Treatment and prevention of cancer using HER3 antigen-binding molecules

Abstract

The present disclosure provides the use of an antigen-binding molecule that binds to HERS for the treatment or prevention of cancer comprising (i) at least one gene that encodes a positive regulator of HERS-mediated signaling that does not comprise an activating mutation, or (ii) at least one gene that encodes a negative regulator of HERS-mediated signaling that does not comprise an inactivating mutation.

Description

[Technical field]

[0001] This application claims priority from US63 / 232,883, filed August 13, 2021, the contents and elements of which are incorporated herein by reference for all purposes.

[0002] FIELD OF THEINVENTION The present disclosure relates to the field of molecular biology, and more specifically to antibody technology and methods of medical treatment and prevention. [Background technology]

[0003] HER3 activation has emerged as a key mechanism for both tumor progression and acquired resistance to standard therapies in multiple indications. HER3-targeting approaches to date have not demonstrated the expected clinical efficacy. Suboptimal inhibition of HER3-mediated signaling is one possible explanation. Summary of the Invention [Means for solving the problem]

[0004] In a first aspect, the disclosure provides an antigen-binding molecule that binds HER3 for use in a method of treating or preventing a HER3-associated cancer in a subject, where the HER3-associated cancer (i) comprises at least one gene encoding a positive regulator of HER3-mediated signaling that does not comprise an activating mutation; or (ii) comprises at least one gene encoding a negative regulator of HER3-mediated signaling that does not comprise an inactivating mutation.

[0005] Also provided is the use of an antigen binding molecule that binds HER3 in the manufacture of a medicament for use in treating or preventing a HER3-associated cancer in a subject, where the HER3-associated cancer (i) contains at least one gene encoding a positive regulator of HER3-mediated signaling that does not contain an activating mutation; or (ii) contains at least one gene encoding a negative regulator of HER3-mediated signaling that does not contain an inactivating mutation.

[0006] Also provided is a method of treating or preventing a HER3-associated cancer in a subject, the method comprising administering to the subject a therapeutically or prophylactically effective amount of an antigen binding molecule that binds to HER3, wherein the HER3-associated cancer (i) contains at least one gene encoding a positive regulator of HER3-mediated signaling that does not contain an activating mutation; or (ii) contains at least one gene encoding a negative regulator of HER3-mediated signaling that does not contain an inactivating mutation.

[0007] In some embodiments according to various aspects of the disclosure, the HER3-associated cancer (i) does not contain an activating mutation in a gene selected from KRAS, PIK3CA, PIK3CB, PIK3CD, ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GRB2, GAB2, PTPN11, SHP2, SOS1, HRAS, NRAS, RAF1, MAP2K1, MAP2K2, MAPK1, MYC, RPS6KA1, RPS6, MKNK1, CREB1, MTOR, PDK1, AKT1, AKT2, AKT3, JAK2, STAT3, STAT5, and BRAF; or (ii) does not contain an inactivating mutation in a gene selected from PTEN, PPP2CA, PIK3R1, PIK3R2, NF1, BAD, and PHLPP1. In some embodiments according to various aspects of the disclosure, the HER3-associated cancer (i) does not contain an activating mutation in a gene selected from KRAS, PIK3CA, PIK3CB, PIK3CD, ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GRB2, GAB2, PTPN11, SHP2, SOS1, HRAS, NRAS, RAF1, MAP2K1, MAP2K2, MAPK1, MYC, RPS6KA1, RPS6, MKNK1, CREB1, MTOR, PDK1, AKT1, AKT2, AKT3, JAK2, STAT3, and STAT5; or (ii) does not contain an inactivating mutation in a gene selected from PTEN, PPP2CA, PIK3R1, PIK3R2, NF1, BAD, and PHLPP1.

[0008] In some embodiments, the HER3-associated cancer does not (i) contain an activating mutation for KRAS; or (ii) contain an activating mutation for PIK3CA; or (iii) contain an activating mutation for BRAF; or (iv) contain an inactivating mutation for PTEN. In some embodiments, the HER3-associated cancer does not (i) contain an activating mutation for KRAS; or (ii) contain an activating mutation for PIK3CA; or (iii) contain an inactivating mutation for PTEN.

[0009] In some embodiments, the HER3-associated cancer is characterized as having (i) no activating mutations for KRAS and no activating mutations for PIK3CA; or (ii) no activating mutations for KRAS and no inactivating mutations for PTEN; or (iii) no activating mutations for PIK3CA and no inactivating mutations for PTEN; or (iv) no activating mutations for KRAS and no activating mutations for BRAF; or (v) no activating mutations for PIK3CA and no activating mutations for BRAF; or (vi) no activating mutations for BRAF and no inactivating mutations for PTEN; or (vii) no activating mutations for KRAS and no activating mutations for PIK3CA. or (viii) does not contain activating mutations for PIK3CA, does not contain activating mutations for BRAF, and does not contain inactivating mutations for PTEN; or (ix) does not contain activating mutations for BRAF, does not contain activating mutations for KRAS, and does not contain inactivating mutations for PTEN; or (x) does not contain activating mutations for KRAS, does not contain activating mutations for PIK3CA, and does not contain inactivating mutations for PTEN; or (xi) does not contain activating mutations for KRAS, does not contain activating mutations for BRAF, does not contain activating mutations for PIK3CA, and does not contain inactivating mutations for PTEN. In some embodiments, the HER3-associated cancer (i) does not contain an activating mutation for KRAS and does not contain an activating mutation for PIK3CA; or (ii) does not contain an activating mutation for KRAS and does not contain an inactivating mutation for PTEN; or (iii) does not contain an activating mutation for PIK3CA and does not contain an inactivating mutation for PTEN; or (iv) does not contain an activating mutation for KRAS, does not contain an activating mutation for PIK3CA, and does not contain an inactivating mutation for PTEN.

[0010] In some embodiments, the HER3-associated cancer does not contain a mutation that results in upregulation of HER3-mediated signaling.

[0011] In some embodiments, a HER3-associated cancer comprises cells that express NRG1 at a level higher than the level expressed by a comparable non-cancerous cell.

[0012] Also provided is an antigen-binding molecule that binds HER3 for use in a method of treating or preventing a HER3-associated cancer in a subject, where the HER3-associated cancer contains a mutation that results in upregulation of HER3-mediated signaling, and the method further comprises the step of administering an antagonist of HER3-mediated signaling.

[0013] Also provided is the use of an antigen binding molecule that binds HER3 in the manufacture of a medicament for use in treating or preventing a HER3-associated cancer in a subject, wherein the HER3-associated cancer contains a mutation that results in upregulation of HER3-mediated signaling, and the method further comprises the step of administering an antagonist of HER3-mediated signaling.

[0014] Also provided is a method of treating or preventing a HER3-associated cancer in a subject, the method comprising administering to the subject a therapeutically or prophylactically effective amount of an antigen binding molecule that binds HER3, wherein the HER3-associated cancer comprises a mutation that results in upregulation of HER3-mediated signaling, and the method further comprises administering an antagonist of HER3-mediated signaling.

[0015] In some embodiments, the mutation that results in upregulation of HER3-mediated signaling is an activating mutation in a gene encoding a positive regulator of HER3-mediated signaling or an inactivating mutation in a gene encoding a negative regulator of HER3-mediated signaling.

[0016] In some embodiments according to various aspects of the disclosure, the HER3-associated cancer comprises (i) an activating mutation in a gene selected from KRAS, PIK3CA, PIK3CB, PIK3CD, ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GRB2, GAB2, PTPN11, SHP2, SOS1, HRAS, NRAS, RAF1, MAP2K1, MAP2K2, MAPK1, MYC, RPS6KA1, RPS6, MKNK1, CREB1, MTOR, PDK1, AKT1, AKT2, AKT3, JAK2, STAT3, STAT5, and BRAF; and / or (ii) an inactivating mutation in a gene selected from PTEN, PPP2CA, PIK3R1, PIK3R2, NF1, BAD, and PHLPP1. In some embodiments according to various aspects of the disclosure, the HER3-associated cancer comprises (i) an activating mutation in a gene selected from KRAS, PIK3CA, PIK3CB, PIK3CD, ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GRB2, GAB2, PTPN11, SHP2, SOS1, HRAS, NRAS, RAF1, MAP2K1, MAP2K2, MAPK1, MYC, RPS6KA1, RPS6, MKNK1, CREB1, MTOR, PDK1, AKT1, AKT2, AKT3, JAK2, STAT3 and STAT5; and / or (ii) an inactivating mutation in a gene selected from PTEN, PPP2CA, PIK3R1, PIK3R2, NF1, BAD and PHLPP1.

[0017] In some embodiments, the HER3-associated cancer comprises one or more of the following: an activating mutation for KRAS, an activating mutation for PIK3CA, an activating mutation for BRAF, or an inactivating mutation for PTEN. In some embodiments, the HER3-associated cancer comprises one or more of the following: an activating mutation for KRAS, an activating mutation for PIK3CA, or an inactivating mutation for PTEN.

[0018] Also provided is a method of selecting a subject for treatment with an antigen-binding molecule that binds HER3, comprising the steps of: (a) analyzing a cancer in a subject to determine whether the cancer contains: (i) at least one gene encoding a positive regulator of HER3-mediated signaling that does not contain an activating mutation; or (ii) at least one gene encoding a negative regulator of HER3-mediated signaling that does not contain an inactivating mutation; (b) if the subject's cancer is determined to not contain such a mutation in step (a), selecting the subject for treatment with an antigen binding molecule that binds HER3. A method is also provided, including:

[0019] In some embodiments, the method comprises: (c) administering to the subject selected for treatment in step (b) an antigen-binding molecule that binds HER3. Further includes:

[0020] Also provided is a method of selecting a subject for treatment with (i) an antagonist of HER3-mediated signaling and (ii) an antigen binding molecule that binds HER3, comprising: (a) analyzing a subject's cancer to determine whether the cancer contains a mutation that results in upregulation of HER3-mediated signaling; (b) if the subject's cancer is determined to contain such a mutation in step (a), selecting the subject for treatment with (i) an antagonist of HER3-mediated signaling and (ii) an antigen binding molecule that binds HER3. A method is also provided, including:

[0021] In some embodiments, the method comprises: (c) administering (i) an antagonist of HER3-mediated signaling and (ii) an antigen-binding molecule that binds HER3 to a subject selected for treatment in step (b). Further includes:

[0022] In some embodiments according to various aspects of the disclosure, the HER3 associated cancer is selected from solid tumors, breast cancer, breast cancer, ductal carcinoma, gastric cancer, gastric cancer, gastric adenocarcinoma, colorectal cancer, colorectal adenocarcinoma, head and neck cancer, head and neck squamous cell carcinoma, lung cancer, non-small cell lung cancer, lung adenocarcinoma, squamous cell lung carcinoma, ovarian cancer, ovarian cancer, serous ovarian adenocarcinoma, 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, esophageal cancer, esophageal adenocarcinoma, liver cancer, hepatocellular carcinoma, cholangiocarcinoma, uterine cancer, endometrial cancer, thyroid cancer, thyroid cancer, pheochromocytoma, paraganglioma, bladder cancer, bladder urothelial carcinoma, prostate cancer, prostate adenocarcinoma, sarcoma, and thymoma.

[0023] In some embodiments according to various aspects of the disclosure, the antigen binding molecule that binds to HER3 is selected from 10D1F, seribantumab, elgemtumab, patritumab, GSK2849330, lumletuzumab, CDX-3379, AV-203, varsetamab, TK-A3, TK-A4, MP-EV20, 1A5-3D4, 9F7-F11, 16D3-C1, NG33, A5, F4, huHER3-8, REGN1400, and xenoctuzumab.

[0024] 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 and 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.

[0025] In some embodiments, the antigen-binding molecule comprises: (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.

[0026] In some embodiments, the antigen-binding molecule comprises: 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.

[0027] In some embodiments, the antigen-binding molecule comprises: 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. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0028] The present disclosure is based on the unexpected observation by the present inventors that cancers that lack mutations in genes encoding mediators of HER3-mediated signal transduction respond exceptionally well to treatment with anti-HER3 antibodies.In particular, cancers that have homozygous wild-type genotypes for KRAS, PIK3CA and PTEN, or cancers that have homozygous wild-type genotypes for KRAS, PIK3CA, BRAF and PTEN, are highly responsive to anti-HER3 antibody treatment.

[0029] HER3 and HER3-Mediated Signaling 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 transmembrane ErbB receptor tyrosine kinase with an N-terminal extracellular region (SEQ ID NO: 9) that contains two leucine-rich subdomains (domains I and III, shown in SEQ ID NO: 15 and 17, respectively) and two cysteine-rich subdomains (domains II and IV, shown in SEQ ID NO: 16 and 18, respectively). Domain II contains a β-hairpin dimerization loop (SEQ ID NO: 19), which 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 juxtamembrane 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 "homolog" 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 homologs 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 that 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 species of the reference protein (e.g., human HER3 isoforms 1-5 are all isoforms of each other). A "homolog" 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. Homologs 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 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 can 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 a functional property / activity of a reference HER3 (e.g., human HER3 isoform 1), for example, 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 (type 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, a 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 heteromultimerization of the receptor (i.e., with other ErBB receptors, e.g., HER2, EGFR) and the resulting autophosphorylation by the protein kinase domain of tyrosine residues in the cytoplasmic region. HER3 lacks kinase activity and does not form stable homodimers. Thus, HER3 must be transphosphorylated by binding to a kinase-active heterodimeric partner (e.g., EGFR or HER2) to cause 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 a dimerization arm within domain II to allow 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 active heterodimers. The traditional model of activation is ligand-dependent, i.e., the equilibrium shifts when HER3 in the open conformation is stabilized by binding to its ligands, such as neuregulins (NRGs), e.g., NRG1 (also known as heregulin, HRG) or NRG2. In addition, the presence of either dimerization partner in sufficient concentration will transiently bind and stabilize 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 multimeric ErBB family member receptor complexes containing HER3. "Signal transduction" refers to signal transduction and other cellular processes that control cellular activity. HER3-mediated signal transduction can be mediated by HER3 receptor-containing complexes, such as heteromultimeric complexes that contain HER3 and other HER receptors (e.g., HER2, EGFR). HER3-mediated signal transduction can be ligand-dependent, e.g., caused by the binding of NRG (e.g., NRG1, NRG2), or can be ligand-independent.

