Inhibitory antibodies of cytokine interleukin 11 dependent signaling pathways

Abstract

The present invention relates to inhibitory antibodies of cytokine interleukin 11 (IL-11) dependent signaling pathways. Further disclosed are methods for the production of said antibodies, uses in diagnostic, treatment and prevention of IL-11 associated disorders.

Description

[0001] Inhibitory antibodies of cytokine interleukin 11 dependent signaling pathways

[0002] The present invention relates to inhibitory antibodies of cytokine interleukin 11 (IL-11 ) dependent signaling pathways. Further disclosed are methods for the production of said antibodies, uses in diagnostic, treatment and prevention of IL-11 associated disorders.

[0003] Background

[0004] Interleukin 11 (IL-11 ), also known as Adipogenesis Inhibitory Factor (AGIF), is a cytokine that belongs to the IL-6 family. In humans, IL-11 (UniProtKB - P20809) is a 19 kDa monomeric protein composed of 199 amino acids, encoded by the IL-11 gene. It is primarily produced by stromal cells and has orthologs (homology >88%) in various model organisms, including mice, rats, and cynomolgus monkeys.

[0005] IL-11 is part of a family of cytokines, including IL-6, LIF, OSM, and IL-27, that utilize the IL-6 Signal Transducer (IL6ST, also known as glycoprotein 130 or gp130) subunit in their receptor complexes creating a composite receptor consisting of a cytokinespecific receptor sub-unit (e.g. IL-11 RA) and a non-specific signal-mediating sub-unit (gp130). While the binding site on IL-11 RA is estimated to possess an affinity of 2-22 nM to IL-11 (Hilton et al., 1994; Karow et al., 1996; Schleinkofer et al., 2001 ; Lee et al., 2008), the affinity of the complete receptor to IL-11 was measured to be in the sub- nanomolar range (400-800 pM; Hilton et al. 1994).

[0006] The binding of IL-11 to its native receptor complex, which exists in a 2:2:2 configuration (IL-11 RA, gp130, and IL-11 ), triggers intracellular signaling pathways. These signaling pathways often involve the activation of transcription factors such as STAT3 and STAT1 or operate through the PI3K / Akt and RAS / ERK cascades — processes that centrally regulate cell growth, proliferation, and survival, thereby playing a pivotal role in the development and maintenance of pathological conditions such as chronic inflammation, autoimmunity, or cancer. IL-11 expression has been detected in a variety of pathologically affected tissues. For example, it is present in the synovial tissue and fluid of patients with Rheumatoid Arthritis (RA), where IL-11 contributes to inflammation and joint damage. In patients with Multiple Sclerosis (MS), IL-11 is suspected to induce Th17 cell responses, contributing to autoimmune and inflammatory processes. Elevated levels of IL-11 are also associated with airway inflammation and hyperresponsiveness in asthma.

[0007] Furthermore, IL-11 is implicated in exacerbating intestinal inflammation in Inflammatory Bowel Diseases (IBD) such as Crohn's disease and ulcerative colitis. It is upregulated in Systemic Sclerosis (SSc), where it contributes to the fibrotic and inflammatory processes characteristic of the disease. IL-11 is also linked to lung tissue damage and chronic inflammation in Chronic Obstructive Pulmonary Disease (COPD), and it plays a role in the inflammatory responses seen in chronic conditions like psoriasis and periodontitis.

[0008] In oncology, IL-11 has been associated with the promotion of tumor growth, metastasis, and cancer-associated fibrosis, making it an attractive therapeutic target, especially in solid tumors. IL-11 also plays a significant role in the signaling and development of Non-Alcoholic Steatohepatitis (NASH), contributing to hepatocyte death, fibrosis, inflammation, and fat accumulation in the liver.

[0009] In general, elevated IL-11 levels are linked to fibrosis in organs such as the heart and kidneys, contributing to disease progression and organ failure. IL-11 drives chronic fibrotic responses by activating fibroblasts and promoting fibrogenic protein synthesis through noncanonical ERK-dependent autocrine signaling.

