squamous cell carcinoma. In some embodiments, the cancer is an NRG1 and HER3 expressing / overexpressing esophageal squamous cell carcinoma. In some embodiments, the cancer is an NRG1 and HER3 expressing / overexpressing hypopharyngeal squamous cell carcinoma.

[0224] In some embodiments, the cancer being treated / prevented is an EGFR-expressing / overexpressing cancer. In some embodiments, the cancer being treated / prevented is an EGFR-expressing / overexpressing squamous cell carcinoma. In some embodiments, the cancer is an EGFR-expressing / overexpressing head and neck squamous cell carcinoma. In some embodiments, the cancer is an EGFR-expressing / overexpressing esophageal squamous cell carcinoma. In some embodiments, the cancer is an EGFR-expressing / overexpressing tongue squamous cell carcinoma.

[0225] In some embodiments, the cancer being treated / prevented is EGFR-expressing / overexpressing colorectal cancer. In some embodiments, the cancer being treated / prevented is EGFR-expressing / overexpressing colon adenocarcinoma.

[0226] In some embodiments, the cancer may be a recurrent cancer. As used herein, a "recurrent" cancer refers to a cancer that has responded to treatment (e.g., a first-line cancer treatment) but has subsequently reappeared / progressed, for example, after a period of remission. For example, a recurrent cancer may be a cancer whose growth / progression has been inhibited by treatment (e.g., a first-line cancer treatment) and has subsequently grown / progressed.

[0227] In some embodiments, the cancer may be a refractory cancer. As used herein, "refractory" cancer refers to a cancer that has not responded to treatment (e.g., a first-line treatment for cancer). For example, a refractory cancer may be a cancer whose growth / progression has not been inhibited by treatment (e.g., a first-line treatment for cancer). In some embodiments, a refractory cancer may be a cancer in which a subject undergoing cancer treatment has not shown a partial or complete response to the treatment.

[0228] In some embodiments, the cancer is a cancer comprising cells that express / overexpress an EGFR family member (e.g., HER3, EGFR, HER2, or HER4), a cancer comprising cells that express / overexpress HER3, a cancer comprising cells that express / overexpress EGFR, a cancer comprising cells that express / overexpress HER3 and EGFR, a cancer comprising cells with a mutation that results in increased expression of a ligand for HER3, a cancer comprising cells with a mutation that results in increased expression of a ligand for EGFR, a cancer comprising cells with an NRG gene fusion, a solid tumor, a hematologic cancer, a squamous cell carcinoma, an EGFR-amplified squamous cell carcinoma, breast cancer, breast cancer, invasive breast cancer, ductal carcinoma, metastatic breast cancer, triple-negative breast cancer, HER2-positive breast cancer, gastric cancer, gastric adenocarcinoma, gastrointestinal adenocarcinoma, colorectal cancer, metastatic colorectal cancer, colon cancer, colorectal cancer, colorectal adenocarcinoma, colon adenocarcinoma, head and neck cancer, head and neck squamous Epithelial carcinoma (HNSCC), lung cancer, non-small cell lung cancer, lung adenocarcinoma, invasive mucinous lung adenocarcinoma, lung squamous cell carcinoma (LUSC), ovarian cancer, ovarian cancer, serous ovarian adenocarcinoma, serous ovarian cystadenocarcinoma, fallopian tube cancer, kidney cancer, renal cell carcinoma, renal clear cell carcinoma, renal cell adenocarcinoma, papillary renal cell carcinoma, pancreatic cancer, pancreatic adenocarcinoma, pancreatic ductal adenocarcinoma, cervical cancer, cervical squamous cell carcinoma, skin cancer, melanoma, oral cancer, oropharyngeal cancer, esophageal cancer, esophageal squamous cell carcinoma (ESCC), esophageal adenocarcinoma, liver cancer, hepatocellular carcinoma, biliary ductal carcinoma, gallbladder cancer, uterine cancer, endometrial cancer, endometrial carcinoma, uterine carcinosarcoma, thyroid cancer, thyroid cancer, thyroid carcinoma, pheochromocytoma, paraganglioma, bladder cancer, bladder urothelial carcinoma, prostate cancer, prostate adenocarcinoma, retinoblastoma, sarcoma, soft tissue sarcoma, peritoneal cancer, thymoma, neuroendocrine tumor, nasopharyngeal neuroendocrine tumor, squamous cell carcinoma of the skin, astrocytoma, low-grade astrocytoma, high-grade astrocytoma, anaplastic astrocytoma, and glioblastoma multiforme.

[0229] In some embodiments, cancer is selected from squamous cell carcinoma or squamous cell carcinoma (SCC).SCC can originate from stratified squamous epithelium in any anatomical location.For example, SCC can be non-melanoma skin cancer, head and neck cancer (HNSCC), esophageal cancer (ESCC) or non-small cell lung cancer (sqNSCLC).

[0230] In some embodiments, the cancer is selected from cancer comprising cells that express / overexpress HER3, cancer comprising cells that express / overexpress EGFR, cancer comprising cells that express / overexpress HER3 and EGFR, squamous cell carcinoma, EGFR-amplified squamous cell carcinoma, esophageal cancer, esophageal squamous cell carcinoma, head and neck cancer, head and neck squamous cell carcinoma, colorectal cancer, metastatic colorectal cancer, colon adenocarcinoma, pancreatic cancer, pancreatic adenocarcinoma, pancreatic ductal adenocarcinoma, lung cancer, and lung squamous cell carcinoma.

[0231] Treating cancer according to the methods of the present disclosure achieves one or more of the following treatment effects: reducing the number of cancer cells in a subject, reducing the size of a cancerous tumor / lesion in a subject, inhibiting (e.g., preventing or slowing) the growth of cancer cells in a subject, inhibiting (e.g., preventing or slowing) the growth of a cancerous tumor / lesion in a subject, inhibiting (e.g., preventing or slowing) the development / progression of cancer (e.g., to a later stage or metastasis), reducing the severity of cancer symptoms in a subject, extending the subject's survival (e.g., progression-free survival or overall survival), reducing the number or activity correlates of cancer cells in a subject, and / or reducing the cancer burden in a subject.

[0232] To determine the response to treatment, the subject can be evaluated according to the Revised Criteria for Response Assessment: Lugano classification (e.g., as described in Cheson et al., J Clin Oncol (2014) 32:3059-3068, incorporated herein by reference). In some embodiments, the subject treated according to the methods of the present disclosure achieves one of a complete response, a partial response, or stable disease.

[0233] Prevention can refer to preventing cancer from occurring and / or preventing cancer from getting worse, eg, preventing cancer from progressing to a later stage (eg, metastasis). In some embodiments, administration of a combination / composition according to the present disclosure may be associated with one or more of the following: inhibiting cancer development / progression, delaying / preventing the onset of cancer, reducing / delaying / preventing tumor growth, reducing / delaying / preventing tissue invasion, reducing / delaying / preventing metastasis, reducing the severity of one or more symptoms of cancer, reducing the number of cancer cells, reducing the cancer burden, reducing the tumor size / volume, and / or increasing the survival (e.g., progression-free survival or overall survival) of a subject with cancer.

[0234] According to various embodiments of the present disclosure, methods of treating and / or preventing cancer according to the present disclosure may include inhibiting tumor growth, reducing tumor size / volume, and / or extending survival of a subject with cancer.

[0235] According to various aspects of the present disclosure, methods are provided for one or more of the following or including (e.g., in the context of treating / preventing cancer, e.g., a cancer as described herein) one or more of the following: Killing cells that express HER3 and / or EGFR; increasing ADCC of cells expressing HER3 and / or EGFR; inhibiting tumor growth and / or reducing tumor size / volume (e.g., in HER3- and / or EGFR-expressing cancers); Prolonging survival in subjects with cancer (e.g., cancers that express HER3 and / or EGFR); inhibiting tumor growth and / or reducing tumor size / volume (e.g., of HER3- and / or EGFR-expressing cancers) to a degree greater than the tumor growth inhibition / reduction in tumor size / volume observed when the component agents of the combination / composition are used alone; prolonging survival of subjects with cancer (e.g., HER3- and / or EGFR-expressing cancers) to an extent greater than the survival observed when the component drugs of the combination / composition are used alone; synergistically inhibiting tumor growth and / or synergistically reducing tumor size / volume (e.g., of HER3- and / or EGFR-expressing cancers) compared to the tumor growth inhibition / reduction in tumor size / volume observed when the component agents of the combination / composition are used alone; and / or A synergistic extension of survival in subjects with cancer (e.g., a cancer that expresses HER3 and / or EGFR) compared to the extension of survival observed when the component drugs of the combination / composition are used alone.

[0236] Also provided are agents according to the present disclosure for use in such methods, and the use of agents according to the present disclosure in the manufacture of compositions (e.g., medicaments) for use in such methods. It will be understood that the methods typically include the step of administering to a subject an antigen-binding molecule that binds to HER3 and an antigen-binding molecule that binds to EGFR.