[0043] HER3-mediated signaling proceeds in cells via the MAPK / ERK and PI3K / AKT / mTOR pathways to promote cell survival and proliferation. HER3-mediated signaling 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 phospho-tyrosine residues for recruiting 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, and thereby the activation of, Ras family GTPases, such as H-Ras, N-Ras, and K-Ras. 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 the translation of mRNA into protein through phosphorylation of RSK and the resulting 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 through its SH2 domain. Assembly of p85 triggers allosteric activation of the lipid kinase p100α subunit of PI3K. Activated PI3K leads to the conversion of PIP2 to PIP3, which recruits AKT to be 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 inhibition of p85 on the p110 catalytic subunit of PI3K, respectively (Cuevas et al., J Biol Chem. 2001 Jul 20;276(29):27455-6). SHP2 mediates the direct dephosphorylation of RAS (Bunda et al., Nat Commun. 2015 Nov). 30;6():8859), and has been shown to activate RAS by inhibition of 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, and the tumor-promoting effects of SHP2 are partly related to Akt signaling (Cao et al., Pathol. Res. Pract. (2019) 215:152-621).

[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 of 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 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] cancer The present disclosure relates to the treatment and prevention of cancer.

[0049] A cancer according to the present disclosure may be any unwanted cell proliferation (or any disease that manifests itself by unwanted cell proliferation), a neoplasm, or a tumor. A cancer may be benign or malignant. A 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 (and / or originate from) any organ / tissue.

[0050] The cancer can be, for example, a cancer of 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, and / or white blood cells.

[0051] The cancer may be or may include one or more tumors. The cancer may be glioma, medulloblastoma, meningioma, neurofibroma, ependymoma, schwannoma, neurofibrosarcoma, astrocytoma and oligodendroglioma, melanoma, mesothelioma, myeloma, lymphoma, non-Hodgkin's lymphoma (NHL), Hodgkin's lymphoma, cutaneous T-cell lymphoma (CTCL), leukemia, chronic myeloid leukemia (CML), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), myelodysplastic syndrome (MDS), liver cancer, epidermoid carcinoma, prostate cancer, breast cancer, lung cancer, NSCLC, colon cancer, ovarian cancer, pancreatic cancer, thymic cancer, blood cancer or sarcoma.

[0052] In some embodiments, the cancer according to the present disclosure is selected from solid tumors, breast cancer, breast cancer, ductal carcinoma, gastric cancer, gastric cancer, gastric adenocarcinoma, colorectal cancer, colorectal adenocarcinoma, head and neck cancer, squamous cell carcinoma of the head and neck (SCCHN), lung cancer, non-small cell lung cancer, lung adenocarcinoma, squamous cell lung carcinoma, ovarian cancer, ovarian cancer, serous ovarian adenocarcinoma, 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, esophageal cancer, esophageal adenocarcinoma, liver cancer, hepatocellular carcinoma, cholangiocarcinoma, uterine cancer, endometrial cancer, thyroid cancer, thyroid cancer, pheochromocytoma, paraganglioma, bladder cancer, bladder urothelial carcinoma, prostate cancer, prostate adenocarcinoma, sarcoma, and thymoma.

[0053] In some embodiments, the cancer according to the present disclosure is selected from gastric cancer (e.g., gastric cancer, gastric adenocarcinoma, gastrointestinal adenocarcinoma), head and neck cancer (e.g., head and neck squamous cell carcinoma), breast cancer, ovarian cancer (e.g., ovarian cancer), lung cancer (e.g., NSCLC, lung adenocarcinoma, squamous cell lung carcinoma), melanoma, prostate cancer, oral cancer (e.g., oropharyngeal cancer), renal cancer (e.g., renal cell carcinoma) or colorectal cancer (e.g., colorectal carcinoma), esophageal cancer, pancreatic cancer, solid cancers, and liquid cancers (i.e., blood cancers).

[0054] In some embodiments, the cancer 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 overexpresses an EGFR family member and / or a ligand for an EGFR family member. Overexpression of an EGFR family member and / or a ligand for an EGFR family member can be determined by detecting a higher expression level by cancer cells / tumor tissue than the expression level by comparable non-cancerous cells / non-tumor tissue.

[0055] 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.

[0056] In some embodiments, the cancer according to the present disclosure is a HER3-associated cancer.

[0057] HER3 and its association with and role in cancer are reviewed, for example, in Karachaliou et al., BioDrugs. (2017) 31(1):63-73, and Zhang et al., Acta Biochimica et Biophysica Sinica (2016) 48(1):39-48, both of which are incorporated by reference in their entireties.

[0058] As used herein, "HER3-associated" cancer refers to a cancer in which HER3 gene and / or protein expression is a risk factor (e.g., positively associated) for the occurrence, development, progression, or severity of symptoms of cancer. In some embodiments, the cancer overexpresses HER3. In some embodiments, the cancer may include cells that have elevated HER3 gene and / or protein expression, for example, compared to expression levels by comparable non-cancerous cells (e.g., non-cancerous cells of the same type, e.g., non-cancerous cells derived from the same organ / tissue).

[0059] In a preferred embodiment, the HER3-associated cancer may comprise cells that express HER3. In some embodiments, the cancer comprises cells that express HER3 protein. The cancer comprises cells that express HER3 protein may be referred to as "HER3 positive" cancer. In some embodiments, the cancer comprises cells that express HER3 protein on the cell surface (i.e., in or on the cell membrane).

[0060] In some embodiments, the HER3-associated cancer may be selected from ovarian cancer, breast cancer, prostate cancer, gastric cancer, bladder cancer, pancreatic cancer, lung cancer, melanoma, colorectal cancer, squamous cell carcinoma, and oral cancer.

[0061] Elevated plasma levels of NRG1 are associated with de novo resistance to treatment with the anti-EGFR antibody cetuximab in patients with colorectal cancer (Yonesaka et al., Transl Med. (2011) 3(99). Patients who responded to cetuximab treatment showed significantly lower expression of NRG1 in pretreatment tumor samples. Liu et al., Journal of Clinical Oncology (2016) 34(36):4345-4353 report that in patients with advanced platinum-resistant or refractory ovarian cancer, NRG1-expressing cancers respond well to treatment with seribantumab.

[0062] In some embodiments, the cancer being treated / prevented comprises cells expressing a ligand for HER3 (e.g., NRG1 and / or NRG2). In some embodiments, the cancer being treated / prevented comprises cells expressing a higher expression level of NRG1 and / or NRG2 than the expression level by a comparable non-cancerous cell / non-tumor tissue. In some embodiments, the cancer comprises a mutation that results in increased (gene and / or protein) expression of a ligand for HER3 (e.g., NRG1 and / or NRG2) compared to a comparable cell carrying only the wild-type allele.

[0063] Mutations that cause increased expression of a ligand for HER3 are described, for example, in WO2021 / 048274A1, which is incorporated herein by reference in its entirety. In some embodiments, the mutation that causes increased expression of a ligand for HER3 is an NRG gene fusion. In some embodiments, the ligand for HER3 is a product of an NRG gene fusion (i.e., a polypeptide encoded by 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.

[0064] NRG gene fusions are described, for example, in WO2021 / 048274A1 (incorporated by reference hereinabove). In some embodiments, the NRG gene fusions include 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-NRG1, P Selected from ARP8-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, MCPH1-NRG1 and SLC12A2-NRG2.

[0065] Aspects and embodiments of the present disclosure relate to therapeutic and prophylactic interventions for the treatment / prevention of cancers characterized by the absence or presence of certain variants in genes encoding factors involved in HER3-mediated signaling.

[0066] Mutations, alleles and genotypes As used herein, "mutation" refers to a difference to the most common nucleotide sequence of a given gene. A mutation may be or may include an insertion, deletion, substitution, or larger translocation / rearrangement of a nucleotide sequence relative to the most common nucleotide sequence of the gene.

[0067] A mutation that "results in" upregulation of signaling (e.g., HER3-mediated signaling, signaling through the MAPK / ERK pathway, signaling through the PI3K / AKT / mTOR pathway) can be a mutation that is known or predicted to cause increased levels of the relevant signaling in a cell that has one or more copies of the mutation-containing allele compared to the level of signaling by a comparable cell that lacks a copy of the mutation-containing allele (e.g., a comparable cell that is homozygous for the wild-type allele).

[0068] HER3-mediated signal transduction can be analyzed, for example, by using assays for correlates of HER3-mediated signal transduction, such as cell proliferation and / or phosphorylation of one or more signal transduction molecules of PI3K / AKT / mTOR and / or MAPK / ERK signal transduction pathways.For example, the level of PI3K / AKT / mTOR and / or MAPK / ERK signal transduction can be analyzed by detecting and quantifying the phosphorylation level of one or more components of PI3K / AKT / mTOR and / or MAPK / ERK pathways.Such analysis can be carried out in vitro in cell-based assays of HER3-mediated signal transduction, for example, as described in Example 4.3 of section 8.9 of WO2019185878A1.

[0069] The most common version of a nucleotide sequence of a given gene may be referred to as the wild-type allele of the gene. A version of a nucleotide sequence of a given gene that contains a mutation may be referred to as a mutant allele of the gene. It will be understood that the nucleotide sequence of a mutant allele of a given gene has a nucleotide sequence that is not identical to the nucleotide sequence of the wild-type allele.

[0070] As used herein, an "activating mutation" refers to a mutation that results in an increased expression level of the associated gene and / or an increased activity of the gene product. Conversely, an "inactivating mutation" refers to a mutation that results in a decreased expression level of a given gene and / or a decreased activity of the gene product.

[0071] In some embodiments, the mutation that leads to upregulation of HER3-mediated signaling is an activating mutation in a gene that encodes a positive regulator of HER3-mediated signaling. In some embodiments, the mutation that leads to upregulation of HER3-mediated signaling is an inactivating mutation in a gene that encodes a negative regulator of HER3-mediated signaling.

[0072] Herein, a cancer that comprises cells with certain characteristics may be referred to simply as a cancer that comprises those characteristics. In embodiments herein, it will be understood that a cancer that comprises cells with certain characteristics may be or comprise one or more tumors that comprise cells with those characteristics. That is, when a cancer is described as having a given mutation state, allele, or genotype, it will be understood that the cells of the cancer have the associated mutation state, allele, or genotype. By way of illustration, when a cancer is described as comprising a given mutation, the cancer comprises cells that comprise the mutation. Similarly, when a cancer is described as being homozygous for a given allele, the cancer comprises cells that are homozygous for the allele.

[0073] When a cancer is described as having a given mutation state, allele or genotype, one or more cells of the cancer have the relevant mutation state, allele or genotype.In some embodiments, when a cancer is described as having a given mutation state, allele or genotype, a majority (i.e., more than 50%) of the cells of the cancer have the relevant mutation state, allele or genotype.In some embodiments, one of 60% or more, 65% or more, 70% or more, 75% or more, 80% or more, 85% or more, 90% or more, 95% or more, or 100% of the cells of the cancer have the relevant mutation state, allele or genotype.

[0074] In some embodiments, a cancer that contains a given mutation / allele / genotype can be a cancer in which more than 10% of the cancer's cells (e.g., one of 20% or more, 50% or more, 40% or more, 50% or more, 60% or more, 65% or more, 70% or more, 75% or more, 80% or more, 85% or more, 90% or more, 95% or more, or 100%) contain the mutation / allele / genotype. In some embodiments, a cancer that does not contain a given mutation / allele / genotype can be a cancer in which less than 25% of the cancer's cells (e.g., one of 20% or less, 15% or less, 10% or less, 5% or less, 1% or less, or none) contain the mutation / allele / genotype.

[0075] As used herein, when a cell is described as containing a given mutation, it will be understood that one or both alleles of the relevant gene contain such mutation (i.e., the cell is heterozygous or homozygous for the mutation / mutant allele). Conversely, when a cell is described as not containing a given mutation, it will be understood that neither allele of the relevant gene contains such mutation (i.e., the cell is neither homozygous nor heterozygous for the mutation / mutant allele).

[0076] In some embodiments, an activating mutation in accordance with the present disclosure can increase transcription of the gene; can increase the level of RNA encoded by the gene; can decrease the degradation of RNA encoded by the gene; can increase the level of the protein encoded by the gene; can increase (can promote) normal splicing of pre-mRNA encoded by the gene; can increase translation of mRNA encoding the protein encoded by the gene; can increase (can promote) normal post-translational processing of the protein encoded by the gene; can increase (can promote) normal trafficking of the protein encoded by the gene; can decrease the degradation of the protein encoded by the gene; can increase the level of function of the protein encoded by the gene; and / or can confer novel properties to the protein encoded by the gene.

[0077] In some embodiments, an inactivating mutation in accordance with the present disclosure can reduce transcription of the gene; can reduce the level of RNA encoded by the gene; can increase the degradation of RNA encoded by the gene; can reduce the level of protein encoded by the gene; can reduce (can disrupt) normal splicing of pre-mRNA encoded by the gene; can reduce translation of mRNA encoding the protein encoded by the gene; can reduce (can disrupt) normal post-translational processing of the protein encoded by the gene; can reduce (can disrupt) normal transport of the protein encoded by the gene; can increase the degradation of the protein encoded by the gene; and / or can reduce the level of function of the protein encoded by the gene.

[0078] As used herein, a "positive regulator" of signaling through a given pathway refers to an agent whose expression / activity generally positively contributes to (i.e., enhances, potentiates) signaling through the associated pathway. An increased level of expression and / or activity of a positive regulator can result in an increased level of signaling through the associated pathway (e.g., as determined by analysis of correlates of such signaling). A decreased level of expression and / or activity of a positive regulator can result in a decreased level of signaling through the associated pathway.

[0079] A "negative regulator" of signaling through a given pathway refers to an agent whose expression / activity generally negatively contributes to (i.e., inhibits, antagonizes) signaling through the relevant pathway. An increase in the level of expression and / or activity of a negative regulator can result in a decrease in the level of signaling through the relevant pathway (e.g., as determined by analysis of correlates of such signaling). A decrease in the level of expression and / or activity of a negative regulator can result in an increase in the level of signaling through the relevant pathway.

[0080] Cancers that are not homozygous / heterozygous / do not contain a mutation that results in upregulation of HER3-mediated signaling The aspects and embodiments of the present disclosure relate to cancers that are not homozygous, heterozygous or do not contain one or more mutations that lead to upregulation of HER3-mediated signal transduction.Such cancers may be considered to be particularly sensitive / sensitive (and therefore likely to respond well) to therapeutic / prophylactic intervention (e.g., as monotherapy) with antigen-binding molecules that bind to HER3.

[0081] In some aspects and embodiments according to the present disclosure, the cancer to be treated / prevented is not homozygous for the mutation that leads to the upregulation of HER3-mediated signaling.In some embodiments, the cancer is not heterozygous for the mutation that leads to the upregulation of HER3-mediated signaling.In some embodiments, the cancer does not contain the mutation that leads to the upregulation of HER3-mediated signaling.