[0010] Given its multifaceted roles in disease pathogenesis and its specific upregulation in various pathological tissues, targeting IL-11 signaling presents a promising therapeutic strategy for a wide array of diseases, particularly those involving acute or chronic inflammation, fibrosis, and abnormal cell proliferation. Thus, compounds that inhibit IL- 11 signaling, either by specifically binding to the IL-11 receptor (IL-11 RA) or to the ligand (IL-11 ), are actively being developed to treat or mitigate disease progression and improve outcomes in the above conditions.

[0011] Thus, there is a need for antibodies that specifically inhibit the IL-11 -dependent signaling cascade, particularly those that bind the IL-11 ligand.

[0012] Such antibodies are provided herein.

[0013] Detailed description

[0014] The present invention relates to antibodies having specificity for the cytokine interleukin 11 (IL-11 ) and uses thereof.

[0015] Inventive Antibodies

[0016] The first aspect of the invention relates to a monoclonal antibody or antigen-binding fragment thereof which binds to Interleukin-11 (IL-11 ) comprising

[0017] (a) a variable heavy chain (VH) region comprising complementarity-determining regions (CDRs) CDR-H1 , CDR-H2 and CDR-H3 (according to Kabat), wherein

[0018] (i) the CDR-H1 comprises an amino acid sequence according to SEQ ID NO: 1 , 12, 20, or 28, or an amino acid sequence comprising a deletion, insertion and / or substitution, particularly a conservative substitution, of 1 or 2 amino acids,

[0019] (ii) the CDR-H2 comprises an amino acid sequence according to SEQ ID NO: 2, 13, 21 , or 29, or an amino acid sequence comprising a deletion, insertion and / or substitution, particularly a conservative substitution, of 1 or 2 amino acids, (iii) the CDR-H3 comprises an amino acid sequence according to SEQ ID NO: 3, 14, 22, or 30, or an amino acid sequence comprising a deletion, insertion and / or substitution, particularly a conservative substitution, of 1 or 2 amino acids, and optionally

[0020] (b) a variable light chain (VL) region, particularly a VL region comprising complementarity-determining regions (CDRs) CDR-L1 , CDR-L2 and CDR-L3, wherein

[0021] (i) the CDR-L1 comprises an amino acid sequence according to SEQ ID NO: 7, 16, 24, or 32, or an amino acid sequence comprising a deletion, insertion and / or substitution, particularly a conservative substitution, of 1 or 2 amino acids

[0022] (ii) the CDR-L2 comprises an amino acid sequence according to SEQ ID NO: 8, 17, 25, or 33, or an amino acid sequence comprising a deletion, insertion and / or substitution, particularly a conservative substitution, of 1 or 2 amino acids,

[0023] (iii) the CDR-L3 comprises an amino acid sequence according to SEQ ID NO: 9, 18, 26, or 34, or an amino acid sequence comprising a deletion, insertion and / or substitution, particularly a conservative substitution, of 1 or 2 amino acids.

[0024] According to a second aspect of the invention, the anti-IL-11 antibody of the present disclosure comprises:

[0025] (a) a variable heavy chain (VH) region comprising complementarity-determining regions (CDRs) CDR-H1 , CDR-H2 and CDR-H3 (according to IMGT), wherein

[0026] (i) the CDR-H1 comprises an amino acid sequence according to SEQ ID NO: 36, 42, 48, or 54, or an amino acid sequence comprising a comprising a deletion, insertion and / or substitution, particularly a conservative substitution, of

[0027] 1 or 2 amino acids, (ii) the CDR-H2 comprises an amino acid sequence according to SEQ

[0028] ID NO: 37, 43, 49, or 55, or an amino acid sequence comprising a deletion, insertion and / or substitution, particularly a conservative substitution, of 1 or 2 amino acids,

[0029] (iii) the CDR-H3 comprises an amino acid sequence according to SEQ ID NO: 38, 44, 50, or 56, or an amino acid sequence comprising a comprising a deletion, insertion and / or substitution, particularly a conservative substitution, of 1 or 2 amino acids, and optionally

[0030] (b) a variable light chain (VL) region, particularly a VL region comprising complementarity-determining regions (CDRs) CDR-L1 , CDR-L2 and CDR-L3, wherein

[0031] (i) the CDR-L1 comprises an amino acid sequence according to SEQ ID NO: 39, 45, 51 , or 57, or an amino acid sequence comprising a comprising a deletion, insertion and / or substitution, particularly a conservative substitution, of 1 or 2 amino acids,