[0237] Similarly, one or more of the following may be observed in a subject after therapeutic or prophylactic intervention in accordance with the present disclosure (e.g., compared to pre-intervention levels / numbers / proportions, etc.): Killing cells that express HER3 and / or EGFR; increasing ADCC of cells expressing HER3 and / or EGFR; inhibiting tumor growth and / or reducing tumor size / volume (e.g., in HER3- and / or EGFR-expressing cancers); Prolonging survival in subjects with cancer (e.g., cancers that express HER3 and / or EGFR); inhibiting tumor growth and / or reducing tumor size / volume (e.g., of HER3- and / or EGFR-expressing cancers) to a degree greater than the tumor growth inhibition / reduction in tumor size / volume observed when the component agents of the combination / composition are used alone; prolonging survival of subjects with cancer (e.g., HER3- and / or EGFR-expressing cancers) to an extent greater than the survival observed when the component drugs of the combination / composition are used alone; synergistically inhibiting tumor growth and / or synergistically reducing tumor size / volume (e.g., of HER3- and / or EGFR-expressing cancers) compared to the tumor growth inhibition / reduction in tumor size / volume observed when the component agents of the combination / composition are used alone; and / or A synergistic extension of survival in subjects with cancer (e.g., a cancer that expresses HER3 and / or EGFR) compared to the extension of survival observed when the component drugs of the combination / composition are used alone.

[0238] In some embodiments, therapeutic / prophylactic interventions according to the present disclosure may be described as being "associated with" one or more of the effects described in the preceding paragraphs. Those skilled in the art can readily assess such properties using techniques routinely practiced in the art.

[0239] In some embodiments, therapeutic / prophylactic intervention with an antigen-binding molecule that binds HER3 and an antigen-binding molecule that binds EGFR according to the present disclosure provides an improved treatment effect compared to the effect observed when either agent is used as monotherapy. In some embodiments, intervention with an antigen-binding molecule that binds HER3 and an antigen-binding molecule that binds EGFR provides a synergistic (i.e., super-increased) therapeutic and / or prophylactic effect compared to the level of the associated effect observed when either agent is used alone.

[0240] The administration of the agents, pharmaceutical combinations, and pharmaceutical compositions of the present disclosure is preferably in a "therapeutically effective" or "prophylactically effective" amount, which is sufficient to provide a therapeutic or prophylactic benefit to the subject. The actual amount administered, and the rate and time course of administration, will depend on the nature and severity of the disease / condition and the specific product being administered. Prescribing treatment, such as determining dosage, is within the responsibility of general practitioners and other medical professionals and typically takes into account the disease / disorder being treated, the condition of the individual subject, the delivery site, the method of administration, and other factors known to practitioners. Examples of the techniques and protocols mentioned above can be found in Remington's "The Science and Practice of Pharmacy" (Ed. A. Adejare), 23rd Edition (2020), Academic Press.

[0241] Administration of the articles of the present disclosure can be, for example, parenteral, systemic, topical, intracavitary, intravascular, intravenous, intraarterial, intramuscular, intrathecal, intraocular, intraconjunctival, intratumoral, subcutaneous, intradermal, oral, or transdermal. Administration can also be by injection, infusion, or ingestion.

[0242] In some aspects and embodiments, the articles of the present disclosure may be administered to a target tissue / organ (e.g., a tissue / organ affected by a disease / condition) affected by a condition (e.g., a tissue / organ where symptoms of the disease / condition are manifested). In some aspects and embodiments, the articles of the present disclosure may be administered into the blood (i.e., intravenously / intra-arterially) by injection or infusion (e.g., via a cannula), or may be administered subcutaneously or orally. In some aspects and embodiments, the articles of the present disclosure may be administered to a tumor.

[0243] When two or more agents (e.g., an antigen-binding molecule that binds to HER3 and an antigen-binding molecule that binds to EGFR) are administered in combination, the agents can be administered either simultaneously or sequentially.

[0244] Concurrent administration refers to the administration of two or more agents (e.g., an antigen-binding molecule that binds to HER3 and an antigen-binding molecule that binds to EGFR) together, for example, as a pharmaceutical composition containing both agents (i.e., as a combined preparation), or shortly after each other (e.g., within 1, 4, 6, 8, or 12 hours), optionally via the same route of administration, e.g., into the same artery, vein, or other blood vessel.

[0245] Sequential administration refers to the separate administration of one agent followed by another agent after a given time interval. It is not required that the agents be administered by the same route, although in some embodiments this is the case. The time interval can be any time interval.

[0246] In some embodiments, therapeutic or prophylactic intervention according to the present disclosure comprises (i) administering to a subject with cancer (e.g., a cancer described herein) an antigen-binding molecule that binds to HER3, and (ii) administering to the subject an antigen-binding molecule that binds EGFR. In some embodiments, (i) and (ii) are performed simultaneously. In some embodiments, (i) and (ii) are performed sequentially (e.g., (i) may be followed by (ii), or (ii) may be followed by (i)).

[0247] Multiple doses of drugs, pharmaceutical combinations, and pharmaceutical compositions can be provided.The multiple doses can be separated by a predetermined time interval, which can be selected to be one of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or 31 days, or 1, 2, 3, 4, 5, or 6 months.For example, doses can be given every 7, 14, 21, or 28 days (± 3, 2, or 1 day).

[0248] In some embodiments, therapeutic or prophylactic intervention according to the present disclosure may further include administering another agent for the treatment / prevention of a related disease / condition. For example, therapeutic or prophylactic intervention according to the present disclosure may further include administering a chemotherapeutic agent. In some embodiments, therapeutic or prophylactic intervention according to the present disclosure includes (i) administering an antigen-binding molecule that binds to HER3 to a subject with cancer (e.g., a cancer described herein), (ii) administering an antigen-binding molecule that binds EGFR to the subject, and (iii) administering a chemotherapeutic agent to the subject. In some embodiments, two or more of (i), (ii), and (iii) are performed simultaneously (e.g., (i), (ii), and (iii) may be performed simultaneously, or (i) and (ii) may be performed simultaneously, or (iii) may be performed sequentially either before or after (i) and (ii)). In some embodiments, at least one of (i), (ii) and (iii) is carried out sequentially (e.g., (i), (ii) and (iii) are carried out sequentially). For example, (i) may be followed by (ii) and then by (iii); (ii) may be followed by (i) and then by (iii); (iii) may be followed by (i) and then by (ii); or (iii) may be followed by (ii) and then by (i). Preferably, the chemotherapeutic agent is a microtubule targeting agent, for example, a taxane. More preferably, the chemotherapeutic agent is a taxane, for example, paclitaxel, docetaxel or cabazitaxel. Most preferably, the chemotherapeutic agent is docetaxel.

[0249] Chemotherapy refers to the treatment of cancer with drugs (chemotherapeutic agents). Chemotherapeutic agents may be chemical entities, such as small molecule drugs, antibiotics, DNA intercalators, protein inhibitors (e.g., kinase inhibitors), or biological agents, such as antibodies, antibody fragments, aptamers, nucleic acids (e.g., DNA, RNA), peptides, polypeptides, or proteins. Chemotherapeutic agents may be formulated as pharmaceutical compositions or medicaments. Formulations may include one or more chemotherapeutic agents in combination with one or more pharmaceutically acceptable diluents, excipients, or carriers.

[0250] The chemotherapeutic agent may be administered by one or more routes of administration, for example, parenterally, by intravenous injection, orally, subcutaneously, intradermally, intraperitoneally, or intratumorally. Chemotherapy can be administered according to a treatment regime.A treatment regime can be a predetermined timetable, plan, scheme or schedule of chemotherapy administration, which can be created by a doctor or medical professional and can be tailored to the patient who needs treatment.A treatment regime can indicate one or more of the following: the type of chemotherapy to be administered to a patient; the dose of each drug; the administration interval; the length of each treatment; and if any, the number and nature of any treatment holidays.In the case of combination therapy, a single treatment regime can be provided that indicates the method of administration of each drug.