[0082] In some embodiments, the cancer is not homozygous for an activating mutation in a gene that encodes a positive regulator of HER3-mediated signal transduction.In some embodiments, the cancer is not heterozygous for an activating mutation in a gene that encodes a positive regulator of HER3-mediated signal transduction.In some embodiments, the cancer does not comprise an activating mutation in a gene that encodes a positive regulator of HER3-mediated signal transduction.

[0083] In some embodiments, the cancer is not homozygous for an inactivating mutation in a gene that codes for a negative regulator of HER3-mediated signal transduction.In some embodiments, the cancer is not heterozygous for an inactivating mutation in a gene that codes for a negative regulator of HER3-mediated signal transduction.In some embodiments, the cancer does not comprise an inactivating mutation in a gene that codes for a negative regulator of HER3-mediated signal transduction.

[0084] In some embodiments, the cancer to be treated / prevented is not homozygous for a mutation that leads to upregulation of signaling through the MAPK / ERK pathway. In some embodiments, the cancer is not heterozygous for a mutation that leads to upregulation of signaling through the MAPK / ERK pathway. In some embodiments, the cancer does not contain a mutation that leads to upregulation of signaling through the MAPK / ERK pathway.

[0085] In some embodiments, the cancer is not homozygous for an activating mutation in a gene encoding a positive regulator of signaling through the MAPK / ERK pathway. In some embodiments, the cancer is not heterozygous for an activating mutation in a gene encoding a positive regulator of signaling through the MAPK / ERK pathway. In some embodiments, the cancer does not comprise an activating mutation in a gene encoding a positive regulator of signaling through the MAPK / ERK pathway.

[0086] In some embodiments, the cancer is not homozygous for an inactivating mutation in a gene encoding a negative regulator of signaling through the MAPK / ERK pathway. In some embodiments, the cancer is not heterozygous for an inactivating mutation in a gene encoding a negative regulator of signaling through the MAPK / ERK pathway. In some embodiments, the cancer does not comprise an inactivating mutation in a gene encoding a negative regulator of signaling through the MAPK / ERK pathway.

[0087] In some embodiments, the cancer to be treated / prevented is not homozygous for a mutation that leads to upregulation of signaling through the PI3K / AKT / mTOR pathway. In some embodiments, the cancer is not heterozygous for a mutation that leads to upregulation of signaling through the PI3K / AKT / mTOR pathway. In some embodiments, the cancer does not contain a mutation that leads to upregulation of signaling through the PI3K / AKT / mTOR pathway.

[0088] In some embodiments, the cancer is not homozygous for an activating mutation in a gene that encodes a positive regulator of signaling through the PI3K / AKT / mTOR pathway. In some embodiments, the cancer is not heterozygous for an activating mutation in a gene that encodes a positive regulator of signaling through the PI3K / AKT / mTOR pathway. In some embodiments, the cancer does not comprise an activating mutation in a gene that encodes a positive regulator of signaling through the PI3K / AKT / mTOR pathway.

[0089] In some embodiments, the cancer is not homozygous for an inactivating mutation to a gene encoding a negative regulator of signaling through the PI3K / AKT / mTOR pathway. In some embodiments, the cancer is not heterozygous for an inactivating mutation to a gene encoding a negative regulator of signaling through the PI3K / AKT / mTOR pathway. In some embodiments, the cancer does not comprise an inactivating mutation to a gene encoding a negative regulator of signaling through the PI3K / AKT / mTOR pathway.

[0090] As used herein, a positive regulator of HER3-mediated signaling / MAPK / ERK pathway / PI3K / AKT / mTOR pathway can be any factor that positively contributes (i.e., enhances) to signaling through a given pathway, and / or its functional consequences. A negative regulator of HER3-mediated signaling / MAPK / ERK pathway / PI3K / AKT / mTOR pathway can be any factor that negatively contributes (i.e., antagonizes, inhibits) to signaling through a given pathway, and / or its functional consequences.

[0091] In some embodiments, the gene encoding a positive regulator of HER3-mediated signaling may be selected from KRAS, PIK3CA, PIK3CB, PIK3CD, ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GRB2, GAB2, PTPN11, SHP2, SOS1, HRAS, NRAS, RAF1, MAP2K1, MAP2K2, MAPK1, MYC, RPS6KA1, RPS6, MKNK1, CREB1, MTOR, PDK1, AKT1, AKT2, AKT3, JAK2, STAT3 and STAT5. In some embodiments, the gene encoding the positive regulator of HER3-mediated signal transduction can be selected from KRAS, PIK3CA, PIK3CB, PIK3CD, ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GRB2, GAB2, PTPN11, SHP2, SOS1, HRAS, NRAS, RAF1, MAP2K1, MAP2K2, MAPK1, MYC, RPS6KA1, RPS6, MKNK1, CREB1, MTOR, PDK1, AKT1, AKT2, AKT3, JAK2, STAT3, STAT5 and BRAF.In some embodiments, the gene encoding the positive regulator of HER3-mediated signal transduction can be selected from KRAS, PIK3CA and PIK3CB.In some embodiments, the gene encoding the positive regulator of HER3-mediated signal transduction is KRAS. In some embodiments, the gene encoding a positive regulator of HER3-mediated signaling is PIK3CA. In some embodiments, the gene encoding a positive regulator of HER3-mediated signaling is PIK3CB.

[0092] In some embodiments, genes encoding positive regulators of signaling through the MAPK / ERK pathway may be selected from ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GAB2, GRB2, PTPN11, SHP2, SOS1, HRAS, KRAS, NRAS, RAF1, MAP2K1, MAP2K2, MAPK1, MYC, RPS6KA1, RPS6, MKNK1, and CREB1. In some embodiments, the gene encoding the positive regulator of signal transduction through the MAPK / ERK pathway can be selected from ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GAB2, GRB2, PTPN11, SHP2, SOS1, HRAS, KRAS, NRAS, RAF1, MAP2K1, MAP2K2, MAPK1, MYC, RPS6KA1, RPS6, MKNK1, CREB1, and BRAF.In some embodiments, the gene encoding the positive regulator of signal transduction through the MAPK / ERK pathway is KRAS.In some embodiments, the gene encoding the positive regulator of signal transduction through the MAPK / ERK pathway is BRAF.

[0093] In some embodiments, the gene encoding the positive regulator of signal transduction through the PI3K / AKT / mTOR pathway may be selected from ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GAB2, SHP2, CREB1, PIK3CA, PIK3CB, PIK3CD, MTOR, PDK1, AKT1, AKT2, AKT3, JAK2, STAT3 and STAT5. In some embodiments, the gene encoding the positive regulator of signal transduction through the PI3K / AKT / mTOR pathway is PIK3CA. In some embodiments, the gene encoding the positive regulator of signal transduction through the PI3K / AKT / mTOR pathway is PIK3CB.

[0094] In some embodiments, the gene encoding a negative regulator of HER3-mediated signaling may be selected from PTEN, PPP2CA, PIK3R1, PIK3R2, NF1, BAD and PHLPP1. In some embodiments, the gene encoding a negative regulator of signaling through the PI3K / AKT / mTOR pathway may be selected from PTEN, PPP2CA, PIK3R1, PIK3R2, BAD and PHLPP1. In some embodiments, the gene encoding a negative regulator of signaling through the MAPK / ERK pathway may be NF1. In some embodiments, the gene encoding a negative regulator of HER3-mediated signaling is PTEN. In some embodiments, the gene encoding a negative regulator of signaling through the PI3K / AKT / mTOR pathway is PTEN.

[0095] In some embodiments, the cancer does not include cells that are homozygous for an allele of a gene encoding a positive regulator of HER3-mediated signaling that includes an activating mutation. In some embodiments, the cancer does not include cells that encode an allele of a gene encoding a positive regulator of HER3-mediated signaling that includes an activating mutation. In some embodiments, in the cells of the cancer, one or both copies of the gene encoding a positive regulator of HER3-mediated signaling are wild-type alleles. In some embodiments, the cancer does not include cells that encode a mutant allele of a gene encoding a positive regulator of HER3-mediated signaling. In some embodiments, the cancer does not include cells that are homozygous for an allele of a gene encoding a negative regulator of HER3-mediated signaling that includes an inactivating mutation. In some embodiments, the cancer does not include cells that encode an allele of a gene encoding a negative regulator of HER3-mediated signaling that includes an inactivating mutation. In some embodiments, in the cells of the cancer, one or both copies of the gene encoding a negative regulator of HER3-mediated signaling are wild-type alleles. In some embodiments, the cancer does not include cells that encode a mutant allele of a gene encoding a negative regulator of HER3-mediated signaling.

[0096] In some embodiments, the cancer does not include cells that are homozygous for an allele of a gene encoding a positive regulator of signaling through the MAPK / ERK pathway that comprises an activating mutation. In some embodiments, the cancer does not include cells that encode an allele of a gene encoding a positive regulator of signaling through the MAPK / ERK pathway that comprises an activating mutation. In some embodiments, in the cells of the cancer, one or both copies of the gene encoding a positive regulator of signaling through the MAPK / ERK pathway are wild-type alleles. In some embodiments, the cancer does not include cells that encode a mutant allele of a gene encoding a positive regulator of signaling through the MAPK / ERK pathway. In some embodiments, the cancer does not include cells that are homozygous for an allele of a gene encoding a negative regulator of signaling through the MAPK / ERK pathway that comprises an inactivating mutation. In some embodiments, the cancer does not include cells that encode an allele of a gene encoding a negative regulator of signaling through the MAPK / ERK pathway that comprises an inactivating mutation. In some embodiments, in the cells of the cancer, one or both copies of the gene encoding a negative regulator of signaling through the MAPK / ERK pathway are wild-type alleles. In some embodiments, the cancer does not include cells encoding mutant alleles of the gene encoding a negative regulator of signaling through the MAPK / ERK pathway.

[0097] In some embodiments, the cancer does not include cells that are homozygous for an allele of a gene encoding a positive regulator of signaling through the PI3K / AKT / mTOR pathway that comprises an activating mutation. In some embodiments, the cancer does not include cells that encode an allele of a gene encoding a positive regulator of signaling through the PI3K / AKT / mTOR pathway that comprises an activating mutation. In some embodiments, in the cells of the cancer, one or both copies of the gene encoding a positive regulator of signaling through the PI3K / AKT / mTOR pathway are wild type alleles. In some embodiments, the cancer does not include cells that encode a mutant allele of a gene encoding a positive regulator of signaling through the PI3K / AKT / mTOR pathway. In some embodiments, the cancer does not include cells that are homozygous for an allele of a gene encoding a negative regulator of signaling through the PI3K / AKT / mTOR pathway that comprises an inactivating mutation. In some embodiments, the cancer does not include cells that encode an allele of a gene encoding a negative regulator of signaling through the PI3K / AKT / mTOR pathway that comprises an inactivating mutation. In some embodiments, in the cells of the cancer, one or both copies of the gene encoding a negative regulator of signaling through the PI3K / AKT / mTOR pathway are wild-type alleles. In some embodiments, the cancer does not include cells encoding mutant alleles of the gene encoding a negative regulator of signaling through the PI3K / AKT / mTOR pathway.

[0098] In some embodiments, cancer does not include cells that are homozygous for the allele of KRAS that comprises activating mutation.In some embodiments, cancer does not include cells that encode the allele of KRAS that comprises activating mutation.In some embodiments, in the cell of cancer, one or both copies of KRAS are wild type alleles.In some embodiments, cancer does not include cells that encode the mutant allele of KRAS.

[0099] Activating mutations for KRAS are described, for example, in Hobbs and Der, Cancer Discov. (2019) 9(6):696-698, which is incorporated herein by reference in its entirety. Activating mutations for KRAS include mutations for G12 (e.g., G12A, G12D, G12D, G12R, G12C, G12S and G12V), mutations for G13 (e.g., G13D, G13C), mutations for Q61 (e.g., Q61H, Q61L, Q61K, Q61R), mutations for A146 (e.g., A146T, A146V) and mutations for K117 (e.g., K117N). In some embodiments, the activating mutation for KRAS according to the present disclosure is G12C.

[0100] In some embodiments, cancer does not include cells that are homozygous for the allele of PIK3CA that comprises activating mutation.In some embodiments, cancer does not include cells that encode the allele of PIK3CA that comprises activating mutation.In some embodiments, in the cell of cancer, one or both copies of PIK3CA are wild type alleles.In some embodiments, cancer does not include cells that encode the mutant allele of PIK3CA.

[0101] Activating mutations for PIK3CA are described, for example, in Ligresti et al., Cell Cycle. (2009) 8(9):1352-1358, which is incorporated herein by reference in its entirety. Activating mutations for PIK3CA include mutations for H1047 (e.g., H1047R, H1047L), mutations for E542 (e.g., E542K, E542Q), mutations for E545 (e.g., E545K), mutations for P449 (e.g., P449T) and mutations for Q546 (e.g., Q546R). In some embodiments, the activating mutation for PIK3CA according to the present disclosure is Q546R or P449T.

[0102] In some embodiments, cancer does not include cells that are homozygous for the allele of PIK3CB that comprises activating mutation.In some embodiments, cancer does not include cells that encode the allele of PIK3CB that comprises activating mutation.In some embodiments, in the cell of cancer, one or both copies of PIK3CB are wild type alleles.In some embodiments, cancer does not include cells that encode the mutant allele of PIK3CB.

[0103] Activating mutations for PIK3CB are described, for example, in Nakanishi et al., Cancer Res. (2016) 76(5):1193-203, which is incorporated herein by reference in its entirety. Activating mutations for PIK3CB include mutations for D1067 (e.g., D1067Y, D1067A, D1067V). In some embodiments, the activating mutation for PIK3CB according to the present disclosure is D1067Y.

[0104] In some embodiments, the cancer does not include cells that are homozygous for the allele of BRAF that comprises an activating mutation.In some embodiments, the cancer does not include cells that encode the allele of BRAF that comprises an activating mutation.In some embodiments, the cancer does not include cells that encode the mutant allele of BRAF.

[0105] Activating mutations for BRAF are described, for example, in Van Cutsem et al., Journal of Clinical Immunology (2011) 29(15):2011-2019, which is incorporated herein by reference in its entirety. Activating mutations for BRAF include mutations for V600 (e.g., V600E or V600K), T119 (e.g., T119S) and L597 (e.g., L597R). In some embodiments, the activating mutation according to the present disclosure is V600E, V600K, T119S, or L597R.