[0032] (ii) the CDR-L2 comprises an amino acid sequence according to SEQ ID NO: 40, 46, 52, or 58, or an amino acid sequence comprising a deletion, insertion and / or substitution, particularly a conservative substitution, of 1 or 2 amino acids,

[0033] (iii) the CDR-L3 comprises an amino acid sequence according to SEQ ID NO: 41 , 47, 53, or 59, or an amino acid sequence comprising a deletion, insertion and / or substitution, particularly a conservative substitution, of 1 or 2 amino acids. The specific assignment of the amino acid sequences according to the IMGT system to the inventive antibodies can be seen in Table 8.

[0034] Preferably, these antibodies bind to an epitope on IL-11 , which either represents or overlaps with the natural binding site for the IL-11 receptor (IL-11 RA). This binding site on human IL-11 comprises amino acids 105-114, as shown in Figure 6.

[0035] As used herein, the term ‘interleukin 1 T (IL-11 ) has its common meaning in the art and refers specifically to the human cytokine interleukin 11 , a 199 amino acid protein encoded by interleukin 11 gene (IL-11 ) and is in detailed described as UniprotKB entry IL- 11 _HUMAN (UniProtKB Acc: P20809). Herein, the gene name IL-11 might be used for the protein IL-11 and vice versa. In the context of the present disclosure, the term interleukin 11 also includes chimeric polypeptides and homologues as well as fragments and precursors of human interleukin 11.

[0036] The sequence of native human Interleukin 11 corresponds to SEQ ID NO:60 (UniprotKB Acc P20809).

[0037] The term "antibody" as used herein refers to immunoglobulin molecules and immunologically active portions of immunoglobulin molecules, i.e., molecules that contain an antigen binding site that immune-specifically binds an antigen. As such, the term antibody encompasses not only whole antibody molecules, but also antibody fragments as well as variants (including derivatives) of antibodies and antibody fragments. As used herein the term "antibody" or "immunoglobulin" have the same meaning and will be used equally in the present disclosure. Also the terms “inventive antibody” and “inventive antibodies” can be used interchangeably herein.

[0038] The antibodies claimed herein are monoclonal antibodies. The terms "monoclonal antibody" or "monoclonal antibody composition" as used herein refer to a preparation of antibody molecules of a single molecular composition. A monoclonal antibody composition displays a single binding specificity and affinity for a particular epitope.

[0039] The claimed antibodies are preferably isolated antibodies, in particular isolated monoclonal antibodies. An "isolated antibody", as used herein, refers to an antibody that is substantially free, preferably free, of other antibodies having different antigenic specificities (e.g., an isolated antibody that specifically binds to IL-11 is substantially free of antibodies that specifically bind to other antigens than IL-11 ). An isolated antibody that specifically binds to IL-11 may, however, have cross-reactivity to other antigens, such as related IL-11 molecules from other species. Moreover, an isolated antibody may be substantially free of other cellular material and / or chemicals.

[0040] In natural antibodies, two heavy chains are linked to each other by disulfide bonds and each heavy chain is linked to a light chain by a disulfide bond. There are two types of light chain, lambda (1 ) and kappa (k). There are five main heavy chain classes (or isotypes) which determine the functional activity of an antibody molecule: IgM, IgD, IgG, IgA and IgE. Each chain contains distinct sequence domains. The light chain includes two domains, a variable domain (VL) and a constant domain (CL).

[0041] The variable regions of both light (VL) and heavy (VH) chains determine binding recognition and specificity to the antigen. The constant region domains of the light (CL) and heavy (CH) chains confer important biological properties such as antibody chain association, secretion, trans-placental mobility, complement binding, and binding to Fc receptors (FcR). The light and heavy chains of an immunoglobulin each have three CDRs, designated CDR-L1 , CDR-L2, CDR-L3 and CDR-H1 , CDR-H2, CDR-H3, respectively.

[0042] An antigen-binding site, therefore, typically includes six CDRs, comprising the CDRs set from each of a heavy and a light chain V region. Framework Regions (FRs) refer to amino acid sequences interposed between CDRs. Accordingly, the variable regions of the light and heavy chains typically comprise 4 framework regions and 3 CDRs of the following sequence: FR1 -CDR1 -FR2-CDR2-FR3-CDR3-FR4.