[0251] Chemotherapeutic agents include abemaciclib, abiraterone acetate, avitrexate (methotrexate), Abraxane (paclitaxel albumin-stabilized nanoparticles), ABVD, ABVE, ABVE-PC, AC, acalabrutinib, AC-T, Adcetris (brentuximab vedotin), ADE, Ado-trastuzumab emtansine, Adriamycin (doxorubicin hydrochloride), afatinib dimaleate, and afi Nitol (everolimus), Aquinzeo (netupitant and palonosetron hydrochloride), Aldara (imiquimod), aldesleukin, Alecensa (alectinib), alectinib, alemtuzumab, Alimta (pemetrexed disodium), Alicopa (copanlisib hydrochloride), Alkeran for injection (melphalan hydrochloride), Alkeran tablets (melphalan), Aloxi (palonosetron hydrochloride), Alunbrig (brigatinib) , Ambochlorin (chlorambucil), Ambochlorin (chlorambucil), Amifostine, Aminolevulinic acid, Anastrozole, Aprepitant, Aredia (pamidronate disodium), Arimidex (anastrozole), Aromasin (exemestane), Alanon (nelarabine), Arsenic trioxide, Arzera (ofatumumab), Asparaginase Erwini Acrisanthimy, atezolizumab, Avastin (bevacizumab), avelumab, axiconib butagene ciloreucel, axitinib, azacitidine, Bavencio (avelumab), BEACOPP, Besenam (carmustine), Beleodac (belinostat), belinstat, bendamustine hydrochloride, BEP, Besponsa (inotuzumab ozogamicin), bevacizumab, bexarotene, Bexar (tositumomab and iodine I 131 tositumomab), bicalutamide, BiCNU (carmustine), bleomycin, blinatumomab, Bilincyto (blinatumomab), bortezomib, Bosulif (bosutinib), bosutinib, brentuximab vedotin, brigatinib, BuMel, busulfan, Busulfex (busulfan), cabazitaxel, Cabometyx (cabozantinib-S-malate), cabozantinib-S-malate, CAF, Calquence (acalabrutinib), Cambus (alemtuzumab),Camptosar (irinotecan hydrochloride), capecitabine, CAPOX, Carlac (topical fluorouracil), carboplatin, carboplatin-taxol, carfilzomib, Carumbris (carmustine), carmustine, carmustine implant, Casodex (bicalutamide), CEM, ceritinib, Cerbidine (daunorubicin hydrochloride), Cervarix (recombinant HPV bivalent vaccine), cetuximab, CEV, chlorambucil, chlorambucil-prednisone, CHOP, cisplatin, cladribine, Clafen (cyclophosphamide) d), clofarabine, Clofarex (clofarabine), Chloral (clofarabine), CMF, cobimetinib, Cometriq (cabozantinib-S-malate), copanlisib hydrochloride, COPDAC, COPP, COPP-ABV, Cosmegen (dactinomycin), Cotellic (cobimetinib), crizotinib, CVP, cyclophosphamide, Cyfos (ifosfamide), Cyramza (ramucirumab), cytarabine, cytarabine liposome, Cytosar-U (cytarabine), Cytoxan (cyclophosphamide), dabrafenib, daca Rubadin, Dacogen (decitabine), dactinomycin, daratumumab, Darzalex (daratumumab), dasatinib, daunorubicin hydrochloride, daunorubicin hydrochloride and cytarabine liposome, decitabine, defibrotide sodium, Defitelio (defibrotide sodium), degarelix, denileukin diftitox, denosumab, DepoCyt (cytarabine liposome), dexamethasone, dexrazoxane hydrochloride, dinutuximab, docetaxel, Doxil (doxorubicin hydrochloride liposome), doxorubicin hydrochloride , doxorubicin hydrochloride liposome, Dox-SL (doxorubicin hydrochloride liposome), DTIC-Dome (dacarbazine), durvalumab, Efudex (topical fluorouracil), ERYTECH (rasburicase), Elence (epirubicin hydrochloride), elotuzumab, Eloxatin (oxaliplatin), eltrombopag olamine, Emend (aprepitant), Empliciti (elotuzumab), enasidenib mesylate, enzalutamide, epirubicin hydrochloride, EPOCH, Erbitux (cetuximab), eribulin mesylate,Erivedge (vismodegib), erlotinib hydrochloride, Erwinase (asparaginase erwinia chrysanthemum), Ethiol (amifostine), Etopophos (etoposide phosphate), etoposide, etoposide phosphate, Evacet (doxorubicin hydrochloride liposomal), everolimus, Evista (raloxifene hydrochloride), Evomela (melphalan hydrochloride), exemestane, 5-FU (fluorouracil injection), 5-FU (fluorouracil - topical), Fairston (toremifene), Farydak (panobinostat), febuxostat Solodex (fulvestrant), FEC, Femara (letrozole), filgrastim, Fludara (fludarabine phosphate), fludarabine phosphate, Fluoroplex (fluorouracil-topical), fluorouracil injection, fluorouracil-topical, flutamide, Folex (methotrexate), Folex PFS (methotrexate), FOLFIRI, FOLFIRI-bevacizumab, FOLFIRI-cetuximab, FOLFIRINOX, FOLFOX, Folotin (pralatrexate), FU-LV, fulvestrant , Gardasil (recombinant HPV quadrivalent vaccine), Gardasil 9 (recombinant HPV nonvalent vaccine), Gazyva (obinutuzumab), gefitinib, gemcitabine hydrochloride, gemcitabine-cisplatin, gemcitabine-oxaliplatin, gemtuzumab ozogamicin, Gemzar (gemcitabine hydrochloride), Giotrif (afatinib dimaleate), Glivec (imatinib mesylate), Gliadel (carmustine implant), Gliadel wafer (carmustine implant), glucarpidase, goserelin acetate, Halaven (eribulin mesylate) silate), Hemandiol (propranolol hydrochloride), Herceptin (trastuzumab), HPV bivalent vaccine, recombinant, HPV nonvalent vaccine, recombinant, HPV quadrivalent vaccine, recombinant, Hycamtin (topotecan hydrochloride), Hydrea (hydroxyurea), hydroxyurea, Hyper-CVAD, Ibrance (palbociclib), ibritumomab tiuxetan, ibrutinib, ICE, Iclusig (ponatinib hydrochloride), Idamycin (idarubicin hydrochloride), idarubicin hydrochloride, idelalisib, Idhifa (enasidenib mesylate),Ifex (ifosfamide), ifosfamide, Ifosfamidam (ifosfamide), IL-2 (aldesleukin), imatinib mesylate, Imbruvica (ibrutinib), Imfinzi (durvalumab), imiquimod, Imlijiq (talimogene laherparepvec), Inlyta (axitinib), inotuzumab ozogamicin, interferon alfa-2b, recombinant, interleukin-2 (aldesleukin), Intron A (recombinant interferon alfa-2b), iodine I 131 Tositumomab and tositumomab, ipilimumab, Iressa (gefitinib), irinotecan hydrochloride, irinotecan hydrochloride liposomal, Istodax (romidepsin), ixabepilone, ixazomib citrate, Ixempra (ixabepilone), Jakavi (ruxolitinib), JEB, Jevtana (cabazitaxel), Kadcyla (Ado-trastuzumab emtansine), keoxifene (La Roxifene hydrochloride), Kepivance (palifermin), Keytruda (pembrolizumab), Kisqali (ribociclib), Kymriah (tisagenlecleucel), Kyprolis (carfilzomib), lanreotide acetate, lapatinib ditosylate, Raltruvo (olaratumab), lenalidomide, lenvatinib mesylate, Lenvima (lenvatinib mesylate), letrozole, leucovorin calcium, Leukeran (clear drug), Lorambucil), Leuprolide acetate, Leustatin (Cladribine), Levran (Aminolevulinic acid), Linfolidine (Chlorambucil), Lipodox (Doxorubicin hydrochloride liposomal), Lomustine, Lonsurf (Trifluridine and Tipiracil hydrochloride), Lupron (Leuprolide acetate), Lupron Depot (Leuprolide acetate), Lupron Depot-Ped (Leuprolide acetate), Rim Parza (olaparib), Marquibo (vincristine sulfate liposomal), Maturan (procarbazine hydrochloride), mechlorethamine hydrochloride, megestrol acetate, Mekinist (trametinib), melphalan, melphalan hydrochloride, mercaptopurine, mesna, Methnex (mesna), metazolastone (temozolomide), methotrexate, methotrexate LPF (methotrexate), methylnaltrexone bromide.Mexate (methotrexate), Mexate-AQ (methotrexate), midostaurin, mitomycin C, mitoxantrone hydrochloride, Mitozytrex (mitomycin C), MOPP, Mozobil (plelixafor), Mustargen (mechlorethamine hydrochloride), Mutamycin (mitomycin C), Myleran (busulfan), Myrosal (azacitidine), Mylotarg (gemtuzumab ozogamicin), nanoparticle paclitaxel (paclitaxel albumin-stabilized nanoparticle formulation), Navelbine (vinorelbine tartrate), necitumumab, ne Larabine, Neosar (cyclophosphamide), neratinib maleate, Nerlynx (neratinib maleate), netupitant and palonosetron hydrochloride, Neulasta (pegfilgrastim), Neupogen (filgrastim), Nexavar (sorafenib tosylate), Nilandrone (nilutamide), nilotinib, nilutamide, Ninlaro (ixazomib citrate), niraparibut tosylate monohydrate, nivolumab, Nolvadex (tamoxifen citrate), N-Plate (romiplostim), obinutuzumab, Odomzo (sonidegib) ), OEPA, ofatumumab, OFF, olaparib, olaratumab, omacetaxine mepesuxinate, Oncaspar (peguaspargase), ondansetron hydrochloride, Onibide (irinotecan hydrochloride liposomal), Ontak (denileukin diftitox), Opdivo (nivolumab), OPPA, osimertinib, oxaliplatin, paclitaxel, paclitaxel albumin-stabilized nanoparticle formulation, PAD, palbociclib, palifermin, palonosetron hydrochloride, palonosetron hydrochloride and netupitant, pamidronate disodium tumour, panitumumab, panobinostat, Paraplat (carboplatin), Paraplatin (carboplatin), pazopanib hydrochloride, PCV, PEB, pegaspargase, pegfilgrastim, peginterferon alfa-2b, PEG-Intron (peginterferon alfa-2b), pembrolizumab, pemetrexed disodium, Perjeta (pertuzumab), pertuzumab, Platinol (cisplatin), Platinol-AQ (cisplatin), plerixafor, pomalidomide, Pomalyst (pomalidomide), ponatinib hydrochloride,Portraza (necitumumab), pralatrexate, prednisone, procarbazine hydrochloride, Proleukin (aldesleukin), Pralia (denosumab), Promacta (eltrombopag olamine), propranolol hydrochloride, Provenzi (sipuleucel-T), Purintoll (mercaptopurine), Prixan (mercaptopurine), [input, None], radium-223 dichloride, raloxifene hydrochloride, ramucirumab, rasburicase, R-CHOP, R-CVP, recombinant human papillomavirus (HPV) bivalent vaccine, recombinant human papillomavirus (HPV) nonvalent vaccine, recombinant human papillomavirus (HPV) quadrivalent vaccine, recombinant interferon alfa-2b, regorafenib, Relistol (methylnaltrexone bromide), R-EPOCH, Revlimid (lenalidomide), Rheumatrex (methotrexate), ribociclib, R-ICE, Rituxan (rituximab), Rituxan Nhycera (rituximab and human hyaluronidase), rituximab, rituximab and human hyaluronidase, rolapitant hydrochloride, romidepsin, romiplostim, rubidomycin (daunorubicin hydrochloride), rubraca (rucaparib camsylate), rucaparib camsylate, ruxolitinib phosphate, Ridapt (midostaurin), sclerosol intrapleural aerosol (talc), siltuximab, sipuleucel-T, somatuline depot (lanreotide acetate), sonidegib, sorafenib tosylate, Sprycel (dasatinib), STANFORD V, Sterile Talc Powder (Talc), Steritalc (Talc), Stivarga (Regorafenib), Sunitinib Malate, Sutent (Sunitinib Malate), Silatron (Peginterferon alfa-2b), Silvant (Siltuximab), Synribo (Omacetaxine Mepesuccinate), Tabloid (Thioguanine), TAC, Tafinlar (Dabrafenib), Tagrisso (Osimertinib), Talc, Talimogene Laherparepvec, Tamoxifen Citrate, Tarabin PFS (Cytarabine) , Tarceva (erlotinib hydrochloride), Targretin (bexarotene), Tasigna (nilotinib), Taxol (paclitaxel), Taxotere (docetaxel), Tecentriq (atezolizumab), Temodar (temozolomide), temozolomide, temsirolimus, thalidomide, Thalomid (thalidomide), thioguanine, thiotepa, tisagenlecleucel, Tolak (fluorouracil-topical), topotecan hydrochloride, toremifene, Tolicel (temsirolimus), tositumomab, and iodine I131 Tositumomab, Totect (dexrazoxane hydrochloride), TPF, trabectedin, trametinib, trastuzumab, Treanda (bendamustine hydrochloride), trifluridine and tipiracil hydrochloride, Trisenox (arsenic trioxide), Tykerb (lapatinib ditosylate), Unituxin (dinutuximab), uridine triacetate, VAC, valrubicin, Valstar (valrubicin), vandetanib, VAMP, Varubi (rolapitant hydrochloride), Vectibix (panitumumab), VeIP, Velban (vinblastine) sulfate), Velcade (bortezomib), Versal (vinblastine sulfate), vemurafenib, Venclexta (venetoclax), venetoclax, Verzenio (abemaciclib), Viadur (leuprolide acetate), Vidaza (azacitidine), vinblastine sulfate, Vincasal PFS (vincristine sulfate), vincristine sulfate, vincristine sulfate liposomal, vinorelbine tartrate, VIP, vismodegib, Vistogard (uridine triacetate), voraxazepam ( Voraxaze) (glucarpidase), vorinostat, Votrient (pazopanib hydrochloride), Vixeos (daunorubicin hydrochloride and cytarabine liposomal), Wellcovorin (leucovorin calcium), Xalkori (crizotinib), Xeloda (capecitabine), XELIRI, XELOX, Xgeva (denosumab), Xofigo (radium-223 dichloride), Xtandi (enzalutamide), Yervoy (ipilimumab), Yescarta (axicabtagene siloxane), Yondelis (trabectedin), Zaltrap ( The drug may be selected from Ziv-aflibercept, Zarxio (filgrastim), Zedula (niraparibut tosylate monohydrate), Zelboraf (vemurafenib), Zevalin (ibritumomab tiuxetan), Zinecard (dexrazoxane hydrochloride), Ziv-aflibercept, Zofran (ondansetron hydrochloride), Zoladex (goserelin acetate), zoledronic acid, Zolinza (vorinostat), Zometa (zoledronic acid), Zydelig (idelalisib), Zykadia (ceritinib), and Zytiga (abiraterone acetate).

