IL-11RA Antibody

Abstract

Antigen-binding molecules capable of binding to IL-11Rα, and medical treatment and prevention methods using the same are provided.

Description

[0001] This application claims priority to GB1809700.6 filed on June 13, 2018, the contents and elements of which are incorporated herein by reference for all purposes. Field of the Invention

[0002] The present invention relates to the field of molecular biology, and more specifically to antibody technology. The present invention also relates to medical treatment and prevention methods. Specifically, an antigen-binding molecule capable of binding to IL-11Rα is provided. Background of the Invention

[0004] IL-11-mediated signaling has been shown to stimulate hematopoiesis, osteoclast activity, neurogenesis, inhibit adipogenesis, reduce the expression of proinflammatory cytokines, regulate extracellular matrix (ECM) metabolism, and mediate normal growth control of gastrointestinal epithelial cells. The physiological role of interleukin-11 (IL-11) remains unclear. IL-11 / IL-11R signaling is closely associated with the activation of hematopoietic cells and platelet production, but is also thought to have proinflammatory, anti-inflammatory, and proangiogenic properties and to be important in tumorigenesis. SUMMARY OF THE INVENTION

[0006] In a first aspect, the present invention provides an optionally isolated antigen binding molecule capable of binding to IL-11Rα, wherein the antigen binding molecule comprises:

[0007] (i) a heavy chain variable region (VH) comprising the following CDRs:

[0008] HC-CDR1 having the amino acid sequence of SEQ ID NO: 18

[0009] HC-CDR2 having the amino acid sequence of SEQ ID NO: 52

[0010] an HC-CDR3 having the amino acid sequence of SEQ ID NO: 21; and

[0011] (ii) a light chain variable region (VL) comprising the following CDRs:

[0012] LC-CDR1 having the amino acid sequence of SEQ ID NO: 22

[0013] LC-CDR2 having the amino acid sequence of SEQ ID NO: 23

[0014] LC-CDR3 having the amino acid sequence of SEQ ID NO: 24.

[0015] In some embodiments, the antigen binding molecule comprises:

[0016] (i) a heavy chain variable region (VH) comprising the following CDRs:

[0017] HC-CDR1 having the amino acid sequence of SEQ ID NO: 18

[0018] HC-CDR2 having the amino acid sequence of SEQ ID NO: 19

[0019] an HC-CDR3 having the amino acid sequence of SEQ ID NO: 21; and

[0020] (ii) a light chain variable region (VL) comprising the following CDRs:

[0021] LC-CDR1 having the amino acid sequence of SEQ ID NO: 22

[0022] LC-CDR2 having the amino acid sequence of SEQ ID NO: 23

[0023] LC-CDR3 having the amino acid sequence of SEQ ID NO: 24.

[0024] In some embodiments, the antigen binding molecule comprises:

[0025] (i) a heavy chain variable region (VH) comprising the following CDRs:

[0026] HC-CDR1 having the amino acid sequence of SEQ ID NO: 18

[0027] HC-CDR2 having the amino acid sequence of SEQ ID NO: 20

[0028] an HC-CDR3 having the amino acid sequence of SEQ ID NO: 21; and

[0029] (ii) a light chain variable region (VL) comprising the following CDRs:

[0030] LC-CDR1 having the amino acid sequence of SEQ ID NO: 22

[0031] LC-CDR2 having the amino acid sequence of SEQ ID NO: 23

[0032] LC-CDR3 having the amino acid sequence of SEQ ID NO: 24.

[0033] In some embodiments, the antigen binding molecule comprises:

[0034] A VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 7, 8, 9, 10, 11, or 12; and

[0035] A VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 13, 14, 15, 16, or 17.

[0036] In some embodiments, the antigen binding molecule comprises a polypeptide comprising or consisting of an amino acid sequence that has at least 70%, preferably one of 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: 70. In some embodiments, the antigen binding molecule comprises a polypeptide comprising or consisting of an amino acid sequence that has at least 70%, preferably one of 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: 73. In some embodiments, the antigen binding molecule comprises a polypeptide comprising or consisting of an amino acid sequence that has at least 70%, preferably one of 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:74. In some embodiments, the antigen binding molecule comprises a polypeptide comprising or consisting of an amino acid sequence that has 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:70 or 73, and a polypeptide comprising or consisting of an amino acid sequence that has 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:74.

[0037] In some embodiments, the antigen binding molecule is capable of inhibiting IL-11 mediated signaling.

[0038] The present invention also provides an optionally isolated antigen-binding molecule comprising (i) an antigen-binding molecule described herein, and (ii) an antigen-binding molecule capable of binding an antigen other than IL-11Rα.

[0039] In some embodiments, the antigen binding molecule is capable of inhibiting the interaction between IL-11Rα or a complex comprising IL-11Rα and an interacting partner of IL-11Rα or a complex comprising IL-11Rα.

[0040] The present invention also provides a chimeric antigen receptor (CAR) comprising the antigen binding molecules described herein.

[0041] The present invention also provides one or more optionally isolated nucleic acids encoding the antigen binding molecules or CARs described herein.

[0042] The present invention also provides one or more expression vectors comprising one or more nucleic acids described herein.

[0043] The present invention also provides a cell comprising an antigen binding molecule, a CAR, one or more nucleic acids, or one or more expression vectors described herein.

[0044] The present invention also provides a method comprising culturing a cell comprising one or more nucleic acids or one or more expression vectors described herein under conditions suitable for expressing the antigen binding molecule or CAR from the nucleic acid or expression vector.

[0045] The present invention also provides a composition comprising the antigen binding molecule described herein, CAR, one or more nucleic acids, one or more expression vectors or cells.

[0046] The present invention also provides the antigen binding molecules, CARs, one or more nucleic acids, one or more expression vectors, cells or compositions described herein for use in medical treatment or prevention methods.

[0047] The present invention also provides an antigen binding molecule, CAR, one or more nucleic acids, one or more expression vectors, cells or compositions described herein for use in a method for treating or preventing fibrosis, a disease characterized by fibrosis, cancer, inflammation or a disease characterized by inflammation.

[0048] The present invention also provides the use of the antigen binding molecules, CARs, one or more nucleic acids, one or more expression vectors, cells or compositions described herein in the preparation of a medicament for use in a method for treating or preventing fibrosis, a disease characterized by fibrosis, cancer, inflammation or a disease characterized by inflammation.

[0049] The present invention also provides a method for treating or preventing fibrosis, a disease characterized by fibrosis, cancer, inflammation, or a disease characterized by inflammation, the method comprising administering to a subject a therapeutically or prophylactically effective amount of an antigen binding molecule, CAR, one or more nucleic acids, one or more expression vectors, cells or compositions described herein.

[0050] The present invention also provides a method for inhibiting IL-11-mediated signal transduction, comprising contacting a cell expressing IL-11Rα with the antigen-binding molecule described herein.

[0051] The present invention also provides an optionally isolated in vitro complex comprising an antigen binding molecule as described herein bound to IL-11Rα or a complex comprising IL-11Rα.

[0052] The present invention also provides a method comprising contacting a sample containing or suspected of containing IL-11Rα or a complex containing IL-11Rα with the antigen-binding molecule described herein, and detecting formation of a complex between the antigen-binding molecule and IL-11Rα or a complex containing IL-11Rα.

[0053] The present invention also provides a method for selecting or stratifying subjects for treatment with an IL-11Rα targeting agent, the method comprising contacting a sample from the subject with an antigen-binding molecule described herein in vitro, and detecting formation of a complex between the antigen-binding molecule and IL-11Rα or a complex comprising IL-11Rα.

[0054] The present invention also provides the use of the antigen binding molecules described herein as in vitro or in vivo diagnostic or prognostic agents.

[0055] Details

[0056] The present invention relates to novel IL-11Rα binding molecules with improved properties compared to known anti-IL-11Rα antibodies. Compared to IL-11Rα binding antigen-binding molecules disclosed in the prior art, the IL-11Rα binding molecules of the present invention have a desirable combination of biophysical and functional properties.

[0057] Interleukin-11 and IL-11 receptor

[0058] Interleukin-11 (IL-11), also known as adipogenesis inhibitory factor, is a pleiotropic cytokine and a member of the IL-6 family of cytokines, which includes IL-6, IL-11, IL-27, IL-31, oncostatin, leukemia inhibitory factor (LIF), cardiotrophin-1 (CT-1), cardiotrophin-like cytokine (CLC), ciliary neurotrophic factor (CNTF), and neuronal protein (NP-1).

[0059] Interleukin 11 (IL-11) is expressed in a variety of mesenchymal cell types. The IL-11 genomic sequence has been mapped to the centromeric regions of chromosomes 19 and 71 and is transcribed with a canonical signal peptide that ensures efficient secretion from cells. The IL-11 activating protein complex, cJun / AP-1, is located in its promoter sequence and is crucial for the basal transcriptional regulation of IL-11 (Du and Williams, Blood 1997, Vol 89: 3897-3908). The immature form of human IL-11 is a 199 amino acid polypeptide, while the mature form of IL-11 encodes a protein of 178 amino acid residues (Garbers and Scheller., Biol. Chem. 2013; 394(9): 1145-1161). The amino acid sequence of human IL-11 is available under UniProt accession number P20809 (P20809.1 GI:124294; SEQ ID NO:1). Recombinant human IL-11 (oprelvekin) is also commercially available. IL-11 from other species, including mouse, rat, pig, cow, several teleost fish, and primates, has also been cloned and sequenced.

[0060] In this specification, "IL-11" refers to IL-11 from any species, and includes isoforms, fragments, variants or homologs of IL-11 from any species. As used herein, a "fragment," "variant," or "homolog" of a protein can 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 with the amino acid sequence of a reference protein. In some embodiments, fragments, variants, isoforms, and homologs of a reference protein can be characterized by their ability to perform the function performed by the reference protein.

[0061] "Fragment" generally refers to a portion of a reference protein. "Variant" generally refers to a protein with an amino acid sequence that contains substitutions, insertions, deletions or other modifications of one or more amino acids relative to the amino acid sequence of the reference protein, but retains a considerable degree of sequence identity (e.g., at least 60%) with the amino acid sequence of the reference protein. "Isoform" generally refers to a variant of a reference protein expressed by the same species as the reference protein. "Homolog" generally refers to a variant of a reference protein produced by a different species compared to the species of the reference protein. Homologs include orthologs. A "fragment" can have any length (in terms of amino acid number), although it can optionally be at least 20% of the length of the reference protein (i.e., the protein from which the fragment is derived), and can 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. The minimum length of an IL-11 fragment is 10 amino acids, and the maximum length is one of 15, 20, 25, 30, 40, 50, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 195 amino acids.

[0062] In some embodiments, the IL-11 is IL-11 from a mammal (e.g., primate (rhesus monkey, macaque, non-human primate, or human) and / or rodent (e.g., rat or murine) IL-11). The isoform, fragment, variant, or homolog of IL-11 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 with the amino acid sequence of an immature or mature IL-11 isoform from a given species (e.g., human). In some embodiments, the IL-11 of the present invention 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 with SEQ ID NO: 1.

[0063] The isoforms, fragments, variants or homologs of IL-11 may optionally be characterized by the ability to bind to an IL-11 receptor (e.g., IL-11Rα, gp130 and / or a complex comprising IL-11Rα and gp130, preferably from the same species) and stimulate signal transduction in cells expressing IL-11Rα and gp130 (e.g., as described in Curtis et al., Blood, 1997, 90(11); or Karpovich et al., Mol. Hum. Reprod. 2003 9(2):75-80).

[0064] IL-11 signals through a homodimer of the ubiquitously expressed glycoprotein 130 (gp130; also known as glycoprotein 130, IL-6ST, IL-6-beta, or CD130). Gp130 is a transmembrane protein that forms a subunit of the type I cytokine receptor with the IL-6 receptor family. Specificity is acquired through the separate interleukin-11 receptor subunit α (IL-11Rα), and although the initial cytokine binding event to the α receptor leads to the final formation of a complex with gp130, it is not directly involved in signal transduction.

[0065] Human gp130 (including a 22-amino acid signal peptide) is a 918-amino acid protein. The mature form is 866 amino acids long, consisting of a 597-amino acid extracellular domain, a 22-amino acid transmembrane domain, and a 277-amino acid intracellular domain. The extracellular domain of the protein contains the cytokine binding module (CBM) of gp130. The CBM of gp130 includes the gp130 Ig-like domain D1 and the fibronectin type III domains D2 and D3. The amino acid sequence of human gp130 is available under UniProt accession number P40189-1 (SEQ ID NO: 2).

[0066] Human IL-11Rα is a 422 amino acid polypeptide (UniProt Q14626) that shares ~85% nucleotide and amino acid sequence identity with murine IL-11Rα (Du and Williams, Blood, Vol. 89, No. 11, June 1, 1997). Two isoforms of IL-11Rα, HCR1 (SEQ ID NO: 3) and HCR2 (SEQ ID NO: 4), have been reported, differing in their cytoplasmic domains; HCR2 lacks the C-terminal 32 amino acids of isoform HCR1 (Du and Williams, supra). The IL-11 receptor α chain (IL-11Rα) shares many structural and functional similarities with the IL-6 receptor α chain (IL-6Rα). The extracellular domain displays 24% amino acid identity, including a characteristic conserved Trp-Ser-X-Trp-Ser (WSXWS) motif. The short cytoplasmic domain lacks the box 1 and box 2 regions required for activation of the JAK / STAT signaling pathway.

[0067] The receptor binding site on murine IL-11 has been mapped and three sites have been identified - sites I, II and III. Binding to gp130 is reduced by substitutions in the site II region and by substitutions in the site III region. Site III mutants show no detectable agonist activity and possess IL-11Rα antagonist activity (Cytokine Inhibitors, Chapter 8; edited by Gennaro Ciliberto and Rocco Savino, Marcel Dekker, Inc. 2001).

[0068] In this specification, the IL-11 receptor / receptor for IL-11 (IL-11R) refers to a polypeptide or polypeptide complex capable of binding to IL-11 and / or a complex comprising IL-11. In some embodiments, the IL-11 receptor is capable of binding to IL-11 and / or a complex comprising IL-11 and inducing signal transduction in cells expressing the IL-11 receptor. A "complex comprising IL-11" can be a non-covalent complex of IL-11 and a polypeptide capable of non-covalently binding to IL-11.

[0069] The IL-11 receptor may be from any species and includes subtypes, fragments, variants or homologs of the IL-11 receptor from any species. In a preferred embodiment, the species is human (Homo sapiens).

[0070] In some embodiments, the IL-11 receptor (IL-11R) may be IL-11Rα. In some embodiments, the IL-11 receptor may be a polypeptide complex comprising IL-11Rα. In some embodiments, the IL-11 receptor may be a polypeptide complex comprising IL-11Rα and gp130. In some embodiments, the IL-11 receptor may be gp130 or a complex comprising IL-11-bound gp130.

[0071] As used herein, "IL-11Rα" refers to IL-11Rα from any species, and includes isoforms, fragments, variants, or homologs of IL-11Rα from any species. The minimum length of an IL-11Rα fragment is 10 amino acids, and the maximum length is one of 15, 20, 25, 30, 40, 50, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 250, 300, 400, or 415 amino acids.

[0072] In some embodiments, the IL-11Rα is an IL-11Rα from a mammal (e.g., a primate (rhesus monkey, macaque, non-human primate, or human) and / or rodent (e.g., rat or murine) IL-11). The isoform, fragment, variant, or homolog of IL-11Rα can optionally be characterized as having at least 70%, preferably 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 96%, 97%, 98%, 99%, or 100% amino acid sequence identity to the amino acid sequence of an immature or mature IL-11Rα isoform from a given species (e.g., human). In some embodiments, the IL-11Rα of the present disclosure 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 SEQ ID NO: 3 or 4.

[0073] The isoforms, fragments, variants or homologs of IL-11Rα may optionally be characterized by their ability to bind IL-11 and / or complexes comprising IL-11 (preferably from the same species) and stimulate signal transduction in cells expressing IL-11Rα and gp130 (as described in Curtis et al., Blood, 1997, 90(11) or Karpovich et al., Mol. Hum. Reprod. 2003, 9(2):75-80).

[0074] IL-11 / IL-11R signaling

[0075] IL-11 binds to IL-11Rα with low affinity (Kd ~10 nmol / L), and the interaction between these binding partners alone is insufficient to transduce biological signals. The production of a high-affinity receptor (Kd ∼400 to 800 pmol / L) capable of signal transduction requires co-expression of IL-11Rα and gp130 (Curtis et al. (Blood 1997 Dec 1;90(11):4403-12; Hilton et al., EMBO J 13:4765, 1994; Nandurkar et al., Oncogene 12:585, 1996). Binding of IL-11 to cell surface IL-11Rα induces heterodimerization, tyrosine phosphorylation, activation of gp130, and downstream signaling. This occurs primarily through the mitogen-activated protein kinase (MAPK) cascade and the Janus kinase / signal transducer and activator of transcription (Jak / STAT) pathway (Garbers and Scheller, supra).

[0076] In principle, soluble IL-11Rα can also form a biologically active soluble complex with IL-11 (Pflanz et al., 1999 FEBS Lett, 450, 117-122), which raises the possibility that, similar to IL-6, IL-11, under certain circumstances, binds to soluble IL-11Rα before binding to cell surface gp130 (Garbers and Scheller, supra). Curtis et al. (Blood 1997 Dec 1; 90(11): 4403-12) described the expression of a soluble murine IL-11Rα chain (sIL-11R) and examined signaling in cells expressing gp130. In the presence of gp130 but without the transmembrane IL-11R, sIL-11R-mediated IL-11-dependent differentiation of M1 leukemia cells and proliferation of Ba / F3 cells, along with early intracellular events including phosphorylation of gp130, STAT3, and SHP2, resemble signaling through the transmembrane IL-11R. It has recently been demonstrated that IL-11 binding to soluble IL-11Rα can activate signaling through membrane-bound gp130 (Lokau et al., 2016 Cell Reports 14, 1761–1773). This so-called IL-11 trans-signaling may be a very important component of IL-11-mediated signaling and may even be the most common form of IL-11-mediated signaling, because, whereas expression of IL-11Rα is restricted to a relatively small number of cell types, gp130 is expressed in a wide variety of cell types.

[0077] As used herein, "IL-11 signaling," "IL-11-mediated signaling," and "IL-11 / IL-11R signaling" refer to signaling mediated by the binding of IL-11 or a fragment thereof having the function of a mature IL-11 molecule, or a complex comprising IL-11 or a fragment thereof having the function of a mature IL-11 molecule, to the IL-11 receptor.

[0078] As used herein, "IL-11 trans-signaling" refers to signal transduction triggered by the binding of IL-11 bound to IL-11Rα and gp130. IL-11 can bind to IL-11Rα as a non-covalent complex. gp130 is membrane-bound and is expressed by cells that signal after the IL-11:IL-11Rα complex binds to gp130. In some embodiments, IL-11Rα can be soluble IL-11Rα. In some embodiments, soluble IL-11Rα is a soluble (secreted) isoform of IL-11Rα (e.g., lacking a transmembrane domain). In some embodiments, soluble IL-11Rα is a release product of proteolytic cleavage of the extracellular domain of cell membrane-bound IL-11Rα. In some embodiments, IL-11Rα may be cell membrane bound, and signaling through gp130 may be triggered by binding of IL-11 bound to cell membrane bound IL-11Rα, referred to as "IL-11 cis signaling."

[0079] IL-11-mediated signaling has been shown to stimulate hematopoiesis and platelet production, stimulate osteoclast activity, stimulate neurogenesis, inhibit adipogenesis, reduce proinflammatory cytokine expression, regulate extracellular matrix (ECM) metabolism, and mediate normal growth control of gastrointestinal epithelial cells (Du and Williams, supra).

[0080] The physiological role of interleukin 11 (IL-11) remains unclear. IL-11 is most closely associated with the activation of hematopoietic cells and the production of platelets, but is also considered to have pro-inflammatory, as well as anti-inflammatory, pro-angiogenic properties, and is important for tumor formation. It is known that TGFβ1 or tissue damage can induce IL-11 expression (Zhu, M. et al. PLOS ONE 10, (2015); Yashiro, R. et al. J. Clin. Periodontol. 33, 165-71 (2006); Obana, M. et al. Circulation 121, 684-91 (2010); Tang, W et al. J. Biol. Chem. 273, 5506-13 (1998)).

[0081] IL-11 is an important post-transcriptional regulator of TGFβ-mediated signaling. TGFβ1 has been shown to stimulate the AP-1 promoter region of IL-11, and TGFβ-induced IL-11 secretion has been shown to induce activation of ERK p42 / 44 and p38 MAP kinase in intestinal myofibroblasts (Bamba et al., Am J Physiol Gastrointest Liver Physiol. (2003) (285(3): G529-38). MAP kinase inhibitors can significantly reduce TGFβ-induced IL-11 secretion, and p38 MAP kinase-mediated mRNA stabilization has been shown to be crucial for TGFβ-induced IL-11 secretion.

[0082] IL-11 mediated signaling has recently been shown to play a key role in the fibrotic process of various tissues. See, for example, WO 2017 / 103108 A1 and Schafer et al. (2017) Nature 552: 110–115, both of which are incorporated herein by reference in their entirety.

[0083] WO 2017 / 103108 A1 (hereby incorporated by reference in its entirety) reports the profibrotic effects of IL-11 mediated signaling and establishes the therapeutic role of IL-11 mediated signaling antagonists in the treatment / prevention of fibrosis. Figure 7 A and 7B demonstrate that incubation of primary human atrial fibroblasts with recombinant human IL-11 increases collagen deposition by fibroblasts, a well-established fibrogenic process. Treatment with neutralizing anti-IL-11 antibodies (but not isotype control antibodies) has been shown to abrogate collagen production induced by TGFβ1 stimulation of fibroblasts (a known pro-fibrotic stimulus). Example 3 and Figure 10 of WO 2017 / 103108 A1 further demonstrate the ability of neutralizing anti-IL-11 antibodies to abrogate the increased collagen production by human atrial fibroblasts in response to various other pro-fibrotic stimuli (ANG2, PDGF, ET-1). Examples 5.2 and 6 of WO 2017 / 103108 A1 Figure 20A-20E provides further data supporting the profibrotic effect of IL-11 in cardiac tissue. The results showed that human atrial fibroblasts significantly increased the production of extracellular matrix components (collagen, periostin) and the expression of profibrotic markers (αSMA, IL-6, MMP2, TIMP1) after treatment with human IL-11 protein, and the production of these factors increased in response to treatment with the profibrotic stimulant TGFβ1. Example 5.3.1 and Figures 38A to 38D also show that human primary liver fibroblasts increased the production of extracellular matrix components and the expression of fibrotic markers in response to treatment with human IL-11, as well as the ability of neutralizing anti-IL-11 antibodies to eliminate the fibrotic effects of TGFβ1 stimulation. WO 2017 / 103108 A1 Figure 22A to 2 2F and Figure 23A and 23B showed that TGFβ1-mediated fibrosis could be inhibited by treatment with a neutralizing anti-IL-11 antibody, and Figure 24 It was shown that decoy receptor molecules that bind to IL-11, neutralizing anti-IL-11Rα antibodies, and oligonucleotides encoding siRNA for antisense knockdown of IL-11 and IL-11RA gene expression are similarly able to inhibit TGFβ1-mediated conversion of fibroblasts to myofibroblasts (fibrotic effector cells). Figure 32A and 32B Further data are provided showing the use of decoy IL-11 receptors to inhibit TGFβ1-mediated fibrotic responses. Example 5.3.3 of WO 2017 / 103108 A1 and Figure 21 Figures B and 21C present in vivo data demonstrating that IL-11 is profibrotic in multiple tissues. Injection of mice with recombinant mouse IL-11 resulted in increases in the relative weights of the heart, kidneys, lungs, and liver ( Figure 21 B), which is associated with increased collagen content in these tissues ( Figure 21 C). In Examples 7.2 and 7.3 of WO2017 / 103108 A1 and Figures 27A to 27D Further in vivo data supporting the pro-fibrotic effects of IL-11 are provided in Figures 31 and 38. These experiments show that mice deficient in IL-11RA are protected from cardiac and renal tissue fibrosis induced by fibrogenic stimuli, indicating that signaling through the IL-11 receptor is an important mediator of the fibrotic process. Figure 31A and 31Breported that 7 days after trabeculectomy, more fibrosis was detected in ocular sections obtained from wild-type mice than from IL-11RA knockout mice. Thus, WO 2017 / 103108 A1 provides extensive data from in vitro and in vivo studies demonstrating that IL-11 / IL-11R signaling is a key mediator of fibrosis in a wide range of tissues, as determined by analysis of various markers of fibrotic response, and demonstrates that inhibition of IL-11-mediated signaling reduces fibrosis.

[0084] Antigen-binding molecules capable of binding to IL-11Rα

[0085] The present invention provides antigen-binding molecules capable of binding to IL-11Rα.

[0086] "Antigen binding molecules" refer to molecules that can bind to a target antigen, including monoclonal antibodies, polyclonal antibodies, monospecific and multispecific antibodies (e.g., bispecific antibodies) and antibody fragments (e.g., Fv, scFv, Fab, scFab, F(ab')2, Fab2, diabodies, triabodies, scFv-Fc, miniantibodies, single domain antibodies (e.g., VhH), etc.), as long as they exhibit binding to the relevant target molecule. "Antibodies" include fragments and derivatives thereof, including synthetic antibodies and fragments. The antibodies used herein are polypeptides that can specifically bind to the relevant target molecule (i.e., the antigen for which the antibody is specific). The antibodies and antigen binding molecules of the present invention can be provided in an isolated form.

[0087] In view of the contemporary technology related to monoclonal antibody technology, antibodies can be prepared against most antigens. The antigen binding portion can be part of an antibody (e.g., a Fab fragment) or a synthetic antibody fragment (e.g., a single-chain Fv fragment [ScFv]). Suitable monoclonal antibodies against the selected antigen can be prepared by known techniques, such as those disclosed in "Monoclonal Antibodies: A manual of techniques", H Zola (CRC Press, 1988) and "Monoclonal Hybridoma Antibodies: Techniques and Applications", JGR Hurrell (CRC Press, 1982). Neuberger et al. (1988, 8th International Symposium on Biotechnology Part 2 (8 th Chimeric antibodies are discussed in International Biotechnology Symposium Part 2), 792-799).

[0088] Monoclonal antibodies (mAbs) can be used in the methods of the present invention and are a homogeneous population of antibodies that specifically target a single epitope on an antigen.

[0089] Antigen-binding fragments of antibodies, such as Fab and Fab2 fragments, can also be used / provided as genetically engineered antibodies and antibody fragments. H ) and variable lightness (V L ) domains are involved in antigen recognition, a fact first recognized by early protease digestion experiments. This was further confirmed by the discovery of "humanization" of rodent antibodies. Rodent variable domains can be fused to human constant domains, resulting in antibodies that retain the antigenic specificity of the rodent parent antibody (Morrison et al (1984) Proc. Natl. Acad. Sd. USA 81, 6851-6855).

[0090] In some embodiments, the antigen-binding molecules of the present invention are fully human antibodies / antibody fragments. Fully human antibodies / antibody fragments are encoded by human nucleic acid sequences. Fully human antibodies / antibody fragments do not contain non-human amino acid sequences.

[0091] The two most commonly used techniques for producing fully human antibodies are (i) phage display, in which human antibody genes are expressed in phage display libraries, and (ii) producing antibodies in transgenic mice engineered to have human antibody genes (described in Park and Smolen, Advances in Protein Chemistry (2001) 56: 369-421). In short, in human antibody gene phage display technology, genes encoding VH and VL chains are produced by PCR amplification and cloning from "natural" human lymphocytes and assembled into a library from which they can be expressed as disulfide-linked Fab fragments or single-chain Fv (scFv) fragments. Fab- or scFv-encoding genes are fused to the surface coat protein of filamentous phage, and Fab or scFv that can bind to the target of interest can then be identified by screening the library with antigen. Molecular evolution or affinity maturation procedures can be used to enhance the affinity of Fab / scFv fragments. In transgenic mouse technology, mice in which the endogenous murine Ig loci have been replaced by their human homologs by homologous recombination are immunized with antigen, and monoclonal antibodies are prepared by conventional hybridoma technology to produce fully human monoclonal antibodies.

[0092] In some embodiments, the antigen-binding molecules of the present invention are murine antibodies / antibody fragments. In some embodiments, antibodies / antibody fragments can be prepared by phage display using human primary antibody gene libraries.

[0093] In some embodiments, the antigen binding molecules of the present invention are mouse / human chimeric antibodies / antibody fragments (e.g., antigen binding molecules comprising mouse variable domains and human constant regions). In some embodiments, the antigen binding molecules are humanized antibodies / antibody fragments (e.g., antigen binding molecules comprising mouse CDRs and human framework and constant regions).

[0094] Mouse / human chimeric antigen binding molecules can be prepared from mouse monoclonal antibodies by a chimerization process, for example, as described in "Human Monoclonal Antibodies: Methods and Protocols," Michael Steinitz (ed.), Methods in Molecular Biology 1060, Springer Protocols, Humana Press (2014), Chapter 8 thereof, particularly Chapter 8, Section 3.

[0095] Humanized antigen-binding molecules can be prepared from mouse antibodies by a humanization process, for example, as described in "Human Monoclonal Antibodies: Methods and Protocols", Michael Steinitz (ed.), Methods in Molecular Biology 1060, Springer Protocols, Humana Press (2014), Chapter 7 thereof, in particular Chapter 7, Section 3.1, "Antibody Humanization".

[0096] The antigen binding molecules of the present invention include a portion that can be combined with one or more target antigens. In some embodiments, the portion that can be combined with the target antigen includes an antibody heavy chain variable region (VH) and an antibody light chain variable region (VL) that can specifically bind to the target antigen. In some embodiments, the portion that can be combined with the target antigen includes an aptamer that can be combined with the target antigen or is composed of an aptamer, and the aptamer (e.g., nucleic acid aptamer) can be combined with the target antigen (as reviewed in Zhou and Rossi, Nat Rev Drug Discov 2017 16(3):181-202). In some embodiments, the portion capable of binding to the target antigen comprises or consists of an antigen-binding peptide / polypeptide, such as a peptide aptamer, a thioredoxin, a monomer, 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 avidin, an armadillo repeat protein (ArmRP), an OBody, or a fibronectin—as reviewed in Reverdatto et al., Curr Top Med Chem. 2015; 15(12): 1082-1101, which is incorporated herein by reference in its entirety (see also, e.g., Boersma et al., J Biol Chem (2011) 286: 41273-85 and Emanuel et al., Mabs (2011) 3: 38-48).

[0097] The antigen-binding molecules of the present invention generally comprise an antigen-binding domain comprising the VH and VL of an antibody capable of specifically binding to a target antigen. The antigen-binding domain formed by VH and VL may also be referred to herein as an Fv region.

[0098] Antigen binding molecules can be or can comprise antigen binding polypeptides or antigen binding polypeptide complexes.Antigen binding molecules can comprise more than one polypeptide, which together form an antigen binding domain.The polypeptides can be covalently or non-covalently bound.In some embodiments, the polypeptides form a portion of a larger polypeptide comprising the polypeptide (e.g., in the case of scFv comprising VH and VL, or in the case of scFab comprising VH-CH1 and VL-CL).

[0099] An antigen binding molecule may refer to a non-covalent or covalent complex of more than one polypeptide (eg, 2, 3, 4, 6, or 8 polypeptides), such as an IgG-like antigen binding molecule comprising two heavy chain polypeptides and two light chain polypeptides.

[0100] Antigen binding molecules of the present invention can be designed and prepared using sequences of monoclonal antibodies (mAbs) that bind to IL-11Rα. Antigen binding regions of antibodies, such as single-chain variable fragments (scFv), Fab, and F(ab')2 fragments, can also be used / provided. An "antigen binding region" is any fragment of an antibody that is capable of binding to a target specific for a given antibody.

[0101] Antibodies typically 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. Together, the six CDRs define the antibody's paratope, which is the part of the antibody that binds to its target antigen.

[0102] The VH and VL regions contain framework regions (FRs) on both sides of each CDR, which provide 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.

[0103] There are several different conventions for defining antibody CDRs and FRs, such as the conventions described by Kabat et al., Protein Sequences of Immunological Significance, 5th ed., U.S. National Institutes of Health, Public Health Service, Bethesda, Maryland (1991), Chothia et al., J. Mol. Biol. 196:901-917 (1987), and VBASE2, as described in Retter et al., Nucl. Natl. Acad. Sci. (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 Kabat system.

[0104] In some embodiments, the antigen-binding molecule comprises a CDR of an antigen-binding molecule capable of binding to IL-11Rα. In some embodiments, the antigen-binding molecule comprises a FR of an antigen-binding molecule capable of binding to IL-11Rα. In some embodiments, the antigen-binding molecule comprises a CDR and a FR of an antigen-binding molecule capable of binding to IL-11Rα. That is, in some embodiments, the antigen-binding molecule comprises a VH region and a VL region of an antigen-binding molecule capable of binding to IL-11Rα.

[0105] In some embodiments, the antigen binding molecule comprises a VH region and a VL region, which are or are derived from the VH / VL regions of an IL-11Rα binding antibody clone described herein, i.e., the anti-IL-11Rα antibody clone BSO-9A7 (comprising 9A7 VH, 9A7 VH 1, 9A7 VH2, 9A7 VH 3, 9A7 VH 4, or 9A7 VH 5 and 9A7 VL, 9A7 VL 1, 9A7 VL 2, 9A7 VL 3, or 9A7 VL 4).

[0106] In some embodiments, the antigen binding molecule comprises a VL region comprising an amino acid sequence that has at least 70% sequence identity, 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 shown in SEQ ID NO: 13, 14, 15, 16 or 17, wherein the position corresponding to position 91 is not arginine, and / or wherein the position corresponding to position 105 is not methionine.

[0107] In some embodiments, the antigen binding molecule comprises a VL region comprising an amino acid sequence that has at least 70% sequence identity, 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 shown in SEQ ID NO: 13, 14, 15, 16 or 17, wherein the position corresponding to position 91 is serine, and / or wherein the position corresponding to position 105 is leucine or isoleucine.

[0108] In some embodiments, the antigen-binding molecule comprises a VL region that has less than 100% sequence identity with the amino acid sequence of SEQ ID NO: 59. In some embodiments, the antigen-binding molecule does not comprise a VL region that comprises or consists of the amino acid sequence of SEQ ID NO: 59. In some embodiments, the antigen-binding molecule does not comprise a peptide / polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 59.

[0109] In some embodiments, the antigen-binding molecule comprises a LC-CDR3 that has less than 100% sequence identity with the amino acid sequence of SEQ ID NO: 60. In some embodiments, the antigen-binding molecule does not comprise a LC-CDR3 that comprises or consists of the amino acid sequence of SEQ ID NO: 60. In some embodiments, the antigen-binding molecule does not comprise a peptide / polypeptide that comprises or consists of the amino acid sequence of SEQ ID NO: 60.