[0106] In some embodiments, the cancer does not include cells that are homozygous for the allele of PTEN that comprises inactivating mutation.In some embodiments, the cancer does not include cells that encode the allele of PTEN that comprises inactivating mutation.In some embodiments, in the cells of the cancer, one or both copies of PTEN are wild type alleles.In some embodiments, the cancer does not include cells that encode the mutant allele of PTEN.

[0107] Inactivating mutations to PTEN are described, for example, in Chang et al., Biomolecules. (2019) 9(11):713, which is incorporated herein by reference in its entirety. Inactivating mutations to PTEN include mutations to R130, mutations to R173, mutations to R233, mutations to K267, and mutations to N323.

[0108] In some embodiments, the cancer contains (i) at least one gene encoding a positive regulator of HER3-mediated signaling that does not contain an activating mutation; or (ii) at least one gene encoding a negative regulator of HER3-mediated signaling that does not contain an inactivating mutation.

[0109] In some embodiments, the cancer contains (i) at least one gene encoding a positive regulator of HER3-mediated signaling that is not homozygous for an activating mutation; or (ii) at least one gene encoding a negative regulator of HER3-mediated signaling that is not homozygous for an inactivating mutation.

[0110] In some embodiments, the cancer is (i) not homozygous for an activating mutation in a gene selected from KRAS, PIK3CA, PIK3CB, PIK3CD, ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GRB2, GAB2, PTPN11, SHP2, SOS1, HRAS, NRAS, RAF1, MAP2K1, MAP2K2, MAPK1, MYC, RPS6KA1, RPS6, MKNK1, CREB1, MTOR, PDK1, AKT1, AKT2, AKT3, JAK2, STAT3, and STAT5; or (ii) not homozygous for an inactivating mutation in a gene selected from PTEN, PPP2CA, PIK3R1, PIK3R2, NF1, BAD, and PHLPP1.

[0111] In some embodiments, the cancer does not (i) contain an activating mutation in a gene selected from KRAS, PIK3CA, PIK3CB, PIK3CD, ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GRB2, GAB2, PTPN11, SHP2, SOS1, HRAS, NRAS, RAF1, MAP2K1, MAP2K2, MAPK1, MYC, RPS6KA1, RPS6, MKNK1, CREB1, MTOR, PDK1, AKT1, AKT2, AKT3, JAK2, STAT3, and STAT5; or (ii) contain an inactivating mutation in a gene selected from PTEN, PPP2CA, PIK3R1, PIK3R2, NF1, BAD, and PHLPP1.

[0112] In some embodiments, the cancer is (i) not homozygous for an activating mutation in a gene selected from KRAS, PIK3CA, PIK3CB, PIK3CD, ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GRB2, GAB2, PTPN11, SHP2, SOS1, HRAS, NRAS, RAF1, MAP2K1, MAP2K2, MAPK1, MYC, RPS6KA1, RPS6, MKNK1, CREB1, MTOR, PDK1, AKT1, AKT2, AKT3, JAK2, STAT3, STAT5, and BRAF; or (ii) not homozygous for an inactivating mutation in a gene selected from PTEN, PPP2CA, PIK3R1, PIK3R2, NF1, BAD, and PHLPP1.

[0113] In some embodiments, the cancer does not (i) contain an activating mutation in a gene selected from KRAS, PIK3CA, PIK3CB, PIK3CD, ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GRB2, GAB2, PTPN11, SHP2, SOS1, HRAS, NRAS, RAF1, MAP2K1, MAP2K2, MAPK1, MYC, RPS6KA1, RPS6, MKNK1, CREB1, MTOR, PDK1, AKT1, AKT2, AKT3, JAK2, STAT3, STAT5, and BRAF; or (ii) contain an inactivating mutation in a gene selected from PTEN, PPP2CA, PIK3R1, PIK3R2, NF1, BAD, and PHLPP1.

[0114] It will be understood that in this specification, reference to "gene" means "at least one gene" and includes "one or more genes". Thus, the "gene" selected from a given list may be one gene in the list (depending on the number of genes listed in the list), or it may be one of 2, 3, 4, 5, 6, 7, 9, 10 or more, or all of the genes listed in the list. For clarity, a cancer satisfies the conditions of the previous paragraph, provided that at least one of the genes listed in item (i) lacks an activating mutation or at least one of the genes listed in item (ii) lacks an inactivating mutation.

[0115] In some embodiments, the cancer is: (i) are not homozygous for an activating mutation for KRAS and are not homozygous for an activating mutation for PIK3CA; (ii) are not homozygous for an activating mutation for KRAS and are not homozygous for an inactivating mutation for PTEN; (iii) not homozygous for an activating mutation for KRAS but not homozygous for an activating mutation for BRAF; (iv) not homozygous for an activating mutation for PIK3CA but not homozygous for an inactivating mutation for PTEN; (v) not homozygous for an activating mutation for PIK3CA but not homozygous for an activating mutation for BRAF; (vi) not homozygous for an inactivating mutation to PTEN but not homozygous for an activating mutation to BRAF; (vii) not homozygous for an activating mutation for KRAS, not homozygous for an activating mutation for PIK3CA, and not homozygous for an inactivating mutation for PTEN; (viii) not homozygous for an activating mutation for KRAS, not homozygous for an activating mutation for PIK3CA, and not homozygous for an activating mutation for BRAF; (ix) not homozygous for an activating mutation for KRAS, not homozygous for an inactivating mutation for PTEN, and not homozygous for an activating mutation for BRAF; (x) is not homozygous for an activating mutation in PIK3CA, is not homozygous for an inactivating mutation in PTEN, and is not homozygous for an activating mutation in BRAF; or (xi) not homozygous for an activating mutation for KRAS, not homozygous for an activating mutation for PIK3CA, not homozygous for an inactivating mutation for PTEN, and not homozygous for an activating mutation for BRAF.

[0116] In some embodiments, the cancer is: (i) did not contain activating mutations for KRAS and did not contain activating mutations for PIK3CA; (ii) did not contain activating mutations for KRAS and did not contain inactivating mutations for PTEN; (iii) did not contain activating mutations for KRAS and did not contain activating mutations for BRAF; (iv) did not contain activating mutations for PIK3CA and did not contain inactivating mutations for PTEN; (v) did not contain activating mutations for PIK3CA and did not contain activating mutations for BRAF; (vi) did not contain inactivating mutations for PTEN and did not contain activating mutations for BRAF; (vii) no activating mutations for KRAS, no activating mutations for PIK3CA, and no inactivating mutations for PTEN; (viii) no activating mutations for KRAS, no activating mutations for PIK3CA, and no activating mutations for BRAF; (ix) no activating mutations for KRAS, no inactivating mutations for PTEN, and no activating mutations for BRAF; (x) no activating mutations for PIK3CA, no inactivating mutations for PTEN, and no activating mutations for BRAF; (xi) no activating mutations for KRAS, no activating mutations for PIK3CA, no inactivating mutations for PTEN, and no activating mutations for BRAF.

[0117] Cancers containing / heterozygous / homozygous for mutations that result in upregulation of HER3-mediated signaling The aspects and embodiments of the present disclosure relate to cancer that comprises, is heterozygous for, or is homozygous for one or more mutations that lead to upregulation of HER3-mediated signal transduction.Such cancers may be considered to be less sensitive / resistant (and therefore less likely to respond well) to therapeutic / prophylactic intervention (e.g., as monotherapy) by antigen-binding molecules that bind to HER3.

[0118] In some aspects and embodiments according to the present disclosure, particularly those relating to combined intervention using an inhibitor of HER3-mediated signal transduction and an antigen-binding molecule that binds to HER3, the cancer to be treated / prevented comprises a mutation that leads to upregulation of HER3-mediated signal transduction.In some embodiments, the cancer is heterozygous for the mutation that leads to upregulation of HER3-mediated signal transduction.In some embodiments, the cancer is homozygous for the mutation that leads to upregulation of HER3-mediated signal transduction.

[0119] In some embodiments, the cancer comprises an activating mutation in the gene that codes for the positive regulator of HER3-mediated signal transduction.In some embodiments, the cancer is heterozygous for the activating mutation in the gene that codes for the positive regulator of HER3-mediated signal transduction.In some embodiments, the cancer is homozygous for the activating mutation in the gene that codes for the positive regulator of HER3-mediated signal transduction.

[0120] In some embodiments, the cancer comprises an inactivating mutation in the gene that codes for the negative regulator of HER3-mediated signal transduction.In some embodiments, the cancer is heterozygous for the inactivating mutation in the gene that codes for the negative regulator of HER3-mediated signal transduction.In some embodiments, the cancer is homozygous for the inactivating mutation in the gene that codes for the negative regulator of HER3-mediated signal transduction.

[0121] In some embodiments, the cancer comprises a mutation that leads to upregulation of signaling through the MAPK / ERK pathway. In some embodiments, the cancer is heterozygous for the mutation that leads to upregulation of signaling through the MAPK / ERK pathway. In some embodiments, the cancer is homozygous for the mutation that leads to upregulation of signaling through the MAPK / ERK pathway.

[0122] In some embodiments, the cancer comprises an activating mutation in a gene encoding a positive regulator of signaling through the MAPK / ERK pathway. In some embodiments, the cancer is heterozygous for an activating mutation in a gene encoding a positive regulator of signaling through the MAPK / ERK pathway. In some embodiments, the cancer is homozygous for an activating mutation in a gene encoding a positive regulator of signaling through the MAPK / ERK pathway.

[0123] In some embodiments, the cancer comprises an inactivating mutation in a gene encoding a negative regulator of signaling through the MAPK / ERK pathway. In some embodiments, the cancer is heterozygous for an inactivating mutation in a gene encoding a negative regulator of signaling through the MAPK / ERK pathway. In some embodiments, the cancer is homozygous for an inactivating mutation in a gene encoding a negative regulator of signaling through the MAPK / ERK pathway.

[0124] In some embodiments, the cancer comprises a mutation that leads to upregulation of signaling through the PI3K / AKT / mTOR pathway. In some embodiments, the cancer is heterozygous for a mutation that leads to upregulation of signaling through the PI3K / AKT / mTOR pathway. In some embodiments, the cancer is homozygous for a mutation that leads to upregulation of signaling through the PI3K / AKT / mTOR pathway.

[0125] In some embodiments, the cancer comprises an activating mutation in a gene that encodes a positive regulator of signaling through the PI3K / AKT / mTOR pathway. In some embodiments, the cancer is heterozygous for an activating mutation in a gene that encodes a positive regulator of signaling through the PI3K / AKT / mTOR pathway. In some embodiments, the cancer is homozygous for an activating mutation in a gene that encodes a positive regulator of signaling through the PI3K / AKT / mTOR pathway.

[0126] In some embodiments, the cancer comprises an inactivating mutation in a gene encoding a negative regulator of signaling through the PI3K / AKT / mTOR pathway. In some embodiments, the cancer is heterozygous for an inactivating mutation in a gene encoding a negative regulator of signaling through the PI3K / AKT / mTOR pathway. In some embodiments, the cancer is homozygous for an inactivating mutation in a gene encoding a negative regulator of signaling through the PI3K / AKT / mTOR pathway.

[0127] In some embodiments, the cancer comprises cells encoding a mutant allele of a gene encoding a negative regulator of HER3-mediated signaling. In some embodiments, in the cells of the cancer, one or both copies of the gene encoding a negative regulator of HER3-mediated signaling are not wild-type alleles. In some embodiments, the cancer comprises cells encoding an allele of a gene encoding a negative regulator of HER3-mediated signaling that includes an inactivating mutation. In some embodiments, the cancer comprises cells that are homozygous for an allele of a gene encoding a negative regulator of HER3-mediated signaling that includes an inactivating mutation. In some embodiments, the cancer comprises cells encoding a mutant allele of a gene encoding a positive regulator of HER3-mediated signaling. In some embodiments, in the cells of the cancer, one or both copies of a gene encoding a positive regulator of HER3-mediated signaling are not wild-type alleles. In some embodiments, the cancer comprises cells encoding an allele of a gene encoding a positive regulator of HER3-mediated signaling that includes an activating mutation. In some embodiments, the cancer comprises cells that are homozygous for an allele of a gene encoding a positive regulator of HER3-mediated signaling that includes an activating mutation.

[0128] In some embodiments, the cancer comprises cells encoding a mutant allele of a gene encoding a negative regulator of signaling through the MAPK / ERK pathway. In some embodiments, in the cells of the cancer, one or both copies of the gene encoding a negative regulator of signaling through the MAPK / ERK pathway are not wild-type alleles. In some embodiments, the cancer comprises cells encoding an allele of a gene encoding a negative regulator of signaling through the MAPK / ERK pathway that comprises an inactivating mutation. In some embodiments, the cancer comprises cells that are homozygous for an allele of a gene encoding a negative regulator of signaling through the MAPK / ERK pathway that comprises an inactivating mutation. In some embodiments, the cancer comprises cells encoding a mutant allele of a gene encoding a positive regulator of signaling through the MAPK / ERK pathway. In some embodiments, in the cells of the cancer, one or both copies of the gene encoding a positive regulator of signaling through the MAPK / ERK pathway are not wild-type alleles. In some embodiments, the cancer comprises cells encoding an allele of a gene encoding a positive regulator of signaling through the MAPK / ERK pathway that comprises an activating mutation, hi some embodiments, the cancer comprises cells that are homozygous for an allele of a gene encoding a positive regulator of signaling through the MAPK / ERK pathway that comprises an activating mutation.

[0129] In some embodiments, the cancer comprises cells encoding a mutant allele of a gene encoding a negative regulator of signaling through the PI3K / AKT / mTOR pathway. In some embodiments, in the cells of the cancer, one or both copies of the gene encoding a negative regulator of signaling through the PI3K / AKT / mTOR pathway are not wild-type alleles. In some embodiments, the cancer comprises cells encoding an allele of a gene encoding a negative regulator of signaling through the PI3K / AKT / mTOR pathway that comprises an inactivating mutation. In some embodiments, the cancer comprises cells that are homozygous for an allele of a gene encoding a negative regulator of signaling through the PI3K / AKT / mTOR pathway that comprises an inactivating mutation. In some embodiments, the cancer comprises cells encoding a mutant allele of a gene encoding a positive regulator of signaling through the PI3K / AKT / mTOR pathway. In some embodiments, in the cells of the cancer, one or both copies of the gene encoding a positive regulator of signaling through the PI3K / AKT / mTOR pathway are not wild-type alleles. In some embodiments, the cancer comprises cells encoding an allele of a gene encoding a positive regulator of signaling through the PI3K / AKT / mTOR pathway that comprises an activating mutation. In some embodiments, the cancer comprises cells that are homozygous for an allele of a gene encoding a positive regulator of signaling through the PI3K / AKT / mTOR pathway that comprises an activating mutation.