[0252] subject The subject according to the embodiments described herein can be any animal or human. The subject is preferably a mammal, more preferably a human. The subject can be a non-human mammal, but more preferably a human. The subject can be male or female. The subject can be a patient. The subject may have been diagnosed with a disease or condition (e.g., cancer, e.g., a cancer described herein) for which treatment is required, may be suspected of having such a disease / condition, or may be at risk of developing / suffering from such a disease / condition.

[0253] In some embodiments, the subject treated according to the therapeutic or prophylactic methods disclosed herein is a subject who has or is at risk of developing cancer, e.g., a cancer described herein. In embodiments according to the present disclosure, a subject may be selected for treatment according to the method based on their characterization for certain markers of such disease / condition.

[0254] In some embodiments, patients may be selected for the treatments described herein based, for example, on the detection of a cancer that expresses / overexpresses HER3 and / or EGFR in a sample obtained from the subject (e.g., a biopsy of a tumor).

[0255] kit The present disclosure also provides kits of parts. Kits according to the present disclosure may include components for performing all or part of the methods described herein.

[0256] The kit can have at least one container having a predetermined amount of a combination or composition described herein. In some aspects of the present disclosure, a kit of parts is provided.In some embodiments, the kit may include an antigen-binding molecule that binds to HER3 as described herein and an antigen-binding molecule that binds to EGFR as described herein.The antigen-binding molecule that binds to HER3 and the antigen-binding molecule that binds to EGFR may be provided in a predetermined amount.The antigen-binding molecule that binds to HER3 and the antigen-binding molecule that binds to EGFR may be provided in separate containers or in the same container.

[0257] In some embodiments, the kit comprises a pharmaceutical combination or a pharmaceutical composition according to the present disclosure. The kit may be provided together with instructions for administering the antigen-binding molecule that binds to HER3, the antigen-binding molecule that binds to EGFR, the pharmaceutical combination, or the pharmaceutical composition to a patient to treat a particular disease / condition (e.g., a disease / condition described herein, e.g., cancer).

[0258] The kit may further comprise the reagents, buffers and / or standards required for carrying out the method according to the present disclosure.The kit according to the present disclosure may comprise instructions for use, for example, in the form of a manual or leaflet.The instructions may comprise a protocol for carrying out any one or more of the methods described herein.

[0259] Sequence identity As used herein, "sequence identity" refers to the percentage of nucleotides / amino acid residues in a subject sequence that are identical to nucleotides / amino acid residues in a reference sequence, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity between the sequences. Pairwise multiple sequence alignment for determining the percent sequence identity between two or more amino acid or nucleic acid sequences can be achieved by various means known to those skilled in the art, for example, using commercially available computer software such as ClustalOmega (Soding, J. 2005, Bioinformatics 21, 951-960), T-coffee (Notredame et al., 2000, J. Mol. Biol. (2000), 302, 205-217), Kalign (Lassmann and Sonnhammer 2005, BMC Bioinformatics, 6 (298)), and MAFFT (Katoh and Standley 2013, Molecular Biology and Evolution, 30 (4), 772-780) software. When using such software, it is preferable to use default parameters, for example, gap penalties and extension penalties.

[0260] array

[0261] [Table 3-1]

[0262] [Table 3-2]

[0263] [Table 3-3]

[0264] [Table 3-4]

[0265] [Table 3-5]

[0266] [Table 3-6]

[0267] [Table 3-7]

[0268] [Table 3-8]

[0269] [Table 3-9] The present disclosure includes combinations of the described embodiments and preferred features unless such combinations are expressly disallowed or explicitly avoided.

[0270] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described. Aspects and embodiments of the present disclosure will now be illustrated, by way of example, with reference to the accompanying drawings. Further aspects and embodiments will be apparent to those skilled in the art. All documents mentioned in this text are incorporated herein by reference.