[0110] In some embodiments, the antigen binding molecule comprises a VH region according to one of the following (1) to (3):

[0111] (1) (9A7 VH, 9A7 VH 1, 9A7 VH 2, 9A7 VH 3, 9A7 VH 4, 9A7 VH 5) VH area containing the following CDRs:

[0112] HC-CDR1 having the amino acid sequence of SEQ ID NO: 18

[0113] HC-CDR2 having the amino acid sequence of SEQ ID NO: 52

[0114] an HC-CDR3 having the amino acid sequence of SEQ ID NO: 21,

[0115] or a variant thereof, wherein one or two or three amino acids in one or more of HC-CDR1, HC-CDR2 or HC-CDR3 are substituted with another amino acid.

[0116] (2) (9A7 VH, 9A7 VH 1, 9A7 VH 2, 9A7 VH 3, 9A7 VH 4) VH area containing the following CDRs:

[0117] HC-CDR1 having the amino acid sequence of SEQ ID NO: 18

[0118] HC-CDR2 having the amino acid sequence of SEQ ID NO: 19

[0119] an HC-CDR3 having the amino acid sequence of SEQ ID NO: 21,

[0120] or a variant thereof, wherein one or two or three amino acids in one or more of HC-CDR1, HC-CDR2 or HC-CDR3 are substituted with another amino acid.

[0121] (3) (9A7 VH 5) VH region comprising the following CDRs:

[0122] HC-CDR1 having the amino acid sequence of SEQ ID NO: 18

[0123] HC-CDR2 having the amino acid sequence of SEQ ID NO: 20

[0124] an HC-CDR3 having the amino acid sequence of SEQ ID NO: 21,

[0125] or a variant thereof, wherein one or two or three amino acids in one or more of HC-CDR1, HC-CDR2 or HC-CDR3 are substituted with another amino acid.

[0126] In some embodiments, the antigen binding molecule comprises a VH region according to one of the following (4) to (9):

[0127] (4) (9A7 VH) VH region comprising the following FRs:

[0128] HC-FR1 having the amino acid sequence of SEQ ID NO: 25

[0129] HC-FR2 having the amino acid sequence of SEQ ID NO: 29

[0130] HC-FR3 having the amino acid sequence of SEQ ID NO: 33

[0131] HC-FR4 having the amino acid sequence of SEQ ID NO: 37,

[0132] or a variant thereof, wherein one or two or three amino acids in one or more of HC-FR1, HC-FR2, HC-FR3 or HC-FR4 are substituted with another amino acid.

[0133] (5) (9A7 VH 1) VH region comprising the following FRs:

[0134] HC-FR1 having the amino acid sequence of SEQ ID NO: 26

[0135] HC-FR2 having the amino acid sequence of SEQ ID NO: 30

[0136] HC-FR3 having the amino acid sequence of SEQ ID NO: 34

[0137] HC-FR4 having the amino acid sequence of SEQ ID NO: 38,

[0138] or a variant thereof, wherein one or two or three amino acids in one or more of HC-FR1, HC-FR2, HC-FR3 or HC-FR4 are substituted with another amino acid.

[0139] (6) (9A7 VH 2) VH region comprising the following FRs:

[0140] HC-FR1 having the amino acid sequence of SEQ ID NO: 27

[0141] HC-FR2 having the amino acid sequence of SEQ ID NO: 30

[0142] HC-FR3 having the amino acid sequence of SEQ ID NO: 35

[0143] HC-FR4 having the amino acid sequence of SEQ ID NO: 38,

[0144] or a variant thereof, wherein one or two or three amino acids in one or more of HC-FR1, HC-FR2, HC-FR3 or HC-FR4 are substituted with another amino acid.

[0145] (7) (9A7 VH 3) VH region comprising the following FRs:

[0146] HC-FR1 having the amino acid sequence of SEQ ID NO: 28

[0147] HC-FR2 having the amino acid sequence of SEQ ID NO: 31

[0148] HC-FR3 having the amino acid sequence of SEQ ID NO: 35

[0149] HC-FR4 having the amino acid sequence of SEQ ID NO: 38,

[0150] or a variant thereof, wherein one or two or three amino acids in one or more of HC-FR1, HC-FR2, HC-FR3 or HC-FR4 are substituted with another amino acid.

[0151] (8)(9A7 VH 4) VH region comprising the following FRs:

[0152] HC-FR1 having the amino acid sequence of SEQ ID NO: 28

[0153] HC-FR2 having the amino acid sequence of SEQ ID NO: 31

[0154] HC-FR3 having the amino acid sequence of SEQ ID NO: 36

[0155] HC-FR4 having the amino acid sequence of SEQ ID NO: 38,

[0156] or a variant thereof, wherein one or two or three amino acids in one or more of HC-FR1, HC-FR2, HC-FR3 or HC-FR4 are substituted with another amino acid.

[0157] (9)(9A7 VH 5) has a VH region comprising the following FRs:

[0158] HC-FR1 having the amino acid sequence of SEQ ID NO: 28

[0159] HC-FR2 having the amino acid sequence of SEQ ID NO: 32

[0160] HC-FR3 having the amino acid sequence of SEQ ID NO: 36

[0161] HC-FR4 having the amino acid sequence of SEQ ID NO: 38,

[0162] or a variant thereof, wherein one or two or three amino acids in one or more of HC-FR1, HC-FR2, HC-FR3 or HC-FR4 are substituted with another amino acid.

[0163] In some embodiments, the antigen-binding molecule comprises a VH region comprising a CDR according to one of (1) to (3) above and a FR according to one of (4) to (9) above.

[0164] In some embodiments, the antigen binding molecule comprises a VH region according to one of the following (10) to (15):

[0165] (10) (9A7 VH) A VH region comprising CDRs according to (2) and FRs according to (4).

[0166] (11) (9A7 VH 1) A VH region comprising CDRs according to (2) and FRs according to (5).

[0167] (12) (9A7 VH 2) A VH region comprising CDRs according to (2) and FRs according to (6).

[0168] (13) (9A7 VH 3) A VH region comprising CDRs according to (2) and FRs according to (7).

[0169] (14) (9A7 VH 4) A VH region comprising CDRs according to (2) and FRs according to (8).

[0170] (15) (9A7 VH 5) comprises a VH region comprising CDRs according to (3) and FRs according to (9).

[0171] In some embodiments, the antigen binding molecule comprises a VH region according to one of the following (16) to (21):

[0172] (16) (9A7 VH) comprises a VH region of an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 7, more preferably at least one of 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity.

[0173] (17) (9A7 VH 1) comprises a VH region of an amino acid sequence having at least 70% sequence identity, more preferably at least one of 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: 8.

[0174] (18) (9A7 VH 2) comprises a VH region of an amino acid sequence having at least 70% sequence identity, more preferably at least one of 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: 9.

[0175] (19) (9A7 VH 3) comprises a VH region of an amino acid sequence having at least 70% sequence identity, more preferably at least one of 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: 10.

[0176] (20) (9A7 VH 4) comprises a VH region of an amino acid sequence having at least 70% sequence identity, more preferably at least one of 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: 11.

[0177] (21) (9A7 VH 5) comprises a VH region of an amino acid sequence having at least 70% sequence identity, more preferably at least one of 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: 12.

[0178] In some embodiments, the antigen binding molecule comprises a VL region according to (22):

[0179] (22) (9A7 VL, 9A7 VL 1, 9A7 VL 2, 9A7 VL 3, 9A7 VL 4) VL area containing the following CDRs:

[0180] LC-CDR1 having the amino acid sequence of SEQ ID NO: 22

[0181] LC-CDR2 having the amino acid sequence of SEQ ID NO: 23

[0182] an LC-CDR3 having the amino acid sequence of SEQ ID NO: 24;

[0183] or a variant thereof, wherein one or two or three amino acids in one or more of LC-CDR1, LC-CDR2 or LC-CDR3 are substituted with another amino acid.

[0184] In some embodiments, the antigen binding molecule comprises a VL region according to one of the following (23)-(27):

[0185] (23)(9A7 VL) contains the following FR VL region:

[0186] LC-FR1 having the amino acid sequence of SEQ ID NO: 39

[0187] LC-FR2 having the amino acid sequence of SEQ ID NO: 41

[0188] LC-FR3 having the amino acid sequence of SEQ ID NO: 45

[0189] LC-FR4 having the amino acid sequence of SEQ ID NO: 50,

[0190] or a variant thereof, wherein one or two or three amino acids in one or more of LC-FR1, LC-FR2, LC-FR3 or LC-FR4 are substituted with another amino acid.

[0191] (24)(9A7 VL 1) contains the following FR VL region:

[0192] LC-FR1 having the amino acid sequence of SEQ ID NO: 40

[0193] LC-FR2 having the amino acid sequence of SEQ ID NO: 42

[0194] LC-FR3 having the amino acid sequence of SEQ ID NO: 46

[0195] LC-FR4 having the amino acid sequence of SEQ ID NO: 51,

[0196] or a variant thereof, wherein one or two or three amino acids in one or more of LC-FR1, LC-FR2, LC-FR3 or LC-FR4 are substituted with another amino acid.

[0197] (25)(9A7 VL 2) contains the following FR VL region:

[0198] LC-FR1 having the amino acid sequence of SEQ ID NO: 40

[0199] LC-FR2 having the amino acid sequence of SEQ ID NO: 43

[0200] LC-FR3 having the amino acid sequence of SEQ ID NO: 47

[0201] LC-FR4 having the amino acid sequence of SEQ ID NO: 51,

[0202] or a variant thereof, wherein one or two or three amino acids in one or more of LC-FR1, LC-FR2, LC-FR3 or LC-FR4 are substituted with another amino acid.

[0203] (26)(9A7 VL 3) contains the following FR VL region:

[0204] LC-FR1 having the amino acid sequence of SEQ ID NO: 40

[0205] LC-FR2 having the amino acid sequence of SEQ ID NO: 44

[0206] LC-FR3 having the amino acid sequence of SEQ ID NO: 48

[0207] LC-FR4 having the amino acid sequence of SEQ ID NO: 51,

[0208] or a variant thereof, wherein one or two or three amino acids in one or more of LC-FR1, LC-FR2, LC-FR3 or LC-FR4 are substituted with another amino acid.

[0209] (27)(9A7 VL 4) contains the following FR VL region:

[0210] LC-FR1 having the amino acid sequence of SEQ ID NO: 40

[0211] LC-FR2 having the amino acid sequence of SEQ ID NO: 44

[0212] LC-FR3 having the amino acid sequence of SEQ ID NO: 49

[0213] LC-FR4 having the amino acid sequence of SEQ ID NO: 51,

[0214] or a variant thereof, wherein one or two or three amino acids in one or more of LC-FR1, LC-FR2, LC-FR3 or LC-FR4 are substituted with another amino acid.

[0215] In some embodiments, the antigen binding molecule comprises a VL region comprising the CDR according to (22) above and the FR according to one of (23) to (27) above.

[0216] In some embodiments, the antigen binding molecule comprises a VL region according to one of the following (28) to (32):

[0217] (28) A VL region comprising CDRs according to (22) and FRs according to (23).

[0218] (29) (9A7 VL 1) comprises a VL region comprising CDRs according to (22) and FRs according to (24).

[0219] (30) (9A7 VL 2) comprises a VL region comprising CDRs according to (22) and FRs according to (25).

[0220] (31) (9A7 VL 3) comprises a VL region comprising CDRs according to (22) and FRs according to (26).

[0221] (32) (9A7 VL 4) comprises a VL region comprising CDRs according to (22) and FRs according to (27).

[0222] In some embodiments, the antigen binding molecule comprises a VL region according to one of the following (33) to (37):

[0223] (33) (9A7 VL) comprises a VL region of an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 13, more preferably at least one of 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity.

[0224] (34) (9A7 VL 1) comprises a VL region of an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 14, more preferably at least one of 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity.

[0225] (35) (9A7 VL 2) comprises a VL region of an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 15, more preferably at least one of 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity.

[0226] (36) (9A7 VL 3) comprises a VL region of an amino acid sequence having at least 70% sequence identity, more preferably at least one of 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: 16.

[0227] (37) (9A7 VL 4) comprises a VL region of an amino acid sequence having at least 70% sequence identity, more preferably at least one of 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: 17.

[0228] In some embodiments, the antigen-binding molecule comprises the VH region according to any one of (1) to (21) above and the VL region according to any one of (22) to (37) above.

[0229] In some embodiments, the antigen binding molecule comprises a VH region according to (10) or (16) and a VL region according to (28) or (33).

[0230] In some embodiments, the antigen-binding molecule comprises a VH region according to (14) or (20) and a VL region according to (32) or (37). In some embodiments, the antigen-binding molecule comprises a VH region according to (14) and a VL region according to (32). In some embodiments, the antigen-binding molecule comprises a VH region according to (20) and a VL region according to (37).

[0231] In some embodiments, the antigen binding molecule comprises a VH region according to (13) or (19) and a VL region according to (30) or (35). In some embodiments, the antigen binding molecule comprises a VH region according to (13) or (19) and a VL region according to (31) or (36). In some embodiments, the antigen binding molecule comprises a VH region according to (13) or (19) and a VL region according to (32) or (37). In some embodiments, the antigen binding molecule comprises a VH region according to (14) or (20) and a VL region according to (30) or (35). In some embodiments, the antigen binding molecule comprises a VH region according to (14) or (20) and a VL region according to (31) or (36).

[0232] In embodiments according to the present invention where one or more amino acids are substituted with another amino acid, the substitution may be a conservative substitution, such as a substitution according to the table below. In some embodiments, amino acids in the same block in the middle column are substituted. In some embodiments, amino acids in the same row in the rightmost column are substituted:

[0233]

[0234] In some embodiments, the substitutions may be functionally conservative. That is, in some embodiments, the substitutions may not affect (or may not substantially affect) one or more functional properties (e.g., target binding) of the antigen binding molecule comprising the substitutions, compared to an equivalent unsubstituted molecule.

[0235] In some embodiments, substitutions relative to a reference VH or VL sequence can be concentrated in one or more specific regions of the VH or VL sequence. For example, changes that deviate from a reference VH or VL sequence can be concentrated in one or more framework regions (FR1, FR2, FR3, and / or FR4).

[0236] The VH and VL regions of an antibody's antigen-binding region together constitute the Fv region. In some embodiments, the antigen-binding molecules of the present invention comprise or consist of an Fv region that binds to IL-11Rα. In some embodiments, the VH and VL regions of the Fv are provided as a single polypeptide connected by a linker region, i.e., a single-chain Fv (scFv).

[0237] In some embodiments, the antigen binding molecules of the present invention 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 the heavy chain constant sequence of IgG (e.g., IgG1, IgG2, IgG3, IgG4), IgA (e.g., IgA1, IgA2), IgD, IgE, or IgM.

[0238] In some embodiments, the immunoglobulin heavy chain constant sequence is the human immunoglobulin G1 constant sequence (IGHG1; UniProt: P01857-1, v1; SEQ ID NO: 53). Positions 1 to 98 of SEQ ID NO: 53 form the CH1 region (SEQ ID NO: 54). Positions 99 to 110 of SEQ ID NO: 53 form the hinge region between the CH1 and CH2 regions (SEQ ID NO: 55). Positions 111 to 223 of SEQ ID NO: 53 form the CH2 region (SEQ ID NO: 56). Positions 224 to 330 of SEQ ID NO: 53 form the CH3 region (SEQ ID NO: 57).

[0239] In some embodiments, the CH1 region comprises or consists of SEQ ID NO: 54, or comprises or consists of an amino acid sequence that has 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 SEQ ID NO: 54. In some embodiments, the CH1-CH2 hinge region comprises or consists of SEQ ID NO: 55, or comprises or consists of an amino acid sequence that has 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 SEQ ID NO: 55. In some embodiments, the CH2 region comprises or consists of SEQ ID NO: 56, or comprises or consists of an amino acid sequence that has 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 SEQ ID NO: 56. In some embodiments, the CH3 region comprises or consists of SEQ ID NO: 57, or comprises or consists of an amino acid sequence that has 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 SEQ ID NO: 57.

[0240] In some embodiments, the immunoglobulin heavy chain constant sequence is the human immunoglobulin G4 constant sequence (IGHG1; UniProt: P01861, v1; SEQ ID NO: 61). Positions 1 to 98 of SEQ ID NO: 61 form the CH1 region (SEQ ID NO: 62). Positions 99 to 110 of SEQ ID NO: 61 form the hinge region between the CH1 and CH2 regions (SEQ ID NO: 63). Positions 111 to 220 of SEQ ID NO: 61 form the CH2 region (SEQ ID NO: 64). Positions 221 to 327 of SEQ ID NO: 61 form the CH3 region (SEQ ID NO: 65).

[0241] In some embodiments, the CH1 region comprises or consists of SEQ ID NO: 62, or comprises or consists of an amino acid sequence that has 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 SEQ ID NO: 62. In some embodiments, the CH1-CH2 hinge region comprises or consists of SEQ ID NO: 63, or comprises or consists of an amino acid sequence that has 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 SEQ ID NO: 63. In some embodiments, the CH2 region comprises or consists of the sequence of SEQ ID NO: 64, or comprises or consists of an amino acid sequence that has 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 SEQ ID NO: 64. In some embodiments, the CH3 region comprises or consists of the sequence of SEQ ID NO: 65, or comprises or consists of an amino acid sequence that has 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 SEQ ID NO: 65.

[0242] In some embodiments, the immunoglobulin heavy chain constant sequence is a human immunoglobulin G 4 constant sequence (IGHG1; UniProt: P01861, v1) comprising amino acid substitutions that confer improved properties to the antigen-binding molecules of the present invention. In some embodiments, the immunoglobulin heavy chain constant region is a human IgG4 comprising the substitutions S241P and / or L248E. The S241P mutation acts as a hinge stabilizer, while the L248E mutation further reduces the already low ADCC effector function of IgG4 (Davies and Sutton, Immunol Rev. 2015 Nov; 268(1): 139–159; Angal et al. Mol Immunol. 1993 Jan; 30(1): 105-8). Lower ADCC activity is advantageous for possible subcutaneous administration of antibodies.

[0243] In some embodiments, the immunoglobulin heavy chain constant sequence is a human immunoglobulin G4 constant sequence (IGHG1; UniProt: P01861, v1) comprising the substitution S241P (numbering according to the Kabat system), as set forth in SEQ ID NO:66. Positions 1 to 98 of SEQ ID NO:66 form the CH1 region (SEQ ID NO:62). Positions 99 to 110 of SEQ ID NO:66 form the hinge region between the CH1 and CH2 regions (SEQ ID NO:67). Positions 111 to 220 of SEQ ID NO:66 form the CH2 region (SEQ ID NO:64). Positions 221 to 327 of SEQ ID NO:66 form the CH3 region (SEQ ID NO:65).

[0244] In some embodiments, the CH1 region comprises or consists of SEQ ID NO: 62, or comprises or consists of an amino acid sequence that has 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 SEQ ID NO: 62. In some embodiments, the CH1-CH2 hinge region comprises or consists of SEQ ID NO: 67, or comprises or consists of an amino acid sequence that has 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 SEQ ID NO: 67. In some embodiments, the CH2 region comprises or consists of the sequence of SEQ ID NO: 64, or comprises or consists of an amino acid sequence that has 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 SEQ ID NO: 64. In some embodiments, the CH3 region comprises or consists of the sequence of SEQ ID NO: 65, or comprises or consists of an amino acid sequence that has 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 SEQ ID NO: 65.

[0245] In some embodiments, the immunoglobulin heavy chain constant sequence is a human immunoglobulin G4 constant sequence (IGHG1; UniProt: P01861, v1) comprising substitutions S241P and L248E (numbering according to the Kabat system), as set forth in SEQ ID NO: 68. Positions 1 to 98 of SEQ ID NO: 68 form the CH1 region (SEQ ID NO: 62). Positions 99 to 110 of SEQ ID NO: 68 form the hinge region between the CH1 and CH2 regions comprising the S241P substitution (SEQ ID NO: 67). Positions 111 to 220 of SEQ ID NO: 68 form the CH2 region comprising the L248E substitution (SEQ ID NO: 69). Positions 221 to 327 of SEQ ID NO: 68 form the CH3 region (SEQ ID NO: 65).

[0246] In some embodiments, the CH1 region comprises or consists of SEQ ID NO: 62, or comprises or consists of an amino acid sequence that has 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 SEQ ID NO: 62. In some embodiments, the CH1-CH2 hinge region comprises or consists of SEQ ID NO: 67, or comprises or consists of an amino acid sequence that has 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 SEQ ID NO: 67. In some embodiments, the CH2 region comprises or consists of the sequence of SEQ ID NO: 69, or comprises or consists of an amino acid sequence that has 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 SEQ ID NO: 69. In some embodiments, the CH3 region comprises or consists of the sequence of SEQ ID NO: 65, or comprises or consists of an amino acid sequence that has 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 SEQ ID NO: 65.

[0247] In some embodiments, the antigen binding molecules of the present invention comprise one or more regions of an immunoglobulin light chain constant sequence. In some embodiments, the immunoglobulin light chain constant sequence is a human immunoglobulin kappa constant sequence (IGKC; Cκ; UniProt: P01834-1, v2; SEQ ID NO: 58). In some embodiments, the immunoglobulin light chain constant sequence is a human immunoglobulin lambda constant sequence (IGLC; Cλ), such as IGLC1, IGLC2, IGLC3, IGLC6, or IGLC7 (SEQ ID NO: 75, 76, 77, 78, or 79). In some embodiments, the CL region comprises or consists of the sequence shown in SEQ ID NO: 58, or comprises or consists of an amino acid sequence 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 with the amino acid sequence shown in SEQ ID NO: 58. In some embodiments, the CL region comprises or consists of the sequence shown in SEQ ID NO:75, 76, 77, 78 or 79, or comprises or consists of an amino acid sequence 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 with the amino acid sequence shown in SEQ ID NO:75, 76, 77, 78 or 79.

[0248] The VL and light chain constant (CL) regions of the antigen-binding region of an antibody, as well as the VH region and heavy chain constant 1 (CH1) region, together constitute the Fab region. In some embodiments, the antigen-binding molecule comprises a Fab region comprising VH, CH1, VL, and CL (e.g., Cκ or Cλ). In some embodiments, the Fab region comprises a polypeptide comprising VH and CH1 (e.g., a VH-CH1 fusion polypeptide), and a polypeptide comprising VL and CL (e.g., a VL-CL fusion polypeptide). In some embodiments, the Fab region comprises a polypeptide comprising VH and CL (e.g., a VH-CL fusion polypeptide), and a polypeptide comprising VL and CH (e.g., a VL-CH1 fusion polypeptide); that is, in some embodiments, the Fab region is a CrossFab region. In some embodiments, the VH, CH1, VL, and CL regions of a Fab or CrossFab are provided as a single polypeptide connected by a connecting region, i.e., a single-chain Fab (scFab) or a single-chain CrossFab (scCrossFab).

[0249] In some embodiments, the antigen-binding molecules of the present invention comprise or consist of a Fab region that binds to IL-11Rα.

[0250] In some embodiments, the antigen binding molecules described herein comprise or consist of intact antibodies that bind to IL-11Rα. As used herein, "intact antibodies" refer to antibodies having a structure substantially similar to that of immunoglobulins (Ig). For example, various types of immunoglobulins and their structures are described in Schroeder and Cavacini, J Allergy Clin Immunol. (2010) 125(202): S41-S52, which is incorporated herein by reference in its entirety.

[0251] G-type immunoglobulins (i.e., IgG) are glycoproteins of approximately 150 kDa, consisting of two heavy chains and two light chains. From N-terminus to C-terminus, the heavy chain comprises VH, followed by a heavy chain constant region comprising three constant domains (CH1, CH2, and CH3). Similarly, the light chain comprises VL, followed by CL. Based 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. Light chains can be either kappa (κ) or lambda (λ).

[0252] In some embodiments, the antigen binding molecules described herein comprise or consist of IgG (e.g., IgG1, IgG2, IgG3, IgG4), IgA (e.g., IgA1, IgA2), IgD, IgE, or IgM that binds to IL-11Rα.

[0253] In some embodiments, the antigen-binding molecules of the present invention bind to IL-11Rα at least monovalently. Binding valency refers to the number of binding sites in an antigen-binding molecule for a given antigenic determinant. Thus, in some embodiments, the antigen-binding molecule comprises at least one IL-11Rα binding site.

[0254] In some embodiments, the antigen binding molecule comprises multiple IL-11Rα binding sites, such as 2, 3, or 4 binding sites. The binding sites can be the same or different. In some embodiments, the antigen binding molecule is, for example, bivalent, trivalent, or tetravalent for IL-11Rα.

[0255] Aspects of the present invention relate to multispecific antigen binding molecules. "Multispecific" refers to an antigen binding molecule that exhibits specific binding to more than one target. In some embodiments, the antigen binding molecule is a bispecific antigen binding molecule. In some embodiments, the antigen binding molecule comprises at least two different antigen binding domains (i.e., at least two antigen binding domains, e.g., comprising different VH and VL).

[0256] In some embodiments, the antigen binding molecule binds IL-11Rα and another target (eg, an antigen other than IL-11Rα) and is thus at least bispecific. The term "bispecific" refers to an antigen binding molecule that is able to specifically bind to at least two different antigenic determinants.

[0257] It should be understood that the antigen-binding molecules of the present invention (e.g., multispecific antigen-binding molecules) can include antigen-binding molecules that can bind to the specific target of the antigen-binding molecule. For example, antigen-binding molecules that can bind to IL-11Rα and antigens other than IL-11Rα can include: (i) antigen-binding molecules that can bind to IL-11Rα, and (ii) antigen-binding molecules that can bind to antigens other than IL-11Rα.

[0258] It should also be understood that the antigen-binding molecules of the present invention (e.g., multispecific antigen-binding molecules) can comprise an antigen-binding polypeptide or antigen-binding polypeptide complex that can bind to the specific target of the antigen-binding molecule. For example, the antigen-binding molecules of the present invention can include, for example, (i) an antigen-binding polypeptide complex that can bind to IL-11Rα, which comprises a light chain polypeptide (comprising a VL-CL structure) and a heavy chain polypeptide (comprising a VH-CH1-CH2-CH3 structure), and ii) an antigen-binding polypeptide complex that can bind to an antigen other than IL-11Rα, which comprises a light chain polypeptide (comprising a VL-CL structure) and a heavy chain polypeptide (comprising a VH-CH1-CH2-CH3 structure).

[0259] In some embodiments, a component antigen binding molecule of a larger antigen binding molecule (e.g., a multispecific antigen binding molecule) can be referred to, for example, as an "antigen binding domain" or "antigen binding region" of the larger antigen binding molecule.

[0260] In some embodiments, the antigen binding molecule comprises an antigen binding molecule capable of binding to IL-11Rα and an antigen binding molecule capable of binding to an antigen other than IL-11Rα. In some embodiments, the antigen other than IL-11Rα is an immune cell surface molecule. In some embodiments, the antigen other than IL-11Rα is a cancer cell antigen. In some embodiments, the antigen other than IL-11Rα is a receptor molecule, such as a cell surface receptor. In some embodiments, the antigen other than IL-11Rα is a cell signaling molecule, such as a cytokine, chemokine, interferon, interleukin, or lymphokine. In some embodiments, the antigen other than IL-11Rα is a growth factor or hormone.

[0261] Cancer cell antigens are antigens expressed or overexpressed by cancer cells. Cancer cell antigens can be any peptide / polypeptide, glycoprotein, lipoprotein, polysaccharide, glycolipid, lipid or its fragment. The expression of cancer cell antigens may be related to cancer. Cancer cell antigens can be abnormally expressed by cancer cells (e.g., cancer cell antigens can be abnormally positioned and expressed), or can be expressed by cancer cells with abnormal structures. Cancer cell antigens may be able to trigger an immune response. In some embodiments, the antigen is expressed on the cell surface of cancer cells (i.e., the cancer cell antigen is a cancer cell surface antigen). In some embodiments, a portion of the antigen bound to the antigen binding molecules described herein is displayed on the outer surface of the cancer cell (i.e., extracellular). Cancer cell antigens may be cancer-associated antigens. In some embodiments, the cancer cell antigen is an antigen whose expression is related to the development, progression or severity of cancer symptoms. Antigens associated with cancer may be related to the cause or pathology of cancer, or may be abnormally expressed due to cancer. In some embodiments, the cancer cell antigen is an antigen whose expression is upregulated (e.g., at the RNA and / or protein level) by cancer cells compared to the expression level of corresponding non-cancerous cells (e.g., non-cancerous cells derived from the same tissue / cell type). In some embodiments, the cancer associated antigen may be preferentially expressed by cancer cells, but not by corresponding non-cancerous cells (e.g., non-cancerous cells derived from the same tissue / cell type). In some embodiments, the cancer associated antigen may be the product of a mutated oncogene or a mutated tumor suppressor gene. In some embodiments, the cancer associated antigen may be an overexpressed cellular protein, a cancer antigen produced by an oncovirus, a carcinoembryonic antigen, or a product of a cell surface glycolipid or glycoprotein.

[0262] Immune cell surface molecules can be any peptide / polypeptide, glycoprotein, lipoprotein, polysaccharide, glycolipid, lipid or its fragment on the cell surface or surface expression of immune cell.In some embodiments, a part for the immune cell surface molecules combined by the antigen binding molecules of the present invention is on the outer surface of immune cell (i.e., extracellular). Immune cell surface molecules can be expressed on the cell surface of any immune cell.In some embodiments, immune cell can be a cell of hematopoietic origin, such as neutrophil, eosinophil, basophil, dendritic cell, lymphocyte or monocyte. Lymphocyte can be, such as T cell, B cell, natural killer (NK) cell, NKT cell or innate lymphoid cell (ILC) or its precursor (such as thymocyte or B precursor cell).In some embodiments, immune cell surface molecules can be costimulatory molecules (such as CD28, OX40, 4-1BB, ICOS or CD27) or its part. In some embodiments, the immune cell surface molecule can be a checkpoint molecule (e.g., PD-1, CTLA-4, LAG-3, TIM-3, TIGIT, or BTLA) or a ligand thereof.

[0263] The multispecific antigen-binding molecules of the present invention may be provided in any suitable form, such as those described in Brinkmann and Kontermann MAbs (2017) 9(2): 182-212, which is incorporated herein by reference in its entirety. Suitable forms include those described in Brinkmann and Kontermann MAbs (2017) 9(2): 182-212. Figure 2Formats shown in: antibody conjugates, such as IgG2, F(ab')2 or CovX-antibodies; IgG or IgG-like molecules, such as IgG, chimeric IgG, κλ-antibody common HC; CH1 / CL fusion proteins, such as scFv2-CH1 / CL, VHH2-CH1 / CL; "variable domain only" bispecific antigen binding molecules, such as tandem scFv (taFV), triabodies, diabodies (Db), dsDb, Db(kih), DART, scDB, dsFv-dsFv, tandAb, three-head antibodies, tandem dAb / VHH, trivalent dAb.VHH; non-Ig fusion proteins, such as scFv2-albumin, scDb-albumin, taFv-albumin, taFv-toxin, miniantibodies, DNL-Fab2, DNL-Fab2-scFv, DNL-Fab2-IgG-cytokine 2, ImmTAC (TCR-scFv); modified F c and CH3 fusion proteins, such as scFv-Fc (kih), scFv-Fc (CH3 charge pair), scFv-Fc (EW-RVT), scFv-fc (HA-TF), scFv-Fc (SEED antibody), taFv-Fc (kih), scFv-Fc (kih) -Fv, Fab-Fc (kih) -scFv, Fab-scFv-Fc (kih), Fab-scFv-Fc (BE AT), Fab-scFv-Fc (SEED antibody), DART-Fc, scFv-CH3 (kih), TriFabs; Fc fusions, such as diabodies, scDb-Fc, taFv-Fc, scFv-Fc-scFv, HCAb-VHH, Fab-scFv-Fc, scFv4-Ig, scFv2-Fcab; CH3 fusions, such as diabodies, scDb-CH3; IgE / IgM CH2 fusions, such as scFv-EHD2-scFv, scFvMHD2-scFv; Fab fusion proteins, such as Fab-scFv (diabodies), Fab-scFv2 (tribodies), Fab-Fv, Fab-dsFv, Fab-VHH, orthogonal Fab-Fab; non-Ig fusion proteins, such as DNL-Fab3, DNL-Fab2-scFv, DNL-Fab2-IgG-cytokine 2;Asymmetric IgG or IgG-like molecules, such as IgG(kih), IgG(kih) universal LC, ZW1 IgG universal LC, Biclonics universal LC, CrossMab, CrossMab(kih), scFab-IgG(kih), Fab-scFab-IgG(kih), orthogonal Fab IgG(kih), DuetMab, CH3 charge pair + CH1 / CL charge pair, hinge / CH3 charge pair, SEED-antibody, bispecific antibody, four-in-one CrossMab(kih), LUZ-Y universal LC; LUZ-Y scFab-IgG, FcFc*; appended and Fc-modified IgG, such as IgG(kih)-Fv, IgG HA-TF-Fv, IgG (kih) scFab, scFab-Fc (kih) -scFv2, scFab-Fc (kih) -scFv, half DVD-Ig, DVI-Ig (four in one), CrossMab-Fab; modified Fc and CH3 fusion proteins, such as Fab-Fc (kih) -scFv, Fab-scFv-Fc (kih), Fab-scFv-Fc (BEAT), Fab-scFv-Fc-SEED antibody, TriFab; additional IgG-HC fusion proteins, such as IgG-HC, scFv, IgG-dAb, IgG-taFV, IgG-CrossFab, IgG-orthogonal Fab, IgG-(CαCβ) Fab, scFv-HC-Ig G, tandem Fab-IgG (orthogonal Fab) Fab-IgG (CαCβFab), Fab-IgG (CR3), Fab-hinge-IgG (CR3); additional IgG-LC fusions, such as IgG-scFv (LC), scFv (LC)-IgG, dAb-IgG; additional IgG-HC and LC fusions, such as DVD-Ig, TVD-Ig, CODV-Ig, scFv4-IgG, Zy antibodies; Fc fusions, such as Fab-scFv-Fc, scFv4-Ig; F(ab')2 fusions, such as F(ab')2-scFv2; CH1 / CL fusion protein scFv2-CH1-hinge / CL; modified IgG, such as DAF (two-in-one IgG), DutaMab, Mab; 2 ; and non-Ig fusions, such as DNL-Fab4-IgG.

[0264] Technicians are able to design and prepare bispecific antigen-binding molecules. Methods for producing bispecific antigen-binding molecules include chemical cross-linking of antigen-binding molecules or antibody fragments, for example, by having reducible disulfide bonds or non-reducible thioether bonds, as described in Segal and Bast in 2001 in "Generation of Bispecific Antigen-Binding Molecules", Current Protocols in Immunology 14: IV: 2.13: 2.13.1–2.13.16, which is incorporated herein by reference in its entirety. For example, N-succinimidyl-3-(-2-pyridyldithio)-propionate (SPDP) can be used for chemical cross-linking, such as Fab fragments via hinge region SH-groups to produce disulfide-linked bispecific F(ab)2 heterodimers.