[0043] The sequences of the CDR variants may differ from the disclosed CDR sequences by amino acid deletion, insertion or substitution of 1 or 2 amino acids. Amino acid substitution is preferred, particularly a conservative substitution of 1 or 2 amino acids. In the context of the present disclosure, conservative substitutions may be defined by substitutions within the classes of amino acids reflected as follows:

[0044] Aliphatic residues I, L, V, and M

[0045] Cycloalkenyl-associated residues F, H, W, and Y

[0046] Hydrophobic residues A, C, F, G, H, I, L, M, R, T, V, W, and Y

[0047] Negatively charged residues D and E

[0048] Polar residues C, D, E, H, K, N, Q, R, S, and T

[0049] Positively charged residues H, K, and R

[0050] Small residues A, C, D, G, N, P, S, T, and V

[0051] Very small residues A, G, and S

[0052] Residues involved in turn A, C, D, E, G, H, K, N, Q, R, S, P, and formation T

[0053] Flexible residues Q, T, K, S, G, P, D, E, and R

[0054] More conservative substitutions groupings include valine-leucine-isoleucine, phenylalanine-tyrosine, lysine-arginine, alanine-valine, and asparagine-glutamine.

[0055] As used herein, the term "antibody" collectively refers to immunoglobulins or immunoglobulin-like molecules including by way of example and without limitation, IgA, IgD, IgE, IgG and IgM, combinations thereof, and similar molecules produced during an immune response in any vertebrate, for example, in mammals such as humans, goats, rabbits and mice, as well as nonmammalian species, such as shark immunoglobulins. In specific embodiments, an antibody provided herein is an antibody fragment, and more particularly any protein including an antigen-binding domain of an antibody as disclosed herein. Antibody fragments include, but are not limited to, Fv, Fab, F(ab’)2, Fab’, dsFv, scFv, sc(Fv)2 and diabodies. The terms “antibody fragment” and “antigenbinding fragment” can be used interchangeably herein.

[0056] The Fv fragment is the N-terminal part of the Fab fragment of an immunoglobulin and consists of the variable portions of one light chain and one heavy chain. The specificity of the antibody resides in the structural complementarity between the antibody combining site and the antigenic determinant.

[0057] As used herein, the term “Fab” denotes an antibody fragment having a molecular weight of about 50,000 and antigen binding activity, in which about a half of the N- terminal side of H chain and the entire L chain, among fragments obtained by treating IgG with a protease, papaine, are bound together through a disulfide bond.

[0058] The term “F(ab')2” refers to an antibody fragment having a molecular weight of about 100,000 and antigen binding activity, which is slightly larger than the Fab bound via a disulfide bond of the hinge region, among fragments obtained by treating IgG with a protease, pepsin.

[0059] The term “Fab”’ refers to an antibody fragment having a molecular weight of about 50,000 and antigen binding activity, which is obtained by cutting a disulfide bond of the hinge region of the F(ab')2.

[0060] A single chain Fv ("scFv") polypeptide is a covalently linked VH:VL heterodimer which is usually expressed from a gene fusion including VH and VL encoding genes linked by a peptide-encoding linker. The human scFv fragment of the disclosure includes CDRs that are held in appropriate conformation, preferably by using gene recombination techniques.

[0061] The residues in antibody variable domains are conventionally numbered according to a system devised by Kabat et al. This system is set forth in Kabat et al., 1987, in Sequences of Proteins of Immunological Interest, US Department of Health and Human Services, NIH, USA (hereafter “Kabat et al.”). This numbering system is mainly used in the present specification and in particular if not indicated otherwise. However, beside Kabat there are further frequently used numbering schemes e.g. Chotia, IMGT, Martin or the AHO numbering scheme that allow the precise definition of an antibody position in the variable domain of an antibody. The Kabat residue designations do not always correspond directly with the linear numbering of the amino acid residues in SEQ ID sequences. The actual linear amino acid sequence may contain fewer or additional amino acids than in the strict Kabat numbering corresponding to a shortening of, or insertion into, a structural component, whether framework or complementarity determining region (CDR), of the basic variable domain structure. The correct Kabat numbering of residues may be determined for a given antibody by alignment of residues of homology in the sequence of the antibody with a “standard” Kabat numbered sequence.