[0271] Throughout this specification, including the claims which follow, unless the context requires otherwise, the word "comprise", and variations such as "comprises" and "comprising", will be understood to imply the inclusion of a stated integer or step or group of integers or steps, and not the exclusion of any other integer or step or group of integers or steps.

[0272] It should be noted that, as used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. As used herein, ranges may be expressed as ranging from "about" one particular value and / or to "about" another particular value. When such a range is expressed, another embodiment includes the range from the one particular value and / or to the other particular value. Similarly, when values ​​are expressed as approximations, by use of the antecedent "about," it will be understood that the particular value forms another embodiment.

[0273] When a nucleic acid sequence is disclosed or referred to herein, its reverse complement is also expressly contemplated. The methods described herein may preferably be performed in vitro. The term "in vitro" is intended to encompass procedures performed on cells in culture, while the term "in vivo" is intended to encompass procedures on / on intact multicellular organisms.

[0274] Embodiments and experiments illustrating the principles of the present disclosure will now be discussed with reference to the accompanying figures. [Brief explanation of the drawings]

[0275] [Figure 1A]Graphs showing the effects of treating mice bearing a KYSE-150 cell-derived mouse model of esophageal squamous cell carcinoma with 10D1F (HMBD-001), cetuximab, 10D1F in combination with cetuximab, or vehicle control (PBS). Figure 1A shows the tumor volume over time for mice in different treatment groups. Figure 1B shows the body weight over time for mice in different treatment groups. [Figure 1B] Graphs showing the effects of treating mice bearing a KYSE-150 cell-derived mouse model of esophageal squamous cell carcinoma with 10D1F (HMBD-001), cetuximab, 10D1F in combination with cetuximab, or vehicle control (PBS). Figure 1A shows the tumor volume over time for mice in different treatment groups. Figure 1B shows the body weight over time for mice in different treatment groups. [Figure 2A] 2A and 2B are graphs showing the effect of treating mice with 10D1F (HMBD-001), cetuximab, 10D1F in combination with cetuximab, or vehicle control (PBS) in an OE21 cell-derived mouse model of esophageal squamous cell carcinoma. Figure 2A shows the tumor volume over time for mice in different treatment groups. Figure 2B shows the body weight over time for mice in different treatment groups. [Figure 2B] 2A and 2B are graphs showing the effect of treating mice with 10D1F (HMBD-001), cetuximab, 10D1F in combination with cetuximab, or vehicle control (PBS) in an OE21 cell-derived mouse model of esophageal squamous cell carcinoma. Figure 2A shows the tumor volume over time for mice in different treatment groups. Figure 2B shows the body weight over time for mice in different treatment groups. [Figure 3A] 3A and 3B are graphs showing the effect of treating mice with 10D1F (HMBD-001), cetuximab, 10D1F in combination with cetuximab, or vehicle control (PBS) in a LIM1215 cell-derived mouse model of colon adenocarcinoma. Figure 3A shows the tumor volume over time for mice in different treatment groups. Figure 3B shows the body weight over time for mice in different treatment groups. [Figure 3B]3A and 3B are graphs showing the effect of treating mice with 10D1F (HMBD-001), cetuximab, 10D1F in combination with cetuximab, or vehicle control (PBS) in a LIM1215 cell-derived mouse model of colon adenocarcinoma. Figure 3A shows the tumor volume over time for mice in different treatment groups. Figure 3B shows the body weight over time for mice in different treatment groups. [Figure 4A] 4A and 4B are graphs showing the effects of treatment of mice bearing a CAL-27 cell-derived mouse model of tongue squamous cell carcinoma with 10D1F (HMBD-001), cetuximab, 10D1F in combination with cetuximab, or vehicle control (PBS). Figure 4A shows tumor volume over time for mice in different treatment groups. Figure 4B shows body weight over time for mice in different treatment groups. [Figure 4B] 4A and 4B are graphs showing the effects of treatment of mice bearing a CAL-27 cell-derived mouse model of tongue squamous cell carcinoma with 10D1F (HMBD-001), cetuximab, 10D1F in combination with cetuximab, or vehicle control (PBS). Figure 4A shows tumor volume over time for mice in different treatment groups. Figure 4B shows body weight over time for mice in different treatment groups. [Figure 5A] 5A and 5B are graphs showing the effect of treating mice with 10D1F (HMBD-001), cetuximab, 10D1F combined with cetuximab, or vehicle control (PBS) in a FaDu cell-derived mouse model of hypopharyngeal squamous cell carcinoma. Figure 5A shows the tumor volume over time for mice in different treatment groups. Figure 5B shows the body weight over time for mice in different treatment groups. [Figure 5B] 5A and 5B are graphs showing the effect of treating mice with 10D1F (HMBD-001), cetuximab, 10D1F combined with cetuximab, or vehicle control (PBS) in a FaDu cell-derived mouse model of hypopharyngeal squamous cell carcinoma. Figure 5A shows the tumor volume over time for mice in different treatment groups. Figure 5B shows the body weight over time for mice in different treatment groups. [Figure 6A]6A and 6B are graphs showing the effect of treating mice with 10D1F (HMBD-001), cetuximab, 10D1F in combination with cetuximab, or vehicle control (PBS) in a BxPC-3 cell-derived mouse model of pancreatic ductal adenocarcinoma. Figure 6A shows the tumor volume over time for mice in different treatment groups. Figure 6B shows the body weight over time for mice in different treatment groups. [Figure 6B] 6A and 6B are graphs showing the effect of treating mice with 10D1F (HMBD-001), cetuximab, 10D1F in combination with cetuximab, or vehicle control (PBS) in a BxPC-3 cell-derived mouse model of pancreatic ductal adenocarcinoma. Figure 6A shows the tumor volume over time for mice in different treatment groups. Figure 6B shows the body weight over time for mice in different treatment groups. [Figure 7A] 7A and 7B are graphs showing the effect of treating mice bearing a human patient-derived mouse model of lung squamous cell carcinoma (CTG-2552) with 10D1F (HMBD-001), cetuximab, 10D1F combined with cetuximab, or an isotype control. Figure 7A shows the tumor volume over time for mice in different treatment groups. Figure 7B shows the body weight over time for mice in different treatment groups. [Figure 7B] 7A and 7B are graphs showing the effect of treating mice bearing a human patient-derived mouse model of lung squamous cell carcinoma (CTG-2552) with 10D1F (HMBD-001), cetuximab, 10D1F combined with cetuximab, or an isotype control. Figure 7A shows the tumor volume over time for mice in different treatment groups. Figure 7B shows the body weight over time for mice in different treatment groups. [Figure 8A]8A and 8B are graphs showing the effect of treating mice bearing a cell line-derived xenograft (CDX) model of lung squamous cell carcinoma with 10D1F (HMBD-001), cetuximab (5 mg / kg and 10 mg / kg), docetaxel, 10D1F combined with cetuximab (5 mg / kg and 10 mg / kg), 10D1F combined with docetaxel, cetuximab (5 mg / kg and 10 mg / kg) combined with docetaxel, 10D1F combined with cetuximab (5 mg / kg and 10 mg / kg) and docetaxel, or vehicle control (PBS). Figure 8A shows tumor volume over time for mice in different treatment groups (5 mg / kg cetuximab in the cetuximab-containing treatment group). Figure 8B shows tumor volume over time for mice in different treatment groups (10 mg / kg cetuximab in the cetuximab-containing treatment group), and Figure 8C shows body weight over time for mice in different treatment groups. [Figure 8B] 8A and 8B are graphs showing the effect of treating mice bearing a cell line-derived xenograft (CDX) model of lung squamous cell carcinoma with 10D1F (HMBD-001), cetuximab (5 mg / kg and 10 mg / kg), docetaxel, 10D1F combined with cetuximab (5 mg / kg and 10 mg / kg), 10D1F combined with docetaxel, cetuximab (5 mg / kg and 10 mg / kg) combined with docetaxel, 10D1F combined with cetuximab (5 mg / kg and 10 mg / kg) and docetaxel, or vehicle control (PBS). Figure 8A shows tumor volume over time for mice in different treatment groups (5 mg / kg cetuximab in the cetuximab-containing treatment group). Figure 8B shows tumor volume over time for mice in different treatment groups (10 mg / kg cetuximab in the cetuximab-containing treatment group), and Figure 8C shows body weight over time for mice in different treatment groups. [Figure 8C]8A and 8B are graphs showing the effect of treating mice bearing a cell line-derived xenograft (CDX) model of lung squamous cell carcinoma with 10D1F (HMBD-001), cetuximab (5 mg / kg and 10 mg / kg), docetaxel, 10D1F combined with cetuximab (5 mg / kg and 10 mg / kg), 10D1F combined with docetaxel, cetuximab (5 mg / kg and 10 mg / kg) combined with docetaxel, 10D1F combined with cetuximab (5 mg / kg and 10 mg / kg) and docetaxel, or vehicle control (PBS). Figure 8A shows tumor volume over time for mice in different treatment groups (5 mg / kg cetuximab in the cetuximab-containing treatment group). Figure 8B shows tumor volume over time for mice in different treatment groups (10 mg / kg cetuximab in the cetuximab-containing treatment group), and Figure 8C shows body weight over time for mice in different treatment groups. [Example]

[0276] Example 1 Characterization of 10D1F in WO2019 / 185878A1 and WO2021 / 048274A1 A HER3-binding antibody clone designated 10D1F is described in WO2019 / 185878A1 (incorporated by reference in its entirety).