[0265] Other methods for producing bispecific antigen-binding molecules include fusing antibody-producing hybridomas, for example, with polyethylene glycol to produce a quadrivalent cell capable of secreting the bispecific antibody, as described in DM and Bast, BJ, 2001, "Generation of Bispecific Antigen-Binding Molecules," Current Protocols in Immunology, 14: IV: 2.13: 2.13.1–2.13.16.

[0266] The bispecific antigen-binding molecules of the present invention can also be produced recombinantly, for example, by expression from a nucleic acid construct encoding a polypeptide of the antigen-binding molecule, for example, as described in Antibody Engineering: Methods and Protocols, 2nd Edition (Humana Press, 2012), Chapter 40: Generation of Bispecific Antibodies: Diabodies and Tandem scFv or in French (Hornig and ), How to make bispecific antigen binding molecules, Methods in Molecular Medicine 2000, 40: 333-339, which is incorporated herein by reference in its entirety. For example, light chain and heavy chain variable domains encoding two antigen binding fragments (i.e., light chain and heavy chain variable domains of an antigen binding fragment capable of binding to VISTA, and light chain and heavy chain variable domains of an antigen binding fragment capable of binding to another target protein) can be prepared by molecular cloning techniques, and a DNA construct comprising a sequence encoding a suitable linker or dimerization domain between the antigen binding fragments. Recombinant bispecific antibodies can be produced by expressing (e.g., in vitro) a construct in a suitable host cell (e.g., a mammalian host cell), and the expressed recombinant bispecific antibodies can then be optionally purified.

[0267] In some embodiments, the antigen-binding molecules of the present invention comprise an Fc region. The Fc region is composed of the CH2 and CH3 regions of one polypeptide and the CH2 and CH3 regions of another polypeptide. The CH2 and CH3 regions from the two polypeptides together form the Fc region.

[0268] In some embodiments, the antigen binding molecules of the present invention include an Fc region containing modifications that promote the association of the Fc region in one or more CH2 and CH3 regions. The recombinant co-expression and subsequent association of the constituent polypeptides of the antigen binding molecules result in several possible combinations. In order to increase the yield of the desired polypeptide combination in the antigen binding molecules in recombinant production, it is advantageous to introduce modifications in the Fc region that promote the association of the desired combination of heavy chain polypeptides. The modification can promote, for example, hydrophobic and / or electrostatic interactions between the CH2 and / or CH3 regions of different polypeptide chains. Suitable modifications are described in, for example, Ha et al., Front.Immnol (2016) 7: 394, which is incorporated herein by reference in its entirety. In some embodiments, the antigen binding molecules of the present invention include an Fc region comprising paired substitutions in the CH3 region of the Fc region in one of the following forms, as shown in Table 1 of Ha et al., Immnol (2016) 7: 394: KiH, KiH s-s , HA-TF, ZW1, 7.8.60, DD-KK, EW-RVT, EW-RVT s-s , SEED or A107.

[0269] peptides

[0270] The present invention also provides polypeptide components of antigen binding molecules. The polypeptides can be provided in isolated or substantially purified form.

[0271] The antigen-binding molecule of the present invention may be or may comprise a complex of polypeptides.

[0272] In the present specification, when a polypeptide comprises more than one domain or region, it should be understood that the multiple domains / regions are preferably present in the same polypeptide chain. That is, a polypeptide comprising more than one domain or region is a fusion polypeptide comprising these domains / regions.

[0273] In some embodiments, the polypeptides of the present invention comprise or consist of a VH as described herein. In some embodiments, the polypeptides of the present invention comprise or consist of a VL as described herein.

[0274] In some embodiments, the polypeptide further comprises one or more antibody heavy chain constant regions (CH). In some embodiments, the polypeptide further comprises one or more antibody light chain constant regions (CL). In some embodiments, the polypeptide comprises the CH1, CH2, and / or CH3 regions of an immunoglobulin (Ig).

[0275] In some embodiments, the polypeptide comprises one or more regions of an immunoglobulin heavy chain constant sequence. In some embodiments, the polypeptide comprises a CH1 region as described herein. In some embodiments, the polypeptide comprises a CH1-CH2 hinge region as described herein. In some embodiments, the polypeptide comprises a CH2 region as described herein. In some embodiments, the polypeptide comprises a CH3 region as described herein.

[0276] In some embodiments, the polypeptide comprises a CH3 region comprising any one of the amino acid substitutions / combinations of amino acid substitutions shown in Table 1 of Ha et al., Front. Immnol (2016) 7:394 (incorporated above by reference).

[0277] In some embodiments, the CH2 and / or CH3 region of the polypeptide comprises one or more amino acid substitutions to promote association of the polypeptide with another polypeptide comprising a CH2 and / or CH3 region.

[0278] In some embodiments, the polypeptide comprises one or more regions of an immunoglobulin light chain constant sequence. In some embodiments, the polypeptide comprises a CL region as described herein.

[0279] In some embodiments, the polypeptide of the present invention comprises one of the following structures from the N-terminus to the C-terminus:

[0280] (i)VH

[0281] (ii) VL

[0282] (iii) VH-CH1

[0283] (iv)VL-CL

[0284] (v)VL-CH1

[0285] (vi) VH-CL

[0286] (vii) VH-CH1-CH2-CH3

[0287] (viii)VL-CL-CH2-CH3

[0288] (ix)VL-CH1-CH2-CH3

[0289] (x)VH-CL-CH2-CH3.

[0290] The present invention also provides antigen-binding molecules composed of the polypeptides of the present invention. In some embodiments, the antigen-binding molecules of the present invention comprise a polypeptide combination of one of the following:

[0291] (A)VH+VL

[0292] (B)VH-CH1+VL-CL

[0293] (C)VL-CH1+VH-CL

[0294] (D)VH-CH1-CH2-CH3+VL-CL

[0295] (E)VH-CL-CH2-CH3+VL-CH1

[0296] (F)VL-CH1-CH2-CH3+VH-CL

[0297] (G)VL-CL-CH2-CH3+VH-CH1

[0298] (H)VH-CH1-CH2-CH3+VL-CL-CH2-CH3

[0299] (I) VH-CL-CH2-CH3+VL-CH1-CH2-CH3.

[0300] In some embodiments, the antigen-binding molecule comprises a plurality of polypeptides in the combination shown in (A) to (I) above. For example, referring to (D) above, in some embodiments, the antigen-binding molecule comprises two polypeptides comprising a VH-CH1-CH2-CH3 structure, and two polypeptides comprising a VL-CL structure.

[0301] In some embodiments, the antigen binding molecules of the present invention comprise one of the following polypeptide combinations:

[0302] (J) VH (anti-IL-11Rα) + VL (anti-IL-11Rα)

[0303] (K) VH (anti-IL-11Rα)-CH1 + VL (anti-IL-11Rα)-CL

[0304] (L) VL (anti-IL-11Rα)-CH1 + VH (anti-IL-11Rα)-CL

[0305] (M) VH (anti-IL-11Rα)-CH1-CH2-CH3 + VL (anti-IL-11Rα)-CL

[0306] (N)VH(anti-IL-11Rα)-CL-CH2-CH3+VL(anti-IL-11Rα)-CH1

[0307] (O)VL(anti-IL-11Rα)-CH1-CH2-CH3+VH(anti-IL-11Rα)-CL

[0308] (P)VL(anti-IL-11Rα)-CL-CH2-CH3+VH(anti-IL-11Rα)-CH1

[0309] (Q) VH (anti-IL-11Rα)-CH1-CH2-CH3 + VL (anti-IL-11Rα)-CL-CH2-CH3

[0310] (R) VH (anti-IL-11Rα)-CL-CH2-CH3 + VL (anti-IL-11Rα)-CH1-CH2-CH3.

[0311] Wherein: "VH (anti-IL-11Rα)" refers to a VH of an antigen-binding molecule capable of binding to IL-11Rα as described herein, for example, as defined in any one of (1) to (21); "VL (anti-IL-11Rα)" refers to a VL of an antigen-binding molecule capable of binding to IL-11Rα as described herein, for example, as defined in any one of (22) to (37).

[0312] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 70%, preferably one of 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 one of SEQ ID NOs: 7 to 17.

[0313] In some embodiments, the antigen-binding molecules of the present invention comprise a polypeptide comprising 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 with the amino acid sequence of SEQ ID NO: 70, or consisting thereof. In some embodiments, the antigen-binding molecules of the present invention comprise a polypeptide comprising 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 with the amino acid sequence of SEQ ID NO: 71, or consisting thereof. In some embodiments, the antigen-binding molecules of the present invention comprise a polypeptide 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 with the amino acid sequence of SEQ ID NO: 72. In some embodiments, the antigen-binding molecules of the present invention comprise a polypeptide 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 with the amino acid sequence of SEQ ID NO: 73. In some embodiments, the antigen binding molecule of the present invention comprises a polypeptide comprising or consisting of an amino acid sequence having at least 70%, preferably at least 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: 74. Linkers and Additional Sequences

[0314] In some embodiments, the antigen binding molecules and polypeptides of the present invention comprise one or more linker sequences between the amino acid sequences. A linker sequence can be provided at one or both ends of one or more of the VH, VL, CH1-CH2 hinge region, CH2 region, and CH3 region of the antigen binding molecules / polypeptide.

[0315] Linker sequences are known to those skilled in the art and are described, for example, in Chen et al., Adv Drug Deliv Rev (2013) 65(10): 1357-1369, which is incorporated herein by reference in its entirety. In some embodiments, the linker sequence can be a flexible linker sequence. Flexible linker sequences allow relative movement of the amino acid sequences connected by the linker sequence. Flexible linkers are known to those skilled in the art and several have been identified in Chen et al., Adv Drug Deliv Rev (2013) 65(10): 1357-1369. Flexible linker sequences typically contain a high proportion of glycine and / or serine residues.

[0316] In some embodiments, the linker sequence comprises at least one glycine residue and / or at least one serine residue. In some embodiments, the linker sequence is composed of glycine and serine residues. In some embodiments, the linker sequence has 1-2, 1-3, 1-4, 1-5 or 1-10 amino acid length.

[0317] The antigen binding molecules and polypeptide of the present invention can additionally include other amino acids or amino acid sequences.For example, the antigen binding molecules and polypeptide can include amino acid sequences to promote expression, folding, transport, processing, purification or detection of antigen binding molecules / polypeptides.For example, the antigen binding molecules / polypeptides can include sequences encoding His (such as 6XHis), Myc, GST, MBP, FLAG, HA, E or biotin tags optionally at the N- or C-terminal ends of antigen binding molecules / polypeptides.In some embodiments, the antigen binding molecules / polypeptides include detectable moieties, such as fluorescence, luminescence, immunoassay, radiation, chemistry, nucleic acid or enzyme labeling.

[0318] The antigen binding molecules and polypeptides of the present invention may further comprise a signal peptide (also referred to as a leader sequence or signal sequence). A signal peptide is typically composed of a sequence of 5-30 hydrophobic amino acids that form a single α helix. Secreted proteins and proteins expressed on the cell surface typically comprise a signal peptide.

[0319] The signal peptide may be present at the N-terminus of the antigen-binding molecule / polypeptide and may be present in newly synthesized antigen-binding molecules / polypeptides. The signal peptide provides for efficient transport and secretion of the antigen-binding molecule / polypeptide. The signal peptide is typically removed by cleavage and is therefore not included in the mature antigen-binding molecule / polypeptide secreted by the cell expressing the antigen-binding molecule / polypeptide.

[0320] Signal peptides are known for many proteins and are recorded in databases such as GenBank, UniProt, Swiss-Prot, TrEMBL, Protein Information Resource, Protein Data Bank, Ensembl, and InterPro, and / or can be identified / predicted using amino acid sequence analysis tools such as SignalP (Petersen et al., 2011 Nature Methods 8:785-786) or Signal-BLAST (Frank and Sippl, 2008 Bioinformatics 24:2172-2176).

[0321] Labels and conjugates

[0322] In some embodiments, the antigen binding molecules of the invention further comprise a detectable moiety or chemical moiety.

[0323] In some embodiments, the antigen binding molecules include detectable moieties, such as fluorescent markers, phosphorescent markers, luminescent markers, immune detectable markers (such as epitope markers), radioactive markers, chemistry, nucleic acid or enzyme markers. The antigen binding molecules can be covalently or non-covalently labeled with detectable moieties.

[0324] Fluorescent labels include, for example, fluorescein, rhodamine, allophycocyanin, eosin and NDB, green fluorescent protein (GFP) chelates of rare earth elements (e.g., europium (Eu), terbium (Tb), and samarium (Sm)), tetramethylrhodamine, Texas Red, 4-methylumbelliferone, 7-amino-4-methylcoumarin, Cy3, and Cy5. Radioactive labels include radioisotopes such as iodine. 123 ,iodine 125 ,iodine 126 ,iodine 131 ,iodine 133 ,bromine 77 ,technetium 99m 、Indium 111 ,indium 113m ,gallium 67 ,gallium 68 ,ruthenium 95 ,ruthenium 97 ,ruthenium 103 ,ruthenium 105 ,mercury 207 ,mercury 203 ,rhenium 99m ,rhenium 101 ,rhenium 105 ,scandium 47 ,tellurium 121m ,tellurium122m ,tellurium 125m ,thulium 165 ,thulium 167 ,thulium 168 ,copper 67 ,fluorine 18 ,yttrium 90 ,palladium 100 ,bismuth 217 and antimony 211 Luminescent labels include radioluminescence, chemiluminescence (e.g., apidinyl esters, luminol, isoluminol), and bioluminescent labels. Immunodetectable labels include haptens, peptides / polypeptides, antibodies, receptors, and ligands, such as biotin, avidin, streptavidin, or digoxigenin. Nucleic acid labels include aptamers. Enzyme labels include, for example, peroxidase, alkaline phosphatase, glucose oxidase, β-galactosidase, and luciferase.

[0325] In some embodiments, the antigen binding molecules of the present invention are conjugated to a chemical moiety. The chemical moiety can be a moiety for providing a therapeutic effect. Antibody-drug conjugates are reviewed in Parslow et al., Biomedicines. 2016 Sep; 4(3): 14. In some embodiments, the chemical moiety can be a drug moiety (e.g., a cytotoxic agent). In some embodiments, the drug moiety can be a chemotherapeutic agent. In some embodiments, the drug moiety is selected from calicheamicin, DM1, DM4, monomethyl autamicin E (MMAE), monomethyl autamicin F (MMAF), SN-38, doxorubicin, ducathiamycin, D6.5, and PBD.

[0326] Functional properties of antigen-binding molecules

[0327] The antigen binding molecules described herein can be characterized by reference to certain functional properties. In some embodiments, the antigen binding molecules described herein can have one or more of the following properties:

[0328] a) specific binding to IL-11Rα (eg, human IL-11 and / or mouse IL-11Rα);

[0329] b) binds to IL-11Rα (e.g., human IL-11Rα) with a binding affinity of EC50 = less than 1000 ng / ml, e.g., as determined by ELISA;

[0330] c) inhibiting the interaction between IL-11Rα and IL-11;

[0331] d) inhibiting the interaction between IL-11Rα and gp130;

[0332] e) inhibiting the interaction between the IL-11Rα:gp130 receptor complex and IL-11;

[0333] f) inhibit the interaction between the IL-11:IL-11Rα complex and gp130;

[0334] g) inhibiting IL-11 / IL-11R signaling;

[0335] h) inhibiting IL-11-mediated signaling;

[0336] i) inhibiting signaling mediated by IL-11 binding to the IL-11Rα:gp130 receptor complex;

[0337] j) inhibiting signal transduction mediated by the binding of the IL-11:IL-11Rα complex to gp130 (i.e., IL-11 trans-signaling);

[0338] k) inhibiting fibroblast proliferation;

[0339] 1) inhibiting the production of myofibroblasts from fibroblasts;

[0340] m) reversal / regression of myofibroblast generation from fibroblasts;

[0341] n) inhibiting the production of myofibroblasts from astrocytes, such as hepatic or pancreatic stellate cells;

[0342] o) reversal / regression of myofibroblast generation from astrocytes, such as hepatic or pancreatic stellate cells;

[0343] p) inhibiting the migratory and / or invasive behavior of fibroblasts, astrocytes or myofibroblasts (i.e., inhibiting migration and / or invasion);

[0344] q) suppressing the presence of immune cells in organs;

[0345] r) inhibit pathological processes mediated by IL-11 / IL-11R signaling;

[0346] s) inhibiting fibrosis;

[0347] t) reversal / regression of fibrosis;

[0348] u) inhibiting gene or protein expression of one or more of collagen, fibronectin, periostin, IL-6, IL-11, αSMA (ACTA2), TIMP1, MMP2, TNFα, CCL2 in fibroblasts or stellate cells, e.g., after stimulation with profibrotic factors;

[0349] v) inhibiting extracellular matrix production by fibroblasts or astrocytes;

[0350] w) inhibiting the proliferation and / or survival of cancer cells;

[0351] x) inhibiting the development and / or progression of cancer in vivo;

[0352] y) inhibiting tumor growth;

[0353] z) Killing cells expressing / containing IL-11Rα or a complex containing IL-11Rα.

[0354] As used herein, "inhibit" refers to a reduction, decrease, or attenuation relative to a control condition. For example, inhibition of a process by an antigen-binding molecule refers to the extent / degree of reduction, decrease, or attenuation of the process in the absence of the antigen-binding molecule and / or in the presence of an appropriate control antigen-binding molecule.

[0355] Inhibition may also be referred to herein as neutralization or antagonism. That is, an IL-11Rα binding antigen binding molecule that can inhibit a function or process (e.g., an interaction mediated by IL-11Rα or a complex containing IL-11Rα, signaling, or other activity) can be referred to as a "neutralizing" or "antagonistic" antigen binding molecule of the relevant function or process. For example, an antigen binding molecule that can inhibit IL-11-mediated signaling can be referred to as an antigen binding molecule that can neutralize IL-11-mediated signaling, or can be referred to as an antagonist of IL-11-mediated signaling.

[0356] A skilled artisan is able to determine appropriate control conditions for a given assay. For example, a control antigen-binding molecule can be an antigen-binding molecule directed against a target protein that is known not to have the effect associated with the property being investigated in the assay. The control antigen-binding molecule can be of the same isotype as the anti-IL-11Rα antigen-binding molecule being analyzed, for example, having the same constant region.

[0357] The antigen binding molecules described herein preferably show specific binding to IL-11R alpha. As used herein, "specific binding" refers to selective binding to an antigen and can be distinguished from non-specific binding to a non-target antigen. Compared to binding to other non-target molecules, the antigen binding molecules that specifically bind to the target molecule preferably have a greater affinity for binding to the target molecule and / or a longer duration. In some embodiments, the binding affinity of the antibody / fragment of the present invention to IL-11R alpha may be greater than to one or more members of the IL-6 receptor family. In some embodiments, the binding affinity of the antigen binding molecules of the present invention to IL-11R alpha is greater than to one or more of IL-6R alpha, leukemia inhibitory factor receptor (LIFR), oncostatin M receptor (OSMR) and ciliary neurotrophic factor receptor alpha (CNTFR alpha).

[0358] In some embodiments, inhibition of IL-11-mediated signaling is achieved by disrupting IL-11-mediated cis signaling but not disrupting IL-11-mediated trans signaling, for example, by inhibiting the gp130-mediated cis complex involving membrane-bound IL-11Rα. In some embodiments, inhibition of IL-11-mediated signaling is achieved by disrupting IL-11-mediated trans signaling but not disrupting IL-11-mediated cis signaling, i.e., by inhibiting the gp130-mediated trans signaling complex, such as IL-11 bound to soluble IL-11Rα or IL-6 bound to soluble IL-6R. In some embodiments, inhibition of IL-11-mediated signaling is achieved by disrupting both IL-11-mediated cis signaling and IL-11-mediated trans signaling.

[0359] The ability of a given polypeptide to specifically bind to a given molecule can be determined by assays known in the art, such as ELISA, surface plasmon resonance (SPR; see, e.g., Hearty et al., Methods Mol Biol (2012) 907: 411-442), biolayer interferometry (see, e.g., Lad et al., (2015) J Biomol Screen 20(4): 498-507), flow cytometry, or by radiolabeled antigen binding assay (RIA) enzyme-linked immunosorbent assay. By such assays, binding to a given molecule can be measured and quantified. In some embodiments, the binding can be a response detected in a given assay.

[0360] In some embodiments, the extent of binding of the antigen binding molecule to the non-target molecule is less than about 10% of the extent of binding of the antibody to the target molecule as measured, for example, by ELISA, SPR, biolayer interferometry, or RIA. Alternatively, binding specificity can be reflected as binding affinity, where the dissociation constant (K) for the antigen binding molecule to bind to IL-11Rα is D ) is greater than the K of the antigen-binding molecule to the non-target molecule D At least 0.1 orders of magnitude (i.e. 0.1×10 n , where n is an integer representing an order of magnitude). This may optionally be at least one of 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.5 or 2.0.

[0361] In some embodiments, the antigen binding molecules show binding to human IL-11Rα. In some embodiments, the antigen binding molecules show binding to mouse IL-11Rα. In some embodiments, the antigen binding molecules show binding to human IL-11Rα and mouse IL-11Rα. That is, in some embodiments, the antigen binding molecules have cross-reactivity to human IL-11Rα and mouse IL-11Rα. In some embodiments, the antigen binding molecules of the present invention show cross-reactivity to IL-11Rα of non-human primates.

[0362] In some embodiments, the antigen binding molecules of the present invention bind to the K of IL-11Rα D 5 μM or less, preferably one of ≤1 μM, ≤500 nM, ≤100 nM, ≤75 nM, ≤50 nM, ≤40 nM, ≤30 nM, ≤20 nM, ≤15 nM, ≤12.5 nM, ≤10 nM, ≤9 nM, ≤8 nM, ≤7 nM, ≤6 nM, ≤5 nM, ≤4 nM, ≤3 nM, ≤2 nM, ≤1 nM, ≤500 pM.

[0363] In some embodiments, the antigen binding molecules of the present invention bind to the K of IL-11Rα (e.g., human IL-11Rα). D is ≤20 nM, ≤15 nM, ≤12.5 nM, ≤10 nM, ≤9 nM, ≤8 nM, ≤7 nM or ≤6 nM, for example, as determined by SPR analysis. In some embodiments, the antigen binding molecule binds to IL-11Rα with a K of ≤1.5. D is ≥1 nM and ≤20 nM, such as ≥1 nM and ≤15 nM, or ≥1 nM and ≤10 nM. In some embodiments, the antigen binding molecule binds to IL-11Rα with a K of D is ≥5 nM and ≤20 nM, such as ≥5 nM and ≤15 nM, or ≥5 nM and ≤10 nM.

[0364] In some embodiments, the antigen binding molecules of the present invention bind to IL-11Rα with a binding affinity (e.g., as determined by ELISA) of EC50 = 1000 ng / ml or lower, preferably one of ≤900 ng / ml, ≤800 ng / ml, ≤700 ng / ml, ≤600 ng / ml, ≤500 ng / ml, ≤400 ng / ml, ≤300 ng / ml, ≤200 ng / ml, ≤100 ng / ml, ≤90 ng / ml, ≤80 ng / ml, ≤70 ng / ml, ≤60 ng / ml, ≤50 ng / ml, ≤40 ng / ml, ≤30 ng / ml, ≤20 ng / ml, ≤15 ng / ml, ≤10 ng / ml, ≤7.5 ng / ml, ≤5 ng / ml, ≤2.5 ng / ml or ≤1 ng / ml.

[0365] The binding affinity of antigen binding molecules to IL-11R α can be determined by ELISA in vitro analysis. Suitable assays are well known in the art and can be performed by technical personnel, such as "Antibody Engineering", Volume 1, (2nd Edition), Springer Protocol, Springer (2010), Part V, Pages 657-665. For example, the affinity of antigen binding molecules combined with IL-11R α can be analyzed according to the method described in the experimental examples herein.

[0366] The ability of an antigen binding molecule to inhibit the interaction between two proteins can be determined by, for example, analyzing the interaction in the presence of the antigen binding molecule, or after incubation of one or both interaction partners with the antigen binding molecule. An example of a suitable assay for determining whether a given antigen binding molecule is able to inhibit the interaction between two interaction partners is a competitive ELISA assay.

[0367] Antigen binding molecules that can inhibit an interaction (e.g., between IL-11Rα and IL-11, or between IL-11Rα and gp130, or between IL-11Rα:gp130 and IL-11, or between IL-11:IL-11Rα and gp130) can be identified by observing a decrease / reduction in the level of interaction between the interaction partners in the presence of the antigen binding molecule, or after incubation of one or both of the interaction partners with the antigen binding molecule, compared to the level of interaction achieved in the absence of the antigen binding molecule (or in the presence of an appropriate control antigen binding molecule). Suitable assays can be performed in vitro, for example, using recombinant interaction partners or using cells expressing the interaction partner. Cells expressing the interaction partner may be endogenous or may be expressed by introducing nucleic acid into the cell. For the purpose of such an assay, one or both of the interaction partner and / or antigen binding molecule may be labeled or used in conjunction with a detectable entity to detect and / or measure the level of interaction.

[0368] The ability of an antigen binding molecule to inhibit the interaction between two binding partners can also be determined by analyzing the downstream functional consequences of such interaction, such as receptor signaling. For example, downstream functional consequences of the interaction between IL-11Rα:gp130 and IL-11 or between IL-11:IL-11Rα and gp130 may include fibroblast proliferation, generation of fibroblasts into myofibroblasts, or expression of one or more of the following genes or proteins: collagen, fibronectin, periostin, IL-1, IL-11, αSMA, TIMP1, MMP2.

[0369] The fibroblasts disclosed herein can be derived from any tissue, including liver, lung, kidney, heart, blood vessels, eyes, skin, pancreas, spleen, intestine (e.g., large or small intestine), brain, and bone marrow. In a specific embodiment, for analyzing antigen binding molecules, the fibroblasts can be cardiac fibroblasts (e.g., atrial fibroblasts), skin fibroblasts, lung fibroblasts, kidney fibroblasts, or liver fibroblasts. The fibroblasts can be characterized by expression of one or more genes or proteins of COL1A, ACTA2, prolyl 4-hydroxylase, MAS516, and FSP1.

[0370] Gene expression can be measured by various means known to those skilled in the art, such as by quantitative real-time PCR (qRT-PCR) or by reporter gene-based methods to measure the level of mRNA. Similarly, protein expression can be measured by various methods well known in the art, such as antibody-based methods, Western blotting, immunohistochemistry, immunocytochemistry, flow cytometry, ELIS, ELISPOT or reporter gene-based methods.

[0371] In some embodiments, the antigen binding molecules of the present invention can inhibit the protein expression of one or more fibrosis markers, such as one or more of the following proteins: collagen, fibronectin, periostin, IL-6, IL-11, αSMA, TIMP1, MMP2.

[0372] The ability of an antigen-binding molecule to inhibit the interaction between IL-11Rα:gp130 and IL-11 can be analyzed by, for example, stimulating fibroblasts with TGFβ1, incubating the cells in the presence of the antigen-binding molecule, and analyzing the proportion of cells with an αSMA-positive phenotype after a determined period of time. In such examples, inhibition of the interaction between IL-11Rα:gp130 and IL-11 is identified by observing a lower proportion of cells with an αSMA-positive phenotype compared to a positive control condition in which cells are treated with TGFβ1 in the absence of the antigen-binding molecule (or in the presence of a suitable control antigen-binding molecule) or in the presence of a suitable control antigen-binding molecule.

[0373] Such assays are also suitable for analyzing the ability of antigen binding molecules to inhibit IL-11-mediated signaling.

[0374] In some embodiments, the antigen binding molecules of the invention are capable of inhibiting the interaction between IL-11Rα and IL-11 to less than 100% of the level of interaction between IL-11Rα and IL-11 in the absence of the antigen binding molecule (or in the presence of an appropriate control antigen binding molecule), such as one of 99% or less, 95% or less, 90% or less, 85% or less, 80% or less, 75% or less, 70% or less, 65% or less, 60% or less, 55% or less, 50% or less, 45% or less, 40% or less, 35% or less, 30% or less, 25% or less, 20% or less, 15% or less, 10% or less, 5% or less, or 1% or less. In some embodiments, the antigen binding molecules of the present invention are capable of inhibiting the interaction between IL-11Rα and IL-11 to less than 1-fold the level of interaction between IL-11Rα and IL-11 in the absence of the antigen binding molecule (or in the presence of an appropriate control antigen binding molecule), such as one of the following: ≤0.99-fold, ≤0.95-fold, ≤0.9-fold, ≤0.85-fold, ≤0.8-fold, ≤0.75-fold, ≤0.7-fold, ≤0.65-fold, ≤0.6-fold, ≤0.55-fold, ≤0.5-fold, ≤0.45-fold, ≤0.4-fold, ≤0.35-fold, ≤0.3-fold, ≤0.25-fold, ≤0.2-fold, ≤0.15-fold, ≤0.1-fold.

[0375] In some embodiments, the antigen binding molecules of the invention are capable of inhibiting the interaction between IL-11Rα and gp130 to less than 100% of the level of interaction between IL-11Rα and gp130 in the absence of the antigen binding molecule (or in the presence of an appropriate control antigen binding molecule), such as one of 99% or less, 95% or less, 90% or less, 85% or less, 80% or less, 75% or less, 70% or less, 65% or less, 60% or less, 55% or less, 50% or less, 45% or less, 40% or less, 35% or less, 30% or less, 25% or less, 20% or less, 15% or less, 10% or less, 5% or less, or 1% or less. In some embodiments, the antigen binding molecules of the present invention are capable of inhibiting the interaction between IL-11Rα and gp130 to less than 1-fold the level of interaction between IL-11Rα and gp130 in the absence of the antigen binding molecule (or in the presence of an appropriate control antigen binding molecule), such as one of the following: ≤0.99-fold, ≤0.95-fold, ≤0.9-fold, ≤0.85-fold, ≤0.8-fold, ≤0.75-fold, ≤0.7-fold, ≤0.65-fold, ≤0.6-fold, ≤0.55-fold, ≤0.5-fold, ≤0.45-fold, ≤0.4-fold, ≤0.35-fold, ≤0.3-fold, ≤0.25-fold, ≤0.2-fold, ≤0.15-fold, ≤0.1-fold.

[0376] In some embodiments, the antigen binding molecules of the invention are capable of inhibiting the interaction between IL-11Rα:gp130 and IL-11 to less than 100% of the level of interaction between IL-11Rα:gp130 and IL-11 in the absence of the antigen binding molecule (or in the presence of an appropriate control antigen binding molecule), such as one of: 99% or less, 95% or less, 90% or less, 85% or less, 80% or less, 75% or less, 70% or less, 65% or less, 60% or less, 55% or less, 50% or less, 45% or less, 40% or less, 35% or less, 30% or less, 25% or less, 20% or less, 15% or less, 10% or less, 5% or less, or 1% or less. In some embodiments, the antigen binding molecules of the present invention are capable of inhibiting the interaction between IL-11Rα:gp130 and IL-11 to less than 1-fold the level of interaction between IL-11Rα:gp130 and IL-11 in the absence of the antigen binding molecule (or in the presence of an appropriate control antigen binding molecule), such as one of the following: ≤0.99-fold, ≤0.95-fold, ≤0.9-fold, ≤0.85-fold, ≤0.8-fold, ≤0.75-fold, ≤0.7-fold, ≤0.65-fold, ≤0.6-fold, ≤0.55-fold, ≤0.5-fold, ≤0.45-fold, ≤0.4-fold, ≤0.35-fold, ≤0.3-fold, ≤0.25-fold, ≤0.2-fold, ≤0.15-fold, ≤0.1-fold.

[0377] In some embodiments, the antigen binding molecules of the invention are capable of inhibiting the interaction between the IL-11:IL-11Rα complex and gp130 to less than 100% of the level of interaction between the IL-11:IL-11Rα complex and gp130 in the absence of the antigen binding molecule (or in the presence of an appropriate control antigen binding molecule), such as one of: 99% or less, 95% or less, 90% or less, 85% or less, 80% or less, 75% or less, 70% or less, 65% or less, 60% or less, 55% or less, 50% or less, 45% or less, 40% or less, 35% or less, 30% or less, 25% or less, 20% or less, 15% or less, 10% or less, 5% or less, or 1% or less. In some embodiments, the antigen binding molecule is capable of inhibiting the interaction between the IL-11:IL-11Rα complex and gp130 to less than 1-fold the level of interaction between the IL-11:IL-11Rα complex and gp130 in the absence of the antigen binding molecule (or in the presence of an appropriate control antigen binding molecule), such as one of the following: ≤0.99-fold, ≤0.95-fold, ≤0.9-fold, ≤0.85-fold, ≤0.8-fold, ≤0.75-fold, ≤0.7-fold, ≤0.65-fold, ≤0.6-fold, ≤0.55-fold, ≤0.5-fold, ≤0.45-fold, ≤0.4-fold, ≤0.35-fold, ≤0.3-fold, ≤0.25-fold, ≤0.2-fold, ≤0.15-fold, ≤0.1-fold.

[0378] You can also use 3 Inhibition of IL-11-mediated signaling can be analyzed by H-thymidine incorporation and / or Ba / F3 cell proliferation assays, such as those described in Curtis et al. Blood, 1997, 90(11) and Karpovich et al. Mol. Hum. Reprod. 2003 9(2):75-80. Ba / F3 cells co-express IL-11Rα and gp130.

[0379] As used herein, IL-11-mediated signal transduction and / or processes mediated by IL-11-mediated signal transduction include signal transduction mediated by fragments of IL-11 or IL-11Rα and polypeptide complexes comprising IL-11, IL-11Rα, or fragments thereof. IL-11-mediated signal transduction can be signal transduction mediated by human IL-11 or IL-11Rα and / or mouse IL-11 or IL-11Rα. IL-11-mediated signal transduction may occur after IL-11 or a complex comprising IL-11 binds to a receptor to which IL-11 or the complex binds.

[0380] In some embodiments, the antibodies and fragments of the present invention are capable of inhibiting the biological activity of IL-11, IL-11Rα, or a complex containing IL-11 or IL-11Rα. In some embodiments, the antibodies / fragments bind to IL-11Rα in a region critical for binding to IL-11 or gp130, thereby disrupting binding and / or IL-11-mediated signaling.

[0381] In some embodiments, the antigen binding molecules of the present invention are antagonists of one or more signaling pathways activated by signaling through receptors comprising IL-11Rα and / or gp130, e.g., IL-11Rα:gp130. In some embodiments, the antigen binding molecules are capable of inhibiting signaling through one or more receptor complexes comprising IL-11Rα and / or gp130, e.g., IL-11Rα:gp130.