[0062] According to preferred embodiments, the antibody or antigen-binding fragment thereof may comprise

[0063] (i) a VH region comprising the CDR-H1 of SEQ ID NO:1 , the CDR-H2 of SEQ ID NO:2, and the CDR-H3 of SEQ ID NO:3, and optionally

[0064] (ii) a VL region comprising the CDR-L1 of SEQ ID NO:7, the CDR-L2 of the SEQ ID NO:8 and the CDR-L3 of the SEQ ID NO:9.

[0065] The antibodies 18A2, HC7LC8 and HC10LC8 have a VH region comprising CDRs as defined above.

[0066] The antibodies 18A2, HC10LC8 and HC7LC8 have a VL region comprising CDRs as defined above. The antibody or antigen-binding fragment thereof may comprise

[0067] (a) a VH region comprising an amino acid sequence according to SEQ ID NO: 4, 5 or 6 or an amino acid sequence having an identity thereof of at least 85%, at least 90%, at least 95% or at least 99%, and optionally

[0068] (b) a VL region, particularly a VL region comprising an amino acid sequence according to SEQ ID NO: 10 or 11 or an amino sequence having an identity thereof of at least 85%, at least 90%, at least 95% or at least 99%.

[0069] The antibody 18A2 shows a VH region according to SEQ ID NO:6 and a VL region according to SEQ ID NO: 11 .

[0070] The antibody HC7LC8 shows a VH region according to SEQ ID NO:4 and a VL region according to SEQ ID NO: 10.

[0071] The antibody HC10LC8 shows a VH region according to SEQ ID NO:5 and a VL region according to SEQ ID NO: 10.

[0072] The antibody or antigen-binding fragment thereof may comprise

[0073] (a) a VH region comprising an amino acid sequence according to SEQ ID NO: 4 (corresponding to the VH region HC7) or an amino acid sequence having an identity thereof of at least 85%, at least 90%, at least 95% or at least 99%, and optionally

[0074] (b) a VL region, particularly a VL region comprising an amino acid sequence according to SEQ ID NO: 10 (corresponding to the VL region LC8) or an amino sequence having an identity thereof of at least 85%, at least 90%, at least 95% or at least 99%.

[0075] The antibody or antigen-binding fragment thereof may comprise

[0076] (a) a VH region comprising an amino acid sequence according to SEQ ID NO: 5 (corresponding to the VH region HC10) or an amino acid sequence having an identity thereof of at least 85%, at least 90%, at least 95% or at least 99%, and optionally (b) a VL region, particularly a VL region comprising an amino acid sequence according to SEQ ID NO: 10 (corresponding to the VL region LC8) or an amino sequence having an identity thereof of at least 85%, at least 90%, at least 95% or at least 99%.

[0077] The antibody or antigen-binding fragment thereof may comprise

[0078] (i) a VH region comprising the CDR-H1 of SEQ ID NO: 12, the CDR-H2 of SEQ ID NO: 13, and the CDR-H3 of SEQ ID NO: 14, and optionally

[0079] (ii) a VL region comprising the CDR-L1 of SEQ ID NO: 16, the CDR-L2 of the SEQ ID NO: 17 and the CDR-L3 of the SEQ ID NO: 18.

[0080] The antibody 12B3 has a VH and a VL region comprising CDRs as defined above.

[0081] The antibody or antigen-binding fragment thereof may comprise

[0082] (a) a VH region comprising an amino acid sequence according to SEQ ID NO: 15 (corresponding to the VH region 12B3) or an amino acid sequence having an identity thereof of at least 85%, at least 90%, at least 95% or at least 99%, and optionally

[0083] (b) a VL region, particularly a VL region comprising an amino acid sequence according to SEQ ID NO: 19 (corresponding to the VL region 12B3) or an amino sequence having an identity thereof of at least 85%, at least 90%, at least 95% or at least 99%.

[0084] The antibody 12B3 has a VH and a VL region comprising SEQ ID NO: 15 and 19.