[0130] In some embodiments, the cancer comprises cells that encode a mutant allele of KRAS.In some embodiments, in the cells of the cancer, one or both copies of KRAS are not wild-type alleles.In some embodiments, the cancer comprises cells that encode an allele of KRAS that comprises activating mutation.In some embodiments, the cancer comprises cells that are homozygous for the allele of KRAS that comprises activating mutation.

[0131] In some embodiments, the cancer comprises cells that encode mutant alleles of PIK3CA.In some embodiments, in the cells of the cancer, one or both copies of PIK3CA are not wild-type alleles.In some embodiments, the cancer comprises cells that encode alleles of PIK3CA that comprise activating mutations.In some embodiments, the cancer comprises cells that are homozygous for alleles of PIK3CA that comprise activating mutations.

[0132] In some embodiments, the cancer comprises cells that encode mutant alleles of PIK3CB.In some embodiments, in the cells of the cancer, one or both copies of PIK3CB are not wild-type alleles.In some embodiments, the cancer comprises cells that encode alleles of PIK3CB that comprise activating mutations.In some embodiments, the cancer comprises cells that are homozygous for alleles of PIK3CB that comprise activating mutations.

[0133] In some embodiments, the cancer comprises cells that encode mutant alleles of BRAF.In some embodiments, in the cells of the cancer, one or both copies of BRAF are not wild type alleles.In some embodiments, the cancer comprises cells that encode BRAF alleles that contain activating mutations.In some embodiments, the cancer comprises cells that are homozygous for BRAF alleles that contain activating mutations.

[0134] In some embodiments, the cancer comprises cells that encode a mutant allele of PTEN. In some embodiments, in the cells of the cancer, one or both copies of PTEN are not wild-type alleles. In some embodiments, the cancer comprises cells that encode an allele of PTEN that comprises an inactivating mutation. In some embodiments, the cancer comprises cells that are homozygous for an allele of PTEN that comprises an inactivating mutation.

[0135] In some embodiments, the cancer contains (i) at least one gene encoding a positive regulator of HER3-mediated signaling that comprises an activating mutation; or (ii) at least one gene encoding a negative regulator of HER3-mediated signaling that comprises an inactivating mutation.

[0136] In some embodiments, the cancer contains (i) at least one gene encoding a positive regulator of HER3-mediated signaling that is homozygous for an activating mutation; or (ii) at least one gene encoding a negative regulator of HER3-mediated signaling that is homozygous for an inactivating mutation.

[0137] In some embodiments, the cancer comprises (i) an activating mutation in a gene selected from KRAS, PIK3CA, PIK3CB, PIK3CD, ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GRB2, GAB2, PTPN11, SHP2, SOS1, HRAS, NRAS, RAF1, MAP2K1, MAP2K2, MAPK1, MYC, RPS6KA1, RPS6, MKNK1, CREB1, MTOR, PDK1, AKT1, AKT2, AKT3, JAK2, STAT3, and STAT5; or (ii) an inactivating mutation in a gene selected from PTEN, PPP2CA, PIK3R1, PIK3R2, NF1, BAD, and PHLPP1. In some embodiments, the cancer comprises (i) an activating mutation in a gene selected from KRAS, PIK3CA, PIK3CB, PIK3CD, ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GRB2, GAB2, PTPN11, SHP2, SOS1, HRAS, NRAS, RAF1, MAP2K1, MAP2K2, MAPK1, MYC, RPS6KA1, RPS6, MKNK1, CREB1, MTOR, PDK1, AKT1, AKT2, AKT3, JAK2, STAT3, STAT5, and BRAF; or (ii) an inactivating mutation in a gene selected from PTEN, PPP2CA, PIK3R1, PIK3R2, NF1, BAD, and PHLPP1.

[0138] For clarity, a cancer meets the conditions of the previous paragraph, provided that at least one of the genes listed in item (i) contains an activating mutation or at least one of the genes listed in item (ii) contains an inactivating mutation.

[0139] In some embodiments, the cancer is (i) homozygous for an activating mutation in a gene selected from KRAS, PIK3CA, PIK3CB, PIK3CD, ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GRB2, GAB2, PTPN11, SHP2, SOS1, HRAS, NRAS, RAF1, MAP2K1, MAP2K2, MAPK1, MYC, RPS6KA1, RPS6, MKNK1, CREB1, MTOR, PDK1, AKT1, AKT2, AKT3, JAK2, STAT3, and STAT5; or (ii) homozygous for an inactivating mutation in a gene selected from PTEN, PPP2CA, PIK3R1, PIK3R2, NF1, BAD, and PHLPP1. In some embodiments, the cancer is (i) homozygous for an activating mutation in a gene selected from KRAS, PIK3CA, PIK3CB, PIK3CD, ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GRB2, GAB2, PTPN11, SHP2, SOS1, HRAS, NRAS, RAF1, MAP2K1, MAP2K2, MAPK1, MYC, RPS6KA1, RPS6, MKNK1, CREB1, MTOR, PDK1, AKT1, AKT2, AKT3, JAK2, STAT3, STAT5, and BRAF; or (ii) homozygous for an inactivating mutation in a gene selected from PTEN, PPP2CA, PIK3R1, PIK3R2, NF1, BAD, and PHLPP1.

[0140] In some embodiments, the cancer is: (i) contains an activating mutation for KRAS and / or contains an activating mutation for PIK3CA; (ii) contains an activating mutation for KRAS and / or an inactivating mutation for PTEN; (iii) contains an activating mutation for KRAS and / or contains an activating mutation for BRAF; (iv) contains an activating mutation for PIK3CA and / or an inactivating mutation for PTEN; (v) contains an activating mutation for PIK3CA and / or contains an activating mutation for BRAF; (vi) contains an inactivating mutation to PTEN and / or contains an activating mutation to BRAF; (vii) contains an activating mutation for KRAS, and / or contains an activating mutation for PIK3CA, and / or contains an inactivating mutation for PTEN; (viii) contains an activating mutation for KRAS, and / or contains an activating mutation for PIK3CA, and / or contains an activating mutation for BRAF; (ix) contains an activating mutation for KRAS and / or contains an inactivating mutation for PTEN and / or contains an activating mutation for BRAF; (x) contains an activating mutation for PIK3CA and / or contains an inactivating mutation for PTEN and / or contains an activating mutation for BRAF; or (xi) comprises an activating mutation for KRAS, and / or comprises an activating mutation for PIK3CA, and / or comprises an inactivating mutation for PTEN, and / or comprises an activating mutation for BRAF.

[0141] In some embodiments, the cancer is: (i) homozygous for an activating mutation for KRAS and / or homozygous for an activating mutation for PIK3CA; (ii) homozygous for an activating mutation in KRAS and / or homozygous for an inactivating mutation in PTEN; (iii) homozygous for an activating mutation in KRAS and / or homozygous for an activating mutation in BRAF; (iv) homozygous for an activating mutation in PIK3CA and / or homozygous for an inactivating mutation in PTEN; (v) homozygous for an activating mutation in PIK3CA and / or homozygous for an activating mutation in BRAF; (vi) homozygous for an inactivating mutation in PTEN and / or homozygous for an activating mutation in BRAF; (vii) is homozygous for an activating mutation for KRAS and / or is homozygous for an activating mutation for PIK3CA and / or is homozygous for an inactivating mutation for PTEN; (viii) is homozygous for an activating mutation for KRAS and / or is homozygous for an activating mutation for PIK3CA and / or is homozygous for an activating mutation for BRAF; (ix) is homozygous for an activating mutation for KRAS and / or is homozygous for an inactivating mutation for PTEN and / or is homozygous for an activating mutation for BRAF; (x) is homozygous for an activating mutation to PIK3CA and / or is homozygous for an inactivating mutation to PTEN and / or is homozygous for an activating mutation to BRAF; or (xi) is homozygous for an activating mutation for KRAS, and / or is homozygous for an activating mutation for PIK3CA, and / or is homozygous for an inactivating mutation for PTEN, and / or is homozygous for an activating mutation for BRAF.

[0142] antigen binding molecule The present disclosure relates to therapeutic and prophylactic uses of antigen-binding molecules that bind to HER3.

[0143] "Antigen-binding molecule" refers to a molecule that can bind to a target antigen. Antigen-binding molecules include, for example, monoclonal antibodies, polyclonal antibodies, monospecific antibodies and multispecific antibodies (e.g., bispecific antibodies), as well as antibody fragments (e.g., Fv, scFv, Fab, scFab, F(ab')2, Fab2, diabody, triabody, scFv-Fc, minibody, single domain antibody (e.g., VhH), etc.), so long as they exhibit binding to the relevant target molecule.

[0144] Antigen-binding molecules according to the present disclosure also include antibody-derived molecules, e.g., molecules that contain an antigen-binding region / domain derived from an antibody. Antibody-derived antigen-binding molecules may contain an antigen-binding region / domain that comprises or consists of an 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 may include the Fv (e.g., provided as 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) that contain 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 herein 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, e.g., in Dotti et al., Immunol Rev (2014) 257(1), which is incorporated herein by reference in its entirety).

[0145] The antigen-binding molecules of the present disclosure include a moiety capable of binding to a target antigen. In some embodiments, the moiety capable of binding to a target antigen includes 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 a target antigen includes or consists of an aptamer, such as a nucleic acid aptamer (reviewed, for example, 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, e.g., a peptide aptamer, a thioredoxin, a monobody, anticalin, a Kunitz domain, an avimer, a knottin, a fynomer, an atrimer, a DARPin, an affibody, a nanobody (i.e., a single domain antibody (sdAb)), an affilin, an armadillo repeat protein (ArmRP), an OBody, or a 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)).

[0146] The antigen-binding molecule of the present disclosure generally comprises an antigen-binding domain comprising the VH and VL of an antibody capable of specifically binding to a target antigen. In this specification, the antigen-binding domain formed by the VH and VL can also be referred to as the Fv region.

[0147] An antigen-binding molecule may be or may 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 that comprises a VH and a VL, or in the case of an scFab that comprises a VH-CH1 and a VL-CL).

[0148] An antigen-binding molecule may refer to a non-covalent or covalent complex of an IgG-like antigen-binding molecule 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.

[0149] The antigen-binding molecule of the present disclosure can be designed and prepared using the sequence of a monoclonal antibody (mAb) that can bind to HER3. Antigen-binding regions of antibodies such as single chain variable fragments (scFv), Fab and F(ab')2 fragments can also be used / produced. An "antigen-binding region" is any fragment of an antibody that can bind to the target for which a given antibody is specific.

[0150] Antibodies generally contain six complementarity determining regions, 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. The six CDRs together define the paratope of the antibody, which is the portion of the antibody that binds to a target antigen.

[0151] The VH and VL regions comprise, on either side of each CDR, a framework region (FR) that provides a scaffold for the CDRs. From N-terminus 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.

[0152] 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 as described in Lefranc et al., Dev. Comp. Immunol. (2003) 27: 55-77.

[0153] In some embodiments, the antigen-binding molecule comprises the CDR of the antigen-binding molecule that can bind to HER3. In some embodiments, the antigen-binding molecule comprises the FR of the antigen-binding molecule that can bind to HER3. In some embodiments, the antigen-binding molecule comprises the CDR and FR of the antigen-binding molecule that can bind to HER3. That is, in some embodiments, the antigen-binding molecule comprises the VH region and the VL region of the antigen-binding molecule that can bind to HER3.

[0154] In some embodiments, an antigen-binding molecule capable of binding to HER3 in accordance with the present disclosure is any of the embodiments of the antigen-binding molecules described in WO2019185878A1, which is incorporated herein by reference in its entirety, including any of the embodiments of the antigen-binding molecules described in WO2019185878A1 ... Pharmacol. (2017) 79(3):489-495), GSK2849330 (described, e.g., in Clarke et al., Eur J Cancer. (2014) 50:98-9), lumletuzumab (also known as RG7116 and RO-5479599, described, e.g., in Mirschberger et al., Cancer Research (2013) 73(16) 5183-5194), CDX-3379 (also known as KTN3379, described, e.g., in Lee et al., Proc Natl Acad Sci USA. 2015 Oct 27;112(43):13225), AV-203 (also known as CAN-017, described, e.g., 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 and described, for example, in de Vries Schultink et al., Clin Pharmacokinet. (2020) 59: 875-884).

[0155] In some embodiments, the antigen-binding molecule is selected from 10D1F and seribantumab. In some embodiments, the antigen-binding molecule is 10D1F.

[0156] 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.

[0157] In some embodiments, the antigen-binding molecule comprises: (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.

[0158] In some embodiments, the antigen-binding molecule comprises:

[0159] (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.

[0160] (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.

[0161] (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.

[0162] (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: 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: 52.

[0163] (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: 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: 53.

[0164] (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.

[0165] (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.

[0166] (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.

[0167] (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.

[0168] (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: 54.

[0169] (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: 54.

[0170] (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.

[0171] (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.

[0172] (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.

[0173] (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.

[0174] (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: 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: 61.

[0175] (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: 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: 62.

[0176] In embodiments according to the present disclosure in which one or more amino acids are replaced with another amino acid, the substitutions may be conservative substitutions, for example, according to the following table: In some embodiments, amino acids in the same block in the middle column are replaced. In some embodiments, amino acids in the same stretch in the rightmost column are replaced.

[0177] [Table 1] In some embodiments, the substitutions may be functionally conservative, i.e., in some embodiments, the substitutions may not affect (or may not substantially affect) one or more functional properties (e.g., binding to a target) of an antigen-binding molecule that contains the substitution compared to a comparable unsubstituted molecule.

[0178] 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, the VH and VL regions of the Fv are provided as a single polypeptide, i.e., a single chain Fv (scFv), joined by a linker region.

[0179] In some embodiments, the antigen-binding molecules of the present disclosure comprise one or more regions of an immunoglobulin heavy chain constant sequence. In some embodiments, the immunoglobulin heavy chain constant sequence is or is derived from an IgG (e.g., IgG1, IgG2, IgG3, IgG4), IgA (e.g., IgA1, IgA2), IgD, IgE, or IgM heavy chain constant sequence.

[0180] The VL and light chain constant (CL) regions, and the VH and heavy chain constant 1 (CH1) regions of the antigen-binding region of an antibody together constitute a Fab region. In some embodiments, the antigen-binding molecule of the present disclosure comprises or consists of a Fab region that binds to HER3.

[0181] In some embodiments, the antigen-binding molecules described herein comprise or consist of a whole antibody that binds to HER3. As used herein, "whole antibody" refers to an antibody that has 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.