[0277] 10D1F comprises a heavy chain variable region set forth in SEQ ID NO: 36 of WO2019 / 185878A1 (=SEQ ID NO: 33 of the present disclosure) and a light chain variable region set forth in SEQ ID NO: 83 of WO2019 / 185878A1 (=SEQ ID NO: 58 of the present disclosure). 10D1F is also referred to as "10D1_c89" in WO2019 / 185878A1 and may be referred to herein as HMBD-001.

[0278] Example 2.2 of WO2019 / 185878A1 describes a molecule (molecule

[16] ) comprising the VH and VL regions of 10D1F in a human IgG1 / Vκ format (10D1F hIgG1), formed from SEQ ID NO: 206 of WO2019 / 185878A1 (= SEQ ID NO: 75 of the present disclosure) and SEQ ID NO: 207 of WO2019 / 185878A1 (= SEQ ID NO: 76 of the present disclosure).

[0279] Examples 8.1-8.3 and Figures 42-46 of WO2019 / 185878A1 show that 10D1F hIgG1 binds to human HER3 with high affinity and specificity (exhibiting no cross-reactivity with other human EGFR family members), while retaining high affinity binding to cynomolgus monkey, mouse, and rat HER3.

[0280] Example 8.6 and Figures 49A and 49B of WO2019 / 185878A1 demonstrate that 10D1F hIgG1 binds to HER3 in a ligand (NRG)-independent manner and via an epitope on HER3 that is topologically distant from the epitope bound by the anti-HER3 antibodies M-05-74 and M-08-11. Example 8.10 and Figure 78 of WO2021 / 048274A1 demonstrate that 10D1F hIgG1 binds to human HER3 with sub-picomolar affinity in the presence or absence of human NRG1.

[0281] Additionally, Example 3.5 of WO2021 / 048274A1 discloses that antibody clone 10D1 and 10D1-derived clones (including 10D1F) bind to human HER3 in the region corresponding to positions 218-235 of SEQ ID NO: 1 (i.e., SEQ ID NO: 77 of the present disclosure), and that two consensus binding site motifs have been identified within this region (shown in SEQ ID NOs: 78 and 79 of the present disclosure).

[0282] Example 4.1 and Figure 65, as well as Example 8.7 and Figure 52 of WO2019 / 185878A1, demonstrate that 10D1F hIgG1 is highly effective in inhibiting the interaction between HER3 and HER2, and does so in a dose-dependent manner. Example 8.7 and Figure 53 of WO2019 / 185878A1 show that 10D1F hIgG1 inhibits the interaction between HER3 and EGFR in a dose-dependent manner.

[0283] Example 8.8 and Figure 54 of WO2019 / 185878A1 show that 10D1F hIgG1 induces ADCC activity against cells overexpressing HER3 in a dose-dependent manner.

[0284] Example 8.9 and Figures 55, 63, and 64 of WO2019 / 185878A1 demonstrate that 10D1F hIgG1 inhibits HER3-mediated signaling in cells of HER3-expressing cancer cell lines in vitro.

[0285] Example 11 and Figure 71 of WO2019 / 185878A1 show that 10D1F hIgG1 also inhibits HER3-mediated signaling in xenograft tumors derived from human cancer cell lines that express HER3 in vivo. Example 14 and Figure 79 of WO2021 / 048274A1 demonstrate that 10D1F is highly effective in inhibiting the growth of xenograft tumors derived from human cancer cell lines harboring NRG gene fusions.

[0286] Examples 9.3 and 9.4 and Figures 59, 60, 61, 62, 74, and 77 of WO2019 / 185878A1 demonstrate that 10D1F potently inhibits the growth of cancer cells in vitro and the growth of xenograft tumors derived from human cancer cell lines in vivo. Example 10 and Figures 67 and 68 of WO2019 / 185878 A1 demonstrate that 10D1F hIgG1 inhibits the in vitro growth of thyroid cancer cell lines harboring the V600E BRAF mutation.

[0287] Example 12 and Figures 72 and 73 of WO2019 / 185878A1 show that 10D1F hIgG1 is not substantially internalized by cells expressing HER3.

[0288] Example 13 and Figures 75 and 76 of WO2019 / 185878A1 demonstrate the utility of 10D1F hIgG1 for the detection of HER3. Example 8.4 and Figure 47A of WO2019 / 185878A1 show that 10D1F hIgG1 is thermostable, with a melting temperature of 70.0°C as determined by differential scanning fluorimetry.

[0289] Examples 9.1 and 9.2 and Figures 56, 57, 58, and 69, 70 of WO2019 / 185878A1 demonstrate that 10D1F hIgG1 has an advantageous pharmacological and toxicological profile.

[0290] Example 2 Evaluation of the therapeutic effect of combination treatment using 10D1F and cetuximab The therapeutic effect of the combination of 10D1F hIgG1 (i.e., the antibody formed by the polypeptide consisting of SEQ ID NO: 75 and the polypeptide consisting of SEQ ID NO: 76) and cetuximab (i.e., the antibody formed by the polypeptide consisting of SEQ ID NO: 99 and the polypeptide consisting of SEQ ID NO: 100) was investigated in vivo in a variety of different cancers and in human cell and / or patient-derived xenograft models.

[0291] Mice approximately 6–8 weeks old were housed under specific pathogen-free conditions and treated in accordance with the guidelines of the Institutional Animal Care and Use Committee (IACUC). Tumors derived from human cancer cells were established by mixing cells with an equal volume of Matrigel (Corning, USA) and subcutaneously implanting them into the right or left flank of the mice, as indicated. Tumors were approximately 100–300 mm. 3Treatment was initiated when

[0292] ESCC CDX(1): Model: Human cell line-derived model of esophageal squamous cell carcinoma Cell line: KYSE-150 (CVCL_1348). KYSE-150 cells exhibit high expression of NRG1 and HER3 (Meetze et al., Clinical Cancer Research (2015) 21(5):1106-1114).

[0293] 1 × 10 7 The model was established by injecting cells. Treatment Group: Vehicle control (phosphate buffered saline; IP injection, once weekly; n=10).

[0294] HMBD-001 (20 mg / kg body weight IP injection, once weekly; n = 10). cetuximab (10 mg / kg body weight IP injection, once weekly; n = 10). HMBD-001 + cetuximab (IP injection of 20 mg / kg body weight HMBD-001 + 10 mg / kg body weight cetuximab, once weekly; n = 10).

[0295] ESCC CDX(2): Model: Human cell line-derived model of esophageal squamous cell carcinoma Cell line: OE21 (CVCL_2661). OE21 cells express high levels of EGFR dimers (Fichter et al., Int J Cancer (2014) 135(7):1517-1530).

[0296] 1 × 10 6 The model was established by injecting cells. Treatment Group: Vehicle control (phosphate buffered saline; IP injection, twice weekly; n=10).

[0297] HMBD-001 (20 mg / kg body weight IP injection twice weekly; n = 10). cetuximab (10 mg / kg body weight IP injection twice weekly; n = 10). HMBD-001 + cetuximab (IP injection of 20 mg / kg body weight HMBD-001 + 10 mg / kg body weight cetuximab twice weekly; n = 10).

[0298] CRC CDX: Model: Human cell line-derived model of colon adenocarcinoma Cell line: LIM1215 (CVCL_2574). LIM1215 cells express normal levels of EGFR but are sensitive to cetuximab (Misale et al., Nature, (2012) 486(7404):532-536).

[0299] 2 × 10 6 The model was established by injecting cells. Treatment Group: Vehicle control (phosphate buffered saline; IP injection, twice weekly; n=10).

[0300] HMBD-001 (20 mg / kg body weight IP injection twice weekly; n = 11). cetuximab (10 mg / kg body weight IP injection twice weekly; n = 12). HMBD-001 + cetuximab (IP injection of 20 mg / kg body weight HMBD-001 + 10 mg / kg body weight cetuximab twice weekly; n = 12).

[0301] HNSCC CDX(1): Model: Human cell line-derived model of tongue squamous cell carcinoma Cell line: CAL-27 (CVCL_1107). CAL-27 cells overexpress EGFR (Licitra et al., Ann Oncol (2011) 22(8):1886-1893).

[0302] 5 × 10 6 The model was established by injecting cells. Treatment Group: Vehicle control (phosphate buffered saline; IP injection, twice weekly; n=8).

[0303] HMBD-001 (20 mg / kg body weight IP injection twice weekly; n = 8). cetuximab (10 mg / kg body weight IP injection twice weekly; n = 8). HMBD-001 + cetuximab (IP injection of 20 mg / kg body weight HMBD-001 + 10 mg / kg body weight cetuximab twice weekly; n = 8).