[0382] In some embodiments, the antigen binding molecules of the invention are capable of inhibiting IL-11 mediated signaling to less than 100% of the level of IL-11 mediated signaling in the absence of the antigen binding molecule (or in the presence of an appropriate control antigen binding molecule), such as one of 99% or less, 95% or less, 90% or less, 85% or less, 80% or less, 75% or less, 70% or less, 65% or less, 60% or less, 55% or less, 50% or less, 45% or less, 40% or less, 35% or less, 30% or less, 25% or less, 20% or less, 15% or less, 10% or less, 5% or less, or 1% or less. In some embodiments, the antigen binding molecule is capable of reducing IL-11 mediated signaling to less than 1-fold the level of IL-11 mediated signaling in the absence of the antigen binding molecule (or in the presence of an appropriate control antigen binding molecule), such as one of the following: ≤0.99-fold, ≤0.95-fold, ≤0.9-fold, ≤0.85-fold, ≤0.8-fold, ≤0.75-fold, ≤0.7-fold, ≤0.65-fold, ≤0.6-fold, ≤0.55-fold, ≤0.5-fold, ≤0.45-fold, ≤0.4-fold, ≤0.35-fold, ≤0.3-fold, ≤0.25-fold, ≤0.2-fold, ≤0.15-fold, ≤0.1-fold.

[0383] In some embodiments, IL-11-mediated signaling is signaling mediated by IL-11 binding to the IL-11Rα:gp130 receptor. Such signaling can be analyzed, for example, by treating cells expressing IL-11Rα and gp130 with IL-11, or by stimulating IL-11 production in cells expressing IL-11Rα and gp130.

[0384] This can be accomplished by, for example, culturing Ba / F3 cells expressing IL-11Rα and gp130 in the presence of human IL-11 and an IL-11 binding agent, and detecting 3 The IC of antigen-binding molecules inhibiting IL-11-mediated signaling was determined by the incorporation of DNA into H-thymus. 50 .

[0385] In some embodiments, the antigen binding molecules of the invention may exhibit an IC of 10 μg / ml or less in such assays, preferably one of the following: 50 : ≤5μg / ml, ≤4μg / ml, ≤3.5μg / ml, ≤3μg / ml, ≤2μg / ml, ≤1μg / ml, ≤0.9μg / ml, ≤0.8μg / ml, ≤0.7μg / ml, ≤0.6μg / ml or ≤0.5μg / ml.

[0386] In some embodiments, IL-11-mediated signal transduction can be signal transduction mediated by the binding of an IL-11:IL-11Rα complex to gp130. In some embodiments, the IL-11:IL-11Rα complex can be soluble, such as a complex of the extracellular domain of IL-11Rα and IL-11, or a complex of a soluble IL-11Rα isoform / fragment and IL-11. In some embodiments, the soluble IL-11Rα is a soluble (secreted) isoform of IL-11Rα, or a released product of proteolytic cleavage of the extracellular domain of cell membrane-bound IL-11Rα. IL-11-mediated signal transduction mediated by the binding of IL-11 to gp130 is referred to herein as "IL-11 trans-signaling".

[0387] In some embodiments, the IL-11:IL-11Rα complex can be a complex of cell-bound, e.g., cell membrane-bound IL-11Rα and IL-11. Signal transduction mediated by the binding of the IL-11:IL-11Rα complex to gp130 can be analyzed by treating cells expressing gp130 with an IL-11:IL-11Rα complex, e.g., a recombinant fusion protein comprising IL-11 connected to the extracellular domain of IL-11Rα via a peptide linker (e.g., super IL-11 as described herein).

[0388] In some embodiments, the antigen-binding molecules of the present invention can inhibit signal transduction mediated by the binding of IL-11:IL-11Rα complex to gp130, and can also inhibit signal transduction mediated by the binding of IL-11 to IL-11Rα:gp130 receptor.

[0389] In some embodiments, the antigen binding molecules are capable of inhibiting fibroblast proliferation. Fibroblast proliferation can be measured by analyzing cell division over a period of time. This can be done, for example, by in vitro analysis. 3 Cell division of a given fibroblast population can be analyzed by incorporation of H-thymidine or by a CFSE dilution assay (e.g., as described in Fulcher and Wong, Immunol Cell Biol (1999) 77(6):559-564, incorporated herein by reference in its entirety). Proliferating cells (e.g., proliferating fibroblasts) can also be identified by analyzing the incorporation of 5-ethynyl-2'-deoxyuridine (EdU) by a suitable assay, such as that described in Buck et al., Biotechniques. 2008 Jun;44(7):927-9, and Sali and Mitchison, PNAS USA 2008 Feb 19;105(7):2415–2420, both of which are incorporated herein by reference in their entirety.

[0390] The fibroblasts of the present disclosure can be derived from any tissue, including liver, lung, kidney, heart, blood vessels, eyes, skin, pancreas, spleen, intestine (e.g., large or small intestine), brain and bone marrow. In a specific embodiment, for analyzing antigen binding molecules, the fibroblasts can be cardiac fibroblasts (e.g., atrial fibroblasts), skin fibroblasts, lung fibroblasts, kidney fibroblasts or liver fibroblasts.

[0391] In some embodiments, the antigen binding molecules of the invention are capable of inhibiting fibroblast proliferation to less than 100% of the level of fibroblast proliferation in the absence of the antigen binding molecule (or in the presence of an appropriate control antigen binding molecule), such as one of 99% or less, 95% or less, 90% or less, 85% or less, 80% or less, 75% or less, 70% or less, 65% or less, 60% or less, 55% or less, 50% or less, 45% or less, 40% or less, 35% or less, 30% or less, 25% or less, 20% or less, 15% or less, 10% or less, 5% or less, or 1% or less. In some embodiments, the antigen binding molecule is capable of reducing fibroblast proliferation to less than 1-fold the level of fibroblast proliferation in the absence of the antigen binding molecule (or in the presence of an appropriate control antigen binding molecule), such as one of the following: ≤0.99-fold, ≤0.95-fold, ≤0.9-fold, ≤0.85-fold, ≤0.8-fold, ≤0.75-fold, ≤0.7-fold, ≤0.65-fold, ≤0.6-fold, ≤0.55-fold, ≤0.5-fold, ≤0.45-fold, ≤0.4-fold, ≤0.35-fold, ≤0.3-fold, ≤0.25-fold, ≤0.2-fold, ≤0.15-fold, ≤0.1-fold.

[0392] In some embodiments, the antigen binding molecules of the present invention are capable of inhibiting pathological processes mediated by IL-11 / IL-11R signaling, for example, after stimulation with a fibrogenic factor (e.g., TGFβ1). Pathological processes mediated by IL-11 / IL-11R signaling, including fibrosis, can be assessed in vitro or in vivo.

[0393] In some embodiments, the antigen binding molecules of the present invention can inhibit fibrosis. In some embodiments, the antigen binding molecules of the present invention can reverse fibrosis or cause it to disappear. In some embodiments, fibrosis is existing or severe fibrosis. As used herein, inhibiting fibrosis refers to the ability of antigen binding molecules to reduce, inhibit or prevent the development of fibrosis. In some embodiments, the inhibition of fibrosis refers to, for example, a preventive effect that prevents the development of fibrosis. In some embodiments, the inhibition of fibrosis refers to, for example, a therapeutic effect that prevents the development of existing early or late fibrosis or develops to a more advanced stage. As used herein, the reversal or disappearance of fibrosis refers to the ability of antigen binding molecules to improve the fibrotic state from a more advanced state to a non-advanced state or to alleviate the severity of fibrosis itself or its symptoms. The reversal of fibrosis may be relevant to the improvement of the fibrotic state.

[0394] In the experimental examples herein, ACTA2 was measured by, for example, using the Operetta high content imaging system. +ve The inhibition, reversal or regression of fibrosis can be analyzed by measuring the number or percentage of cells, measuring the cellular or organ collagen content by assessing hydroxyproline content, measuring ERK activation / phosphorylation by Western blot, and / or measuring the expression levels of inflammatory markers such as TNFα and CCL2 or fibrosis markers such as TGFβ1, αSMA (ACTA2), TIMP1, COL1A1, COL1A2 or COL3A1 by quantitative PCR. In tissues such as the liver, the inhibition, reversal or regression of fibrosis can be analyzed by, for example, measuring triglyceride content and serum ALT levels.

[0395] Fibrosis can be fibrosis of a specific tissue or several tissues, such as the liver, lung, kidney, heart, blood vessels, eyes, skin, pancreas, spleen, intestine (e.g., large intestine or small intestine), brain, or bone marrow. Fibrosis can be measured by means well known to those skilled in the art, such as by analyzing the gene or protein expression of one or more myofibroblast markers and / or the gene or protein expression of one or more fibrosis markers in a given tissue or tissues.

[0396] Myofibroblast markers may include one or more increased αSMA, vimentin, palladin, cofilin, or desmin. Markers of fibrosis include collagen, fibronectin, periostin, IL-6, IL-11, αSMA, TIMP1, and MMP2, as well as elevated levels of extracellular matrix components, myofibroblast number / ratio, and organ weight.

[0397] Fibrotic inhibition / reversal / disappearance can be measured in vitro or in vivo.For example, it is possible to, by processing the fibroblast derived from this tissue with a fibrinogen stimulant, then analyze whether the antibody can reduce or reverse the myofibroblasts (or such as other fibrosis markers) produced by the fibroblasts to come to in vitro analysis whether the antigen binding molecules can suppress / reverse / disappear the fibrosis in a given tissue. Whether the antigen binding molecules can suppress / reverse / disappear fibrosis can be analyzed in vivo, such as by applying the antigen binding molecules to a subject (such as a subject exposed to a fibrosis stimulant) and analyzing one or more fibrosis markers of tissue.

[0398] In some embodiments, the antigen binding molecules of the invention are capable of inhibiting / reversing / regressing fibrosis to less than 100% of the level of fibrosis in the absence of the antigen binding molecule (or in the presence of an appropriate control antigen binding molecule), such as one of: 99% or less, 95% or less, 90% or less, 85% or less, 80% or less, 75% or less, 70% or less, 65% or less, 60% or less, 55% or less, 50% or less, 45% or less, 40% or less, 35% or less, 30% or less, 25% or less, 20% or less, 15% or less, 10% or less, 5% or less, or 1% or less. In some embodiments, the antigen binding molecule is capable of inhibiting / reversing / regressing fibrosis to less than 1-fold the level of fibrosis in the absence of the antigen binding molecule (or in the presence of an appropriate control antigen binding molecule), such as one of the following: ≤0.99-fold, ≤0.95-fold, ≤0.9-fold, ≤0.85-fold, ≤0.8-fold, ≤0.75-fold, ≤0.7-fold, ≤0.65-fold, ≤0.6-fold, ≤0.55-fold, ≤0.5-fold, ≤0.45-fold, ≤0.4-fold, ≤0.35-fold, ≤0.3-fold, ≤0.25-fold, ≤0.2-fold, ≤0.15-fold, ≤0.1-fold.

[0399] In some embodiments, the antigen binding molecules of the present invention can inhibit the production of myofibroblasts from fibroblasts or astrocytes (e.g., liver or pancreatic astrocytes), such as after fibroblasts or astrocytes are exposed to fibrotic factors. The myofibroblasts produced by fibroblasts or astrocytes can be studied by analyzing myofibroblast markers. The pro-fibrotic factors disclosed herein can be, for example, TGFβ1, IL-11, IL-13, PDGF, ET-1, oncostatin M (OSM) or ANG2 (AngII). In some embodiments, the antigen binding molecules of the present invention can inhibit the activation of fibrotic factors by fibrotic cells or astrocytes.

[0400] In some embodiments, the antigen binding molecules of the present invention can promote astrocyte senescence. Senescence can be measured by detecting the expression of senescence markers such as P16, P21, and P53.

[0401] In some embodiments, the antigen binding molecules can inhibit one or more of collagen, fibronectin, periostin, IL-6, IL-11, αSMA, TIMP1, MMP2, TNFα, CCL2 in fibroblasts, astrocytes or fibroblast / astrocyte-derived cells (e.g., myofibroblasts) gene or protein expression, for example after stimulation with a pro-fibrotic factor. In some embodiments, the antigen binding molecules can inhibit one or more extracellular matrix components in fibroblasts or fibroblast-derived cells (e.g., myofibroblasts) gene or protein expression, for example after stimulation with a pro-fibrotic factor.

[0402] In the experimental examples herein, the Operetta High Content Imaging System was used to measure αSMA protein expression levels to analyze myofibroblasts generated from fibroblasts or astrocytes after fibroblasts were stimulated with TGFβ1.

[0403] In some embodiments, the antigen binding molecules of the invention are capable of inhibiting the production of myofibroblasts from fibroblasts or astrocytes to less than 100% of the level of myofibroblast production by fibroblasts or astrocytes in the absence of the antigen binding molecule (or in the presence of an appropriate control antigen binding molecule), such as one of: 99% or less, 95% or less, 90% or less, 85% or less, 80% or less, 75% or less, 70% or less, 65% or less, 60% or less, 55% or less, 50% or less, 45% or less, 40% or less, 35% or less, 30% or less, 25% or less, 20% or less, 15% or less, 10% or less, 5% or less, or 1% or less. In some embodiments, the antigen binding molecule is capable of inhibiting the production of myofibroblasts by fibroblasts or astrocytes to less than 1-fold the level of myofibroblast production by fibroblasts or astrocytes in the absence of the antigen binding molecule (or in the presence of an appropriate control antigen binding molecule), such as one of the following: ≤0.99-fold, ≤0.95-fold, ≤0.9-fold, ≤0.85-fold, ≤0.8-fold, ≤0.75-fold, ≤0.7-fold, ≤0.65-fold, ≤0.6-fold, ≤0.55-fold, ≤0.5-fold, ≤0.45-fold, ≤0.4-fold, ≤0.35-fold, ≤0.3-fold, ≤0.25-fold, ≤0.2-fold, ≤0.15-fold, ≤0.1-fold.

[0404] In some embodiments, the antigen binding molecules of the present invention are capable of inhibiting gene or protein expression of one or more of collagen, fibronectin, periostin, IL-6, IL-11, αSMA, TIMP1, MMP2, TNFα, CCL2 in fibroblasts, astrocytes or myofibroblasts, for example, after stimulation with pro-fibrotic factors (e.g., TGFβ1). In some embodiments, the antigen binding molecules of the invention are capable of inhibiting gene or protein expression to less than 100% of the gene or protein expression level in the absence of the antigen binding molecule (or in the presence of an appropriate control antigen binding molecule), such as one of: 99% or less, 95% or less, 90% or less, 85% or less, 80% or less, 75% or less, 70% or less, 65% or less, 60% or less, 55% or less, 50% or less, 45% or less, 40% or less, 35% or less, 30% or less, 25% or less, 20% or less, 15% or less, 10% or less, 5% or less, or 1% or less. In some embodiments, the antigen binding molecule is capable of reducing gene or protein expression to less than 1-fold the level of gene or protein expression in the absence of the antigen binding molecule (or in the presence of an appropriate control antigen binding molecule), such as one of the following: ≤0.99-fold, ≤0.95-fold, ≤0.9-fold, ≤0.85-fold, ≤0.8-fold, ≤0.75-fold, ≤0.7-fold, ≤0.65-fold, ≤0.6-fold, ≤0.55-fold, ≤0.5-fold, ≤0.45-fold, ≤0.4-fold, ≤0.35-fold, ≤0.3-fold, ≤0.25-fold, ≤0.2-fold, ≤0.15-fold, ≤0.1-fold.

[0405] In some embodiments, the antigen binding molecules of the present invention can suppress the production of extracellular matrix by fibroblasts or astrocytes (such as astrocytes), such as after stimulating with a pro-fibrotic factor (such as TGF β 1). The generation of extracellular matrix can be by, such as measuring the level of extracellular matrix composition to assess. Extracellular matrix components of the present invention include, such as proteoglycans, heparan sulfate, chondroitin sulfate, keratan sulfate, hyaluronic acid, collagen, periostin, fibronectin, vitronectin, elastin, fibronectin, laminin, Nido root, gelatin and aggrecan. In some embodiments, the antigen binding molecules of the present invention can suppress fibroblasts, astrocytes and / or myofibroblasts to secrete collagen.

[0406] In some embodiments, the antigen binding molecules of the invention are capable of inhibiting the production of extracellular matrix by fibroblasts or astrocytes to less than 100% of the level of extracellular matrix produced by fibroblasts or astrocytes in the absence of the antigen binding molecule (or in the presence of an appropriate control antigen binding molecule), such as one of: 99% or less, 95% or less, 90% or less, 85% or less, 80% or less, 75% or less, 70% or less, 65% or less, 60% or less, 55% or less, 50% or less, 45% or less, 40% or less, 35% or less, 30% or less, 25% or less, 20% or less, 15% or less, 10% or less, 5% or less, or 1% or less. In some embodiments, the antigen binding molecule is capable of reducing the extracellular matrix produced by fibroblasts or astrocytes to less than 1-fold the level of extracellular matrix produced by fibroblasts or astrocytes in the absence of the antigen binding molecule (or in the presence of an appropriate control antigen binding molecule), such as one of the following: ≤0.99-fold, ≤0.95-fold, ≤0.9-fold, ≤0.85-fold, ≤0.8-fold, ≤0.75-fold, ≤0.7-fold, ≤0.65-fold, ≤0.6-fold, ≤0.55-fold, ≤0.5-fold, ≤0.45-fold, ≤0.4-fold, ≤0.35-fold, ≤0.3-fold, ≤0.25-fold, ≤0.2-fold, ≤0.15-fold, ≤0.1-fold.

[0407] In some embodiments, the antigen binding molecules of the present invention can upregulate the gene or protein expression of one or more genes / proteins involved in lipogenesis and / or β-oxidation in hepatocytes. Such genes / proteins can be, for example, ACOX1 (acyl-CoA oxidase 1), SCD1 (stearoyl-CoA desaturase 1), FASN (fatty acid synthase), or SREBF1 (sterol regulatory element binding protein 1). In some embodiments, the antigen binding molecules of the invention are capable of upregulating gene or protein expression to a level greater than 1-fold, such as ≥1.01-fold, ≥1.05-fold, ≥1.1-fold, ≥1.15-fold, ≥1.2-fold, ≥1.25-fold, ≥1.3-fold, ≥1.35-fold, ≥1.4-fold, ≥1.45-fold, ≥1.5-fold, ≥1.55-fold, ≥1.6-fold, ≥1.65-fold, ≥1.7-fold, ≥1.75-fold, ≥1.8-fold, ≥1.85-fold, ≥1.9-fold, ≥1.95-fold, ≥2-fold, ≥2.5-fold, ≥3-fold, ≥3.5-fold, ≥4-fold, ≥4.5-fold, ≥5-fold, ≥6-fold, ≥7-fold, ≥8-fold, ≥9-fold, or ≥10-fold, relative to the level of gene or protein expressed in the absence of the antigen binding molecule (or in the presence of an appropriate control antigen binding molecule).

[0408] In some embodiments, the antigen binding molecules of the present invention can inhibit the migration and / or invasion behavior of fibroblasts, astrocytes or myofibroblasts, i.e., inhibit migration and / or invasion. The migration and invasion of such cells may be crucial in the pathology of fibrosis. The migration of cells can be assessed using, for example, polycarbonate membranes and invasive stimuli such as TGFβ1 or CCL2. The invasion of cells can be detected using, for example, a Boyden chamber invasion assay or an ECM-coated matrigel. In some embodiments, the antigen binding molecules of the invention are capable of inhibiting the migration and / or invasion of fibroblasts, astrocytes or myofibroblasts to less than 100% of the level of migration and / or invasion in the absence of the antigen binding molecule (or in the presence of an appropriate control antigen binding molecule), such as one of 99% or less, 95% or less, 90% or less, 85% or less, 80% or less, 75% or less, 70% or less, 65% or less, 60% or less, 55% or less, 50% or less, 45% or less, 40% or less, 35% or less, 30% or less, 25% or less, 20% or less, 15% or less, 10% or less, 5% or less, or 1% or less. In some embodiments, the antigen binding molecule is capable of reducing migration and / or invasion to less than 1-fold the level of migration and / or invasion in the absence of the antigen binding molecule (or in the presence of an appropriate control antigen binding molecule), such as one of the following: ≤0.99-fold, ≤0.95-fold, ≤0.9-fold, ≤0.85-fold, ≤0.8-fold, ≤0.75-fold, ≤0.7-fold, ≤0.65-fold, ≤0.6-fold, ≤0.55-fold, ≤0.5-fold, ≤0.45-fold, ≤0.4-fold, ≤0.35-fold, ≤0.3-fold, ≤0.25-fold, ≤0.2-fold, ≤0.15-fold, ≤0.1-fold.

[0409] In some embodiments, the antigen binding molecules of the present invention can suppress the presence of immune cells in an organ. In some embodiments, the antigen binding molecules of the present invention can reduce the number of immune cells in an organ. The organ can be an organ susceptible to or suffering from fibrosis, such as the liver or kidney. The immune cell can be a CD45 + cells, such as CD 3+ CD 4+ T cells, CD 3+ CD 8+T cells, B lymphocytes, granulocytes and monocytes. Immune cells can express the mouse monocyte marker LyC6. In some embodiments, the antigen binding molecules of the present invention can reduce the number of immune cells in an organ to less than 100% of the number of immune cells in the absence of the antigen binding molecules (or in the presence of an appropriate control antigen binding molecule), such as one of the following: 99% or less, 95% or less, 90% or less, 85% or less, 80% or less, 75% or less, 70% or less, 65% or less, 60% or less, 55% or less, 50% or less, 45% or less, 40% or less, 35% or less, 30% or less, 25% or less, 20% or less, 15% or less, 10% or less, 5% or less, or 1% or less. In some embodiments, the antigen binding molecule is capable of reducing the number of immune cells in an organ to less than 1-fold the number of immune cells in the organ in the absence of the antigen binding molecule (or in the presence of an appropriate control antigen binding molecule), such as one of the following: ≤0.99-fold, ≤0.95-fold, ≤0.9-fold, ≤0.85-fold, ≤0.8-fold, ≤0.75-fold, ≤0.7-fold, ≤0.65-fold, ≤0.6-fold, ≤0.55-fold, ≤0.5-fold, ≤0.45-fold, ≤0.4-fold, ≤0.35-fold, ≤0.3-fold, ≤0.25-fold, ≤0.2-fold, ≤0.15-fold, ≤0.1-fold.

[0410] In some embodiments, the antigen binding molecules of the present invention can inhibit the proliferation and / or survival of cancer cells. A skilled person can determine whether the release of an antigen binding molecule can inhibit the proliferation and / or survival of cancer cells, for example, by analyzing the effect of the antigen binding molecule on cancer cells. For example, cell proliferation can be detected as described herein, for example, by 3 H Thymus incorporation or CFSE dilution assay. Cell survival can be analyzed by measuring cell viability / cell death markers in cells after treatment with the antigen-binding molecule.

[0411] In some embodiments, the antigen binding molecules of the invention are capable of inhibiting the proliferation and / or survival of cancer cells to less than 100% of the level of proliferation and / or survival of cancer cells in the absence of the antigen binding molecule (or in the presence of an appropriate control antigen binding molecule), such as one of: 99% or less, 95% or less, 90% or less, 85% or less, 80% or less, 75% or less, 70% or less, 65% or less, 60% or less, 55% or less, 50% or less, 45% or less, 40% or less, 35% or less, 30% or less, 25% or less, 20% or less, 15% or less, 10% or less, 5% or less, or 1% or less. In some embodiments, the antigen binding molecule is capable of reducing the proliferation and / or survival of cancer cells to less than 1-fold the level of proliferation and / or survival of cancer cells in the absence of the antigen binding molecule (or in the presence of an appropriate control antigen binding molecule), for example, one of the following: ≤0.99-fold, ≤0.95-fold, ≤0.9-fold, ≤0.85-fold, ≤0.8-fold, ≤0.75-fold, ≤0.7-fold, ≤0.65-fold, ≤0.6-fold, ≤0.55-fold, ≤0.5-fold, ≤0.45-fold, ≤0.4-fold, ≤0.35-fold, ≤0.3-fold, ≤0.25-fold, ≤0.2-fold, ≤0.15-fold, ≤0.1-fold.

[0412] In some embodiments, the antigen binding molecules of the present invention suppress the development and / or progression of cancer in vivo. In some embodiments, the antigen binding molecules cause the killing of cancer cells, for example, by effector immune cells. In some embodiments, the antigen binding molecules cause the minimizing of cancer cell number in vivo, for example, compared with appropriate control conditions. In some embodiments, the antigen binding molecules inhibit tumor growth, for example, by measuring the tumor size / volume that changes over time.

[0413] The ability of the antigen binding molecules of the present invention to inhibit cancer development and / or progression can be analyzed in a suitable in vivo model. The cancer can be a cancer of cells that pathologically involve IL-11-mediated signal transduction and / or expression / comprising IL-11R α or a cell comprising a complex of IL-11R α. Such cancers are included in Xu et al., Cancer Lett. (2016) 373 (2): 156-63 and Johnstone et al., Cytokine & Growth Reviews (2015) 26 (5): 489-498 descriptions, both of which are incorporated herein by reference in their entirety.

[0414] In some embodiments, the antigen binding molecules of the present invention are used and can cause one or more of the following: suppress the development / process of cancer, delay / prevent the onset of cancer, reduce / delay / prevent tumor growth, reduce / delay / prevent metastasis, reduce the severity of cancer symptoms, reduce cancer cell number, reduce tumor size / volume and / or increase survival (such as progression-free survival), such as measured in a suitable cancer in vivo model. In some embodiments, compared with being administered alone, when such as with chemotherapeutics, such as when used in conjunction, such as respectively, simultaneously or sequentially, the antigen binding molecules of the present invention provide additive effects. Additive effects can be any effect as described herein, such as reducing IL-11 signaling, suppressing the development / process of cancer and / or suppressing tumor growth.

[0415] In some embodiments, the antigen binding molecules of the invention are capable of inhibiting tumor growth to less than 100% of the level of tumor growth in the absence of the antigen binding molecule (or in the presence of an appropriate control antigen binding molecule), such as one of 99% or less, 95% or less, 90% or less, 85% or less, 80% or less, 75% or less, 70% or less, 65% or less, 60% or less, 55% or less, 50% or less, 45% or less, 40% or less, 35% or less, 30% or less, 25% or less, 20% or less, 15% or less, 10% or less, 5% or less, or 1% or less. In some embodiments, the antigen binding molecule is capable of reducing tumor growth to less than 1-fold the level of tumor growth in the absence of the antigen binding molecule (or in the presence of an appropriate control antigen binding molecule), such as one of the following: ≤0.99-fold, ≤0.95-fold, ≤0.9-fold, ≤0.85-fold, ≤0.8-fold, ≤0.75-fold, ≤0.7-fold, ≤0.65-fold, ≤0.6-fold, ≤0.55-fold, ≤0.5-fold, ≤0.45-fold, ≤0.4-fold, ≤0.35-fold, ≤0.3-fold, ≤0.25-fold, ≤0.2-fold, ≤0.15-fold, ≤0.1-fold.

[0416] In some embodiments, the antigen binding molecules of the present invention can kill cells expressing / comprising IL-11R alpha or comprising a complex of IL-11R alpha. The killing of cells expressing / comprising IL-11R alpha or comprising a complex of IL-11R alpha can be increased by the effector functions of the antigen binding molecules. In some embodiments, wherein the antigen binding molecules include an Fc region, the antigen binding molecules can kill cells expressing / comprising IL-11R alpha or comprising a complex of IL-11R alpha by one or more of complement dependent cytotoxicity (CDC), antibody dependent cell-mediated cytotoxicity (ADCC) and antibody dependent cellular phagocytosis (ADCP).

[0417] Can be in suitable test, by in the presence of antigen binding molecules, or after the cell of expressing / comprising IL-11R α or the complex comprising IL-11R α is incubated with antigen binding molecules, with the cell killing level detected in the absence of antigen binding molecules (or in the presence of suitable control antigen binding molecules) compared, observe the killing level of the cell of expressing / comprising IL-11R α or the complex comprising IL-11R α to identify the antigen binding molecules that can kill the cell of expressing / comprising IL-11R α or the complex comprising IL-11R α.CDC, ADCC and ADCP tests are well known to technicians.Can also by after exposure to different treatment conditions, detect the number / ratio of survival and / or non-survival cells of expressing / comprising IL-11R α or the complex comprising IL-11R α to measure the killing level of the cell of expressing / comprising IL-11R α or the complex comprising IL-11R α.

[0418] In some embodiments, the antigen binding molecules of the present invention are capable of increasing the killing of cells expressing / containing IL-11Rα or a complex containing IL-11Rα to a level greater than 1-fold, such as one of the following: ≥1.01-fold, ≥1.05-fold, ≥1.1-fold, ≥1.15-fold, ≥1.2-fold, ≥1.25-fold, ≥1.01-fold, ≥1.05-fold, ≥1.15-fold, ≥1.2 ...25-fold, ≥1.01-fold, ≥1.05-fold, ≥1.15-fold, ≥1.15-fold, ≥1.25-fold, ≥1.25-fold times, ≥1.3 times, ≥1.35 times, ≥1.4 times, ≥1.45 times, ≥1.5 times, ≥1.55 times, ≥1.6 times, ≥1.65 times, ≥1.7 times, ≥1.75 times, ≥1.8 times, ≥1.85 times, ≥1.9 times, ≥1.95 times, ≥2 times, ≥2.5 times, ≥3 times, ≥3.5 times, ≥4 times, ≥4.5 times, ≥5 times, ≥6 times, ≥7 times, ≥8 times, ≥9 times or ≥10 times.

[0419] In some embodiments, in comparable experiments, the antigen binding molecules of the present invention are capable of reducing the number of cells expressing / comprising IL-11Rα or a complex comprising IL-11Rα to less than 1-fold, such as one of the following: ≤0.99-fold, ≤0.95-fold, ≤0.9-fold, ≤0.85-fold, ≤0.8-fold, ≤0.75-fold, ≤0.7-fold, ≤0.65-fold, ≤0.6-fold, ≤0.55-fold, ≤0.5-fold, ≤0.45-fold, ≤0.4-fold, ≤0.35-fold, ≤0.3-fold, ≤0.25-fold, ≤0.2-fold, ≤0.15-fold, ≤0.1-fold, relative to the number of cells expressing / comprising IL-11Rα or a complex comprising IL-11Rα after incubation in the absence of the antigen binding molecules (or after incubation in the presence of an appropriate control antigen binding molecule). Cell number and proportion can be determined, for example, by flow cytometry analysis using antibodies capable of detecting cell types.

[0420] In some embodiments, the antigen binding molecules of the present invention have one or more similar or improved properties compared to a reference antibody / antigen-binding fragment thereof that is capable of binding to IL-11Rα.

[0421] In some embodiments, the antigen-binding molecules of the invention exhibit one or more of the following properties compared to a reference antibody / antigen-binding fragment thereof that binds to IL-11Rα:

[0422] (i) binding to IL-11Rα with similar or greater specificity (i.e., similar or reduced cross-reactivity with proteins of the IL-6 cytokine receptor family other than IL-11Rα);

[0423] (ii) binds to IL-11Rα (e.g., human IL-11Rα and / or mouse IL-11Rα) with similar or higher affinity (e.g., similar or lower EC50 as determined by ELISA; similar or lower K as determined by SPR analysis) D );

[0424] (iii) have similar or greater inhibitory effects on the interaction between IL-11Rα and IL-11Rα;

[0425] (iv) have similar or greater inhibitory effects on the interaction between IL-11Rα and gp130;

[0426] (v) have similar or greater inhibitory effects on the interaction between the IL-11Rα:gp130 receptor complex and IL-11;

[0427] (vi) have similar or greater inhibitory effects on the interaction between the IL-11:IL-11Rα complex and gp130;

[0428] (vii) similar or greater inhibitory effects on IL-11 / IL-11R signaling;

[0429] (viii) have similar or greater inhibitory effect on IL-11-mediated signaling (e.g., similar or lower IC50 as determined by ELISA in an appropriate assay);

[0430] (ix) have similar or greater inhibitory effects on signaling mediated by IL-11 binding to the IL-11Rα:gp130 receptor complex;

[0431] (x) have similar or greater inhibitory effects on signaling mediated by the binding of the IL-11:IL-11Rα complex to gp130 (i.e., IL-11 trans-signaling);

[0432] (xi) have similar or greater inhibitory effects on fibroblast proliferation;

[0433] (xii) have similar or greater inhibitory effects on the production of myofibroblasts from fibroblasts;

[0434] (xiii) similar or greater inhibitory reversal / regression effects on myofibroblast production from fibroblasts;

[0435] (xiv) have a similar or greater inhibitory effect on the production of myofibroblasts from astrocytes, such as liver or pancreatic astrocytes;

[0436] (xv) having a similar or greater reversal / regression effect on myofibroblast production from astrocytes, such as liver or pancreatic astrocytes;

[0437] (xvi) having a similar or greater inhibitory effect on the migration and / or invasion behavior of fibroblasts, astrocytes, or myofibroblasts (i.e., inhibiting migration or invasion);

[0438] (xvii) have a similar or greater inhibitory effect on the presence of immune cells in organs;

[0439] (xviii) have similar or greater inhibitory effects on pathological processes mediated by IL-11 / IL-11R signaling;

[0440] (xix) have similar or greater inhibitory effects on fibrosis;

[0441] (xx) have similar or greater reversal / regression effects on fibrosis;

[0442] (xxi) having a similar or greater inhibitory effect on gene or protein expression of one or more of collagen, fibronectin, periostin, IL-6, IL-11, αSMA (ACTA2), TIMP1, MMP2, TNFα, and CCL2 in fibroblasts, for example, after stimulation with profibrotic factors;

[0443] (xxii) have similar or greater inhibitory effects on extracellular matrix production by fibroblasts or astrocytes;

[0444] (xxiii) have similar or greater inhibitory effects on the proliferation and / or survival of cancer cells;

[0445] (xxvi) have similar or greater inhibitory effects on the development and / or progression of cancer in vivo;

[0446] (xxv) have similar or greater inhibitory effects on tumors;

[0447] (xxvi) have similar or greater killing effects on cells expressing / containing IL-11Rα or complexes containing IL-11Rα.