[0182] G-type immunoglobulins (i.e., IgG) are glycoproteins of about 150 kDa that contain two heavy chains and two light chains. From the N-terminus to the C-terminus, the heavy chains contain a VH followed by a heavy chain constant region that contains three constant domains (CH1, CH2, and CH3), and similarly, the light chains contain 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 chains can be kappa (κ) or lambda (λ).

[0183] 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.

[0184] In some embodiments, the antigen-binding molecule comprises: (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:

[0185] The antigen-binding molecule according to the present disclosure may be provided in the form of a composition comprising such an antigen-binding molecule.The antigen-binding molecule may be formulated as a pharmaceutical composition or medicament for clinical use, and may include a pharmaceutically acceptable carrier, diluent, excipient, or adjuvant.The composition may be formulated for local, parenteral, systemic, intracavitary, intravenous, intraarterial, intramuscular, intrathecal, intraocular, intraconjunctival, intratumoral, subcutaneous, intradermal, intrathecal, oral, or transdermal administration routes, which may include injection or infusion.

[0186] Suitable formulations may include antigen-binding molecules in a sterile medium or in an isotonic medium. Medicaments and pharmaceutical compositions may be formulated in a fluid, including in gel form. Fluid formulations may be formulated for administration by injection or infusion (e.g., via a cannula) into the blood, tumor, or selected regions of the human or animal body. In some embodiments, compositions may be formulated for injection or infusion, for example, into blood vessels or tumors.

[0187] Antagonists of HER3-Mediated Signaling Aspects and embodiments of the present disclosure relate to therapeutic / prophylactic intervention with (i) antagonists of HER3-mediated signaling, and (ii) antigen-binding molecules that bind to HER3.

[0188] Although the HER3-binding antigen binding molecules according to the present disclosure may act as antagonists of HER3-mediated signaling, it will be understood that in accordance with aspects and embodiments of the present disclosure relating to combination treatments, components (i) and (ii) of the combination are preferably not identical (i.e., they are different agents).

[0189] Treatment with an antagonist of HER3-mediated signaling according to the present disclosure is particularly contemplated in connection with therapeutic / prophylactic intervention for the treatment / prevention of cancers described hereinabove in the section entitled "Cancers that contain / are heterozygous for / are homozygous for mutations that result in upregulation of HER3-mediated signaling." In accordance with such an embodiment, treatment with an antagonist of HER3-mediated signaling may be effective in restoring the level of signaling to that observed in the absence of the mutation (i.e., the level of signaling by a comparable cell carrying only the wild-type allele).

[0190] Thus, treatment with an antagonist of HER3-mediated signal transduction can be useful for preparing a subject for treatment with the HER3-binding antigen binding molecule described herein.That is, administration of an antagonist of HER3-mediated signal transduction is preferably effective for sensitizing (i.e., making) the cancer susceptible to treatment with the HER3-binding antigen binding molecule, so that treatment of the subject's cancer with the HER3-binding antigen binding molecule is more effective than treatment with the antagonist of HER3-mediated signal transduction.

[0191] It will be understood that the specific antagonist of HER3-mediated signaling utilized in the combination treatment according to the present disclosure can be selected according to the mutation status / genotype of the cancer to be treated.By way of illustration, if the cancer contains an activating mutation to KRAS, the antagonist can be an antagonist of signaling through the MAPK / ERK pathway.Similarly, if the cancer contains an activating mutation to PIK3CA or an inactivating mutation to PTEN, the antagonist can be an antagonist of signaling through the PI3K / AKT / mTOR pathway.

[0192] In some embodiments, the antagonist of HER3-mediated signaling according to the present disclosure is a pan-ErbB inhibitor (e.g., sapitinib or Sym013). In some embodiments, the antagonist of HER3-mediated signaling is an inhibitor of EGFR-mediated signaling (e.g., cetuximab, panitumumab, gefitinib, erlotinib, lapatinib, afatinib, brigatinib, icotinib, osimertinib, zalutumumab, vandetanib, necitumumab, nimotuzumab, dacomitinib, durigotuzumab, or matuzumab). In some embodiments, the antagonist of HER3-mediated signaling is an inhibitor of HER2-mediated signaling (e.g., trastuzumab, pertuzumab, lapatinib, neratinib, afatinib, dacomitinib, MM-111, xenoctuzumab, MCLA-128, or margetuximab). In some embodiments, the antagonist of HER3-mediated signaling is an inhibitor of signaling mediated by HER3 (e.g., seribantumab, lumuletuzumab, elgemtumab, KTN3379, AV-203, GSK2849330, REGN1400, MP-RM-1, EV20, pertuzumab, durigotuzumab, MM-111, xenoctuzumab, istiratumab, MCLA-128, patritumab, EZN-3920, RB200, U3-1402, TX2-121-2, EZN-3920, and miR-205). In some embodiments, the antagonist of HER3-mediated signaling is an inhibitor of signaling mediated by HER4 (eg, lapatinib, ibrutinib, afatinib, dacomitinib, or neratinib).

[0193] In some embodiments, the antagonist of HER3-mediated signaling inhibits downstream effectors of HER3 signaling.Downstream effectors of HER3-mediated signaling include, for example, PI3K, AKT, KRAS, BRAF, MEK / ERK, and mTOR.In some embodiments, the antagonist of HER3-mediated signaling is an inhibitor of MAPK / ERK pathway.In some embodiments, the antagonist of HER3-mediated signaling is an inhibitor of PI3K / ATK / mTOR pathway.

[0194] In some embodiments, the antagonist of HER3-mediated signaling is a PI3K inhibitor (e.g., pictilisib, buparlisib, dactolisib, SAR245409, AZD8186, idelalisib, copanlisib, or duvelisib). In some embodiments, the antagonist of HER3-mediated signaling is an AKT inhibitor (e.g., MK-2206, AZD5363, GSK690693, GSK2110183, ipatasertib, VQD-002, perifosine, or miltefosine). In some embodiments, the antagonist of HER3-mediated signaling is an inhibitor of KRAS (e.g., sotorasib (also known as AMG510), ARS-1620, adagrasib (also known as MRTX849), LY3499446, ARS-3248 / JNJ-74699157, BI2852, or RRSP chimeric toxin). In some embodiments, the antagonist of HER3-mediated signaling is a BRAF inhibitor (e.g., vemurafenib, dabrafenib, SB590885, XL281, RAF265, encorafenib, belbalafenib, PLX8394, LY3009120, LXH254, GDC-0879, PLX-4720, sorafenib, or LGX818). In some embodiments, the antagonist of HER3-mediated signaling is a MEK / ERK inhibitor (e.g., trametinib, cobimetinib, binimetinib, selumetinib, pimasertib, PD-325901, CI-1040, PD035901, or TAK-733). In some embodiments, the antagonist of HER3-mediated signaling is an mTOR inhibitor (e.g., rapamycin, deforolimus, temsirolimus, everolimus, ridaforolimus, or sapanisertib).

[0195] Therapeutic and preventive interventions Aspects and embodiments of the present disclosure relate to therapeutic and prophylactic interventions for the treatment and prevention of cancer as described herein.

[0196] The present disclosure provides an antigen-binding molecule that binds to HER3 for use in a method for treating or preventing a cancer described herein in a subject.Also provides the use of an antigen-binding molecule that binds to HER3 in the manufacture of a medicament for use in treating or preventing a cancer described herein in a subject.Also provides a method for treating or preventing a cancer described herein in a subject, comprising administering to the subject a therapeutically or prophylactically effective amount of an antigen-binding molecule that binds to HER3.

[0197] The present disclosure also provides an antigen-binding molecule that binds to HER3 for use in a method for treating or preventing a cancer described herein in a subject, the method further comprising administering an antagonist of HER3-mediated signaling. Also provides a use of an antigen-binding molecule that binds to HER3 in the manufacture of a medicament for use in treating or preventing a cancer described herein in a subject, the method further comprising administering an antagonist of HER3-mediated signaling. Also provides a method for treating or preventing a cancer described herein in a subject, the method comprising administering to the subject a therapeutically or prophylactically effective amount of an antigen-binding molecule that binds to HER3, and further comprising administering an antagonist of HER3-mediated signaling.

[0198] The present disclosure also provides (i) an antagonist of HER3-mediated signal transduction and (ii) an antigen-binding molecule that binds to HER3 for use in a method for treating or preventing a cancer described herein in a subject. Also provides the use of (i) an antagonist of HER3-mediated signal transduction and (ii) an antigen-binding molecule that binds to HER3 in the manufacture of a medicament for use in treating or preventing a cancer described herein in a subject. Also provides a method for treating or preventing a cancer described herein in a subject, comprising administering to the subject a therapeutically or prophylactically effective amount of (i) an antagonist of HER3-mediated signal transduction and (ii) an antigen-binding molecule that binds to HER3.

[0199] 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.

[0200] The present disclosure also provides an antagonist of HER3-mediated signal transduction for use in a method for treating or preventing a cancer described herein in a subject, the method further comprising administering an antigen-binding molecule that binds to HER3. Also provides a use of an antagonist of HER3-mediated signal transduction in the manufacture of a medicament for use in treating or preventing a cancer described herein in a subject, the method further comprising administering an antigen-binding molecule that binds to HER3. Also provides a method for treating or preventing a cancer described herein in a subject, the method comprises administering a therapeutically or prophylactically effective amount of an antagonist of HER3-mediated signal transduction to the subject, and further comprises administering an antigen-binding molecule that binds to HER3.

[0201] Therapeutic or preventive intervention according to the present disclosure may be effective in reducing the onset or progression of cancer, alleviating one or more symptoms of cancer, or reducing the pathology of cancer. Intervention may be effective in preventing the progression of cancer, for example, preventing the progression of cancer, or slowing the onset rate of cancer. In some embodiments, intervention may result in the improvement of cancer, for example, reducing the symptoms of cancer, or reducing some other correlates of cancer severity / activity. In some embodiments, the method may prevent the onset of later stages of cancer (e.g., more severe stages or metastasis).

[0202] In some embodiments, therapeutic or prophylactic intervention may be aimed at one or more of delaying / preventing the onset / progression of cancer symptoms, reducing the severity of cancer symptoms, reducing cancer cell survival / growth / invasion / metastasis, reducing the number of cancer cells, and / or prolonging the survival of a subject.

[0203] The administration of the articles of the present disclosure (i.e., HER3-binding antigen-binding molecules and antagonists of HER3-mediated signal transduction) is preferably in a "therapeutically effective" or "prophylactically effective" amount, which is an amount sufficient to show 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 article administered. Prescription of treatment, such as the decision about dosage, is within the responsibility of general practitioners and other physicians, 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 Pharmaceutical Sciences, 20th edition, 2000, pub. Lippincott, Williams & Wilkins.

[0204] Administration may be by any suitable route, for example, injection or infusion (e.g., via a cannula). Administration may be to the blood, tumor, or to a selected area of ​​the human or animal body (e.g., an organ / tissue where a cancer symptom is manifested). The particular mode and / or site of administration may be selected depending on where the therapeutic effect is required.

[0205] When two or more agents (eg, an antagonist of HER3-mediated signaling and a HER3-binding antigen binding molecule) are administered in combination, the agents can be administered either simultaneously or sequentially.

[0206] Concomitant administration refers to administration of two or more agents (e.g., an antagonist of HER3-mediated signaling and a HER3-binding antigen binding molecule) together, e.g., as a pharmaceutical composition containing both agents (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.

[0207] 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.

[0208] In some embodiments, a therapeutic or prophylactic intervention according to the present disclosure comprises (i) administering an antagonist of HER3-mediated signaling (e.g., an antagonist of HER3-mediated signaling described herein) to a subject having cancer (e.g., a cancer that contains / is heterozygous for / is homozygous for a mutation that results in upregulation of HER3-mediated signaling as described herein), and (ii) administering to the subject an antigen binding molecule that binds HER3 (e.g., a HER3-binding antigen binding molecule described herein). 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)).

[0209] In some embodiments, the present invention includes further therapeutic or preventive interventions, for example, for the treatment / prevention of cancer. In some embodiments, the therapeutic or preventive interventions are selected from chemotherapy, immunotherapy, radiation therapy, surgery, vaccination, and / or hormonal therapy. In some embodiments, the therapeutic or preventive interventions include leukapheresis. In some embodiments, the therapeutic or preventive interventions include stem cell transplantation.

[0210] Multiple doses of the disclosed article may be provided. The multiple doses may be separated by a predetermined time interval, which may 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. By way of example, doses may be given every 7, 14, 21, or 28 days (± 3, 2, or 1 day). One or more of the doses, or each, may be accompanied by simultaneous or sequential administration of another therapeutic agent.

[0211] Diagnostic and prognostic methods, and patient selection The present disclosure also provides diagnostic, prognostic and predictive methods related to the cancers described herein.

[0212] The method may be performed on a sample obtained from a subject in vitro, or may be performed after processing of a sample obtained from a subject. Once the sample is collected, the method may not be performed within a human or animal body, as the subject is not required to be present for the performance of the in vitro method. However, in some embodiments, the method may be performed in vivo.

[0213] The sample may be taken from any tissue or body fluid. The sample may include or be derived from a volume of blood; a volume of serum from the subject's blood, which may include the fluid portion of blood obtained after removing fibrin clots and blood cells; a tissue sample or biopsy; pleural effusion; cerebrospinal fluid (CSF); or cells isolated from the subject. In some embodiments, the sample may be obtained from or be derived from a tissue or tissues affected by a disease / condition (e.g., a tissue or tissues where a disease symptom is manifested or involved in the pathogenesis of a disease / condition). In some embodiments, the sample may be obtained from or be derived from a cancer, tumor, or cells thereof.

[0214] The method may be performed for the purpose of diagnosing cancer (e.g., cancers described herein). The method may be performed for the purpose of prognosing / predicting a subject's likely response to a therapeutic / prophylactic intervention described herein. The method may be useful, for example, in terms of efficacy, to predict the likely response to a given therapeutic / prophylactic intervention, and thus may be useful for supporting clinical decision-making. The method may be performed for the purpose of identifying / selecting a subject for a therapeutic / prophylactic intervention described herein.

[0215] In some aspects and embodiments, the methods include analyzing the subject's cancer to determine whether the cancer is a cancer described herein, e.g., a cancer that is not homozygous / heterozygous for / does not contain a mutation that results in upregulation of HER3-mediated signaling, or a cancer that contains / is heterozygous for / homozygous for a mutation that results in upregulation of HER3-mediated signaling.