[0304] HNSCC CDX(2): Model: Human cell line-derived model of hypopharyngeal squamous cell carcinoma Cell line: FaDu (CVCL_1218). FaDu cells exhibit high expression of EGFR and NRG1 (Xiao et al., Mol Cancer Ther (2016) 15(4):689-701).

[0305] 1 × 10 injected into the right flank of NCr nude mice 6 The model was established by injecting cells. Treatment Group: Vehicle control (phosphate buffered saline; IP injection, twice weekly; n=10).

[0306] HMBD-001 (20 mg / kg body weight IP injection twice weekly; n = 10). cetuximab (10 mg / kg body weight IP injection twice weekly; n = 10). HMBD-001 + cetuximab (IP injection of 20 mg / kg body weight HMBD-001 + 10 mg / kg body weight cetuximab twice weekly; n = 10).

[0307] PDAC CDX: Model: Human cell line-derived model of pancreatic ductal adenocarcinoma Cell line: BxPC-3 (CVCL_0186).

[0308] 5 × 10 6 The model was established by injecting cells. Treatment Group: Vehicle control (phosphate buffered saline; IP injection, twice weekly; n=11).

[0309] HMBD-001 (20 mg / kg body weight IP injection twice weekly; n = 11). cetuximab (10 mg / kg body weight IP injection twice weekly; n = 11). HMBD-001 + cetuximab (IP injection of 20 mg / kg body weight HMBD-001 + 10 mg / kg body weight cetuximab twice weekly; n = 11).

[0310] LUSC PDX: Model: Human patient-derived model of lung squamous cell carcinoma (CTG-2552) Athymic, Foxn1 nu This model was established by implanting 3-4 tumor fragments into the left flank of nude mice.

[0311] Treatment Group: Isotype control (human IgG1 kappa isotype control; 20 mg / kg body weight IP injection, once weekly; n=10).

[0312] HMBD-001 (20 mg / kg body weight IP injection, once weekly; n = 10). cetuximab (10 mg / kg body weight IP injection, once weekly; n = 10). HMBD-001 + cetuximab (IP injection of 20 mg / kg body weight HMBD-001 + 10 mg / kg body weight cetuximab, once weekly; n = 10).

[0313] Tumor volumes were measured twice weekly using digital calipers and calculated using the formula [L × W × W] / 2. The mean tumor volume in the control group was 1500 mm 3 The study endpoint was reached when the mice reached 100 mg / kg body weight. The body weight of the mice was also monitored.

[0314] The experimental results are shown in Figures 1 to 7. Combination treatment with HMBD-001 and cetuximab achieved greater tumor growth inhibition than either agent alone in CDX models of ESCC, CRC, HNSCC, and PDAC, as well as in LUSC PDX models.

[0315] Discussion—The anti-HER3 antibody 10D1F in combination with an EGFR inhibitor effectively inhibits tumor growth in biomarker-selected preclinical models of squamous cell carcinoma. Squamous cell carcinoma (SCC) originates from stratified squamous epithelium in various anatomical sites, sharing common histological features and genetic mutations. Nonmelanoma skin cancer, head and neck cancer (HNSCC), esophageal cancer (ESCC), and non-small cell lung cancer (sqNSCLC) account for the majority of SCC cases and together represent the largest subtype of cancer. After progression of standard therapies, there is a significant unmet need for effective treatment options. Alterations in receptor tyrosine kinase signaling pathways are commonly found in SCC; for example, EGFR is expressed in 90% of HNSCC, 76% of ESCC, and 82% of sqNSCLC. However, the combination of the EGFR-targeting antibody cetuximab with chemotherapy has provided limited clinical benefit, and even initially sensitive tumors often develop resistance. Studies in SCC suggest that HER3, the primary dimerization partner of HER family members, may be one of the causes of treatment failure due to HER3 heterodimer activation of the PI3K / AKT and MAPK / ERK pathways. In this study, we investigated the possibility of combining HER3 and EGFR inhibitors to improve efficacy and overcome resistance.

[0316] Previous attempts to target HER3 have shown limited clinical efficacy due to suboptimal HER3 inhibition, lack of efficacy, particularly in ligand-independent activation mechanisms, and a lack of biomarker stratification for patients who would benefit from HER3 inhibition. 10D1F is a clinical-stage anti-HER3 antibody that is being evaluated in a first-in-human, open-label, multicenter, dose-escalation, and expansion Phase I / IIa study in patients with HER3-expressing advanced solid tumors (NCT05057013). 10D1F was rationally developed to uniquely block the HER3 heterodimerization interface to potently inhibit all HER3 dimer formation, including ligand-dependent and -independent HER3 dimerization.

[0317] Here, we demonstrate that dual blockade of EGFR and HER3 using cetuximab and 10D1F, respectively, robustly inhibits tumor growth and is superior to either monotherapy. In multiple cell- and patient-derived SCC xenograft models, including models of HNSCC, ESCC, and sqNSCLC, selected based on a novel gene signature that robustly predicts response, 10D1F in combination with cetuximab demonstrated up to 100% tumor growth inhibition and abrogation of PI3K signaling. The combination was well tolerated, with no recurrences observed in selected models, even after chronic treatment for more than 100 days.

[0318] Example 3 Evaluation of the therapeutic efficacy of combination treatment using 10D1F, cetuximab, and docetaxel The therapeutic effect of a combination of 10D1F hIgG1 (i.e., an antibody formed by a polypeptide consisting of SEQ ID NO: 75 and a polypeptide consisting of SEQ ID NO: 76), cetuximab (i.e., an antibody formed by a polypeptide consisting of SEQ ID NO: 99 and a polypeptide consisting of SEQ ID NO: 100), and docetaxel was investigated in vivo in a cell line-derived xenograft (CDX) model of squamous cell carcinoma of the lung (LUSC). Cell line: HARA (KCC-C1). HARA cells exhibit high expression of parathyroid hormone-related protein (PTHrP) and interleukin-1 (IL-1) (Ichinose et al., Cancer Letters (1993) 74(1-2):119-124).

[0319] Mice approximately 6–8 weeks old were housed under specific pathogen-free conditions and treated in accordance with the guidelines of the Institutional Animal Care and Use Committee (IACUC). For tumors derived from human cell lines, cells were mixed with an equal volume of Matrigel (Corning, USA) and inoculated into the right flank of the mice at 2 × 10 6 The tumors were established by subcutaneous transplantation of cells. 3 Treatment was initiated when

[0320] Treatment Group: Vehicle control (phosphate buffered saline; IP injection, once weekly; n=8). HMBD-001 (20 mg / kg body weight IP injection, once weekly; n = 8).

[0321] cetuximab (5 mg / kg body weight IP injection once weekly; n = 8). cetuximab (10 mg / kg body weight IP injection once weekly; n = 8). docetaxel (10 mg / kg body weight IP injection once weekly; n = 8).

[0322] HMBD-001 + cetuximab (IP injection of 20 mg / kg body weight HMBD-001 + 5 mg / kg body weight cetuximab once weekly; n = 8). HMBD-001 + cetuximab (IP injection of 20 mg / kg body weight HMBD-001 + 10 mg / kg body weight cetuximab once weekly; n = 8).

[0323] HMBD-001 + docetaxel (IP injection of 20 mg / kg body weight HMBD-001 + 10 mg / kg body weight docetaxel, once weekly; n = 8). cetuximab plus docetaxel (IP injection of cetuximab at 5 mg / kg body weight plus docetaxel at 10 mg / kg body weight, once weekly; n = 8).

[0324] cetuximab plus docetaxel (IP injection of 10 mg / kg body weight cetuximab plus 10 mg / kg body weight docetaxel once weekly; n = 8). HMBD-001 + cetuximab + docetaxel (IP injection of 20 mg / kg body weight HMBD-001 + 5 mg / kg body weight cetuximab + 10 mg / kg body weight docetaxel, once weekly; n = 8).

[0325] HMBD-001 + cetuximab + docetaxel (IP injection of 20 mg / kg body weight HMBD-001 + 10 mg / kg body weight cetuximab + 10 mg / kg body weight docetaxel, once weekly; n = 8).

[0326] Tumor volumes were measured twice weekly using digital calipers and calculated using the formula [L × W × W) / 2]. Mouse body weights were also monitored. The experimental results are shown in Figure 8. Specifically, Figures 8A and 8B show tumor volume over time for mice in different treatment groups (8A: 5 mg / kg cetuximab in the treatment group containing cetuximab; 8B: 10 mg / kg cetuximab in the treatment group containing cetuximab). Figure 8C shows body weight over time for mice in different treatment groups.