[0448] In some embodiments, "higher specificity" or "higher affinity" or "greater inhibition" or "greater killing" respectively means that the level of specificity, affinity, inhibition or killing is greater than 1 times the level exhibited by a reference antibody / antigen-binding fragment thereof that can bind to IL-11Rα as measured in an appropriate test, for example, ≥1.01 times, ≥1.02 times, ≥1.03 times, ≥1.04 times, ≥1.05 times, ≥1.06 times, ≥1.07 times, ≥1.08 times, ≥1.09 times, ≥1.1 times, ≥1.2 times, ≥1.3 times, ≥1.4 times, ≥1.5 times, ≥1.6 times, ≥1.7 times, ≥1.8 times, ≥1.9 times, ≥1.10 times, ≥1.11 times, ≥1.12 times, ≥1.13 times, ≥1.14 times, ≥1.15 times, ≥1.16 times, ≥1.17 times, ≥1.18 times, ≥1.19 times, ≥20 times, ≥21 times, ≥22 times, ≥23 times, ≥24 times, ≥25 times, ≥26 times, ≥27 times, ≥28 times, ≥29 times, ≥30 times, ≥31 times, ≥32 times, ≥33 times, ≥34 times, ≥35 times, ≥36 times, ≥37 times, ≥38 times, ≥39 times, ≥40 times, ≥41 times, ≥42 times, ≥43 times, ≥44 times, ≥45 times, ≥46 times, ≥47 times, ≥48 times, ≥49 times, ≥50 times, ≥51 times, ≥52 times, ≥53 times, ≥54 times 1.9 times, ≥2 times, ≥2.1 times, ≥2.2 times, ≥2.3 times, ≥2.4 times, ≥2.5 times, ≥2.6 times, ≥2.7 times, ≥2.8 times, ≥2.9 times, ≥3 times, ≥3.5 times, ≥4 times, ≥4.5 times, ≥5 times, ≥6 times, ≥7 times, ≥8 times, ≥9 times, ≥10 times, ≥15 times, ≥20 times, ≥25 times, ≥30 times, ≥35 times, ≥40 times, ≥45 times, ≥50 times, ≥60 times, ≥70 times, ≥80 times, ≥90 times, ≥100 times, ≥200 times, ≥300 times, ≥400 times, ≥500 times, ≥600 times, ≥700 times, ≥800 times, ≥900 times, ≥1000.

[0449] In some embodiments, "similar specificity" or "similar affinity" or "similar inhibition" or "similar killing" respectively means that the level of specificity, affinity, inhibition or killing is ≥0.2 times and ≤5 times, such as ≥0.3 times and ≤4 times, ≥0.4 times and ≤3 times, ≥0.5 times and ≤2 times, ≥0.6 times and ≤1.75 times, ≥0.7 times and ≤1.5 times, ≥0.8 times and ≤1.6 times, ≥0.9 times and ≤2.75 times, ≥0.9 times and ≤1.75 times, ≥0.8 times and ≤2.75 times, ≥0.9 times and ≤1.6 times, ≥0.9 times and ≤2.75 times, ≥0.9 times and ≤1.6 times, ≥0.9 times and ≤2.75 times, ≥0.9 times and ≤1.6 times, ≥0.9 times and ≤1 ... 0.75 times and ≤1.25 times, ≥0.8 times and ≤1.2 times, ≥0.85 times and ≤1.15 times, ≥0.9 times and ≤1.1 times, ≥0.91 times and ≤1.09 times, ≥0.92 times and ≤1.08 times, ≥0.93 times and ≤1.07 times, ≥0.94 times and ≤1.06 times, ≥0.95 times and ≤1.05 times, ≥0.96 times and ≤1.04 times, ≥0.97 times and ≤1.03 times, ≥0.98 times and ≤1.02 times, or ≥0.99 times and ≤1.01 times.

[0450] In some embodiments, the reference antibody capable of binding to IL-11Rα / an antigen-binding fragment thereof may comprise the CDRs of the anti-IL-11Rα antibody clone BSO-9A7. In some embodiments, the reference antibody capable of binding to IL-11Rα / an antigen-binding fragment thereof may comprise or consist of the VH and VL sequences of the anti-IL-11Rα antibody clone BSO-9A7.

[0451] In some embodiments, the reference antibody / antigen-binding fragment thereof capable of binding to IL-11Rα may comprise VH and VL, wherein the VH comprises or consists of the amino acid sequence of SEQ ID NO: 7, and the VL comprises or consists of the amino acid sequence of SEQ ID NO: 59.

[0452] Chimeric antigen receptors (CARs)

[0453] The present invention also provides a chimeric antigen receptor (CAR) comprising the antigen-binding molecule or polypeptide of the present invention.

[0454] CAR is a recombinant receptor that provides antigen binding and T cell activation functions. For example, the structure and engineering of CAR are reviewed in Dotti et al., Immunol Rev (2014) 257 (1), which is incorporated herein by reference in its entirety. CAR comprises an antigen binding region connected to a cell membrane anchoring region and a signaling region. An optional hinge region can provide separation between the antigen binding region and the cell membrane anchoring region and can act as a flexible joint.

[0455] The CAR of the present invention comprises an antigen-binding region, which comprises or consists of an antigen-binding molecule of the present invention, or comprises or consists of a polypeptide of the present invention.

[0456] The cell membrane anchoring region is located between the antigen binding region and the signal transduction region of the CAR, and is used to anchor the CAR to the cell membrane of the cell expressing the CAR, and its antigen binding region is located in the extracellular space, and the signal transduction region in the cell. In some embodiments, the CAR comprises a cell membrane anchoring region, and the cell membrane anchoring region comprises or consists of the following amino acid sequence: a transmembrane region amino acid sequence comprising one of CD3-ζ, CD4, CD8 or CD28, or consisting of or derived therefrom. As used herein, a region “derived from” a reference amino acid sequence includes an amino acid sequence having at least 60%, for example, at least 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with the reference sequence.

[0457] The signaling region of CAR can activate T cells. The signaling region of CAR can include an amino acid sequence of the intracellular domain of CD3-ζ, which provides an immunoreceptor tyrosine-based activation motif (ITAM) for phosphorylation and activation of T cells expressing CAR. Signaling regions containing sequences of other ITAM-containing proteins (e.g., FcγRI) have also been used in CAR (Haynes et al., 2001 J Immunol 166(1):182-187). The signaling region of CAR can also include a costimulatory sequence derived from the signaling region of a costimulatory molecule to promote the activation of T cells expressing CAR after binding to the target protein. Suitable costimulatory molecules include CD28, OX40, 4-1BB, ICOS, and CD27. In some cases, CAR is designed to provide co-stimulation of different intracellular signaling pathways. For example, signaling associated with CD28 costimulation preferentially activates the phosphatidylinositol 3-kinase (P13K) pathway, while 4-1BB-mediated signaling is through the TNF receptor-associated factor (TRAF) adapter protein. Therefore, the signaling region of CAR sometimes includes a costimulatory sequence derived from the signaling region of more than one costimulatory molecule. In some embodiments, the CAR of the present invention includes one or more costimulatory sequences comprising an amino acid sequence comprising one or more intracellular domains of CD28, OX40, 4-1BB, ICOS and CD27 or consisting of or derived from the amino acid sequence thereof, or consisting of.

[0458] The optional hinge region can provide separation between the antigen binding domain and the transmembrane domain and can act as a flexible joint. The hinge region can be derived from IgG1. In some embodiments, the CAR of the present invention comprises a hinge region comprising or consisting of the following amino acid sequence, the amino acid sequence comprising the amino acid sequence of the hinge region of IgG1 or consisting of or derived therefrom.

[0459] Also provided are cells comprising the CAR of the present invention. The CAR of the present invention can be used to generate immune cells expressing CAR, such as CAR-T or CAR-NK cells. CAR can be transformed into immune cells during in vitro culture.

[0460] The antigen binding region of the CAR of the present invention can be provided in any suitable form, such as scFv, scFab, etc.

[0461] Nucleic acids and vectors

[0462] The present invention provides one or more nucleic acids encoding the antigen binding molecules, polypeptides or CARs of the present invention.

[0463] In some embodiments, the nucleic acid is purified or isolated, e.g., from other nucleic acids or naturally occurring biological material. In some embodiments, the one or more nucleic acids comprise or consist of DNA and / or RNA.

[0464] The invention also provides one or more vectors comprising one or more nucleic acids of the invention.

[0465] The nucleotide sequence can be contained in a vector, such as an expression vector. As used herein, a "vector" is a nucleic acid molecule used as a vehicle for transferring exogenous nucleic acid into a cell. The vector can be a vector for expressing nucleic acid in a cell. Such a vector can include a promoter sequence operably linked to the nucleotide sequence encoding the sequence to be expressed. The vector can also include a stop codon and an expression enhancer. Any suitable vector, promoter, enhancer, and stop codon known in the art can be used to express a peptide or polypeptide from the vector of the present invention.

[0466] The term "operably linked" may include situations where a selected nucleotide sequence and a regulatory nucleotide sequence (e.g., a promoter and / or enhancer) are covalently linked (forming an expression cassette) in a manner that places the expression of the nucleotide sequence under the influence or control of the regulatory sequence. Thus, if the regulatory sequence is capable of influencing the transcription of the nucleic acid sequence, the regulatory sequence is operably linked to the selected nucleic acid sequence. The resulting one or more transcripts can then be translated into the desired one or more peptides / polypeptides.

[0467] Suitable vectors include plasmids, binary vectors, DNA vectors, mRNA vectors, viral vectors (e.g., gammaretroviral vectors (e.g., murine leukemia virus (MLV)-derived vectors), lentiviral vectors, adenoviral vectors, adeno-associated viral vectors, vaccinia viral vectors, and herpes viral vectors), transposon-based vectors, and artificial chromosomes (e.g., yeast artificial chromosomes).

[0468] In some embodiments, the vector can be a eukaryotic vector, for example, comprising elements necessary for expressing a protein from the vector in a eukaryotic cell. In some embodiments, the vector can be a mammalian vector, for example, comprising a cytomegalovirus (CMV) or SV40 promoter to drive protein expression.

[0469] The constituent polypeptides of the antigen-binding molecules of the present invention may be encoded by different nucleic acids among multiple nucleic acids, or by different vectors among multiple vectors.

[0470] Cells containing / expressing antigen-binding molecules and polypeptides

[0471] The present invention also provides cells comprising or expressing the antigen binding molecules, polypeptides or CARs of the present invention. A cell is also provided, comprising or expressing one or more nucleic acids, one or more vectors of the present invention.

[0472] The cell can be a eukaryotic cell, such as a mammalian cell. The mammal can be a primate (rhesus monkey, cynomolgus monkey, non-human primate or human) or a non-human mammal (e.g., rabbit, guinea pig, rat, mouse or other rodent (including any rodent), cat, dog, pig, sheep, goat, cattle (including cows, such as dairy cows, or any animal of the order Bos), horse (including any animal of the order Equine), donkey and non-human primate).

[0473] The present invention also provides a method for producing a cell comprising one or more nucleic acids of the present invention or a vector, comprising introducing a nucleic acid of the present invention or multiple nucleic acids, a vector or multiple vectors into a cell. In some embodiments, introducing one or more isolated nucleic acids of the present invention or a vector into a cell comprises transformation, transfection, electroporation or transduction (e.g., retroviral transduction).

[0474] The present invention also provides a method for producing cells expressing / comprising the antigen binding molecules, polypeptides or CARs of the present invention, comprising introducing a nucleic acid or multiple nucleic acids, a vector or multiple vectors of the present invention into cells. In some embodiments, the method further comprises culturing cells under conditions suitable for cell expression of nucleic acids or vectors. In some embodiments, the method is performed in vitro.

[0475] The present invention also provides cells obtained or obtainable by the methods of the present invention.

[0476] Production of antigen-binding molecules and peptides

[0477] The antigen-binding molecules and polypeptides of the present invention can be prepared according to polypeptide production methods known to those skilled in the art.

[0478] Polypeptides can be prepared by chemical synthesis, such as liquid phase or solid phase synthesis. For example, peptides / polypeptides can be synthesized using methods such as those described in Chandrudu et al., Molecules (2013), 18: 4373-4388, which is incorporated herein by reference in its entirety.

[0479] Alternatively, antigen binding molecules and polypeptides can be produced by recombinant expression. Molecular biology techniques suitable for recombinant production of polypeptides are well known in the art, such as those described by Green and Sambrook in Molecular Cloning: A Laboratory Manual (4th Edition), Cold Spring Harbor Press, 2012, and in Nat Methods. (2008); 5 (2): 135-146, both of which are incorporated herein by reference in their entirety. Methods for recombinant production of antigen binding molecules are also described in Frenzel et al., Front Immunol. (2013); 4: 217 and Kunert and Reinhart, Appl Microbiol Biotechnol. (2016) 100: 3451–3461, both of which are incorporated herein by reference in their entirety.

[0480] In some cases, the antigen binding molecules of the present invention are composed of more than one polypeptide chain. In this case, the production of the antigen binding molecules may include the transcription and translation of more than one polypeptide, and the subsequent association of polypeptide chains to form the antigen binding molecules.

[0481] For the recombinant production of the present invention, any cell suitable for expressing the polypeptide can be used. The cell can be a prokaryotic organism or a eukaryotic organism. In some embodiments, the cell is a prokaryotic cell, such as a cell of an archaebacterium or a bacterium. In some embodiments, the bacterium can be a gram-negative bacterium, such as a bacterium of the family Enterobacteriaceae, such as Escherichia coli. In some embodiments, the cell is a eukaryotic cell, such as a yeast cell, a plant cell, an insect cell or a mammalian cell, such as a CHO, HEK (e.g., HEK293), HeLa or COS cell. In some embodiments, the cell is a CHO cell transiently or stably expressing the polypeptide.

[0482] In some cases, the cells are not prokaryotes, as some prokaryotes do not allow the same folding or post-translational modifications as eukaryotic cells. Alternatively, very high expression levels are possible in eukaryotes, and proteins can be more easily purified from eukaryotes using appropriate tags. Specific plasmids can also be used to enhance secretion of proteins into the culture medium.

[0483] In some embodiments, the polypeptide can be produced by cell-free protein synthesis (CFPS), for example using the system described in Zemella et al., Chembiochem (2015) 16(17): 2420-2431, which is incorporated herein by reference in its entirety.

[0484] Production can involve the cultivation or fermentation of eukaryotic cells modified to express one or more polypeptides of interest. The cultivation or fermentation can be carried out in a bioreactor equipped with appropriate nutrients, air / oxygen and / or growth factors. The secreted protein can be collected by separating the culture medium / fermentation broth from the cells, extracting the protein content and isolating the individual proteins to isolate the secreted polypeptide. Cultivation, fermentation and separation techniques are well known to those skilled in the art and are described in, for example, Green and Sambrook, Molecular Cloning: A Laboratory Manual (4th edition; incorporated herein by reference).

[0485] A bioreactor consists of one or more containers in which cells can be cultured. Cultivation in a bioreactor can be continuous, with reactants flowing continuously into the reactor and cultured cells continuously flowing out. Alternatively, culturing can be performed in batches. Bioreactors monitor and control environmental conditions, such as pH, oxygen, flow rates in and out of the container, and agitation within the container, to provide optimal conditions for the cultured cells.

[0486] After culturing cells expressing one or more antigen binding molecules / polypeptides, the one or more polypeptides of interest can be isolated. Any suitable method known in the art for isolating proteins from cells can be used. In order to isolate the polypeptide, it may be necessary to separate the cells from the nutrient medium. If the polypeptide is secreted from the cells, the cells can be separated from the culture medium containing the secreted polypeptide of interest by centrifugation. If the polypeptide of interest is aggregated within the cells, protein separation may include centrifugation to separate the cells from the cell culture medium, treating the cell pellet with a lysis buffer, and, for example, destroying the cells by ultrasonic treatment, rapid freeze-thaw, or osmotic lysis.

[0487] Then it may be necessary to separate one or more polypeptides of interest from the supernatant or culture medium, which may contain other proteins and non-protein components. A common method for separating protein components from supernatant or culture medium is precipitation. Proteins of different solubility can be precipitated under different concentrations of precipitation agents (e.g., ammonium sulfate). For example, water-soluble proteins are extracted under low concentrations of precipitation agents. Therefore, by adding the increased precipitation agents of different concentrations, proteins with different solubility can be distinguished. Subsequently, dialysis can be used to remove ammonium sulfate from the isolated protein.

[0488] Other methods for distinguishing different proteins are known in the art, such as ion exchange chromatography and size chromatography. These can be used as an alternative to precipitation, or can be performed after precipitation.

[0489] Once one or more polypeptides of interest are isolated from the culture, it may be desirable or necessary to concentrate the polypeptides. Many methods for concentrating proteins are known in the art, such as ultrafiltration or lyophilization.

[0490] Composition

[0491] The present invention also provides compositions comprising the antigen-binding molecules, polypeptides, CARs, nucleic acids, expression vectors, and cells described herein.

[0492] Antigen binding molecules as described herein, polypeptide, CAR, nucleic acid, expression vector and cell can be formulated into pharmaceutical compositions or medicines for clinical use, and can include pharmaceutically acceptable carriers, diluents, excipients or adjuvants. Compositions can be formulated for topical, parenteral, systemic, intracavitary, intravenous, intraarterial, intramuscular, intrathecal, intraocular, intraconjunctival, intratumoral, subcutaneous, intradermal, intrathecal, oral or transdermal routes of administration, including injection or infusion.

[0493] Suitable preparations can include antigen binding molecules in sterile or isotonic culture medium. Medicine and pharmaceutical composition can be formulated into fluid forms including gel. Fluid preparations can be formulated to be administered to a selected area of ​​a human or animal body by injection or infusion (e.g., via a catheter).

[0494] In some embodiments, the composition is formulated for injection or infusion, for example, into a blood vessel or a tumor.

[0495] According to the invention described herein, a method for producing a pharmaceutically useful composition is also provided, which production method may include one or more steps selected from the following: producing an antigen binding molecule, polypeptide, CAR, nucleic acid (or multiple thereof), expression vector (or multiple thereof) or cell as described herein; isolating the antigen binding molecule, polypeptide, CAR, nucleic acid (or multiple thereof), expression vector (or multiple thereof) or cell as described herein; and / or mixing the antigen binding molecule, polypeptide, CAR, nucleic acid (or multiple thereof), expression vector (or multiple thereof) or cell as described herein with a pharmaceutically acceptable carrier, adjuvant, excipient or diluent.

[0496] For example, another aspect of the invention described herein relates to a method for formulating or producing a medicament or pharmaceutical composition for treating a disease / disorder (e.g., cancer), the method comprising formulating a pharmaceutical composition or medicament by mixing the antigen binding molecules, polypeptides, CARs, nucleic acids (or multiples thereof), expression vectors (or multiples thereof) or cells described herein with a pharmaceutically acceptable carrier, adjuvant, excipient or diluent.

[0497] Therapeutic and preventive applications

[0498] The antigen binding molecules, polypeptides, CARs, nucleic acids, expression vectors, cells and compositions described herein can be used in methods of treatment and prevention.

[0499] The present invention provides antigen binding molecules as described herein, polypeptide, CAR, nucleic acid (or its multiple), expression vector (or its multiple), cell or composition, for medical treatment or prevention method. Also provided are antigen binding molecules as described herein, polypeptide, CAR, nucleic acid (or its multiple), expression vector (or its multiple), cell or composition in the preparation of a medicine for treating or preventing a disease or illness. Also provided is a method for treating or preventing a disease or illness, including administering to a subject an antigen binding molecule as described herein, polypeptide, CAR, nucleic acid (or its multiple), expression vector (or its multiple), cell or composition for treatment or prevention of an effective amount.

[0500] The method can effectively reduce the development or progression of the disease / condition, alleviate the symptoms of the disease / condition or alleviate the pathology of the disease / condition. The method can effectively prevent the progression of the disease / condition, such as preventing the disease / condition from worsening or slowing down its rate of development. In some embodiments, the method can lead to improvement of the disease / condition, such as alleviating the symptoms of the disease / condition or alleviating some other related factors of the severity / activity of the disease / condition. In some embodiments, the method can prevent the disease / condition from developing to a later stage (such as a chronic phase or metastasis). In some embodiments, the method can effectively reverse or eliminate the disease / condition, such as the pathological condition of the disease / condition can be reversed from a later stage of development to an earlier stage of development. In some embodiments, the method can effectively reverse or eliminate the symptoms of the disease / condition or some other correlations of the severity / activity of the disease / condition. In some embodiments, the method can effectively reverse / eliminate the disease / condition to a state similar to that observed in a subject without the disease / condition.

[0501] It will be understood that the articles of the invention may be used to treat / prevent any disease / disorder that would derive therapeutic or prophylactic benefit from a reduction in the level (e.g., inhibition or antagonism) of IL-11-mediated signaling, or a reduction in the number and / or activity of cells expressing IL-11Rα or a complex containing IL-11Rα.

[0502] For example, the disease / disorder can be a disease / disorder in which IL-11-mediated signaling is pathologically implicated, e.g., an increased level of IL-11-mediated signaling is positively correlated with the onset, development or progression of the disease / disorder, and / or the severity of one or more symptoms of the disease / disorder, or an increased level of IL-11-mediated signaling is a risk factor for the onset, development or progression of the disease / disorder.

[0503] For example, the disease / disorder can be a disease / disorder that pathologically involves cells expressing IL-11Rα, such as a disease / disorder in which an increased number / ratio of cells expressing IL-11Rα is positively correlated with the onset, development or progression of the disease / disorder and / or the severity of one or more symptoms of the disease / disorder, or an increased number / ratio of cells expressing IL-11Rα is a risk factor for the onset, development or progression of the disease / disorder.

[0504] In some embodiments, the disease / disorder to be treated / prevented according to the present invention is a disease / disorder characterized by increased levels of IL-11 mediated signaling or its correlates (e.g., in an organ / tissue where symptoms of the disease / condition are manifest) compared to the levels of IL-11 mediated signaling / correlates in the absence of the disease / disorder.

[0505] In some embodiments, the disease / disorder treated / prevented by the present invention is a disease / disorder characterized by an increase in the number / ratio / activity of cells expressing IL-11Rα compared to the number / ratio / activity of cells expressing IL-11Rα in the absence of the disease / disorder.

[0506] In some embodiments, subjects can be selected for the treatment / prevention methods described herein based on the detection of an increase in the level of IL-11-mediated signaling or its correlation and / or an increase in the number / ratio / activity of cells expressing IL-11Rα, such as in the periphery or in an organ / tissue affected by the disease / disorder (e.g., an organ / tissue in which symptoms of the disease / disorder are apparent). The disease / disorder can affect any tissue or organ or organ system. In some embodiments, the disease / disorder can affect several tissues / organs / organ systems.

[0507] In some embodiments, subjects can be selected for the treatment / prevention methods of the present invention based on the determination of increased levels of IL-11-mediated signaling / its correlations and / or increased number / ratio / activity of cells expressing IL-11Rα in, for example, the periphery or in an organ / tissue relative to the levels of IL-11-mediated signaling / its correlations, or the number / ratio / activity of cells expressing IL-11Rα in healthy subjects.

[0508] The antigen-binding molecules of the present invention are preferably capable of binding to and inhibiting the biological activity of IL-11Rα and molecules / complexes containing IL-11Rα (e.g., IL-11:IL-11Rα complexes). Therefore, the antigen-binding molecules of the present invention can be used to treat or prevent diseases and conditions in which IL-11Rα is associated with the pathology of the disease / condition. That is, it has been found that the antigen-binding molecules of the present invention can be used to treat or prevent diseases and conditions associated with IL-11-mediated signaling.

[0509] In some embodiments, the disease / disorder can be associated with increased expression of IL-11, IL-11Rα and / or gp130 genes or proteins, for example, compared to a control (i.e., non-diseased) state. In some embodiments, the disease / disorder can be associated with increased levels of IL-11-mediated signaling (e.g., IL-11 / IL-11R signaling) compared to a control state. In some embodiments, the disease / disorder can be associated with increased levels of signaling through the ERK and / or STAT3 pathways compared to a control state. In some embodiments, increased expression / activity of IL-11, IL-11Rα and / or gp130 and / or increased levels of IL-11-mediated signaling can be observed in effector cells of the disease / disorder. In some embodiments, increased expression / activity of IL-11, IL-11Rα and / or gp130 and / or increased levels of IL-11-mediated signaling can be observed in cells other than effector cells.

[0510] Signaling through ERK can be measured, for example, using an ERK phosphorylation assay, such as the assay described in Assay Guidance Manual: Phospho-ERK Assays, Kim E. Garbison, Beverly A. Heinz, Mary E. Lajiness, Jeffrey R. Weidner, and G. Sitta Sittampalam, Eli Lilly & Company, Sittampalam GS, Coussens NP, Nelson H, et al., eds. Bethesda (MD): Eli Lilly & Company and National Center for Advancing Translational Sciences; 2004. Signaling through STAT3 can be detected using, for example, an assay for the phosphorylation of STAT3, such as the Phospho-STAT3 (Tyr705) Cellular Assay Kit (Cisbio assay).

[0511] In some embodiments, the treatment is for a disease / disorder for which a reduction in IL-11-mediated signaling is therapeutic. In some embodiments, the treatment is for a disease / disorder associated with excessive ERK and / or STAT3 signaling. In some embodiments, the treatment is for a disease / disorder associated with excessive proliferation or overactivation of fibroblasts or an excess of myofibroblasts.

[0512] In some embodiments, treatment may be directed to preventing or treating a disease / disorder by reducing the number or proportion of myofibroblasts or αSMA-positive fibroblasts.

[0513] In some embodiments, the disease / disorder can be fibrosis, a fibrotic disorder or a disease / disorder characterized by fibrosis. "Fibrosis" as used herein refers to the excessive fibrous connective tissue formed due to extracellular matrix components, such as excessive deposition of collagen. The fibrous connective tissue is characterized by an extracellular matrix (ECM) with a high collagen content. Collagen can be provided in the form of strings or fibers, which can be irregularly arranged or aligned. The ECM of fibrous connective tissue can also include glycosaminoglycans.

[0514] As used herein, "excess fibrous connective tissue" refers to the amount of connective tissue at a given location (e.g., a given tissue or organ, or a portion of a given tissue or organ) that is greater than the amount of connective tissue present at that location in the absence of fibrosis, e.g., under normal, non-pathological conditions. "Excess deposition of extracellular matrix components" as used herein refers to deposition levels of one or more extracellular matrix components that are greater than the deposition levels in the absence of fibrosis, e.g., under normal, non-pathological conditions.

[0515] The cellular and molecular mechanisms of fibrosis are described in Wynn, J. Pathol. (2008) 214(2): 199-210, and Wynn and Ramalingam, Nature Medicine (2012) 18: 1028-1040, which are incorporated herein by reference in their entireties. The major cellular effectors of fibrosis are myofibroblasts that produce a collagen-rich extracellular matrix.

[0516] In response to tissue injury, damaged cells and leukocytes produce profibrotic factors such as TGFβ, IL-13, and PDGF, which activate fibroblasts into myofibroblasts that express αSMA and recruit myofibroblasts to the site of injury. Myofibroblasts produce large amounts of extracellular matrix and are important mediators of wound contracture and closure. However, under conditions of persistent infection or during chronic inflammation, myofibroblasts may become overactivated and recruited, resulting in overproduction of extracellular matrix components, leading to the formation of excessive fibrous connective tissue.

[0517] In some embodiments, fibrosis can be triggered by a pathological condition, such as a condition, infection, or disease state that results in the production of profibrotic factors (e.g., TGFβ1). In some embodiments, fibrosis can be caused by physical injury / irritation, chemical injury / irritation, or environmental injury / irritation. Physical injury / irritation may occur during surgery, such as iatrogenic causes. Chemical injury / irritation may include drug-induced fibrosis, such as after long-term use of drugs such as bleomycin, cyclophosphamide, amiodarone, procainamide, penicillin, gold, and nitrofurantoin (Daba et al., Saudi Med J 2004 June; 25(6):700-6). Environmental injury / irritation may include exposure to asbestos fibers or silica.

[0518] Fibrosis can occur in many tissues of the body. For example, fibrosis can occur in the lungs, liver (e.g., cirrhosis), kidneys, heart, blood vessels, eyes, skin, pancreas, spleen, intestines (e.g., large or small intestine), brain, and bone marrow. Fibrosis can also occur in multiple organs at once.

[0519] In embodiments herein, fibrosis may involve organs of the gastrointestinal system, such as the liver, small intestine, large intestine, or pancreas. In some embodiments, fibrosis may involve organs of the respiratory system, such as the lungs. In embodiments, fibrosis may involve organs of the cardiovascular system, such as the heart or blood vessels. In some embodiments, fibrosis may involve the skin. In some embodiments, fibrosis may involve organs of the nervous system, such as the brain. In some embodiments, fibrosis may involve organs of the urinary system, such as the kidneys. In some embodiments, fibrosis may involve organs of the musculoskeletal system, such as muscle tissue.

[0520] In some preferred embodiments, the fibrosis is cardiac or myocardial fibrosis, hepatic fibrosis, or renal fibrosis. In some embodiments, the cardiac or myocardial fibrosis is associated with dysfunction of the cardiac musculature or electrical properties or thickening of the walls or valves of the heart. In some embodiments, the fibrosis is atrial and / or ventricular fibrosis of the heart. Treating or preventing atrial fibrillation or ventricular fibrosis may help reduce the risk or onset of atrial fibrillation, ventricular fibrillation, or myocardial infarction.

[0521] In some preferred embodiments, liver fibrosis is associated with chronic liver disease or cirrhosis. In some preferred embodiments, kidney fibrosis is associated with chronic kidney disease.

[0522] According to the present invention, diseases / disorders characterized by fibrosis include, but are not limited to, respiratory diseases, such as pulmonary fibrosis, cystic fibrosis, idiopathic pulmonary fibrosis (IPF), progressive massive fibrosis, scleroderma, bronchiolitis obliterans, Hermansky-Pudlak syndrome, asbestosis, silicosis, chronic pulmonary hypertension, AIDS-related pulmonary hypertension, sarcoidosis, lung tumor matrix disease, and asthma; chronic liver disease, primary biliary cirrhosis (PBC), schistosomiasis, cirrhosis, steatohepatitis, nonalcoholic steatohepatitis (NASH), early NASH, late NASH, alcoholic steatohepatitis, steatosis; nonalcoholic fatty liver disease (NAFLD) Cardiovascular diseases, such as hypertrophic cardiomyopathy, dilated cardiomyopathy (DCM), atrial fibrosis, atrial fibrillation, ventricular fibrosis, ventricular fibrillation, myocardial fibrosis, Brugada syndrome, myocarditis, endomyocardial fibrosis, myocardial infarction, fibrosing vasculopathy, hypertensive heart disease, arrhythmogenic right ventricular cardiomyopathy (ARVC), tubulointerstitial and glomerular fibrosis, atherosclerosis, varicose veins, cerebral infarction; nervous system diseases, such as gliosis and Alzheimer's disease; muscular dystrophy, such as Duchenne muscular dystrophy (DMD) or Becker muscular dystrophy (BMD); gastric Intestinal diseases, such as Crohn's disease, microscopic colitis, and primary sclerosing cholangitis (PSC); skin diseases, such as scleroderma, nephrogenic systemic fibrosis, and corneal keloids; arthrofibrosis, Dupuytren's contracture; mediastinal fibrosis, retroperitoneal fibrosis; myelofibrosis; Peyronie's disease; bursitis; kidney diseases (e.g., renal fibrosis, nephrotic syndrome, Alport syndrome, HIV-associated nephropathy, polycystic kidney disease, Fabry disease, diabetic nephropathy, chronic glomerulonephritis, nephritis associated with systemic lupus, renal interstitial fibrosis (IF)); kidney injury, such as acute kidney injury / renal failure; nephrotoxic progressive systemic sclerosis (PSS); chronic graft-versus-host disease ocular diseases / disorders and related processes, such as Grave's ophthalmopathy, epiretinal fibrosis (e.g., diabetic retinopathy (DR)), glaucoma, subretinal fibrosis (e.g., associated with macular degeneration (e.g., dry or dry age-related macular degeneration (AMD)), macular edema, drusen formation, choroidal neovascularization (CNV), post-surgical fibrosis (e.g., posterior capsule after cataract surgery), or glaucoma caused by blebs after trabeculectomy), conjunctival fibrosis, subconjunctival fibrosis; arthritis; pre-fibrotic neoplastic and fibrotic neoplastic diseases; and fibrosis caused by chemical or environmental influences (e.g., cancer chemotherapy, pesticides, radiation / cancer radiotherapy).

[0523] Early NASH, as used herein, refers to the steatosis stage of fatty liver disease, such as nonalcoholic fatty liver disease (NAFLD), which may reach the NASH state where the liver is inflamed. Late NASH, as used herein, refers to the state of persistent liver inflammation, which may include fibrosis.

[0524] It will be appreciated that many of the diseases / conditions listed above are interrelated. For example, ventricular fibrosis can occur after myocardial infarction and is associated with DCM, HCM, and myocarditis.

[0525] In specific embodiments, the disease / disorder can be one of the following: pulmonary fibrosis, atrial fibrillation, ventricular fibrillation, hypertrophic cardiomyopathy (HCM), dilated cardiomyopathy (DCM), non-alcoholic steatohepatitis (NASH), cirrhosis, chronic kidney disease, scleroderma, systemic sclerosis, keloids, cystic fibrosis, Crohn's disease, post-surgical fibrosis, or retinal fibrosis, such as associated with wet age-related macular degeneration (AMD).

[0526] In some embodiments, these methods can effectively reverse or eliminate fibrosis. Fibrosis can be existing or severe fibrosis and can be associated with any disease / disorder described herein. In some embodiments, these methods can effectively reverse or eliminate the disease / disorder provided herein.

[0527] Fibrosis can directly or indirectly lead to disease / disease, and / or increase susceptibility to disease / disease development. For example, more than 80% of hepatocellular carcinomas (HCCs) arise in fibrotic or cirrhotic livers (Affo et al., 2016, Annu Rev Pathol.), suggesting that liver fibrosis plays an important role in the precancerous environment (PME) of the liver.

[0528] Therefore, the antigen binding molecules of the present invention can be used for treating and preventing the disease / disorder related to fibrosis and / or fibrosis being a risk factor.In some embodiments, the disease / disorder related to fibrosis or the disease / disorder with fibrosis as a risk factor is cancer, such as liver cancer (such as hepatocellular carcinoma).

[0529] IL-11-mediated signal transduction is also involved in the pathology of other diseases / disorders, and thus the antigen binding molecules of the present invention can also be used to treat, prevent, alleviate and / or reverse or ablate the symptoms of these diseases / disorders.

[0530] In some embodiments, fibrosis can be associated with, for example, angiogenesis in the eye. In some embodiments, the methods described herein for treating or preventing fibrosis, methods for determining a subject's suitability for such treatment / prevention, and methods for diagnosing / prognosing fibrosis are also applicable to treating / preventing / diagnosing / prognosing angiogenesis, and vice versa. Ocular fibrosis can be associated with choroidal neovascularization (CNV).

[0531] In some embodiments, the antigen binding molecules of the present invention are provided for treating and / or preventing methods for metabolic diseases. That is, by suppressing, for example, the signal transduction of IL-11 mediation in cells, tissues / organs / organ systems / subjects, the present invention provides methods for treating / preventing metabolic diseases. " Metabolic disease " used herein refers to any disease or condition characterized by abnormal metabolism or abnormal metabolism. In this article, "metabolism" refers to that the body converts / processes energy sources, for example, energy and / or for storage through consumption to provide nutritional substances." Normal metabolism " can be the metabolism of healthy subjects without disease, for example, without metabolic disease, or without the symptoms / correlation of metabolic disease.