[0216] In some embodiments, the method includes evaluating the cancer to determine whether it comprises one or more mutations as described above. In some embodiments, the method includes evaluating the cancer to determine whether it comprises cells that comprise one or more copies of an allele that comprises a mutation as described above.

[0217] Cancers identified after such analysis as not containing the mutation and / or not containing cells containing one or more copies of the allele that contains the mutation may be identified as cancers suitable for intervention with an antigen binding molecule that binds HER3 according to the present disclosure.

[0218] Following such analysis, cancers identified as containing a mutation and / or comprising cells that contain one or more copies of an allele that contains a mutation may be identified as cancers suitable for intervention with a combination of (i) an antagonist of HER3-mediated signaling in accordance with the present disclosure and (ii) an antigen binding molecule that binds HER3 in accordance with the present disclosure.

[0219] Aspects and embodiments of the present disclosure also include selecting a subject for therapeutic or prophylactic intervention according to the present disclosure.

[0220] After such analysis, subjects with cancers identified as not containing the mutation and / or not containing cells containing one or more copies of the allele that contains the mutation may be selected for treatment with an antigen binding molecule that binds HER3 according to the present disclosure.

[0221] Following such analysis, subjects with cancers identified as containing a mutation and / or comprising cells that contain one or more copies of an allele that contains a mutation may be selected for treatment with (i) an antagonist of HER3-mediated signaling in accordance with the present disclosure, and (ii) an antigen binding molecule that binds HER3 in accordance with the present disclosure.

[0222] In some embodiments, a subject is selected / not selected for therapeutic intervention according to the present disclosure based on the results of an analysis of the subject's cancer to determine whether the cancer contains cells that contain a mutation described herein and / or one or more copies of an allele that contains a mutation described herein.

[0223] In some embodiments, the methods include selecting a subject for treatment with an antigen binding molecule that binds HER3 according to the present disclosure based on a determination that the subject has a cancer that does not contain a mutation described herein and / or that cells of the subject's cancer have not been determined to contain one or more copies of an allele that contains a mutation described herein.

[0224] In some embodiments, the method includes selecting the subject for treatment with (i) an antagonist of HER3-mediated signaling in accordance with the present disclosure, and (ii) an antigen binding molecule that binds HER3 in accordance with the present disclosure, based on a determination that the subject has a cancer that includes a mutation described herein and / or that cells of the cancer include one or more copies of an allele that includes a mutation described herein.

[0225] In some aspects and embodiments, the method comprises: (a) analyzing the subject's cancer to determine whether the cancer contains a mutation described herein and / or one or more copies of an allele that contains a mutation described herein; and (b) if the subject's cancer is determined to be not homozygous / heterozygous for or not containing a mutation described herein in step (a) (e.g., if the subject's cancer is determined to be a cancer according to the cancer embodiments described in the section entitled "Cancers Not Homozygous / Heterozygous for / Not Containing a Mutation That Results in Upregulation of HER3-Mediated Signaling"), selecting the subject for treatment with an antigen binding molecule that binds HER3 according to the present disclosure. Includes.

[0226] In some embodiments, the method comprises: (c) administering an antigen-binding molecule that binds HER3 according to the present disclosure to the subject selected for treatment in step (b). Further includes:

[0227] In some aspects and embodiments, the method comprises: (a) analyzing the subject's cancer to determine whether the cancer contains a mutation described herein and / or one or more copies of an allele that contains a mutation described herein; and (b) if the subject's cancer is determined to contain or be heterozygous / homozygous for a mutation described herein in step (a) (e.g., if the subject's cancer is determined to be a cancer according to the cancer embodiments described in the section entitled "Cancers containing / heterozygous / homozygous for a mutation that results in upregulation of HER3-mediated signaling"), selecting the subject for treatment with (i) an antagonist of HER3-mediated signaling according to the present disclosure, and (ii) an antigen-binding molecule that binds HER3 according to the present disclosure. Includes.

[0228] In some embodiments, the method comprises: (c) administering (i) an antagonist of HER3-mediated signaling according to the present disclosure, and (ii) an antigen-binding molecule that binds HER3 according to the present disclosure to the subject selected for treatment in step (b). Further includes:

[0229] In some aspects and embodiments, the method comprises: (a) analyzing the subject's cancer to determine whether the cancer contains a mutation described herein, and / or one or more copies of an allele that contains a mutation described herein; (b) if the subject's cancer is determined to contain or be heterozygous / homozygous for a mutation described herein in step (a) (e.g., if the subject's cancer is determined to be a cancer according to the cancer embodiments described in the section entitled "Cancers containing / heterozygous / homozygous for a mutation that results in upregulation of HER3-mediated signaling"), selecting the subject for treatment with (i) an antagonist of HER3-mediated signaling according to the present disclosure, and (ii) an antigen binding molecule that binds HER3 according to the present disclosure; (c) administering an antagonist of HER3-mediated signaling according to the present disclosure to the subject selected for treatment in step (b); and (d) administering an antigen-binding molecule that binds HER3 according to the present disclosure to the subject selected for treatment in step (b). Includes.

[0230] It will be understood that steps (c) and (d) of the preceding paragraph may be performed simultaneously or sequentially.

[0231] The analysis of the cancer may include analyzing the nucleotide sequence of a gene encoding one or more factors involved in HER3-mediated signaling (e.g., in a nucleic acid-containing sample obtained from the cancer) to determine whether the cancer contains a mutation as described herein above (e.g., a mutation resulting in upregulation of HER3-mediated signaling, a mutation resulting in upregulation of signaling through the MAPK / ERK pathway, and / or a mutation resulting in upregulation of signaling through the PI3K / AKT / mTOR pathway; e.g., an activating mutation for a gene encoding a positive regulator of said signaling / signaling pathway, and / or an inactivating mutation for a gene encoding a negative regulator of said signaling / signaling pathway). The analysis may be of cells of the cancer. The analysis may be of a nucleic acid-containing sample / biopsy obtained from the cancer / its cells. Such an analysis is preferably performed in vitro.

[0232] Methods for evaluating the nucleotide sequence of a gene of interest are well known in the art and include, for example, classical chain termination sequencing and next-generation sequencing (NGS) techniques, which are reviewed, for example, in Metzker, ML, Nat Rev Genet (2010) 11(1):31-46, which is incorporated herein by reference in its entirety. Additional methods for evaluating the nucleotide sequence of a gene of interest for the presence / absence of mutations include restriction fragment length polymorphism identification (RFLPI) of genomic DNA, random amplified polymorphism detection (RAPD) of genomic DNA, amplified fragment length polymorphism detection (AFLPD), multiple locus variable number tandem repeat (VNTR) analysis (MLVA), SNP genotyping, multilocus sequence typing, allele-specific oligonucleotide (ASO) probes, and oligonucleotide microarrays or beads. Other suitable methods are described, for example, in Edenberg HJ and Liu Y, Cold Spring Harb Protoc; 2009; doi:10.1101 / pdb.top62, and Tsuchihashi Z and Dracopoli NC, Pharmacogenomics J., 2002, 2:103-110.

[0233] In a further aspect, the disclosure provides methods for determining whether a subject is likely to respond favorably to therapeutic or prophylactic intervention with an antigen binding molecule that binds to HER3.

[0234] The response of a subject / cancer to a given therapeutic or preventive intervention may be assessed 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, which is incorporated herein by reference in its entirety). In some embodiments, a subject / cancer is considered to have "responded" to a given intervention if it achieves one of the following: a complete response, a partial response, or stable disease. In some embodiments, a subject / cancer is considered to have "responded" to a given intervention if it achieves one of the following: a complete response or a partial response.

[0235] In some embodiments, the methods include analyzing the subject's cancer to determine whether the cancer comprises a mutation described herein and / or one or more copies of an allele that comprises a mutation described herein, wherein subjects having a cancer determined to comprise or be heterozygous / homozygous for a mutation described herein (e.g., if the subject's cancer is determined to be a cancer according to the cancer embodiments described in the section entitled "Cancers that comprise / are heterozygous / homozygous for a mutation that results in upregulation of HER3-mediated signaling") are determined to be unlikely to respond well to therapeutic or prophylactic intervention using an antigen binding molecule that binds HER3.

[0236] In some embodiments, the methods include analyzing the subject's cancer to determine whether the cancer comprises a mutation described herein and / or one or more copies of an allele comprising a mutation described herein, and wherein a subject having a cancer determined to be not homozygous / heterozygous for or not containing a mutation described herein (e.g., if the subject's cancer is determined to be a cancer according to the cancer embodiments described in the section entitled "Cancers that are not homozygous / heterozygous for / not containing a mutation that results in upregulation of HER3-mediated signaling") is determined to be likely to respond well to therapeutic or prophylactic intervention using an antigen binding molecule that binds HER3.

[0237] subject The subject according to the embodiment of the present disclosure may be any animal or human. The subject is preferably a mammal, more preferably a human. The subject may be a non-human mammal, but more preferably a human. The subject may be male or female. The subject may be a patient. The subject may have been diagnosed with a disease or condition (e.g., cancer as described herein) that requires treatment, may be suspected of having such a disease / condition, or may be at risk of developing / suffering from such a disease / condition.

[0238] The subject may have cancer.The subject may have (e.g., may have been determined to have) a cancer that is not homozygous, not heterozygous, or does not contain one or more mutations that cause upregulation of HER3-mediated signaling, as described herein.The subject may have (e.g., may have been determined to have) a cancer that contains, is heterozygous, or is homozygous, as described herein, a cancer that causes upregulation of HER3-mediated signaling, as described herein.

[0239] kit The present disclosure also provides kits of parts. Kits according to the present disclosure may include components for carrying out all or part of the methods described herein.

[0240] In some embodiments, the kit may include (i) a means for assessing a cancer to determine whether it contains one or more mutations described herein, and (ii) an antigen-binding molecule that binds to HER3 as described herein. In some embodiments, the kit may include an antagonist of HER3-mediated signaling as described herein.

[0241] The means for assessing a cancer to determine whether it contains one or more mutations described herein may include, for example, one or more oligonucleotides having complementarity to the nucleotide sequences of the genes described herein.

[0242] The kit may include instructions for evaluating cancer to determine whether the cancer comprises one or more mutations described herein.The kit may include instructions for administering to a subject an antigen-binding molecule that binds to HER3 as described herein.The kit may further include instructions for administering to a subject an antagonist of HER3-mediated signal transduction as described herein.

[0243] The kits may further include reagents, buffers and / or standards required for the practice of the methods according to the present disclosure.

[0244] 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 percentage sequence identity between the sequences. Pairwise multiple sequence alignments for the purpose of determining the percent sequence identity between two or more amino acid or nucleic acid sequences can be accomplished 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 preferred to use default parameters, for example, for gap penalties and extension penalties.

[0245] array

[0246] [Table 2-1]

[0247] [Table 2-2]

[0248] [Table 2-3]

[0249] [Table 2-4]

[0250] [Table 2-5]

[0251] [Table 2-6]

[0252] [Table 2-7] The present disclosure includes combinations of the described aspects and preferred features unless such combinations are expressly not permitted or explicitly avoided.

[0253] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

[0254] 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.

[0255] Throughout this specification, including the claims which follow, unless the context requires otherwise, the word "comprise", and variations such as "comprises" and "comprising", are 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.

[0256] 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.

[0257] When a nucleic acid sequence is disclosed herein, the reverse complement thereof is also expressly contemplated.

[0258] The methods described herein may preferably be performed in vitro. The term "in vitro" is intended to encompass procedures performed on cells in culture, whereas the term "in vivo" is intended to encompass procedures on / on an intact multicellular organism.

[0259] BRIEF DESCRIPTION OF THE DRAWINGS Embodiments and experiments illustrating the principles of the present disclosure will now be discussed with reference to the accompanying figures. [Brief description of the drawings]

[0260] [Figure 1A] 1A-1C are bar graphs and graphs showing a comparison of the anti-cancer efficacy of 10D1F / 10D1P against different cancer cell lines as determined by analysis of in vitro and in vivo studies. (1A) Shows the efficacy of 10D1F / 10D1P in inhibiting cell growth of the indicated cancer cell lines in vitro or in vivo in cell line-derived xenograft models. [Figure 1B]1A-1C are bar graphs and graphs showing a comparison of the anti-cancer efficacy of 10D1F / 10D1P against different cancer cell lines as determined by analysis of in vitro and in vivo studies. (1B) Linear regression modeling the relationship between the efficacy determined for treatment of cell lines with 10D1F / 10D1P in vitro and in vivo. [Figure 2A] Graphs showing the response of cancer cell lines with different genotypes (see Example 2) to treatment with (2A) 10D1F / 10D1P, and (2B) seribantumab, as determined by analysis of in vitro and in vivo studies. ****P≦0.0001. [Figure 2B] Graphs showing the response of cancer cell lines with different genotypes (see Example 2) to treatment with (2A) 10D1F / 10D1P, and (2B) seribantumab, as determined by analysis of in vitro and in vivo studies. ****P≦0.0001. [Figure 3A] Graphs showing the response of cancer cell lines with different genotypes (see Example 3) to treatment with (3A) 10D1F / 10D1P, and (3B) seribantumab, as determined by analysis of in vitro and in vivo studies. ****P≦0.0001. [Figure 3B] Graphs showing the response of cancer cell lines with different genotypes (see Example 3) to treatment with (3A) 10D1F / 10D1P, and (3B) seribantumab, as determined by analysis of in vitro and in vivo studies. ****P≦0.0001. EXAMPLES

[0261] In this example, the inventors demonstrate that the degree of response to treatment with anti-HER3 antibodies depends on the mutation status of the cancer of genes that code for regulators of HER3-mediated signal transduction. They show that cancers that lack mutations for KRAS, PIK3CA, BRAF, and PTEN respond exceptionally well to treatment with anti-HER3 antibodies. The example further shows that NRG1 expression predicts positive response to anti-HER3 antibody treatment.

[0262] Example 1 Analysis of the correlation between inhibition of cell proliferation in vitro and inhibition of tumor growth in vivo The inventors investigated whether the results of an analysis of the ability of anti-HER3 antibodies 10D1F hIgG1 (VH=SEQ ID NO:33, VL=SEQ ID NO:58) or 10D1P hIgG1 (VH=SEQ ID NO:21, VL=SEQ ID NO:49) to inhibit cell growth of cancer cell lines in vitro would predict their ability to inhibit tumor growth of cell line-derived xenograft tumors in vivo.

[0263] The cell lines analyzed were N87 (gastric cancer), FaDu (head and neck cancer), OvCAR8 (ovarian cancer), SNU16 (gastric cancer), A549 (lung cancer), HCC95 (lung cancer), AHCN (renal cancer), and 22Rv1 (prostate cancer).