[0327] Combination treatment with HMBD-001, cetuximab, and docetaxel achieved greater tumor growth inhibition in the LUSC CDX model than treatment with either agent alone or any combination of the two agents. Indeed, at both dose levels of cetuximab (5 mg / kg and 10 mg / kg), combination treatment with HMBD-001, cetuximab, and docetaxel achieved an overall reduction in tumor volume 40 days after initial treatment, which was not achieved in any other treatment group. The combination was well tolerated, as indicated by mouse body weight over the treatment period (Figure 8C).

[0328] Discussion—The anti-HER3 antibody 10D1F in combination with an EGFR inhibitor and a taxane (docetaxel) effectively inhibits tumor growth in a cell line-derived xenograft model of lung squamous cell carcinoma (LUSC).

[0329] In this study, we investigated the possibility of combining HER3 and EGFR inhibitors with a taxane (docetaxel) to improve efficacy. Here, we demonstrate that the addition of docetaxel to dual EGFR and HER3 blockade using cetuximab and 10D1F, respectively, robustly inhibited tumor growth and was superior to monotherapy with any of the three drugs individually and any combination therapy using two of the three drugs. Indeed, at both cetuximab dose levels (5 mg / kg and 10 mg / kg), combined treatment with HMBD-001, cetuximab, and docetaxel achieved an overall reduction in tumor volume 40 days after initial treatment, which was not achieved with any other treatment group. In a cell line-derived LUSCUS xenograft model, 10D1F in combination with cetuximab and docetaxel demonstrated greater than 100% tumor growth inhibition and a reduction in overall tumor volume 40 days after initial treatment. This combination was well tolerated, and no recurrences were observed.

Claims

1. An antigen-binding molecule that binds to HER3 for use in a method for treating or preventing cancer, the method comprising the step of administering an antigen-binding molecule that binds to EGFR, wherein the antigen-binding molecule that binds to HER3 binds to a region of HER3 set forth in SEQ ID NO:

77.

2. Use of an antigen-binding molecule that binds to HER3 in the manufacture of a medicament for use in a method for treating or preventing cancer, the method comprising the step of administering an antigen-binding molecule that binds to EGFR, wherein the antigen-binding molecule that binds to HER3 binds to a region of HER3 set forth in SEQ ID NO:

77.

3. A method for treating or preventing cancer, comprising the step of administering to a subject therapeutically or prophylactically effective amounts of (i) an antigen-binding molecule that binds to HER3 and (ii) an antigen-binding molecule that binds to EGFR, wherein the antigen-binding molecule that binds to HER3 binds to a region of HER3 set forth in SEQ ID NO:

77.

4. 4. The antigen-binding molecule for use, use, or method according to any one of claims 1 to 3, wherein the method for treating or preventing cancer further comprises the step of administering docetaxel.

5. A pharmaceutical combination comprising an antigen-binding molecule that binds to HER3 and an antigen-binding molecule that binds to EGFR; wherein the antigen-binding molecule that binds to HER3 binds to a region of HER3 set forth in SEQ ID NO:

77.

6. 6. The pharmaceutical combination of claim 5 for use in a method for treating or preventing cancer.

7. 10. Use of the pharmaceutical combination of claim 5 in the manufacture of a medicament for use in a method of treating or preventing cancer.

8. A method for treating or preventing cancer, comprising administering to a subject a therapeutically or prophylactically effective amount of the pharmaceutical combination of claim 5.

9. 9. The pharmaceutical combination, use or method for use according to any one of claims 6 to 8, wherein the method for treating or preventing cancer further comprises the step of administering docetaxel.

10. An antigen-binding molecule that binds to HER3, (i) the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO: 40 HC-CDR2 having the amino acid sequence of SEQ ID NO: 43 HC-CDR3 having the amino acid sequence of SEQ ID NO: 48 a heavy chain variable (VH) region incorporating (ii) the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 66 LC-CDR2 having the amino acid sequence of SEQ ID NO: 69 LC-CDR3 having the amino acid sequence of SEQ ID NO: 74 A light chain variable (VL) region incorporating 10. The antigen-binding molecule for use, use, or method according to claim 1 , comprising:

11. An antigen-binding molecule that binds to HER3, (i) the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO: 38 HC-CDR2 having the amino acid sequence of SEQ ID NO: 42 HC-CDR3 having the amino acid sequence of SEQ ID NO: 45 a VH region incorporating (ii) the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 63 LC-CDR2 having the amino acid sequence of SEQ ID NO: 67 LC-CDR3 having the amino acid sequence of SEQ ID NO: 70 A VL region incorporating 11. The antigen-binding molecule for use, use, or method according to claim 1 , comprising:

12. An antigen-binding molecule that binds to HER3, a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 33; and A VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:

58.

12. The antigen-binding molecule for use, use, or method according to claim 1 , comprising:

13. An antigen-binding molecule that binds to HER3, A polypeptide comprising or consisting of an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 75, and A polypeptide comprising or consisting of an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:

76.

13. The antigen-binding molecule for use, use, or method according to any one of claims 1 to 12, comprising:

14. The antigen-binding molecule that binds to EGFR is (i) the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO: 92 HC-CDR2 having the amino acid sequence of SEQ ID NO:93 HC-CDR3 having the amino acid sequence of SEQ ID NO:94 a heavy chain variable (VH) region incorporating (ii) the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 96 LC-CDR2 having the amino acid sequence of SEQ ID NO: 97 LC-CDR3 having the amino acid sequence of SEQ ID NO: 98 A light chain variable (VL) region incorporating 14. The antigen-binding molecule for use, use, or method according to any one of claims 1 to 13, comprising:

15. The antigen-binding molecule that binds to EGFR is a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 91; and A VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:

95.

15. The antigen-binding molecule for use, use, or method according to any one of claims 1 to 14, comprising:

16. The antigen-binding molecule that binds to EGFR is A polypeptide comprising or consisting of an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:99; and A polypeptide comprising or consisting of an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:

100.

16. The antigen-binding molecule for use, use, or method according to any one of claims 1 to 15, comprising:

17. The cancer is selected from the group consisting of cancers comprising cells that express / overexpress an EGFR family member, cancers comprising cells that express / overexpress HER3, cancers comprising cells that express / overexpress EGFR, cancers comprising cells that express / overexpress HER3 and EGFR, cancers comprising cells with a mutation that results in increased expression of a ligand for HER3, cancers comprising cells with a mutation that results in increased expression of a ligand for EGFR, cancers comprising cells with an NRG gene fusion, solid cancers, blood cancers, squamous cell carcinoma, EGFR-amplified squamous cell carcinoma, breast cancer, breast cancer, invasive breast cancer, ductal carcinoma, metastatic breast cancer, triple-negative breast cancer, HER2-positive breast cancer, gastric cancer, gastric adenocarcinoma, gastrointestinal adenocarcinoma, colorectal cancer, metastatic colorectal cancer, colon cancer, colorectal adenocarcinoma, colon adenocarcinoma, head and neck cancer, head and neck squamous cell carcinoma, lung cancer, non-small cell lung cancer, lung adenocarcinoma, invasive mucinous lung adenocarcinoma, lung Squamous cell carcinoma, ovarian cancer, ovarian cancer, serous ovarian adenocarcinoma, serous ovarian cystadenocarcinoma, fallopian tube cancer, renal cancer, renal cell carcinoma, renal clear cell carcinoma, renal cell adenocarcinoma, papillary renal cell carcinoma, pancreatic cancer, pancreatic adenocarcinoma, pancreatic ductal adenocarcinoma, cervical cancer, cervical squamous cell carcinoma, skin cancer, melanoma, oral cancer, oropharyngeal cancer, esophageal cancer, esophageal squamous cell carcinoma, esophageal adenocarcinoma, liver cancer, hepatocellular carcinoma, bile duct cancer, gallbladder cancer, uterine cancer, endometrial cancer, endometrial cancer, uterine carcinosarcoma, thyroid cancer, thyroid cancer, 17. The antigen-binding molecule for use, use, or method according to any one of claims 1 to 16, wherein the antigen-binding molecule is selected from: bladder cancer, bladder urothelial carcinoma, prostate cancer, prostate adenocarcinoma, retinoblastoma, sarcoma, soft tissue sarcoma, peritoneal carcinoma, thymoma, neuroendocrine tumor, nasopharyngeal neuroendocrine tumor, squamous cell carcinoma of the skin, astrocytoma, low-grade astrocytoma, high-grade astrocytoma, anaplastic astrocytoma, and glioblastoma multiforme.

18. 18. The antigen-binding molecule for use, use, or method according to any one of claims 1 to 17, wherein the cancer is selected from cancer comprising cells that express / overexpress HER3, cancer comprising cells that express / overexpress EGFR, cancer comprising cells that express / overexpress HER3 and EGFR, squamous cell carcinoma, EGFR-amplified squamous cell carcinoma, esophageal cancer, esophageal squamous cell carcinoma, head and neck cancer, head and neck squamous cell carcinoma, colorectal cancer, metastatic colorectal cancer, colon adenocarcinoma, pancreatic cancer, pancreatic adenocarcinoma, pancreatic ductal adenocarcinoma, lung cancer, and lung squamous cell carcinoma.

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