[0532] A subject with a metabolic disease may exhibit abnormal metabolism. A subject with a metabolic disease may have symptoms / correlates of abnormal metabolism. A subject with a metabolic disease may have been diagnosed as having a metabolic disease. A subject may meet the diagnostic criteria for diagnosing a metabolic disease. In some embodiments, the present invention provides methods for treating / preventing a disease / condition in a subject with a poor prognosis for a metabolic disease.

[0533] In some embodiments, the metabolic disease affects one or more of the liver, pancreas, cardiovascular system, digestive system, excretory system, respiratory system, renal system, reproductive system, circulatory system, muscular system, endocrine system, exocrine system, lymphatic system, immune system, nervous system, and / or skeletal system.

[0534] In some embodiments, the metabolic disease is or includes (e.g., characterized by) obesity, type 2 diabetes (T2D), type 1 diabetes (T1D), prediabetes, overweight, metabolic syndrome, pregnancy-related hyperglycemia (i.e., gestational diabetes), cholestatic liver disease, hyperglycemia, hyperlipidemia, hypertriglyceridemia, hypercholesterolemia, emaciation, cachexia, chemotherapy-related weight loss, pancreatic insufficiency, pancreatitis, acute pancreatitis, chronic pancreatitis, steatosis, non-alcoholic fatty liver disease (NAFLD), non-alcoholic fatty liver (NAFL), non-alcoholic steatohepatitis (NASH), lipodystrophy, excess fat, lipoatrophy, insulin deficiency, insulin resistance, and hyperglucagonemia. In some embodiments, the metabolic disease is or includes obesity. In some embodiments, the metabolic disease is or includes cholestasis, i.e., decreased bile flow from the liver to the duodenum. The disease can be a cholestatic liver disease (Jansen et al., Hepatology (2017) 65(2):722-738 and Pollock and Minuk, J Gastroenterol Hepatol (2017) 32(7):1303-1309, both of which are incorporated herein by reference in their entireties), including, for example, primary biliary cholangitis (PBC) and primary sclerosing cholangitis (PSC).

[0535] Aspects of the present invention relate to treating and / or preventing malfunction and / or insufficiency of cells / tissues / organs / organ systems that play a role in metabolism. Specifically, the present invention contemplates treating and / or preventing malfunction and / or insufficiency of the pancreas / pancreatic tissue / pancreatic cells, as well as treating and / or preventing malfunction and / or insufficiency of liver cells / liver tissue / liver.

[0536] In some embodiments, metabolic disease is or includes "wasting". The term "wasting" as used herein refers to involuntary weight loss, which can be progressive and / or degenerative. Wasting can be defined as muscle loss with or without fat loss, and generally involves significant, usually involuntary weight loss (including skeletal muscle), and may or may not include the loss of adipose tissue. In some cases, adipose tissue loss can occur alone, such as seen in lipodystrophy diseases. Wasting is characterized by a negative balance of protein and energy driven by reduced food intake and metabolic abnormalities (Fearon et al. Lancet Oncol. (2011) 12 (5): 489-95). Wasting can lead to progressive dysfunction, impaired quality of life, and increased risk of morbidity and mortality. In some cases, wasting can lead to weakness (weakness or lack of energy) and / or anemia (lack of red blood cells or hemoglobin in the blood). In some cases, conventional nutritional support or therapeutic interventions attempted so far cannot completely eliminate wasting. Once the body weight reaches 30% of the patient's historical stable weight, death usually occurs (Tisdale, Nature Reviews Cancer, 2, 862-871 (2002)).

[0537] Diseases / disorders characterized by wasting include cachexia (loss of muscle mass unrelated to age), sarcopenia (loss of muscle mass: for example, age-related, disuse, space travel, or denervation), anorexia (protein-energy malnutrition), muscular dystrophy, lipodystrophy (e.g., abnormal or degenerative diseases of adipose tissue), lipoatrophy (age-related loss of subcutaneous fat in the face and other tissues), and myasthenia (muscle wasting in any chronic disease; proposed by Fearon et al., J Cachexia Sarcopenia Muscle. 2011; 2: 1-3). Diseases / disorders characterized by wasting are also referred to herein as "wasting diseases." In some embodiments, the wasting disorders of the present disclosure are cachexia, precachexia, refractory cachexia, sarcopenia, anorexia, lipodystrophy, lipoatrophy, and / or myasthenia. In some embodiments of the various aspects described herein, the wasting disorder is cachexia, precachexia, and / or refractory cachexia.

[0538] Wasting disorders caused by chronic illness can include "mild muscle wasting disorder" (with or without frailty), "moderate muscle wasting disorder" (with or without frailty; sometimes called "precachexia"), or "severe muscle wasting disorder" (sometimes called "cachexia", often with frailty). Cachexia can be defined as involuntary weight loss of >5% from a historical stable weight, a body mass index (BMI) <20 kg / m2 (individuals under 65 years of age) or <22 kg / m2 (individuals over 65 years of age) with any degree of weight loss >2%, or a skeletal muscle index consistent with sarcopenia with any degree of weight loss >2%. Subjects may also show <10% body fat and / or a low blood albumin level of <35 g / l. These criteria may also help determine the "risk" of a population for developing a wasting disorder (Fearon et al. Lancet Oncol. 2011;12(5):489-95).

[0539] A three-step classification of cachexia has been proposed, with severity classified based on the degree of depletion of energy stores and body protein (BMI) and the degree of sustained weight loss.

[0540] 1. Precachexia - when the patient has lost <5% of their body weight but has not yet developed serious complications.

[0541] 2. Cachexia - The syndrome is developing and weight loss exceeds the above parameters, but it may still be treated.

[0542] 3. Refractory cachexia - the point at which the disease no longer responds to treatment or the burdens and risks outweigh the benefits of treatment (Fearon et al., supra). Generally, the refractory stage depends on the overall stage of the underlying disease (as described below) and the patient's condition.

[0543] Metabolic disease may be present in acute or chronic disease environment.Aspects of the present invention provide the treatment / prevention method of the disease / condition relevant to metabolic disease.The disease / condition relevant to metabolic disease comprises the disease / condition that is positively correlated with the outbreak of metabolic disease.In some embodiments, the disease / condition relevant to metabolic disease is to cause / cause / have caused (that is, can cause, cause or have caused) the disease / condition of metabolic disease.

[0544] The disease / condition relevant to metabolic disease also comprises the disease / condition that is caused by metabolic disease and / or increases the weight of (become worse, progress and / or complexity).In some embodiments, the disease / condition relevant to metabolic disease can be positively correlated with the outbreak of metabolic disease, also can increase the weight of by metabolic disease.The disease / condition of " relevant " can be the disease / condition that comprises the metabolic disease associated pathology feature.

[0545] In embodiments of the invention, metabolic diseases or diseases / disorders associated with metabolic diseases may be present in or affect any organ / tissue, such as the heart, liver, kidneys, brain, skin, muscular system, stomach, small intestine, large intestine, pancreas, mouth, salivary glands, pharynx, esophagus, gall bladder, trachea, larynx, bladder, ovaries, uterus, testicles, endocrine gland system (such as pituitary or thyroid), lymphatic system (such as spleen).

[0546] In an embodiment of the present invention, the metabolic disease-related disease / disorder may be one or more of the following: cancer, heart disease, kidney disease, lung disease, liver disease, chronic infection, neurodegenerative disease, acute injury, trauma / wound, postoperative condition or aging.

[0547] According to aspects of the present invention, the methods of treating and / or preventing metabolic diseases of the present invention may include one or more of the following:

[0548] Lowering blood lipid levels;

[0549] Lower blood sugar levels;

[0550] Improve glucose tolerance (e.g., in subjects with impaired glucose tolerance);

[0551] Increased insulin resistance (e.g., in insulin-resistant subjects);

[0552] Enhance pancreatic function;

[0553] Weight loss (e.g., in overweight / obese individuals);

[0554] Reduce body fat;

[0555] Increase lost weight;

[0556] Lower fasting blood sugar levels;

[0557] Lower serum triglyceride levels;

[0558] Lower serum cholesterol levels;

[0559] Improve glucose tolerance;

[0560] Enhance pancreatic function (e.g., exocrine and / or endocrine function);

[0561] Increased growth of pancreatic tissue;

[0562] pancreatic tissue regeneration;

[0563] Increased pancreatic weight;

[0564] Reduce islet cell hyperplasia;

[0565] Reduces glucagon expression;

[0566] Increased insulin expression;

[0567] weight gain (e.g., in subjects with wasting disorders such as cachexia);

[0568] Reduced IL-11 protein expression in the liver;

[0569] Reduced Erk activation in the liver;

[0570] reduction, e.g., of steatosis in the liver;

[0571] Lowering liver triglyceride levels;

[0572] Reduce serum ALT levels;

[0573] Reduce the expression of pro-inflammatory cytokines (e.g., TNFα, CCL2, CCL5, IL-6, CXCL5, and / or CXCL1);

[0574] reducing the expression of profibrotic factors (e.g., IL-11, TIMP1, ACTA2, TGFβ1, MMP2, TIMP2, MMP9, COL1A2, COL1A1, and / or COL3A1);

[0575] Reduce serum TGFβ1 levels;

[0576] Reduce HSC expression / production of IL-11, ACTA2, MMP2, TGFβ1, PDGF, ANG II, bFGF, CCL2, and / or H2O2;

[0577] Inhibition of HSCs leading to the transformation of HSCs into myofibroblasts;

[0578] Reduce the number / proportion of myofibroblasts in the liver;

[0579] Reduces liver hydroxyproline levels;

[0580] Improved liver function;

[0581] Increase the function of organs / tissues affected by metabolic diseases;

[0582] Reduce liver damage; and

[0583] Reduce the number / proportion of CD45+ cells in the liver.

[0584] IL-11-mediated signaling is associated with the occurrence and progression of various cancers. Studies have shown that IL-11-mediated signaling is crucial for promoting the tumorigenesis of chronic gastritis and related gastric cancer, colon cancer, hepatocellular carcinoma, and breast cancer by overactivating STAT3 (Ernst M et al. J Clin Invest. (2008); 118: 1727-1738). IL-11 may promote tumorigenesis by triggering the JAK-STAT intracellular signaling pathway and may also promote metastasis through signaling of the PI3K-AKT-mTORC1 pathway (Xu et al., Cancer Letters (2016) 373(2): 156-163). IL-11 promotes survival, proliferation, invasive angiogenesis, and metastasis through STAT3. The IL-11 / GP130 / JAK / STAT3 signaling axis may be the rate-limiting step in gastrointestinal tumor progression, and elevated IL-11 expression is associated with poor prognosis in breast cancer patients (Johnstone et al., Cytokine & Growth Reviews (2015) 26(5):489-498). IL-11 has also been shown to affect breast cancer stem cell dynamics and tumor heterogeneity (Johnstone et al., Cytokine & Growth Reviews (2015) 26(5):489-498). Recently, IL-11-mediated signaling has been associated with chemoresistance in lung adenocarcinoma; cancer-associated fibroblasts were found to upregulate IL-11 and confer chemoresistance to lung cancer cells by activating the IL-11 / IL-11R / STAT3 anti-apoptotic signaling pathway (Tao et al. 2016, Sci Rep. 6; 6:38408). IL-11-mediated signaling may promote the transformation of fibroblasts into myofibroblasts and promote the production of extracellular matrix in the premalignant environment (PME) and tumor microenvironment (TME).

[0585] In some embodiments, the antigen binding molecules of the present invention are used in methods for treating / preventing cancer. In some embodiments, the cancer may be a cancer that directly or indirectly causes inflammation and / or fibrosis.

[0586] Cancer can be any undesirable cell proliferation (or any disease that itself manifests undesirable cell proliferation), neoplasm or tumor, or an increased risk or susceptibility to undesirable cell proliferation, neoplasm or tumor. Cancer can be benign or malignant, and can be primary or secondary (metastatic). A neoplasm or tumor can be any abnormal growth or proliferation of cells and can be located in any tissue.

[0587] In some embodiments, the antigen binding molecules of the present invention are provided for use in methods of treating / preventing cancer, such as epithelial cell cancer, breast cancer, gastrointestinal cancer (e.g., esophageal cancer, gastric cancer, pancreatic cancer, liver cancer (e.g., HCC), gallbladder cancer, colorectal cancer, anal cancer, gastrointestinal carcinoid) and lung cancer (e.g., non-small cell lung cancer (NSCLC) or small cell lung cancer (SCLC)). In some embodiments, the cancer is a cancer in which acute and / or chronic inflammation is a risk factor. In some embodiments, the cancer is a cancer in which a disease / disorder characterized by fibrosis (e.g., as described herein) is a risk factor.

[0588] In some embodiments, cancer can be associated with an increase in IL-11, IL-11Rα and / or gp130 gene or protein. For example, compared to comparable non-cancerous cells, the expression of IL-11, IL-11Rα and / or gp130 in cancer cells can be increased, or compared to the expression levels of comparable cells in the absence of cancer (e.g., in healthy control subjects), it can be associated with an increase in IL-11, IL-11Rα and / or gp130 expression in other cells (e.g., non-cancerous cells). In some embodiments, it can be determined that cancer cells have increased levels of signaling through the ERK and / or STAT3 pathways compared to comparable non-cancerous cells.

[0589] In some embodiments, cancer can be associated with mutations in IL-11, IL-11Rα, and / or gp130. In some embodiments, such mutations can be associated with increased gene or protein expression levels, or can be associated with increased IL-11 / IL-11R signaling levels, relative to expression / signaling levels observed in the absence of the mutation.

[0590] IL-11 / IL-11R signal transduction is related to diseases / disorders characterized by inflammation. Intra-articular injection of IL-11 has been shown to cause joint inflammation (Wong et al., Cytokine (2005) 29: 72-76), and IL-11 has been shown to have a pro-inflammatory effect at IL-13-mediated tissue inflammation sites (Chen et al., J Immunol (2005) 174: 2305-2313). IL-11 expression has also been observed to increase significantly in chronic skin lesions of atopic dermatitis, and is known to be associated with bronchial inflammation (Toda et al., J Allergy Clin Immunol (2003) 111: 875-881). IL-111-mediated signal transduction is involved in inflammatory bowel disease (IBD) and asthma (Putoczki and Ernst, J Leuko Biol (2010) 88 (6) 1109-1117). IL-11 has also been identified as a risk factor for multiple sclerosis; IL-11 is elevated in the cerebrospinal fluid of patients with clinically isolated syndrome (CIS) compared to control subjects, and serum levels of IL-11 are higher during relapse in patients with relapsing multiple sclerosis. IL-11 may promote the differentiation of CD4+ T cells into a TH17 phenotype - TH17 cells are important cells in the pathogenesis of multiple sclerosis (Zhang et al., Oncotarget (2015) 6(32):32297-32298).

[0591] In some embodiments, the antigen binding molecules of the present invention are provided for treating / preventing a method for treating / preventing a disease / disorder characterized by inflammation. In some embodiments, a disease or disorder characterized by inflammation can be a disease / disorder that directly or indirectly causes cancer and / or fibrosis. Diseases characterized by inflammation include, for example, allergic inflammation, such as allergic asthma and bronchitis, atopic dermatitis, allergic rhinitis and ocular allergic diseases, and autoimmune diseases, such as multiple sclerosis, systemic lupus erythematosus, rheumatoid arthritis, chronic active hepatitis, type 1 diabetes, celiac disease, Grave's disease, uveitis, pemphigus, psoriasis, Crohn's disease, ulcerative colitis, inflammatory bowel disease, anemia and autoimmune thyroiditis.

[0592] In some embodiments, the antigen binding molecules of the present invention are provided for use in methods for treating / preventing hepatotoxicity and diseases / disorders characterized by hepatotoxicity. Hepatotoxicity, as used herein, refers to damage and / or death of liver cells / tissues. Hepatotoxicity may refer to a state in which there is toxic damage to the liver, particularly a state associated with the death of liver cells within the liver. As described below, hepatotoxicity can be determined / diagnosed by detecting one or more correlations of hepatotoxicity. Hepatotoxic damage may cause hepatotoxicity. As used herein, "hepatotoxic damage" refers to any treatment, event, or condition that causes hepatotoxicity. For example, hepatotoxic damage may be caused by chemical / physical treatment / experience or gaseous conditions. In some embodiments, the hepatotoxic damage is chemical, such as in the case of drug-induced liver damage, such as APAP-induced hepatotoxicity. In some embodiments, the hepatotoxic damage is physical, such as in the case of possible surgical damage to liver tissue that produces hepatotoxicity, such as surgery to treat a disease and / or liver transplantation (e.g., hepatotoxicity may have iatrogenic causes). In some embodiments, the hepatotoxic damage is caused by hypoxia, such as ischemia or reperfusion (e.g., IRI may cause hepatotoxic damage).

[0593] Hepatotoxicity can be chemically driven liver damage, such as damage or injury caused by drugs, chemicals, ischemia, reperfusion, sepsis, or herbal or dietary supplements. In some embodiments, hepatotoxicity refers to drug-induced liver injury (DILI). In some embodiments, hepatotoxicity refers to liver damage caused by a hepatotoxin. The hepatotoxin can be alcohol. Hepatotoxicity can also be referred to as toxic hepatitis. Hepatotoxicity can refer to acute and / or chronic hepatotoxicity.

[0594] Hepatotoxicity may be caused directly or indirectly by alcohol consumption, such as chronic alcohol consumption. Hepatotoxicity as mentioned herein may be caused directly or indirectly by fasting, malnutrition, infectious agents (e.g., hepatitis viruses (e.g., hepatitis A, B, C, D, or E), HIV), cancer, or drug interactions.

[0595] Hepatotoxicity may be associated with other disorders, diseases, and conditions. Disorders, diseases, or conditions associated with hepatotoxicity include acute liver injury (ALI), acute liver failure, acute liver disease, chronic liver disease, liver damage, hepatitis, such as viral hepatitis, alcoholic hepatitis, hepatic ischemia-reperfusion injury (IRI), such as "warm" ischemia-reperfusion (WIR), radiation-induced liver disease (RILD), drug-induced liver injury (DILI), autoimmune liver injury, cholestatic liver disease, HIV, and cancer.

[0596] Drug-induced liver injury (DILI) includes intrinsic and idiopathic hepatotoxicity, with idiopathic DILI also including allergic and non-allergic reactions. Intrinsic mechanisms are associated with dose-dependent hepatotoxicity, while idiopathic hepatotoxicity is dose-independent and may occur in an unpredictable manner. Allergic idiopathic hepatotoxicity is also characterized by the presence of typical symptoms and signs of an adaptive immune system response, including fever, skin reactions, eosinophilia, autoantibody formation, and a shorter latency period, especially after re-exposure (Khoury et al., J Clin Transl Hepatol. 2015 Jun 28; 3(2): 99-108).

[0597] In some embodiments, the antigen binding molecules of the present invention can be used for the diagnosis, treatment, and prevention of acetaminophen (APAP)-induced hepatotoxicity. Acetaminophen is also known as N-acetyl-p-aminophenol or paracetamol, or under the brand names Tylenol and Priligy. Acetaminophen poisoning can lead to hepatotoxicity, which is associated with elevated serum concentrations of hepatocyte-derived enzymes (such as aspartate aminotransferase, lactate dehydrogenase, and alanine aminotransferase), lobular degeneration and necrosis, and activation of Kupffer cells (Trepicchio WL et al., Toxicol Pathol. 2001; 29(2): 242-9).

[0598] In some embodiments, the antigen binding molecules of the present invention can be provided for treating / preventing kidney injury, such as acute kidney injury (AKI; acute renal failure) or kidney injury related diseases / illness methods. Kidney injury is characterized by tubular epithelial cell (TEC) damage and / or TEC to epithelial to mesenchymal cell-like phenotype (i.e. EMT) transformation. TEC to the transformation of mesenchymal cell-like phenotype can be characterized by, for example, E-cadherin expression reduction, SNAIL expression increase and / or ACTA2 expression increase. Kidney injury may have any cause, such as including mechanical (i.e. physical) damage or damage, chemical damage or damage, ischemia or genetic susceptibility caused kidney injury. Cause or damage usually lead to impaired renal function, thereby may lead to renal failure. Mechanical damage or injury may include physical damage to experimenter, kidney, TEC or podocyte. It may also include tubular blockage / obstruction, such as urinary tract obstruction. In some embodiments, kidney injury is drug-induced kidney injury or drug-induced acute kidney injury.

[0599] Ischemic injury can be caused by decreased blood flow to the kidneys, which can be caused by many factors, such as low blood pressure, sepsis, blood loss, or the effects of surgery or chemical agents, such as medications or drugs administered to a subject to treat another disease, disorder, or condition. Ischemic kidney injury can be ischemic kidney injury or ischemic acute kidney injury. Kidney injury from crush injuries can be caused by ischemic kidney contraction and vasoconstriction, or by mechanical factors in the renal tubules or the toxic effects of circulating factors, such as myoglobin.

[0600] In some embodiments, kidney injury (which may be AKI) is characterized by damage to the renal tubular epithelial cells (TECs), i.e., the renal tubular epithelial cells, of the kidney, which in some cases may include or result in their death. TECs may be proximal or distal and may be damaged in AKI, and podocytes in the glomeruli may also be damaged. Damage to TECs may also be any type of damage, injury, or injury, such as mechanical, chemical, or ischemic injury, as described above. Damage to TECs is a common cause of kidney injury, especially AKI. The proliferation of TECs provides a mechanism for restoring and recovering renal function, while failure of TEC proliferation can lead to the development and progression of diseases, such as chronic kidney disease and renal failure. Proliferation of podocyte precursors may also occur to restore glomerular function, but is not as good as TEC proliferation. Mechanical damage may include, for example, unilateral ureteral obstruction (UUO).

[0601] In some embodiments, renal injury is nephrotoxicity, which refers to the toxicity of the kidneys. Nephrotoxicity may be the result of certain substances producing toxic effects on renal function and can therefore be considered the result of chemical damage or injury. Just like chemical damage or injury, nephrotoxicity may be a side effect of drug administration to treat a disease or condition that does not occur in the kidneys or occurs simultaneously in the kidneys and one or more other tissues. In some embodiments, nephrotoxicity can be a side effect of a chemotherapeutic agent administered to a subject for the prevention or treatment of cancer. Therefore, nephrotoxicity can be in the form of drug-induced renal injury or drug-induced acute kidney injury. In some embodiments, folic acid can cause renal injury, i.e., folic acid-induced renal injury.

[0602] In some embodiments, antigen binding molecules are provided for diagnosing, treating and / or preventing cisplatin-induced renal injury. This may include acute renal injury caused by cisplatin or nephrotoxicity caused by cisplatin. Cisplatin (dichlorodiaminoplatinum; SP-4-2)-diaminodichloroplatinum (II) is a chemotherapeutic agent widely used in the treatment of various cancers, including head and neck, breast, lung, testicle, ovary, brain and bladder cancer, which is recognized to cause renal injury and renal tubular dysfunction, including renal tubular damage and necrosis (e.g., Oh et al., Electrolyte Blood Press 2014 December; 12 (2): 55-65; PA Arunkumar et al., Asian Pac J Trop Biomed 2012 August 2 (8): 640-644). Other platinum-based chemotherapeutic agents can also cause renal damage.

[0603] It is recognized that subjects with renal injury may also have renal fibrosis due to disease conditions with different etiologies or as a secondary effect of renal injury. In some embodiments, the renal injury diagnosed, treated or prevented is not fibrosis of the kidney, such as renal fibrosis. In some embodiments, the subject does not have fibrosis. In some embodiments, TEC injury occurs in the absence of fibrosis. In some embodiments, fibrosis occurs separately from AKI (e.g., secondary), for example, due to incomplete TEC regeneration. In some embodiments, the damaged TEC in the subject is not a pro-fibrotic TEC. In some embodiments, fibrosis does not occur.

[0604] In some embodiments, the antigen binding molecules of the present invention are provided for use in methods for treating / preventing infection-related diseases / conditions, particularly where infection directly or indirectly leads to fibrosis, cancer, or inflammation. An infection-related disease may be a disease caused or exacerbated by infection with a related infectious agent, or a disease for which infection with an infection-related agent is a risk factor.

[0605] The infection can be any infection or infectious disease, such as a bacterial, viral, fungal or parasitic infection. In certain embodiments, the disease / disorder can be associated with a viral infection. In some embodiments, it may be particularly desirable to treat chronic / persistent infections, such as those associated with inflammation, cancer and / or fibrosis.

[0606] Infection may be chronic, continuous, latent or slow, and may be the result of bacterial, viral, fungal or parasitic infection. In this way, treatment can be provided to patients suffering from bacterial, viral or fungal infection. The example of bacterial infection includes Helicobacter pylori infection and Mycobacterium tuberculosis infection of the lung. The example of viral infection includes EBV, HPV, HIV, hepatitis B or hepatitis C infection.

[0607] The treatment may involve improving, treating or preventing any disease / disorder / condition associated with IL-11 signaling and / or described herein by inhibiting the biological activity of IL-11Rα or a complex comprising IL-11Rα. The treatment may involve reversing or regressing the disease / disorder by inhibiting the biological activity of IL-11Rα or a complex comprising IL-11Rα. Such methods may comprise administering an antibody / fragment / composition of the invention to bind to and inhibit the biological activity of IL-11Rα or a complex comprising IL-11Rα. Inhibiting the biological activity of IL-11Rα or a complex comprising IL-11Rα may be referred to as "neutralizing" herein.

[0608] Treatment methods may optionally include administering a biological adjuvant (e.g., interleukins, cytokines, BCG, monophosphoryl lipid A, etc.) in combination with conventional cancer treatments, such as drugs (e.g., chemotherapy), radiation therapy, or surgery. Medical treatment methods may also involve in vivo, ex vivo, and adoptive immunotherapy, including those using autologous and / or allogeneic cells or immortalized cell lines.

[0609] The treatment can be intended to prevent diseases / conditions associated with overactive / elevated IL-11-mediated signaling. Thus, pharmaceutical compositions or medicaments can be formulated using antibodies, antigen-binding fragments, and polypeptides, and subjects can be treated prophylactically to resist the development of the disease. This can occur before the onset of symptoms of the disease state and / or can be administered to subjects who are considered to be at greater risk for the disease or condition.

[0610] The preparation of the present invention is preferably administered in an amount that is "therapeutically effective" or "preventively effective" sufficient to demonstrate benefit to the subject. The amount actually administered, as well as the rate and time course of administration, will depend on the nature and severity of the disease / disorder and the nature of the preparation. Treatment prescriptions (e.g., decisions on dosage, etc.) are the responsibility of general practitioners and other physicians, and typically take into account the disease / disorder to be treated, the condition of the individual subject, the site of administration, the method of administration, and other factors known to practitioners. Examples of the above techniques and protocols can be found in Remington's Pharmaceutical Sciences (20th edition), published by Lippincott, William & Wilkins Publishers in 2000.

[0611] Antigen binding molecules as described herein, polypeptide, CAR, nucleic acid, expression vector, cell and composition are preferably formulated into medicine together with one or more other pharmaceutically acceptable ingredients well known to those skilled in the art, and the pharmaceutically acceptable ingredients include but are not limited to pharmaceutically acceptable carriers, adjuvants, excipients, diluents, fillers, buffers, preservatives, antioxidants, lubricants, stabilizers, solubilizers, surfactants (such as wetting agents), screening agents, colorants, flavorings and sweeteners. The term "pharmaceutically acceptable" as used herein refers to compounds, ingredients, materials, compositions, dosage forms, etc., which are suitable for contacting the tissue of the object under discussion (such as people) within the scope of reasonable medical judgment, without excessive toxicity, stimulation, allergic reaction or other problems or complications, and with a reasonable benefit / risk ratio. In terms of being compatible with other ingredients of the preparation, each carrier, adjuvant, excipient, etc. must also be "acceptable". Suitable carriers, adjuvants, and excipients can be found in standard pharmaceutical texts, such as Remington's Pharmaceutical Sciences, 18th ed., Mack Publishing, Easton, Pa., 1990; and Handbook of Pharmaceutical Excipients, 2nd ed., 1994.

[0612] Can prepare the preparation that is suitable for disease to be treated / illness to be administered.For example, preparation can be configured to be used for local, parenteral, whole body, intravenous, intraarterial, intramuscular, intrathecal, intraocular, local eye (for example subconjunctival, intravitreal, behind the ball, intracameral), intraconjunctival, subcutaneous, oral or transdermal route administration, can comprise injection.Reagent of the present disclosure can be formulated as fluid or solid form.Can prepare fluid preparation so that by injection or be infused into the selected region of human body or animal body and administer.Injectable preparation can comprise selected reagent in aseptic or isotonic medium.

[0613] The preparation can be prepared by any method known in the pharmaceutical field. Such methods include the step of combining the active compound with a carrier constituting one or more auxiliary ingredients. Generally, the preparation is prepared by uniformly and tightly combining the active compound with a carrier (e.g., a liquid carrier, a finely divided solid carrier, etc.) and then shaping the product.

[0614] The present invention also provides methods for producing pharmaceutically useful compositions, such methods of production may include one or more steps selected from the group consisting of: isolating the antibodies or antigen-binding fragments described herein; and / or mixing the isolated antibodies or antigen-binding fragments described herein with a pharmaceutically acceptable carrier, adjuvant, excipient, or diluent. For example, another aspect of the present invention relates to a method for formulating or producing a medicament or pharmaceutical composition for use in a medical treatment method, comprising formulating a pharmaceutical composition or medicament by mixing the antibodies or antigen-binding fragments described herein with a pharmaceutically acceptable carrier, adjuvant, excipient, or diluent.

[0615] Multiple doses of antigen binding molecules, polypeptides, CARs, nucleic acids (or multiple thereof), expression vectors (or multiple thereof), cells or compositions can be provided. One or more or each dose can be accompanied by another therapeutic agent administered simultaneously or sequentially.

[0616] Multiple doses can be separated by predetermined time intervals, which can be selected from 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 one of 1, 2, 3, 4, 5, or 6 months. For example, a dose can be administered once every 7, 14, 21, or 28 days (plus or minus 3, 2, or 1 day).

[0617] Antigen binding molecules, polypeptides, CARs, nucleic acids, expression vectors, cells, and compositions described herein can be administered alone or in combination with other therapeutic or preventive interventions. Such other therapeutic or preventive interventions may occur before, during, and / or after the treatment encompassed by the present disclosure, and the other therapeutic or preventive interventions may be delivered using the same or different routes of administration as the treatment of the present disclosure.

[0618] In some embodiments, the administration of antigen binding molecules, polypeptides, CARs, nucleic acids, expression vectors, cells, and compositions described herein can be accompanied by reagents (e.g., antibiotics, antivirals, antifungals, or antiparasitics) for treating or preventing infection. In some embodiments, the treatment of antibodies, antigen binding fragments, or compositions of the present invention can be accompanied by reagents (e.g., nonsteroidal anti-inflammatory drugs (NSAIDs)) for treating or preventing inflammation. In some embodiments, the treatment performed by antibodies, antigen binding fragments, or compositions of the present invention can be accompanied by radiotherapy (i.e., treatment with ionizing radiation, such as X-rays or gamma rays) and / or reagents (e.g., chemotherapeutics) for treating or preventing cancer. In some embodiments, the chemotherapeutics can be alkylating agents, such as cisplatin. In some embodiments, antibodies, antigen binding fragments, or compositions of the present invention can be given as part of a combined treatment with immunotherapy.

[0619] Simultaneous administration refers to administering the agent together, for example, as a pharmaceutical composition (i.e., combination preparation) comprising the agents, or immediately after each other, and optionally by the same route of administration, for example, by administering the same artery, vein or other blood vessel. In certain embodiments, after simultaneous administration, two or more agents can be administered by different routes of administration. In some embodiments, simultaneous administration refers to administration at the same time, or within, for example, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 8 hours, 12 hours, 24 hours, 36 hours or 48 hours.

[0620] Sequential administration refers to the administration of one or more agents, followed by the administration of another agent after a given time interval. It is not necessary to administer two medicaments by the same approach, although this is the case in certain embodiments. The time interval can be any time interval, including hours, days, weeks, months, or years. In some embodiments, sequential administration refers to the administration separated by a time interval of at least 10 minutes, 30 minutes, 1 hour, 6 hours, 8 hours, 12 hours, 24 hours, 36 hours, 48 ​​hours, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 1 month, 6 weeks, 2 months, 3 months, 4 months, 5 months, or 6 months.

[0621] Detection method

[0622] The present invention also provides an article of the invention for use in methods for detecting, localizing, or imaging IL-11Rα or a complex comprising IL-11Rα, or a cell expressing / comprising IL-11Rα or a complex comprising IL-11Rα. The antigen binding molecules described herein can be used to bind an antigen binding molecule to IL-11Rα or a complex comprising IL-11Rα. Such methods may involve detecting the bound complex of an antigen binding molecule and IL-11Rα or a complex comprising IL-11Rα.

[0623] Detection of IL-11Rα or a complex comprising IL-11Rα can be used in the diagnosis / prognosis of diseases / disorders that pathologically involve IL-11-mediated signaling and / or expression / inclusion of IL-11Rα or a complex comprising IL-11Rα, in methods for identifying subjects at risk of developing such diseases / disorders, and / or in methods for predicting a subject's response to therapeutic intervention.

[0624] Thus, a method is provided, comprising contacting a sample containing or suspected of containing IL-11Rα or a complex comprising IL-11Rα or expressing / containing IL-11Rα or a complex comprising IL-11Rα with an antigen-binding molecule as described herein, and detecting the formation of a complex between the antigen-binding molecule and the IL-11Rα / complex comprising IL-11Rα. A method is also provided, comprising contacting a sample containing or suspected of containing cells expressing / containing IL-11Rα or a complex comprising IL-11Rα with an antigen-binding molecule as described herein, and detecting the formation of a complex between the antigen-binding molecule and the cells expressing / containing IL-11Rα or a complex comprising IL-11Rα.

[0625] Suitable method forms are well known in the art, including immunoassays, such as sandwich assays, such as ELISA. Said method may include using a detectable portion, such as a fluorescent marker, a phosphorescent marker, a luminescent marker, an immune detectable marker, a radiolabel, a chemistry, a nucleic acid or an enzyme marker to mark an antigen binding molecule or a target, or both. IL-11R α expression can be measured by immunohistochemistry (IHC), such as by a tissue sample obtained by biopsy. In some embodiments, the marker can be selected from: radionucleotides, positron-emitting radionuclides (such as for positron emission tomography (PET)), MRI contrast agents or fluorescent markers.

[0626] Detection techniques are well known to those skilled in the art and can be selected to correspond to the labeled reagent. In vitro or in vivo analysis of processes mediated by IL-11 / IL-11R signaling may involve analysis by positron emission tomography (PET), magnetic resonance imaging (MRI), or fluorescence imaging (e.g., by detecting an appropriately labeled substance).