[0264] For in vitro growth inhibition analysis, cells of different cell lines were treated in triplicate with 10-point serially diluted concentrations of anti-HER3 antibodies (3-fold dilutions starting from the highest concentration of 1500 μg / ml). Cell viability was measured after 3-5 days using a CCK-8 assay (Dojindo). The percentage inhibition of cell growth was calculated by comparison with the CCK-8 assay signal for cells treated with buffer only (PBS). In vitro efficacy was determined to be the percentage inhibition observed at 1500 μg / ml.

[0265] For in vivo growth inhibition analysis, cell line-derived xenograft models were established by subcutaneous injection of cells into the right flank of female NCr nude mice (approximately 6-8 weeks old). Anti-HER3 antibodies were administered by intraperitoneal injection twice weekly at 25 mg / kg body weight per dose. The control treatment group received an equal volume of PBS. Tumor volumes were measured three times weekly using digital calipers and calculated using the formula [L×W2 / 2]. The study endpoint was considered to be reached when tumors in the control arm measured more than 1.5 cm in length. In vivo efficacy was determined as the percentage of tumor growth inhibition at the end of the study.

[0266] Efficacy data from in vitro and in vivo studies were normalized to set the maximum inhibition rate as 100% and the minimum inhibition rate as 0%, with values ​​above or below this range. In vitro-in vivo efficacy correlations were studied by simple linear regression and correlation coefficients (R 2 ) was calculated using GraphPad Prism software.

[0267] The results of the analysis are shown in Figures 1A and 1B. There was a strong correlation between the results obtained in the in vitro and in vivo studies (R 2 = 0.8061), demonstrating that the results of the in vitro cancer cell growth inhibition assay are predictive of efficacy in tumor growth inhibition in vivo (and vice versa).

[0268] Example 2 Analysis of the efficacy of HER3-binding antigen-binding molecules to treat cancer based on the mutation status of regulators of HER3-mediated signaling We used combined data from in vivo and in vitro studies of the anticancer activity of 10D1F / 10D1P or seribantumab to investigate whether the mutational status of cancer cell lines is predictive of efficacy of treatment with the antibodies.

[0269] Cell lines from breast, gastric and non-small cell lung, colorectal and SCCHN, and prostate cancer were included in the analysis. Cell lines were analyzed for the presence of point mutations in KRAS, PIK3CA, and for PTEN loss using public databases such as COSMIC, Cellosaurus, and CCLE. Cell lines were classified as (i) wild-type for KRAS, PIK3CA, and PTEN ("wild-type"); (ii) heterozygous for an activating mutation for KRAS or PIK3CA, or heterozygous for an inactivating mutation for PTEN ("heterozygous"); (iii) homozygous for an activating mutation for KRAS or PIK3CA, or homozygous for an inactivating mutation for PTEN ("homozygous").

[0270] When scoring response to treatment for 10D1F / 10D1P, priority was given to tumor growth inhibition (TGI) data from in vivo studies using 10D1F hIgG1. If such data was not available, TGI data from 10D1P hIgG1 was used. If in vivo data was not available for a cell line, in vitro inhibition data was used.

[0271] The relationship between normalized functional response to treatment with 10D1F / 10D1P or seribantumab and mutation status was analyzed using GraphPad Prism. Unpaired t-tests were performed for comparisons between wild-type, heterozygous, and homozygous groups.

[0272] The genotypes of the cell lines for KRAS, PIK3CA and PTEN, and their normalized functional responses to treatment with 10D1F / 10D1P or seribantumab (determined by analysis in in vitro or in vivo studies described in Example 1) are shown in the table below and the data are represented graphically in Figures 2A and 2B.

[0273] [Table 3] The analysis revealed a very striking pattern in which the response to treatment with anti-HER3 antibodies was dependent on the presence of mutations that lead to upregulation of HER3-mediated signaling. Cell lines in the wild-type group were much more responsive to anti-HER3 antibody treatment compared to cell lines in the heterozygous or homozygous groups. This pattern was observed across the full range of different cancer types analyzed.

[0274] Example 3 Analysis of the efficacy of HER3-binding antigen binding molecules to treat cancer based on PTEN / KRAS / PIK3CA / BRAF mutation status and NRG1 mRNA abundance In further studies, we utilized data from in vivo and in vitro studies of the anti-cancer activity of 10D1F / 10D1P or seribantumab to investigate whether BRAF mutation status could be added to the triple gene signature (PTEN / KRAS / PIK3CA) described in Example 2 to predict the efficacy of anti-HER3 antibody treatment.

[0275] Cell lines derived from breast, gastric and non-small cell lung cancer, colorectal and SCCHN, and prostate cancer were included in the analysis. Cell lines were assessed for the presence of point mutations in KRAS, PIK3CA, and BRAF, and for PTEN loss by empirical sequence analysis. Cell lines were classified as (i) wild-type for KRAS, PIK3CA, PTEN, and BRAF ("wild-type"); (ii) heterozygous for activating mutations for KRAS or PIK3CA or BRAF, or heterozygous for inactivating mutations for PTEN ("heterozygous"); (iii) homozygous for activating mutations for KRAS or PIK3CA or BRAF, or homozygous for inactivating mutations for PTEN ("homozygous"). Expression levels of NRG1 mRNA were also measured for each cell line.

[0276] When scoring response to treatment for 10D1F / 10D1P, priority was given to tumor growth inhibition (TGI) data from in vivo studies using 10D1F hIgG1. If such data was not available, TGI data from 10D1P hIgG1 was used. If in vivo data was not available for a cell line, in vitro inhibition data was used.

[0277] The relationship between normalized functional response to treatment with 10D1F / 10D1P or seribantumab and mutation status was analyzed using GraphPad Prism. Unpaired t-tests were performed for comparisons between wild-type, heterozygous, and homozygous groups.

[0278] The genotypes of the cell lines for KRAS, PIK3CA, PTEN and BRAF as determined by sequence analysis, and their normalized functional response to treatment with 10D1F / 10D1P or seribantumab (determined by analysis in in vitro or in vivo studies described in Example 1) are shown in the table below and the data are represented graphically in Figures 3A and 3B.

[0279] [Table 4] Analysis revealed that the mutational status of BRAF in combination with PIK3CA, KRAS, and PTEN predicted efficacy of anti-HER3 antibody treatment, with cell lines in the wild-type group being much more responsive to anti-HER3 antibody treatment compared to cell lines from the heterozygous or homozygous groups (see Figures 3A and 3B). This pattern was observed across a wide range of preclinical cancer models.

[0280] The data in the table above further demonstrate the association between NRG1 expression and treatment efficacy, with increased NRG1 expression correlating with improved efficacy of anti-HER3 antibody treatment.

[0281] In conclusion, a gene signature including PIK3CA, KRAS, BRAF, and PTEN mutation status consistently and significantly predicts the efficacy of anti-HER3 antibody treatment in preclinical cancer models, and this gene signature can be combined with NRG1 mRNA abundance as an additional predictor.

Claims

1. A pharmaceutical for treating or preventing HER3-associated cancer, the pharmaceutical comprising an antigen-binding molecule that binds to HER3 as an active ingredient, wherein the HER3-associated cancer (i) contains at least one gene encoding a positive regulator of HER3-mediated signaling that does not contain an activating mutation, or (ii) contains at least one gene encoding a negative regulator of HER3-mediated signaling that does not contain an inactivating mutation.

2. HER3-associated cancers include (i) KRAS, PIK3CA, PIK3CB, PIK3CD, ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GRB2, GAB2, PTPN11, SHP2, SOS1, HRAS, NRAS, RAF1, MAP2K1, MAP2K2, MAPK1, MYC, RPS6KA1, RPS6, and MKNK1. , CREB1, MTOR, PDK1, AKT1, AKT2, AKT3, JAK2, STAT3, STAT5, and BRAF, or (ii) does not contain an inactivating mutation in a gene selected from PTEN, PPP2CA, PIK3R1, PIK3R2, NF1, BAD, and PHLPP1.

3. The pharmaceutical agent of claim 1, wherein the HER3-associated cancer (i) does not contain an activating mutation in KRAS, or (ii) does not contain an activating mutation in PIK3CA, or (iii) does not contain an activating mutation in BRAF; or (iv) does not contain an inactivating mutation in PTEN.

4. The HER3-associated cancer (i) does not contain activating mutations for KRAS and does not contain activating mutations for PIK3CA; or (ii) does not contain activating mutations for KRAS and does not contain inactivating mutations for PTEN; or (iii) does not contain activating mutations for PIK3CA and does not contain inactivating mutations for PTEN; or (iv) does not contain activating mutations for KRAS and does not contain activating mutations for BRAF; or (v) does not contain activating mutations for PIK3CA and does not contain activating mutations for BRAF; or (vi) does not contain activating mutations for BRAF and does not contain inactivating mutations for PTEN; or (vii) does not contain activating mutations for KRAS and does not contain activating mutations for PIK3CA. The pharmaceutical composition of claim 1, which does not contain activating mutations for BRAF, or (viii) does not contain activating mutations for PIK3CA, does not contain activating mutations for BRAF, and does not contain inactivating mutations for PTEN; or (ix) does not contain activating mutations for BRAF, does not contain activating mutations for KRAS, and does not contain inactivating mutations for PTEN; or (x) does not contain activating mutations for KRAS, does not contain activating mutations for PIK3CA, and does not contain inactivating mutations for PTEN; or (xi) does not contain activating mutations for KRAS, does not contain activating mutations for BRAF, does not contain activating mutations for PIK3CA, and does not contain inactivating mutations for PTEN.

5. The pharmaceutical agent of claim 1, wherein the HER3-associated cancer does not contain a mutation that results in upregulation of HER3-mediated signaling.

6. The method of claim 1, wherein the HER3-associated cancer comprises cells that express NRG1 at a level higher than the expression level by comparable non-cancerous cells.

7. A pharmaceutical for use in a method for treating or preventing a HER3-associated cancer, the pharmaceutical comprising an antigen-binding molecule that binds to HER3 as an active ingredient, the HER3-associated cancer comprising a mutation that results in upregulation of HER3-mediated signaling, and the method further comprising the step of administering an antagonist of HER3-mediated signaling.

8. The pharmaceutical agent of claim 7, wherein the mutation that results in upregulation of HER3-mediated signaling is an activating mutation in a gene encoding a positive regulator of HER3-mediated signaling or an inactivating mutation in a gene encoding a negative regulator of HER3-mediated signaling.

9. HER3-associated cancers include (i) KRAS, PIK3CA, PIK3CB, PIK3CD, ERBB3, ERBB2, ERBB4, EGFR, IGF1R, NRG1, NRG2, EGF, IRS2, GRB2, GAB2, PTPN11, SHP2, SOS1, HRAS, NRAS, RAF1, MAP2K1, MAP2K2, MAPK1, MYC, RPS6KA1, RPS6, and MKNK.

8. The pharmaceutical composition of claim 7, comprising (i) an activating mutation in a gene selected from: PIK3R1, PIK3R2, NF1, BAD, and PHLPP1; and / or (ii) an inactivating mutation in a gene selected from: PIK3R1, PIK3R2, NF1, BAD, and PHLPP1.

10. The pharmaceutical agent of claim 1 or claim 7, wherein the HER3-associated cancer comprises one or more of an activating mutation in KRAS, an activating mutation in PIK3CA, an activating mutation in BRAF, or an inactivating mutation in PTEN.

11. 1. A method for selecting a subject to be treated with an antigen-binding molecule that binds to HER3, comprising: (a) analyzing the subject's cancer to determine whether the cancer (i) contains at least one gene encoding a positive regulator of HER3-mediated signaling that does not contain an activating mutation; or (ii) contains at least one gene encoding a negative regulator of HER3-mediated signaling that does not contain an inactivating mutation; (b) if the subject's cancer is determined to not contain such a mutation in step (a), selecting the subject to be treated with an antigen-binding molecule that binds to HER3; A method comprising:

12. 1. A method for selecting a subject to be treated with (i) an antagonist of HER3-mediated signaling and (ii) an antigen-binding molecule that binds to HER3, comprising: (a) analyzing the subject's cancer to determine whether the cancer contains a mutation that results in upregulation of HER3-mediated signaling; (b) if the subject's cancer is determined to contain such a mutation in step (a), selecting the subject to be treated with (i) an antagonist of HER3-mediated signaling and (ii) an antigen-binding molecule that binds to HER3; A method comprising:

13. The medicament of claim 1 or claim 7, or the method of claim 11 or claim 12, wherein the HER3-associated cancer is selected from solid tumors, breast cancer, breast cancer, ductal breast cancer, gastric cancer, gastric adenocarcinoma, colorectal cancer, colorectal adenocarcinoma, head and neck cancer, head and neck squamous cell carcinoma, lung cancer, non-small cell lung cancer, lung adenocarcinoma, squamous cell lung carcinoma, ovarian cancer, ovarian cancer, serous ovarian adenocarcinoma, renal cancer, renal cell carcinoma, clear cell renal carcinoma, renal cell adenocarcinoma, papillary renal cell carcinoma, pancreatic cancer, pancreatic adenocarcinoma, pancreatic ductal adenocarcinoma, cervical cancer, cervical squamous cell carcinoma, skin cancer, melanoma, esophageal cancer, esophageal adenocarcinoma, liver cancer, hepatocellular carcinoma, cholangiocarcinoma, uterine cancer, endometrial cancer, thyroid cancer, thyroid cancer, pheochromocytoma, paraganglioma, bladder cancer, bladder urothelial carcinoma, prostate cancer, prostate adenocarcinoma, sarcoma, and thymoma.

14. The pharmaceutical composition of claim 1 or 7, or the method of claim 11 or 12, wherein the antigen-binding molecule that binds to HER3 is selected from 10D1F, seribantumab, elgemtumab, patritumab, GSK2849330, lumletuzumab, CDX-3379, AV-203, varicetamab, TK-A3, TK-A4, MP-EV20, 1A5-3D4, 9F7-F11, 16D3-C1, NG33, A5, F4, huHER3-8, REGN1400, and xenoctuzumab.

15. 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 The medicament of claim 1 or claim 7, or the method of claim 11 or claim 12, comprising:

16. The antigen-binding molecule is (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 and 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 The medicament of claim 1 or claim 7, or the method of claim 11 or claim 12, comprising:

17. The antigen-binding molecule 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. The medicament of claim 1 or claim 7, or the method of claim 11 or claim 12, comprising:

18. The antigen-binding molecule 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. The medicament of claim 1 or claim 7, or the method of claim 11 or claim 12, comprising:

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