[0627] Such methods can provide the basis for the diagnostic method of the disease or illness of the complex that needs to detect and / or quantify IL-11R α or comprise IL-11R α.Such methods can be carried out in vitro on subject sample, or after subject sample treatment.Once sample is collected, it is not necessary for the subject to be present in the in vitro diagnostic method to be performed, so the method can be a method not implemented on human or animal body.In some embodiments, there is provided antigen binding molecules of the present invention, polypeptide, CAR, nucleic acid, expression vector, cell or composition, for any diagnosis, detection or quantitative method described herein.

[0628] Such methods may involve detecting or quantifying IL-11Rα or a complex comprising IL-11Rα, or a cell expressing IL-11Rα or a complex comprising IL-11Rα, for example, in a patient sample. If the method includes quantifying a factor of interest, the method may further include comparing the measured amount to a standard or reference value as part of a diagnostic or prognostic assessment. Other diagnostic / prognostic tests may be used in conjunction with those described herein to enhance the accuracy of a diagnosis or prognosis or to confirm the results obtained using the tests described herein.

[0629] Detection of IL-11Rα or a complex comprising IL-11Rα in a sample can be used to diagnose an infectious disease, an autoimmune disease, or a cancerous condition in a subject, diagnose a susceptibility to an infectious disease, an autoimmune disease, or a cancerous condition, or provide a prognosis for an infectious disease, an autoimmune disease, or a cancerous condition. The diagnosis or prognosis may be related to an existing (previously diagnosed) infectious, inflammatory, or autoimmune disease / condition or cancerous condition.

[0630] Detecting an elevated level of IL-11Rα or a complex comprising IL-11Rα in a sample obtained from a subject, or detecting an increased presence or number / ratio of cells expressing / comprising IL-11Rα or a complex comprising IL-11Rα, can diagnose the subject as having a disease / disorder of the present disclosure, or being at risk of developing such a disease / disorder. In such methods, an "increase" in the expression level or number / ratio of cells refers to a level / number / ratio that is greater than the level / number / ratio determined under appropriate control conditions, such as the level / number / ratio detected in a comparable sample obtained from a healthy subject (e.g., a sample of the same type, such as obtained from the same fluid, tissue, organ, etc.).

[0631] Detection of an elevated level of IL-11Rα or a complex containing IL-11Rα, or detection of an increased presence or number / ratio of cells expressing / containing IL-11 or a complex containing IL-11Rα in a sample obtained from a subject can determine that the subject has a worse prognosis compared to a subject in which a lower level of IL-11Rα or a complex containing IL-11Rα or a decreased number / ratio of cells expressing / containing IL-11Rα or a complex containing IL-11Rα is determined in a comparable sample (e.g., a similar sample, such as a sample obtained from the same fluid, tissue, organ, etc.).

[0632] Therefore, the present invention provides a method for selecting / stratifying the subject for treatment of the antigen binding molecules, polypeptides, CARs, nucleic acids, expression vectors, cells or compositions of the present invention. In some embodiments, based on, for example, the detection / quantification of IL-11R α or a complex comprising IL-11R α in a sample obtained by the subject, or expressing IL-11R α or a cell comprising a complex of IL-11R α, to select a subject for the treatment / prevention method of the present invention, or to identify it as a subject who will benefit from such treatment / prevention method. The level of IL-11R α or a complex comprising IL-11R α present in the subject sample can indicate that the subject can respond to the treatment of the antigen binding molecules or compositions of the present invention. The presence of high levels of IL-11R α or a complex comprising IL-11R α in the sample can be used to select a subject for treatment as described herein. Therefore, the antigen binding molecules of the present invention can select a subject for IL-11R α targeted therapy treatment.

[0633] The sample can be obtained from any tissue or body fluid. The sample can include or be derived from: a certain amount of blood; a certain amount of serum extracted from the individual's blood, which may include the liquid portion of the blood obtained after removing the fibrin clot and blood cells; a tissue sample or biopsy; pleural effusion; cerebrospinal fluid (CSF); or cells isolated from the individual. In some embodiments, the sample can be obtained or derived from one or more tissues affected by the disease / disorder (e.g., one or more tissues in which symptoms of the disease are evident, or one or more tissues associated with the onset of the disease / disorder).

[0634] The diagnostic or prognostic methods of the present invention can be performed in vitro on a sample obtained from a subject, or after processing the sample obtained from a subject. Once the sample is collected, the patient does not need to be present for the in vitro diagnostic or prognostic method to be performed, and thus the method can be a method that is not performed on the human or animal body. The term "in vitro" is intended to cover experiments on cultured cells, while the term "in vivo" is intended to cover experiments and / or treatments on intact multicellular organs.

[0635] The diagnostic and prognostic methods of the present invention can be performed on samples obtained from a subject at multiple time points throughout the course of a disease and / or treatment and can be used to monitor the progression of a disease / condition over time, such as the response to a treatment administered to a subject. The characterization results of the methods can be used to inform clinical decisions about when and what therapy to administer to a subject.

[0636] Subjects

[0637] The subject of the various aspects of the invention described herein can be any animal or human. The subject is preferably a mammal, more preferably a human. The subject can be a non-human mammal, but more preferably a human. The subject can be male or female. The subject can be a patient. The subject may have been diagnosed with a disease or condition (e.g., cancer) 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.

[0638] The subject / patient may have a disease / disorder that would benefit from a reduction in IL-11-mediated signaling levels (i.e., inhibition or antagonism thereof) or a reduction in the number and / or activity of cells expressing IL-11Rα or a complex comprising IL-11Rα. The subject / patient may have a disease / disorder as described herein. The subject / patient may have been diagnosed with a disease / disorder in need of treatment as described herein, may be suspected of having such a disease / disorder, or may be at risk of developing such a disease / disorder.

[0639] In embodiments of the present invention, the subject is preferably a human subject. In some embodiments, the subject to be treated in the treatment or prevention methods of the present invention herein is a subject suffering from or at risk of suffering from cancer. In embodiments of the present invention, a subject can be selected for treatment according to the methods based on the characterization of certain markers of the disease / disorder.

[0640] Reagent test kit

[0641] In some aspects of the present invention described herein, there is provided a kit of parts. In some embodiments, the kit may have at least one container having a predetermined amount of antigen binding molecules as described herein, polypeptide, CAR, nucleic acid (or its multiple), expression vector (or its multiple), cell or composition.

[0642] In some embodiments, the kit may contain materials for producing an antigen binding molecule, polypeptide, CAR, nucleic acid (or multiple thereof), expression vector (or multiple thereof), cell or composition as described herein.

[0643] The kit can provide an antigen binding molecule, polypeptide, CAR, nucleic acid (or multiple thereof), expression vector (or multiple thereof), cell or composition, and instructions for administration to a patient for treating a specific disease / disorder.

[0644] In some embodiments, the kit may further include at least one container having a predetermined amount of another therapeutic agent (e.g., an anti-infective agent or a chemotherapeutic agent). In such embodiments, the kit may also include a second drug or pharmaceutical composition so that two drugs or pharmaceutical compositions can be administered simultaneously or separately, thereby providing a combined treatment for a specific disease or condition. The therapeutic agent can also be formulated to be suitable for injection or infusion into a tumor or blood.

[0645] Sequence identity

[0646] As used herein, "sequence identity" refers to the percentage of nucleotides / amino acid residues in a subject sequence that are identical to the nucleotides / amino acid residues in a reference sequence, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percentage of sequence identity between the sequences. For the purpose of determining the percentage of sequence identity between two or more amino acid or nucleic acid sequences, pairwise and multiple sequence alignments can be achieved in a variety of ways known to those skilled in the art, for example, using a library such as ClustalOmega ( J. 2005, Bioinformatics 21, 951-960), T-Coffee (Notredame et al., 2000, J. Mol. Biol. 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) are publicly available computer software. When using such software, it is preferred to use the default parameters (e.g., for gap penalties and extension penalties).

[0647] sequence

[0648]

[0649]

[0650]

[0651]

[0652]

[0653]

[0654] Numbered paragraphs relating to aspects and embodiments of the invention:

[0655] 1. An optionally isolated antigen-binding molecule capable of binding to IL-11Rα, wherein the antigen-binding molecule comprises:

[0656] (i) a heavy chain variable (VH) region comprising the following CDRs:

[0657] HC-CDR1 having the amino acid sequence of SEQ ID NO: 18

[0658] HC-CDR2 having the amino acid sequence of SEQ ID NO: 52

[0659] an HC-CDR3 having the amino acid sequence of SEQ ID NO: 21; and

[0660] (ii) a light chain variable (VL) region comprising the following CDRs:

[0661] LC-CDR1 having the amino acid sequence of SEQ ID NO: 22

[0662] LC-CDR2 having the amino acid sequence of SEQ ID NO: 23

[0663] LC-CDR3 having the amino acid sequence of SEQ ID NO: 24.

[0664] 2. The antigen-binding molecule of paragraph 1, wherein the antigen-binding molecule comprises:

[0665] (i) a heavy chain variable (VH) region comprising the following CDRs:

[0666] HC-CDR1 having the amino acid sequence of SEQ ID NO: 18

[0667] HC-CDR2 having the amino acid sequence of SEQ ID NO: 19

[0668] an HC-CDR3 having the amino acid sequence of SEQ ID NO: 21; and

[0669] (ii) a light chain variable (VL) region comprising the following CDRs:

[0670] LC-CDR1 having the amino acid sequence of SEQ ID NO: 22

[0671] LC-CDR2 having the amino acid sequence of SEQ ID NO: 23

[0672] LC-CDR3 having the amino acid sequence of SEQ ID NO: 24.

[0673] 3. The antigen-binding molecule of paragraph 1, wherein the antigen-binding molecule comprises:

[0674] (i) a heavy chain variable (VH) region comprising the following CDRs:

[0675] HC-CDR1 having the amino acid sequence of SEQ ID NO: 18

[0676] HC-CDR2 having the amino acid sequence of SEQ ID NO: 20

[0677] an HC-CDR3 having the amino acid sequence of SEQ ID NO: 21; and

[0678] (ii) a light chain variable (VL) region comprising the following CDRs:

[0679] LC-CDR1 having the amino acid sequence of SEQ ID NO: 22

[0680] LC-CDR2 having the amino acid sequence of SEQ ID NO: 23

[0681] LC-CDR3 having the amino acid sequence of SEQ ID NO: 24.

[0682] 4. The antigen-binding molecule of any one of paragraphs 1 to 3, wherein the antigen-binding molecule comprises:

[0683] A VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 7, 8, 9, 10, 11, or 12; and

[0684] A VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 13, 14, 15, 16, or 17.

[0685] 5. The antigen binding molecule of any one of paragraphs 1 to 4, wherein the antigen binding molecule is capable of inhibiting IL-11-mediated signaling.

[0686] 6. An optionally isolated antigen-binding molecule comprising (i) the antigen-binding molecule of any one of paragraphs 1 to 5, and (ii) an antigen-binding molecule capable of binding to an antigen other than IL-11Rα.

[0687] 7. The antigen binding molecule according to any one of paragraphs 1 to 6, wherein the antigen binding molecule is capable of inhibiting the interaction between IL-11Rα or a complex comprising IL-11Rα and an interacting partner of IL-11Rα or a complex comprising IL-11Rα.

[0688] 8. A chimeric antigen receptor (CAR) comprising the antigen binding molecule of any one of paragraphs 1-7.

[0689] 9. One or more optionally isolated nucleic acids encoding the antigen binding molecule of any one of paragraphs 1-7 or the CAR of paragraph 8.

[0690] 10. One or more expression vectors comprising one or more nucleic acids of paragraph 9.

[0691] 11. A cell comprising the antigen binding molecule of any one of paragraphs 1-7, the CAR of paragraph 8, the one or more nucleic acids of paragraph 9, or the one or more expression vectors of paragraph 10.

[0692] 12. A method comprising culturing a cell comprising the one or more nucleic acids of paragraph 9 or the one or more expression vectors of paragraph 10 under conditions suitable for expressing an antigen binding molecule or CAR from the one or more nucleic acids or the one or more expression vectors.

[0693] 13. A composition comprising the antigen binding molecule of any one of paragraphs 1-7, the CAR of paragraph 8, the one or more nucleic acids of paragraph 9, the one or more expression vectors of paragraph 10, or the cell of paragraph 11.

[0694] 14. The antigen binding molecule of any one of paragraphs 1-7, the CAR of paragraph 8, the one or more nucleic acids of paragraph 9, the one or more expression vectors of paragraph 10, the cell of paragraph 11 or the composition of paragraph 13, for use in a medical treatment or prevention method.

[0695] 15. The antigen binding molecule of any of paragraphs 1-7, the CAR of paragraph 8, the one or more nucleic acids of paragraph 9, the one or more expression vectors of paragraph 10, the cell of paragraph 11 or the composition of paragraph 13, for use in a method for treating or preventing fibrosis, a disease characterized by fibrosis, cancer, inflammation or a disease characterized by inflammation.

[0696] 16. Use of the antigen binding molecule of any one of paragraphs 1-7, the CAR of paragraph 8, the one or more nucleic acids of paragraph 9, the one or more expression vectors of paragraph 10, the cell of paragraph 11 or the composition of paragraph 13 in the preparation of a medicament for treating or preventing fibrosis, a disease characterized by fibrosis, cancer, inflammation or a disease characterized by inflammation.

[0697] 18. A method for treating or preventing fibrosis, a disease characterized by fibrosis, cancer, inflammation, or a disease characterized by inflammation, comprising administering to a subject a therapeutically or prophylactically effective amount of the antigen binding molecule of any one of paragraphs 1-7, the CAR of paragraph 8, the one or more nucleic acids of paragraph 9, the one or more expression vectors of paragraph 10, the cell of paragraph 11, or the composition of paragraph 13.

[0698] 19. A method for inhibiting IL-11-mediated signal transduction, comprising contacting a cell expressing IL-11Rα with the antigen-binding molecule of any one of paragraphs 1 to 7.

[0699] 20. An optionally isolated in vitro complex comprising the antigen binding molecule of any of paragraphs 1 to 7 bound to IL-11Rα or a complex comprising IL-11Rα.

[0700] 21. A method comprising contacting a sample containing or suspected of containing IL-11Rα or a complex containing IL-11Rα with the antigen-binding molecule of any one of paragraphs 1-7, and detecting formation of a complex between the antigen-binding molecule and IL-11Rα or a complex containing IL-11Rα.

[0701] 22. A method for selecting or stratifying a subject for treatment with an IL-11Rα targeting agent, the method comprising contacting a sample from the subject with the antigen-binding molecule of any one of paragraphs 1-7 in vitro, and detecting formation of a complex between the antigen-binding molecule and IL-11Rα or a complex comprising IL-11Rα.

[0702] 22. Use of the antigen-binding molecule of any one of paragraphs 1 to 7 as an in vitro or in vivo diagnostic or prognostic agent.

[0703] ***

[0704] The present invention includes any combination of the described aspects and preferred features unless such a combination is expressly impermissible or explicitly avoided.

[0705] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

[0706] Aspects and embodiments of the present invention will now be described with reference to the accompanying drawings, by way of example only. Other aspects and embodiments will be apparent to those skilled in the art. All documents mentioned herein are incorporated herein by reference.

[0707] Throughout this specification, including the appended claims, unless the context requires otherwise, the word "comprise" and variations such as "comprises" and "comprising" will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps.

[0708] It must be noted that, as used in the specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Ranges may be expressed herein as from "about" one particular value and / or to "about" another particular value. When such a range is expressed, another embodiment includes from the from particular value and / or to the other particular value. Similarly, when a value is expressed as an approximation by use of the antecedent "about," it will be understood that the particular value forms another embodiment.

[0709] Where a nucleic acid sequence is disclosed herein, its reverse complement is also expressly contemplated.

[0710] The methods described herein may preferably be performed in vitro.The term "in vitro" is intended to encompass procedures performed with cells in culture, whereas the term "in vivo" is intended to encompass procedures performed with / on intact multicellular organisms. Sequence Listing <110> Singapore Health Services Pte Ltd (all countries) National University of Singapore (all countries) Anlefen Biotechnology Co., Ltd. (all countries) Craig, Richard (LS only) <120> IL-11RA antibodies <130> 007487291 <150> GB 1809700.6 <151> 2018-06-13 <160> 79 <170> PatentIn version 3.5 <210> 1 <211> 199 <212> PRT <213> Homo sapiens IL-11 (UniProt: P20809) <400> 1 Met Asn Cys Val Cys Arg Leu Val Leu Val Val Leu Ser Leu Trp Pro 1 5 10 15 Asp Thr Ala Val Ala Pro Gly Pro Pro Pro Gly Pro Pro Arg Val Ser 20 25 30 Pro Asp Pro Arg Ala Glu Leu Asp Ser Thr Val Leu Leu Thr Arg Ser 35 40 45 Leu Leu Ala Asp Thr Arg Gln Leu Ala Ala Gln Leu Arg Asp Lys Phe 50 55 60 Pro Ala Asp Gly Asp His Asn Leu Asp Ser Leu Pro Thr Leu Ala Met 65 70 75 80 Ser Ala Gly Ala Leu Gly Ala Leu Gln Leu Pro Gly Val Leu Thr Arg 85 90 95 Leu Arg Ala Asp Leu Leu Ser Tyr Leu Arg His Val Gln Trp Leu Arg 100 105 110 Arg Ala Gly Gly Ser Ser Leu Lys Thr Leu Glu Pro Glu Leu Gly Thr 115 120 125 Leu Gln Ala Arg Leu Asp Arg Leu Leu Arg Arg Leu Gln Leu Leu Met 130 135 140 Ser Arg Leu Ala Leu Pro Gln Pro Pro Pro Asp Pro Pro Ala Pro Pro 145 150 155 160 Leu Ala Pro Pro Ser Ser Ala Trp Gly Gly Ile Arg Ala Ala His Ala 165 170 175 Ile Leu Gly Gly Leu His Leu Thr Leu Asp Trp Ala Val Arg Gly Leu 180 185 190 Leu Leu Leu Lys Thr Arg Leu 195 <210> 2 <211> 918 <212> PRT <213> Homo sapiens gp130 (UniProt P40189-1) <400> 2 Met Leu Thr Leu Gln Thr Trp Leu Val Gln Ala Leu Phe Ile Phe Leu 1 5 10 15 Thr Thr Glu Ser Thr Gly Glu Leu Leu Asp Pro Cys Gly Tyr Ile Ser 20 25 30 Pro Glu Ser Pro Val Val Gln Leu His Ser Asn Phe Thr Ala Val Cys 35 40 45 Val Leu Lys Glu Lys Cys Met Asp Tyr Phe His Val Asn Ala Asn Tyr 50 55 60 Ile Val Trp Lys Thr Asn His Phe Thr Ile Pro Lys Glu Gln Tyr Thr 65 70 75 80 Ile Ile Asn Arg Thr Ala Ser Ser Val Thr Phe Thr Asp Ile Ala Ser 85 90 95 Leu Asn Ile Gln Leu Thr Cys Asn Ile Leu Thr Phe Gly Gln Leu Glu 100 105 110 Gln Asn Val Tyr Gly Ile Thr Ile Ile Ser Gly Leu Pro Pro Glu Lys 115 120 125 Pro Lys Asn Leu Ser Cys Ile Val Asn Glu Gly Lys Lys Met Arg Cys 130 135 140 Glu Trp Asp Gly Gly Arg Glu Thr His Leu Glu Thr Asn Phe Thr Leu 145 150 155 160 Lys Ser Glu Trp Ala Thr His Lys Phe Ala Asp Cys Lys Ala Lys Arg 165 170 175 Asp Thr Pro Thr Ser Cys Thr Val Asp Tyr Ser Thr Val Tyr Phe Val 180 185 190 Asn Ile Glu Val Trp Val Glu Ala Glu Asn Ala Leu Gly Lys Val Thr 195 200 205 Ser Asp His Ile Asn Phe Asp Pro Val Tyr Lys Val Lys Pro Asn Pro 210 215 220 Pro His Asn Leu Ser Val Ile Asn Ser Glu Glu Leu Ser Ser Ile Leu 225 230 235 240 Lys Leu Thr Trp Thr Asn Pro Ser Ile Lys Ser Val Ile Ile Leu Lys 245 250 255 Tyr Asn Ile Gln Tyr Arg Thr Lys Asp Ala Ser Thr Trp Ser Gln Ile 260 265 270 Pro Pro Glu Asp Thr Ala Ser Thr Arg Ser Ser Phe Thr Val Gln Asp 275 280 285 Leu Lys Pro Phe Thr Glu Tyr Val Phe Arg Ile Arg Cys Met Lys Glu 290 295 300 Asp Gly Lys Gly Tyr Trp Ser Asp Trp Ser Glu Glu Ala Ser Gly Ile 305 310 315 320 Thr Tyr Glu Asp Arg Pro Ser Lys Ala Pro Ser Phe Trp Tyr Lys Ile 325 330 335 Asp Pro Ser His Thr Gln Gly Tyr Arg Thr Val Gln Leu Val Trp Lys 340 345 350 Thr Leu Pro Pro Phe Glu Ala Asn Gly Lys Ile Leu Asp Tyr Glu Val 355 360 365 Thr Leu Thr Arg Trp Lys Ser His Leu Gln Asn Tyr Thr Val Asn Ala 370 375 380 Thr Lys Leu Thr Val Asn Leu Thr Asn Asp Arg Tyr Leu Ala Thr Leu 385 390 395 400 Thr Val Arg Asn Leu Val Gly Lys Ser Asp Ala Ala Val Leu Thr Ile 405 410 415 Pro Ala Cys Asp Phe Gln Ala Thr His Pro Val Met Asp Leu Lys Ala 420 425 430 Phe Pro Lys Asp Asn Met Leu Trp Val Glu Trp Thr Thr Pro Arg Glu 435 440 445 Ser Val Lys Lys Tyr Ile Leu Glu Trp Cys Val Leu Ser Asp Lys Ala 450 455 460 Pro Cys Ile Thr Asp Trp Gln Gln Glu Asp Gly Thr Val His Arg Thr 465 470 475 480 Tyr Leu Arg Gly Asn Leu Ala Glu Ser Lys Cys Tyr Leu Ile Thr Val 485 490 495 Thr Pro Val Tyr Ala Asp Gly Pro Gly Ser Pro Glu Ser Ile Lys Ala 500 505 510 Tyr Leu Lys Gln Ala Pro Pro Ser Lys Gly Pro Thr Val Arg Thr Lys 515 520 525 Lys Val Gly Lys Asn Glu Ala Val Leu Glu Trp Asp Gln Leu Pro Val 530 535 540 Asp Val Gln Asn Gly Phe Ile Arg Asn Tyr Thr Ile Phe Tyr Arg Thr 545 550 555 560 Ile Ile Gly Asn Glu Thr Ala Val Asn Val Asp Ser Ser His Thr Glu 565 570 575 Tyr Thr Leu Ser Ser Leu Thr Ser Asp Thr Leu Tyr Met Val Arg Met 580 585 590 Ala Ala Tyr Thr Asp Glu Gly Gly Lys Asp Gly Pro Glu Phe Thr Phe 595 600 605 Thr Thr Pro Lys Phe Ala Gln Gly Glu Ile Glu Ala Ile Val Val Pro 610 615 620 Val Cys Leu Ala Phe Leu Leu Thr Thr Leu Leu Gly Val Leu Phe Cys 625 630 635 640 Phe Asn Lys Arg Asp Leu Ile Lys Lys His Ile Trp Pro Asn Val Pro 645 650 655 Asp Pro Ser Lys Ser His Ile Ala Gln Trp Ser Pro His Thr Pro Pro 660 665 670 Arg His Asn Phe Asn Ser Lys Asp Gln Met Tyr Ser Asp Gly Asn Phe 675 680 685 Thr Asp Val Ser Val Val Glu Ile Glu Ala Asn Asp Lys Lys Pro Phe 690 695 700 Pro Glu Asp Leu Lys Ser Leu Asp Leu Phe Lys Lys Glu Lys Ile Asn 705 710 715 720 Thr Glu Gly His Ser Ser Gly Ile Gly Gly Ser Ser Cys Met Ser Ser 725 730 735 Ser Arg Pro Ser Ile Ser Ser Ser Asp Glu Asn Glu Ser Ser Gln Asn 740 745 750 Thr Ser Ser Thr Val Gln Tyr Ser Thr Val Val His Ser Gly Tyr Arg 755 760 765 His Gln Val Pro Ser Val Gln Val Phe Ser Arg Ser Glu Ser Thr Gln 770 775 780 Pro Leu Leu Asp Ser Glu Glu Arg Pro Glu Asp Leu Gln Leu Val Asp 785 790 795 800 His Val Asp Gly Gly Asp Gly Ile Leu Pro Arg Gln Gln Tyr Phe Lys 805 810 815 Gln Asn Cys Ser Gln His Glu Ser Ser Pro Asp Ile Ser His Phe Glu 820 825 830 Arg Ser Lys Gln Val Ser Ser Val Asn Glu Glu Asp Phe Val Arg Leu 835 840 845 Lys Gln Gln Ile Ser Asp His Ile Ser Gln Ser Cys Gly Ser Gly Gln 850 855 860 Met Lys Met Phe Gln Glu Val Ser Ala Ala Asp Ala Phe Gly Pro Gly 865 870 875 880 Thr Glu Gly Gln Val Glu Arg Phe Glu Thr Val Gly Met Glu Ala Ala 885 890 895 Thr Asp Glu Gly Met Pro Lys Ser Tyr Leu Pro Gln Thr Val Arg Gln 900 905 910 Gly Gly Tyr Met Pro Gln 915 <21>- 3 <211> 422 <212> PRT <213> Homo sapiens IL-11RA (isoform HCR1, UniProt Q14626-1) <400> 3 Met Ser Ser Ser Cys Ser Gly Leu Ser Arg Val Leu Val Ala Val Ala 1 5 10 15 Thr Ala Leu Val Ser Ala Ser Ser Pro Cys Pro Gln Ala Trp Gly Pro 20 25 30 Pro Gly Val Gln Tyr Gly Gln Pro Gly Arg Ser Val Lys Leu Cys Cys 35 40 45 Pro Gly Val Thr Ala Gly Asp Pro Val Ser Trp Phe Arg Asp Gly Glu 50 55 60 Pro Lys Leu Leu Gln Gly Pro Asp Ser Gly Leu Gly His Glu Leu Val 65 70 75 80 Leu Ala Gln Ala Asp Ser Thr Asp Glu Gly Thr Tyr Ile Cys Gln Thr 85 90 95 Leu Asp Gly Ala Leu Gly Gly Thr Val Thr Leu Gln Leu Gly Tyr Pro 100 105 110 Pro Ala Arg Pro Val Val Ser Cys Gln Ala Ala Asp Tyr Glu Asn Phe 115 120 125 Ser Cys Thr Trp Ser Pro Ser Gln Ile Ser Gly Leu Pro Thr Arg Tyr 130 135 140 Leu Thr Ser Tyr Arg Lys Lys Thr Val Leu Gly Ala Asp Ser Gln Arg 145 150 155 160 Arg Ser Pro Ser Thr Gly Pro Trp Pro Cys Pro Gln Asp Pro Leu Gly 165 170 175 Ala Ala Arg Cys Val Val His Gly Ala Glu Phe Trp Ser Gln Tyr Arg 180 185 190 Ile Asn Val Thr Glu Val Asn Pro Leu Gly Ala Ser Thr Arg Leu Leu 195 200 205 Asp Val Ser Leu Gln Ser Ile Leu Arg Pro Asp Pro Pro Gln Gly Leu 210 215 220 Arg Val Glu Ser Val Pro Gly Tyr Pro Arg Arg Leu Arg Ala Ser Trp 225 230 235 240 Thr Tyr Pro Ala Ser Trp Pro Cys Gln Pro His Phe Leu Leu Lys Phe 245 250 255 Arg Leu Gln Tyr Arg Pro Ala Gln His Pro Ala Trp Ser Thr Val Glu 260 265 270 Pro Ala Gly Leu Glu Glu Val Ile Thr Asp Ala Val Ala Gly Leu Pro 275 280 285 His Ala Val Arg Val Ser Ala Arg Asp Phe Leu Asp Ala Gly Thr Trp 290 295 300 Ser Thr Trp Ser Pro Glu Ala Trp Gly Thr Pro Ser Thr Gly Thr Ile 305 310 315 320 Pro Lys Glu Ile Pro Ala Trp Gly Gln Leu His Thr Gln Pro Glu Val 325 330 335 Glu Pro Gln Val Asp Ser Pro Ala Pro Pro Arg Pro Ser Leu Gln Pro 340 345 350 His Pro Arg Leu Leu Asp His Arg Asp Ser Val Glu Gln Val Ala Val 355 360 365 Leu Ala Ser Leu Gly Ile Leu Ser Phe Leu Gly Leu Val Ala Gly Ala 370 375 380 Leu Ala Leu Gly Leu Trp Leu Arg Leu Arg Arg Gly Gly Lys Asp Gly 385 390 395 400 Ser Pro Lys Pro Gly Phe Leu Ala Ser Val Ile Pro Val Asp Arg Arg 405 410 415 Pro Gly Ala Pro Asn Leu 420 <210> 4 <211> 390 <212> PRT <213> Homo sapiens IL-11RA (isoform HCR2, UniProt Q14626-2) <400> 4 Met Ser Ser Ser Cys Ser Gly Leu Ser Arg Val Leu Val Ala Val Ala 1 5 10 15 Thr Ala Leu Val Ser Ala Ser Ser Pro Cys Pro Gln Ala Trp Gly Pro 20 25 30 Pro Gly Val Gln Tyr Gly Gln Pro Gly Arg Ser Val Lys Leu Cys Cys 35 40 45 Pro Gly Val Thr Ala Gly Asp Pro Val Ser Trp Phe Arg Asp Gly Glu 50 55 60 Pro Lys Leu Leu Gln Gly Pro Asp Ser Gly Leu Gly His Glu Leu Val 65 70 75 80 Leu Ala Gln Ala Asp Ser Thr Asp Glu Gly Thr Tyr Ile Cys Gln Thr 85 90 95 Leu Asp Gly Ala Leu Gly Gly Thr Val Thr Leu Gln Leu Gly Tyr Pro 100 105 110 Pro Ala Arg Pro Val Val Ser Cys Gln Ala Ala Asp Tyr Glu Asn Phe 115 120 125 Ser Cys Thr Trp Ser Pro Ser Gln Ile Ser Gly Leu Pro Thr Arg Tyr 130 135 140 Leu Thr Ser Tyr Arg Lys Lys Thr Val Leu Gly Ala Asp Ser Gln Arg 145 150 155 160 Arg Ser Pro Ser Thr Gly Pro Trp Pro Cys Pro Gln Asp Pro Leu Gly 165 170 175 Ala Ala Arg Cys Val Val His Gly Ala Glu Phe Trp Ser Gln Tyr Arg 180 185 190 Ile Asn Val Thr Glu Val Asn Pro Leu Gly Ala Ser Thr Arg Leu Leu 195 200 205 Asp Val Ser Leu Gln Ser Ile Leu Arg Pro Asp Pro Pro Gln Gly Leu 210 215 220 Arg Val Glu Ser Val Pro Gly Tyr Pro Arg Arg Leu Arg Ala Ser Trp 225 230 235 240 Thr Tyr Pro Ala Ser Trp Pro Cys Gln Pro His Phe Leu Leu Lys Phe 245 250 255 Arg Leu Gln Tyr Arg Pro Ala Gln His Pro Ala Trp Ser Thr Val Glu 260 265 270 Pro Ala Gly Leu Glu Glu Val Ile Thr Asp Ala Val Ala Gly Leu Pro 275 280 285 His Ala Val Arg Val Ser Ala Arg Asp Phe Leu Asp Ala Gly Thr Trp 290 295 300 Ser Thr Trp Ser Pro Glu Ala Trp Gly Thr Pro Ser Thr Gly Thr Ile 305 310 315 320 Pro Lys Glu Ile Pro Ala Trp Gly Gln Leu His Thr Gln Pro Glu Val 325 330 335 Glu Pro Gln Val Asp Ser Pro Ala Pro Pro Arg Pro Ser Leu Gln Pro 340 345 350 His Pro Arg Leu Leu Asp His Arg Asp Ser Val Glu Gln Val Ala Val 355 360 365 Leu Ala Ser Leu Gly Ile Leu Ser Phe Leu Gly Leu Val Ala Gly Ala 370 375 380 Leu Ala Leu Gly Leu Trp 385 390 <210> 5 <211> 516 <212> PRT <213> artificial sequence <220> <223> IL-11:IL-11Ralpha fusion protein <400> 5 Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly 1 5 10 15 Val His Ser Pro Gln Ala Trp Gly Pro Pro Gly Val Gln Tyr Gly Gln 20 25 30 Pro Gly Arg Ser Val Lys Leu Cys Cys Pro Gly Val Thr Ala Gly Asp 35 40 45 Pro Val Ser Trp Phe Arg Asp Gly Glu Pro Lys Leu Leu Gln Gly Pro 50 55 60 Asp Ser Gly Leu Gly His Glu Leu Val Leu Ala Gln Ala Asp Ser Thr 65 70 75 80 Asp Glu Gly Thr Tyr Ile Cys Gln Thr Leu Asp Gly Ala Leu Gly Gly 85 90 95 Thr Val Thr Leu Gln Leu Gly Tyr Pro Pro Ala Arg Pro Val Val Ser 100 105 110 Cys Gln Ala Ala Asp Tyr Glu Asn Phe Ser Cys Thr Trp Ser Pro Ser 115 120 125 Gln Ile Ser Gly Leu Pro Thr Arg Tyr Leu Thr Ser Tyr Arg Lys Lys 130 135 140 Thr Val Leu Gly Ala Asp Ser Gln Arg Arg Ser Pro Ser Thr Gly Pro 145 150 155 160 Trp Pro Cys Pro Gln Asp Pro Leu Gly Ala Ala Arg Cys Val Val His 165 170 175 Gly Ala Glu Phe Trp Ser Gln Tyr Arg Ile Asn Val Thr Glu Val Asn 180 185 190 Pro Leu Gly Ala Ser Thr Arg Leu Leu Asp Val Ser Leu Gln Ser Ile 195 200 205 Leu Arg Pro Asp Pro Pro Gln Gly Leu Arg Val Glu Ser Val Pro Gly 210 215 220 Tyr Pro Arg Arg Leu Arg Ala Ser Trp Thr Tyr Pro Ala Ser Trp Pro 225 230 235 240 Cys Gln Pro His Phe Leu Leu Lys Phe Arg Leu Gln Tyr Arg Pro Ala 245 250 255 Gln His Pro Ala Trp Ser Thr Val Glu Pro Ala Gly Leu Glu Glu Val 260 265 270 Ile Thr Asp Ala Val Ala Gly Leu Pro His Ala Val Arg Val Ser Ala 275 280 285 Arg Asp Phe Leu Asp Ala Gly Thr Trp Ser Thr Trp Ser Pro Glu Ala 290 295 300 Trp Gly Thr Pro Ser Thr Gly Gly Pro Ala Gly Gln Ser Gly Gly Gly 305 310 315 320 Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Val Pro Gly Pro Pro 325 330 335 Pro Gly Pro Pro Arg Val Ser Pro Asp Pro Arg Ala Glu Leu Asp Ser 340 345 350 Thr Val Leu Leu Thr Arg Ser Leu Leu Ala Asp Thr Arg Gln Leu Ala 355 360 365 Ala Gln Leu Arg Asp Lys Phe Pro Ala Asp Gly Asp His Asn Leu Asp 370 375 380 Ser Leu Pro Thr Leu Ala Met Ser Ala Gly Ala Leu Gly Ala Leu Gln 385 390 395 400 Leu Pro Gly Val Leu Thr Arg Leu Arg Ala Asp Leu Leu Ser Tyr Leu 405 410 415 Arg His Val Gln Trp Leu Arg Arg Ala Gly Gly Ser Ser Leu Lys Thr 420 425 430 Leu Glu Pro Glu Leu Gly Thr Leu Gln Ala Arg Leu Asp Arg Leu Leu 435 440 445 Arg Arg Leu Gln Leu Leu Met Ser Arg Leu Ala Leu Pro Gln Pro Pro 450 455 460 Pro Asp Pro Pro Ala Pro Pro Leu Ala Pro Pro Ser Ser Ala Trp Gly 465 470 475 480 Gly Ile Arg Ala Ala His Ala Ile Leu Gly Gly Leu His Leu Thr Leu 485 490 495 Asp Trp Ala Val Arg Gly Leu Leu Leu Leu Lys Thr Arg Leu His His 500 505 510 His His His His 515 <210> 6 <211> 1575 <212> DNA <213> artificial sequence <220> <223> Nucleotide sequence encoding IL-11:IL-11Ralpha fusion protein <400> 6 gaattcccgc cgccaccatg ggctggtcct gcatcatcct gtttctggtg gccacagcca 60 ccggcgtgca ctctccacag gcttggggac ctccaggcgt gcagtatggc cagcctggca 120 gatccgtgaa gctgtgctgt cctggcgtga cagctggcga ccctgtgtcc tggttcagag 180 atggcgagcc caagctgctg cagggcccag attctggact gggccacgaa ctggtgctgg 240 cccaggccga ttctaccgac gagggcacct acatctgcca gaccctggat ggcgccctgg 300 gcggaacagt gacactgcag ctgggctacc ctcccgccag acctgtggtg tcttgtcagg 360 ccgccgacta cgagaacttc agctgcacat ggtcccccag ccagatcagc ggcctgccca 420 ccagatacct gaccagctac cggaagaaaa ccgtgctggg cgccgacagc cagagaagaa gcccttctac aggcccctgg ccctgccctc aggtcctct gggagctgcc aggtgtgg 540 tgcacggcgc cgagttctgg tcccagtacc ggatcaacgt gaccgaagtg aaccccctgg 600 gcgcctccac aagactgctg gatgtgtccc tgcagagcat cctgcggccc gatcctccac 660 agggcctgag agtggaaagc gtgcccggct accccagaag gctgagagcc agctggacat 720 accccgcctc ttggccttgc cagccccact tcctgctgaa gtttcggctg cagtaccggc 780 cagcccagca ccctgcttgg agcacagtgg aacctgccgg cctggaaga gtgatcacag acgccgtggc cggactgcct catgctgtgc gggtgtccgc cagagacttt ctggatgccg900 gcacctggtc tacctggtcc ccagaagcct ggggcacacc ttctactggc ggacctgctg 960 gacagtctgg cggaggcgga ggaagtggcg gaggatcagg gggaggatct gtgcctggac ctcctccagg accccctaga gtgtccccag atcctagggc cgagctggac tctaccgtgc 1080 tgctgaccag atccctgctg gccgacacaa ggcagctggc tgcccagctg agagacaagt 1140 tgctgaccag atccctgctg gccgacacaa ggcagctggc tgcccagctg agagacaagt 1140 tccccgccga cggcgaccac aacctggata gcctgcctac cctggccatg tctgctggcg 1200 tccccgccga cggcgaccac aacctggata gcctgcctac cctggccatg tctgctggcg 1200 cactgggggc tctgcagctg cctggggtgc tgactagact gagagccgac ctgctgagct 1260 cactgggggc tctgcagctg cctggggtgc tgactagact gagagccgac ctgctgagct 1260 acctgcggca tgtgcagtgg ctgagaaggg ctggcggcag cagcctgaaa accctggaac 1320 acctgcggca tgtgcagtgg ctgagaaggg ctggcggcag cagcctgaaa accctggaac 1320 ctgagctggg cacactgcag gccagactgg acagactgct gcgcagactg cagctgctga 1380 ctgagctggg cacactgcag gccagactgg acagactgct gcgcagactg cagctgctga 1380 tgagcagact ggctctgccc cagcctcctc ctgaccctcc tgctcctcca ctggctcctc 1440 tgagcagact ggctctgccc cagcctcctc ctgaccctcc tgctcctcca ctggctcctc 1440 caagctctgc ttggggcgga attagagccg cccacgccat tctgggaggc ctgcacctga 1500 caagctctgc ttggggcgga attagagccg cccacgccat tctgggaggc ctgcacctga 1500 cactggattg ggcagtgcgg ggcctgctgc tgctgaaaac cagactgcac caccaccatc 1560 cactggattg ggcagtgcgg ggcctgctgc tgctgaaaac cagactgcac caccaccatc 1560 accactgata agctt 1575 accactgata agctt 1575 <210> 7<210> 7 <211> 117<211> 117 <212> PRT <212> PRT <213> 人工序列(artificial sequence) <213> Artificial Sequence <220> <220> <223> 9A7 VH <223> 9A7 VH <400> 7 <400> 7 Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Arg Pro Gly Ser Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Arg Pro Gly Ser 1 5 10 15 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr 20 25 30 Trp Met His Trp Leu Lys Gln Arg Pro Val Gln Gly Leu Glu Trp Ile 35 40 45 Gly Asn Ile Gly Pro Ser Asp Ser Lys Thr His Tyr Asn Gln Lys Phe 50 55 60 Lys Asp Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Gln Leu Asn Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Asp Tyr Val Leu Phe Thr Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ala 115 <210> 8 <211> 117 <212> PRT <213> artificial sequence <220> <223> 9A7 VH 1 <400> 8 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr 20 25 30 Trp Met His Trp Leu Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Asn Ile Gly Pro Ser Asp Ser Lys Thr His Tyr Asn Gln Lys Phe 50 55 60 Lys Asp Arg Ala Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Gln Leu Asn Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Asp Tyr Val Leu Phe Thr Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 9 <211> 117 <212> PRT <213> artificial sequence <220> <223> 9A7 VH 2 <400> 9 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr 20 25 30 Trp Met His Trp Leu Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Asn Ile Gly Pro Ser Asp Ser Lys Thr His Tyr Asn Gln Lys Phe 50 55 60 Lys Asp Arg Ala Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Asp Tyr Val Leu Phe Thr Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 10 <211> 117 <212> PRT <213> artificial sequence <220> <223> 9A7 VH 3 <400> 10 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 1 50 55 60 Lys Asp Arg Ala Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Asp Tyr Val Leu Phe Thr Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 11 <211> 117 <212> PRT <213> artificial sequence <220> <223> 9A7 VH 4 <400> 11 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr 20 25 30 Trp Met His Trp Leu Arg Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Asn Ile Gly Pro Ser Asp Ser Lys Thr His Tyr Asn Gln Lys Phe 50 55 60 Lys Asp Arg Val Thr Met Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Asp Tyr Val Leu Phe Thr Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 12 <211> 117 <212> PRT <213> artificial sequence <220> <223> 9A7 VH 5 <400> 12 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr 20 25 30 Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Asn Ile Gly Pro Ser Asp Ser Lys Thr His Tyr Asn Gln Lys Phe 50 55 60 Gln Asp Arg Val Thr Met Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Asp Tyr Val Leu Phe Thr Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 13 <211> 107 <212> PRT <213> artificial sequence <220> <223> 9A7 VL <400> 13 Asp Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Met Thr Pro Gly 1 5 10 15 Asp Ser Val Ser Leu Ser Cys Arg Ala Ser Gln Ser Ile Ser Asn Asn 20 25 30 Leu His Trp Tyr Gln Gln Lys Ser His Glu Ser Pro Arg Leu Leu Ile 35 40 45 Lys Tyr Ala Ser Gln Ser Ile Ser Gly Ile Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Ser Phe Asn Ser Val Glu Thr 65 70 75 80 Glu Asp Phe Gly Val Tyr Phe Cys Gln Gln Ser Tyr Ser Trp Pro Leu 85 90 95 Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys 100 105 <210> 14 <211> 107 <212> PRT <213> Artificial sequence <220> <223> 9A7 VL 1 <400> 14 Asp Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Ile Ser Asn Asn 20 25 30 Leu His Trp Tyr Gln Gln Lys Ser His Glu Ala Pro Arg Leu Leu Ile 35 40 45 Lys Tyr Ala Ser Gln Ser Ile Ser Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Ser Phe Ser Ser Leu Glu Thr 65 70 75 80 Glu Asp Phe Ala Val Tyr Phe Cys Gln Gln Ser Tyr Ser Trp Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 15 <211> 107 <212> PRT <213> Artificial sequence <220> <223> 9A7 VL 2 <400> 15 Asp Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Ile Ser Asn Asn 20 25 30 Leu His Trp Tyr Gln Gln Lys Ser Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45 Lys Tyr Ala Ser Gln Ser Ile Ser Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Thr 65 70 75 80 Glu Asp Phe Ala Val Tyr Phe Cys Gln Gln Ser Tyr Ser Trp Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 16 <211> 107 <212> PRT <213> Artificial sequence <220> <223> 9A7 VL 3 <400> 16 Asp Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Ile Ser Asn Asn 20 25 30 Leu His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45 Lys Tyr Ala Ser Gln Ser Ile Ser Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro 65 70 75 80 Glu Asp Phe Ala Val Tyr Phe Cys Gln Gln Ser Tyr Ser Trp Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 17 <211> 107 <212> PRT <213> artificial sequence <220> <223> 9A7 VL 4 <400> 17 Asp Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Ile Ser Asn Asn 20 25 30 Leu His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45 Lys Tyr Ala Ser Gln Ser Ile Ser Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro 65 70 75 80 Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Ser Tyr Ser Trp Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 18 <211> 5 <212> PRT <213> Artificial sequence <220> <223> 9A7 VH, 9A7 VH 1, 9A7 VH 2, 9A7 VH 3, 9A7 VH 4, 9A7 VH 5 HC-CDR1 <400> 18 Asn Tyr Trp Met His 1 5 <210> 19 <211> 17 <212> PRT <213> Artificial sequence <220> <223> 9A7 VH, 9A7 VH 1, 9A7 VH 2, 9A7 VH 3, 9A7 VH 4 HC-CDR2 <400> 19 Asn Ile Gly Pro Ser Asp Ser Lys Thr His Tyr Asn Gln Lys Phe Lys 1 5 10 15 Asp <210> 20 <211> 17 <212> PRT <213> Artificial sequence <220> <223> 9A7 VH 5 HC-CDR2 <400> 20 Asn Ile Gly Pro Ser Asp Ser Lys Thr His Tyr Asn Gln Lys Phe Gln 1 5 10 15 Asp <210> 21 <211> 8 <212> PRT <213> Artificial sequence <220> <223> 9A7 VH, 9A7 VH 1, 9A7 VH 2, 9A7 VH 3, 9A7 VH 4, 9A7 VH 5 HC-CDR3 <400> 21 Gly Asp Tyr Val Leu Phe Thr Tyr 1 5 <210> 22 <211> 11 <212> PRT <213> Artificial sequence <220> <223> 9A7 VL, 9A7 VL 1, 9A7 VL 2, 9A7 VL 3, 9A7 VL 4 LC-CDR1 <400> 22 Arg Ala Ser Gln Ser Ile Ser Asn Asn Leu His 1 5 10 <210> 23 <211> 7 <212> PRT <213> Artificial sequence <220> <223> 9A7 VL, 9A7 VL 1, 9A7 VL 2, 9A7 VL 3, 9A7 VL 4 LC-CDR2 <400> 23 Tyr Ala Ser Gln Ser Ile Ser 1 5 <210> 24 <211> 9 <212> PRT <213> Artificial sequence <220> <223> 9A7 VL, 9A7 VL 1, 9A7 VL 2, 9A7 VL 3, 9A7 VL 4 LC-CDR3 <400> 24 Gln Gln Ser Tyr Ser Trp Pro Leu Thr 1 5 <210> 25 <211> 30 <212> PRT <213> Artificial sequence <220> <223> 9A7 VH HC-FR1 <400> 25 Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Arg Pro Gly Ser 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr 20 25 30 <210> 26 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> 9A7 VH 1 HC-FR1 <400> 26 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr 20 25 30 <210> 27 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> 9A7 VH 2 HC-FR1 <400> 27 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr 20 25 30 <210> 28 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> 9A7 VH 3, 9A7 VH , 9A7 VH 5 HC-FR1 <400> 28 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 It should be noted that there seems to be a typo in the original text where "9A7 VH 4" is missing a space between "VH" and "4" in the tag <48>. This has been maintained in the translation for consistency with the original.Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr 20 25 30 <210> 29 <211> 14 <212> PRT <213> artificial sequence <220> <223> 9A7 VH HC-FR2 <400> 29 Trp Leu Lys Gln Arg Pro Val Gln Gly Leu Glu Trp Ile Gly 1 5 10 <210> 30 <211> 14 <212> PRT <213> artificial sequence <220> <223> 9A7 VH 1, 9A7 VH 2 HC-FR2 <400> 30 Trp Leu Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile Gly 1 5 10 <210> 31 <211> 14 <212> PRT <213> artificial sequence <220> <223> 9A7 VH 3, 9A7 VH 4 HC-FR2 <400> 31 Trp Leu Arg Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile Gly 1 5 10 <210> 32 <211> 14 <212> PRT <213> Artificial sequence <220> <223> 9A7 VH 5 HC-FR2 <400> 32 Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile Gly 1 5 10 <210> 33 <211> 32 <212> PRT <213> Artificial sequence <220> <223> 9A7 VH HC-FR3 <400> 33 Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr Met Gln 1 5 10 15 Leu Asn Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys Ala Arg 20 25 30 <210> 34 <211> 32 <212> PRT <213> Artificial sequence <220> <223> 9A7 VH 1 HC-FR3 <400> 34 Arg Ala Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr Met Gln 1 5 10 15 Leu Asn Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys Ala Arg 20 25 30 <210> 35 <211> 32 <212> PRT <213> Artificial sequence <220> <223> 9A7 VH 2, 9A7 VH 3 HC - FR3 <400> 35 Arg Ala Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr Met Glu 1 5 10 15 Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg 20 25 30 <210> 36 <211> 32 <212> PRT <213> Artificial sequence <220> <223> 9A7 VH 4, 9A7 VH 5 HC - FR3 <400> 36 Arg Val Thr Met Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr Met Glu 1 5 10 15 Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg 20 25 30 <210> 37 <211> 11 <212> PRT <213> Artificial sequence <220> <223> 9A7 VH HC - FR4 <400> 37 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ala 1 5 10 <210> 38 <211> 11 <212> PRT <213> Artificial sequence <220> <223> 9A7 VH 1, 9A7 VH 2, 9A7 VH 3, 9A7 VH 4, 9A7 VH 5 HC-FR4 <400> 38 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 1 5 10 <210> 39 <211> 23 <212> PRT <213> Artificial sequence <220> <223> 9A7 VL LC-FR1 <400> 39 Asp Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Met Thr Pro Gly 1 5 10 15 Asp Ser Val Ser Leu Ser Cys 20 <210> 40 <211> 23 <212> PRT <213> Artificial sequence <220> <223> 9A7 VL 1, 9A7 VL 2, 9A7 VL 3, 9A7 VL 4 LC-FR1 <400> 40 Asp Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys 20 <210> 41 <211> 15 <212> PRT <213> artificial sequence <220> <223> 9A7 VL LC-FR2 <400> 41 Trp Tyr Gln Gln Lys Ser His Glu Ser Pro Arg Leu Leu Ile Lys 1 5 10 15 <210> 42 <211> 15 <212> PRT <213> artificial sequence <220> <223> 9A7 VL 1 LC-FR2 <400> 42 Trp Tyr Gln Gln Lys Ser His Glu Ala Pro Arg Leu Leu Ile Lys 1 5 10 15 <210> 43 <211> 15 <212> PRT <213> artificial sequence <220> <223> 9A7 VL 2 LC-FR2 <400> 43 Trp Tyr Gln Gln Lys Ser Gly Gln Ala Pro Arg Leu Leu Ile Lys 1 5 10 15 <210> 44 <211> 15 <212> PRT <213> artificial sequence <220> <223> 9A7 VL 3, 9A7 VL 4 LC-FR2 <400> 44 Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Lys 1 5 10 15 <210> 45 <211> 32 <212> PRT <213> Artificial sequence <220> <223> 9A7 VL LC-FR3 <400> 45 Gly Ile Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr 1 5 10 15 Leu Ser Phe Asn Ser Val Glu Thr Glu Asp Phe Gly Val Tyr Phe Cys 20 25 30 <210> 46 <211> 32 <212> PRT <213> Artificial sequence <220> <223> 9A7 VL 1 LC-FR3 <400> 46 Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr 1 5 10 15 Leu Ser Phe Ser Ser Leu Glu Thr Glu Asp Phe Ala Val Tyr Phe Cys 20 25 30 <210> 47 <211> 32 <212> PRT <213> Artificial sequence <220> <223> 9A7 VL 2 LC-FR3 <400> 47 Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr 1 5 10 15 Leu Thr Ile Ser Ser Leu Glu Thr Glu Asp Phe Ala Val Tyr Phe Cys 20 25 30 <210> 48 <211> 32 <212> PRT <213> Artificial sequence <220> <223> 9A7 VL 3 LC-FR3 <400> 48 Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr 1 5 10 15 Leu Thr Ile Ser Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Phe Cys 20 25 30 <210> 49 <211> 32 1 5 10 15 Leu Thr Ile Ser Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Phe Cys 20 25 30 <210> 49 <211> 32 <212> PRT <213> Artificial sequence <220> <223> 9A7 VL 4 LC-FR3 <400> 49 Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr 1 5 10 15 [[ID=

[51] ]Leu Thr Ile Ser Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys 20 25 30 <210> 50 <211> 10 <212> PRT <213> artificial sequence <220> <223> 9A7 VL LC-FR4 <400> 50 Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys 1 5 10 <210> 51 <211> 10 <212> PRT <213> artificial sequence <220> <223> 9A7 VL 1, 9A7 VL 2, 9A7 VL 3, 9A7 VL 4 LC-FR4 <400> 51 Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 1 5 10 <210> 52 <211> 17 <212> PRT <213> artificial sequence <220> <223> 9A7 VH HC-CDR2 consensus sequence <220> <221> MISC_FEATURE <222> (16)..(16) <223> Xaa - King or Queen <400> 52 Asn Ile Gly Pro Ser Asp Ser Lys Thr His Tyr Asn Gln Lys Phe Xaa 1 5 10 15 Asp <210> 53 <211> 330 <212> PRT <213> Constant region of Homo sapiens IgG1 (IGHG1; UniProt:P01857-1, v1) <400> 53 Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys 1 5 10 15 Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 20 25 30 Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 35 40 45 Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 50 55 60 Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr 65 70 75 80 Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys 85 90 95 Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys 100 105 110 Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro 115 120 125 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 130 135 140 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 145 150 155 160 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 165 170 175 Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 180 185 190 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 195 200 205 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 210 215 220 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu 225 230 235 240 Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 245 250 255 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 260 265 270 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 275 280 285 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 290 295 300 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 305 310 315 320 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 325 330 <210> 54 <211> 98 <212> PRT <213> Artificial sequence <22​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​ <211> 12 <212> PRT <213> artificial sequence <220> <223> Hinge IgG1 (positions 99-110 of P01857-1, v1) <400> 55 Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro 1 5 10 <210> 56 <211> 113 <212> PRT <213> artificial sequence <220> <223> CH2 IgG1 (positions 111-223 of P01857-1, v1) <400> 56 Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe 1 5 10 15 Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val 20 25 30 Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe 35 40 45 Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro 50 55 60 Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Tyr Arg Val Val Ser Val Leu Thr 65 70 75 80 Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val 85 90 95 Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala 100 105 110 Lys <210> 57 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> CH3 IgG1 (positions 224 - 330 of P01857 - 1, v1) <400> 57 Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp 1 5 10 15 Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe 20 25 30 Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu<00​​​​​​​​​​​​Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 100 105 <210> 58 <211> 107 <212> PRT <213> artificial sequence <220> <223> Ckappa CL (IGCK; UniProt: P01834-1, v2) <400> 58 Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu 1 5 10 15 Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe 20 25 30 Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln 35 40 45 Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser 50 55 60 Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu 65 70 75 80 Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser 85 90 95 Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 100 105 <210> 59 <211> 107 <212> PRT <213> Artificial sequence <220> <223> 9A7O VL <400> 59 Asp Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Met Thr Pro Gly 1 5 10 15 Asp Ser Val Ser Leu Ser Cys Arg Ala Ser Gln Ser Ile Ser Asn Asn 20 25 30 Leu His Trp Tyr Gln Gln Lys Ser His Glu Ser Pro Arg Leu Leu Ile 35 40 45 Lys Tyr Ala Ser Gln Ser Ile Ser Gly Ile Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Ser Phe Asn Ser Val Glu Thr 65 70 75 80 Glu Asp Phe Gly Val Tyr Phe Cys Gln Gln Arg Tyr Ser Trp Pro Leu 85 90 95 Thr Phe Gly Ala Gly Thr Lys Leu Glu Met Lys 100 105 <210> 60 <211> 九 <212> PRT <213> Artificial sequence <220> <223> 9A7O VL LC-CDR3 <400> 60 Gln Gln Arg Tyr Ser Trp Pro Leu Thr 1 5 <210> 61 <211> 327 <212> PRT <213> Constant region of Homo sapiens IgG4 (IGHG4; UniProt: P01861, v1) <400> 61 Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg 1 5 10 15 Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 20 25 30 Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 35 40 45 Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 50 55 60 Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr 65 70 75 80 Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys 85 90 95 Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Ser Cys Pro Ala Pro 100 105 110 Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 115 120 125 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 130 135 140 Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp 145 150 155 160 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe 165 170 175 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 180 185 190 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu 195 200 205 Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 210 215 220 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys 225 230 235 240 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 245 250 255 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 260 265 270 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 275 280 285 Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser 290 295 300 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 305 310 315 320 Leu Ser Leu Ser Leu Gly Lys 325 <210> 62 <211> 98 <212> PRT <213> artificial sequence <220> <223> CH1 IgG4 (positions 1 - 98 of P01861, v1) <400> 62 Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg 1 5 10 15 Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 20 25 30 Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 35 40 45 Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 50 55 60 Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr 65 70 75 80 Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys 85 90 95 Arg Val <210> 63 <211> 12 <212> PRT <213> artificial sequence <220> <223> Hinge IgG4 (positions 99-110 of P01861, v1) <400> 63 Glu Ser Lys Tyr Gly Pro Pro Cys Pro Ser Cys Pro 1 5 10 <210> 64 <211> 110 <212> PRT <213> artificial sequence <220> <223> CH2 IgG4 (positions 111-220 of P01861, v1) <400> 64 Ala Pro Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 1 5 10 15 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 20 25 30 Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr 35 40 45 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 50 55 60 Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 65 70 75 80 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 85 90 95 Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys 100 105 110 <210> 65 <211> 107​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr 85 90 95 Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys 100 105 <210> 66 <211> 327 <212> PRT <213> Homo sapiens IgG4 constant region (IGHG4; UniProt: P01861, v1; <400> 66 Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg 1 5 10 15 Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 20 25 30 Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 35 40 45 Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 50 55 60 Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr 65 70 75 80 Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys 85 90 95 Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro 100 105 110 Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 115 120 125 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 130 135 140 Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp 145 150 155 160 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe 165 170 175 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 180 185 190 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu 195 200 205 Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 210 215 220 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys 225 230 235 240 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 245 250 255 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 260 265 270 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 275 280 285 Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser 290 295 300 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 305 310 315 320 Leu Ser Leu Ser Leu Gly Lys 325 <210> 67 <211> 12 <212> PRT <213> artificial sequence <220> <223> Hinge IgG4 (positions 99-110 of P01861, v1; S241P) <400> 67 Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro 1 5 10 <210> 68 <211> 327 <212> PRT <213> Homo sapiens IgG4 constant region (IGHG4; UniProt: P01861, v1; <400> 68 Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg 1 5 10 15 Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 20 25 30 Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 35 40 45 Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 50 55 60 Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr 65 70 75 80 Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys 85 90 95 Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro 100 105 110 Glu Phe Glu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 115 120 125 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 130 135 140 Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp 145 150 155 160 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe 165 170 175 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 180 185 190 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu 195 200 205 Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 210 215 220 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys 225 230 235 240 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 245 250 255 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 260 265 270 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 275 280 285 Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser 290 295 300 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 305 310 315 320 Leu Ser Leu Ser Leu Gly Lys 325 <210> 69 <211> 110 <212> PRT <213> Artificial sequence <220> <223> CH2 IgG4 (amino acids 111 - 220 of P01861, v1; L248E) <400> 69 Ala Pro Glu Phe Glu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 1 5 10 15 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 20 25 30 Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr 35 40 45 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 50 55 60 Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 65 70 75 80 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 85 90 95 Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys 100 105 110 <210> 70 <211> 447 <212> PRT <213> Artificial Sequence <220> <223> 9A7 VH4 - Human IgG1 Constant Region (IGHG1; UniProt:P01857 - 1, v1) <400> 70 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr 20 25 30 Trp Met His Trp Leu Arg Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Asn Ile Gly Pro Ser Asp Ser Lys Thr His Tyr Asn Gln Lys Phe 50 55 60 Lys Asp Arg Val Thr Met Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Asp Tyr Val Leu Phe Thr Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu 115 120 125 Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys 130 135 140 Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser 145 150 155 160 Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser 165 170 175 Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser 180 185 190 Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn 195 200 205 Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His 210 215 220 Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val 225 230 235 240 Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr 245 250 255 Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu 260 265 270 Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys 275 280 285 Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser 290 295 300 Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys 305 310 315 320 Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile 325 330 335 Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro 340 345 350 Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu 355 360 365 Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn 370 375 380 Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser 385 390 395 400 Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg 405 410 415 Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu 420 425 430 His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> 71 <211> 444 <212> PRT <213> Artificial Sequence <220> <223> 9A7 VH4 - Human IgG4 Constant Region (IGHG4; UniProt: P01861, v1) <400> 71 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr 20 25 30 Trp Met His Trp Leu Arg Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Asn Ile Gly Pro Ser Asp Ser Lys Thr His Tyr Asn Gln Lys Phe 50 55 60 Lys Asp Arg Val Thr Met Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Asp Tyr Val Leu Phe Thr Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu 115 120 125 Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys 130 135 140 Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser 145 150 155 160 Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser 165 170 175 Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser 180 185 190 Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn 195 200 205 Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro 210 215 220 Ser Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe 225 230 235 240 Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val 245 250 2...

Claims

1. An antigen-binding molecule capable of binding to IL-11Rα, wherein the antigen-binding molecule comprises: (i) a heavy chain variable region (VH) comprising the following CDRs: HC-CDR1 with the amino acid sequence shown in SEQ ID NO: 18 HC-CDR2 with the amino acid sequence shown in SEQ ID NO: 19 HC-CDR3 having an amino acid sequence as shown in SEQ ID NO: 21; and (ii) a light chain variable region (VL) comprising the following CDRs: LC-CDR1 with the amino acid sequence shown in SEQ ID NO: 22 LC-CDR2 with the amino acid sequence shown in SEQ ID NO: 23 The amino acid sequence of LC-CDR3 is shown in SEQ ID NO:

24.

2. The antigen-binding molecule according to claim 1, wherein 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: 7, 8, 9, 10 or 11; and A VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 13, 14, 15, 16 or 17.

3. The antigen-binding molecule according to claim 1 or 2, wherein 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: 11; and A VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:

17.

4. The antigen-binding molecule according to claim 1 or 2, wherein The antigen binding molecule is capable of inhibiting IL-11-mediated signal transduction.

5. An antigen-binding molecule comprising (i) the antigen-binding molecule according to any one of claims 1 to 4, and (ii) an antigen-binding molecule capable of binding to an antigen other than IL-11Rα.

6. The antigen-binding molecule according to claim 1, 2 or 5, wherein The antigen binding molecule is capable of inhibiting the interaction between IL-11Rα or a complex containing IL-11Rα and an interacting partner of IL-11Rα or a complex containing IL-11Rα.

7. A chimeric antigen receptor (CAR) comprising the antigen binding molecule according to any one of claims 1 to 6.

8. A nucleic acid encoding one or more of the antigen binding molecules according to any one of claims 1 to 6 or the CAR according to claim 7.

9. One or more expression vectors comprising one or more nucleic acids of claim 8.

10. A cell comprising the antigen binding molecule of any one of claims 1 to 6, the CAR of claim 7, the one or more nucleic acids of claim 8, or the one or more expression vectors of claim 9, wherein the cell does not include a plant cell.

11. A method comprising culturing a cell comprising one or more nucleic acids of claim 8, or one or more expression vectors of claim 9, under conditions suitable for expressing an antigen binding molecule or CAR from the one or more nucleic acids or one or more expression vectors.

12. A composition comprising the antigen binding molecule of any one of claims 1 to 6, the CAR of claim 7, one or more nucleic acids of claim 8, one or more expression vectors of claim 9 or the cell of claim 10.

13. Use of the antigen binding molecule of any one of claims 1 to 6, the CAR of claim 7, the one or more nucleic acids of claim 8, the one or more expression vectors of claim 9, the cell of claim 10 or the composition of claim 12 for the preparation of a medicament for use in a method for treating cancer.

14. Use of the antigen binding molecule of any one of claims 1 to 6, the CAR of claim 7, the one or more nucleic acids of claim 8, the one or more expression vectors of claim 9, the cell of claim 10 or the composition of claim 12 for the preparation of a medicament for use in a method for treating or preventing fibrosis or a disease characterized by fibrosis selected from the group consisting of: Pulmonary fibrosis, cystic fibrosis, idiopathic pulmonary fibrosis, progressive massive fibrosis, scleroderma, bronchiolitis obliterans, Hermansky-Pudlak syndrome, asbestosis, silicosis, chronic pulmonary hypertension, AIDS-related pulmonary hypertension, sarcoidosis, asthma, fatty hepatitis, cirrhosis, schistosomiasis liver disease, alcoholic fatty hepatitis, fatty degeneration, nonalcoholic fatty liver disease, chronic pancreatitis, pancreatic fibrosis, hypertrophic cardiomyopathy, dilated cardiomyopathy, atrial fibrosis, atrial fibrillation, ventricular fibrosis, ventricular fibrillation, myocardial fibrosis, Brugada syndrome, myocarditis, myocardial endometrial fibrosis, myocardial infarction, fibrosing vascular disease, hypertensive heart disease, arrhythmogenic right ventricular cardiomyopathy, tubulointerstitial and glomerular fibrosis, atherosclerosis, varicose veins, cerebral infarction, gliosis, Alzheimer's disease Alzheimer's disease, muscular dystrophy, Crohn's disease, microscopic colitis, primary sclerosing cholangitis, scleroderma, nephrogenic systemic fibrosis, corneal keloids, arthrofibrosis, Dupuytren's contracture, mediastinal fibrosis, retroperitoneal fibrosis, myelofibrosis, Peyronie's disease, bursitis, renal fibrosis, nephrotic syndrome, Alport syndrome, HIV-associated nephropathy, polycystic kidney disease, Fabry's disease, diabetic nephropathy, chronic glomerulonephritis, nephritis associated with systemic lupus, renal interstitial fibrosis, renal failure, progressive systemic sclerosis, chronic graft-versus-host disease, Grave's ophthalmopathy, supraretinal fibrosis, glaucoma, subretinal fibrosis, macular edema, drusen formation, choroidal neovascularization, postoperative fibrosis, conjunctival fibrosis, subconjunctival fibrosis, arthritis, and fibrotic neoplastic diseases.

15. Use of the antigen binding molecule of any one of claims 1 to 6, the CAR of claim 7, the one or more nucleic acids of claim 8, the one or more expression vectors of claim 9, the cell of claim 10 or the composition of claim 12 for the preparation of a medicament for use in a method for treating or preventing hepatotoxicity or a disease characterized by hepatotoxicity selected from the group consisting of: Acute liver disease, chronic liver disease, hepatitis, liver ischemia-reperfusion injury, radiation-induced liver disease, drug-induced liver injury, autoimmune liver injury, cholestatic liver disease, HIV, and alcohol-induced liver injury.

16. Use of the antigen binding molecule of any one of claims 1 to 6, the CAR of claim 7, the one or more nucleic acids of claim 8, the one or more expression vectors of claim 9, the cell of claim 10 or the composition of claim 12 for the preparation of a medicament for use in a method for treating or preventing metabolic diseases, eating disorders or kidney damage.

17. The use according to claim 16, characterized in that The kidney injury is acute kidney injury or nephrotoxicity.

18. The use according to claim 14, characterized in that The non-alcoholic fatty liver disease is NASH.

19. The use according to claim 18, characterized in that The NASH is early NASH or late NASH.

20. The use according to claim 14, characterized in that The fibrotic tumor disease is a lung tumor matrix disease.

21. The use according to claim 14, characterized in that The cirrhosis is primary biliary cirrhosis.

22. The use according to claim 14, characterized in that The muscular dystrophy is Duchenne muscular dystrophy or Becker muscular dystrophy.

23. The use according to claim 14, characterized in that The retinal fibrosis is diabetic retinopathy.

24. The use according to claim 14, characterized in that The subretinal fibrosis is associated with macular degeneration.

25. The use according to claim 24, characterized in that The macular degeneration is wet age-related macular degeneration or dry age-related macular degeneration.

26. The use according to claim 15, characterized in that The acute liver disease is acute liver injury or acute liver failure.

27. The use according to claim 15, characterized in that The hepatitis is selected from the group consisting of viral hepatitis and alcoholic hepatitis.

28. The use according to claim 15, characterized in that The liver ischemia-reperfusion injury is warm ischemia-reperfusion.

29. A method for inhibiting IL-11-mediated signal transduction, comprising contacting a cell expressing IL-11Rα with the antigen-binding molecule of any one of claims 1 to 6, wherein the method is performed in vitro. 30 . An in vitro complex comprising the antigen-binding molecule of any one of claims 1 to 6 bound to IL-11Rα or a complex comprising IL-11Rα.

31. Use of the antigen binding molecule of any one of claims 1 to 6 for preparing an in vitro or in vivo diagnostic or prognostic agent.

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