[0085] The antibody or antigen-binding fragment thereof may comprise

[0086] (i) a VH region comprising the CDR-H1 of SEQ ID NO: 20, the CDR-H2 of SEQ ID NO: 21 , and the CDR-H3 of SEQ ID NO: 22, and optionally

[0087] (ii) a VL region comprising the CDR-L1 of SEQ ID NO: 24, the CDR-L2 of the SEQ ID NO: 25 and the CDR-L3 of the SEQ ID NO: 26.

[0088] The antibody 14B11 has a VH and a VL region comprising CDRs as defined above. The antibody or antigen-binding fragment thereof may comprise

[0089] (a) a VH region comprising an amino acid sequence according to SEQ ID NO: 23 (corresponding to the VH region 14B11 ) or an amino acid sequence having an identity thereof of at least 85%, at least 90%, at least 95% or at least 99%, and optionally

[0090] (b) a VL region, particularly a VL region comprising an amino acid sequence according to SEQ ID NO: 27 (corresponding to the VL region 14B11 ) or an amino sequence having an identity thereof of at least 85%, at least 90%, at least 95% or at least 99%.

[0091] The antibody 14B11 (“Antibody A”) has a VH and a VL region comprising SEQ ID NO:23 and 27.

[0092] The antibody or antigen-binding fragment thereof may comprise

[0093] (i) a VH region comprising the CDR-H1 of SEQ ID NO: 28, the CDR-H2 of SEQ ID NO: 29, and the CDR-H3 of SEQ ID NO: 30, and optionally

[0094] (ii) a VL region comprising the CDR-L1 of SEQ ID NO: 32, the CDR-L2 of the SEQ ID NO: 33 and the CDR-L3 of the SEQ ID NO: 34.

[0095] The antibody 19C6 (“Antibody C”) has a VH and a VL region comprising CDRs as defined above.

[0096] The antibody or antigen-binding fragment thereof may comprise

[0097] (a) a VH region comprising an amino acid sequence according to SEQ ID NO: 31 or an amino acid sequence having an identity thereof of at least 85%, at least 90%, at least 95% or at least 99%, and optionally

[0098] (b) a VL region, particularly a VL region comprising an amino acid sequence according to SEQ ID NO: 35 or an amino sequence having an identity thereof of at least 85%, at least 90%, at least 95% or at least 99%.

[0099] The antibody 19C6 has a VH and a VL region comprising SEQ ID NO:31 and 35. Further disclosed is antibody-B (19C6-VH 1 14BII-VL), comprising SEQ ID Nos 31 und 27 as defined above.

[0100] The term “identity” as used herein refers to the sequence similarity between two polypeptide molecules or between two nucleic acid molecules. When a position in both compared sequences is occupied by the same base or same amino acid residue, then the respective molecules are identical at that position. The percentage of identity between two sequences corresponds to the number of matching positions shared by the two sequences divided by the number of positions compared and multiplied by 100. Generally, a comparison is made when two sequences are aligned to give maximum identity. The identity may be calculated by alignment using, for example, the GCG (Genetics Computer Group, Program Manual for the GCG Package, Version 7, Madison, Wisconsin) pileup program, or any of sequence comparison algorithms such as BLAST, PASTA or CLUSTALW.

[0101] With regard to inventive antibodies or fragments thereof having an amino sequence having an identity thereof of at least 85% to the disclosed VH and VL sequences it is preferred that the CDR regions 100 % identical to the corresponding CDR regions disclosed as heavy chain CDRs of SEQ ID NOs: 1 -3, or 12-14, or 20-22, or 28-30, or respective light chain CDRs of SEQ ID Nos: 7-9, or 16-18, or 24-26, or 32-34. Preferably 1 , 2, 3, 4 or 5 amino acids have been mutated by amino acid deletion, insertion or substitution, preferably conservative substitution, in the framework regions FR1 , FR2, FR3 and FR4 when compared with corresponding framework regions of the herein disclosed VHA / L sequences such as

[0102] (i) VH sequences according to SEQ ID NO: 4, 5 or 6 (HC7-VH, HC10-VH or rat- 18A2-VH) and VL sequences according to SEQ ID NO: 11 or 11 (LC8-VH or rat- 18A2-VL), or

[0103] (ii) VH sequences according to SEQ ID NO: 15 (12B3-VH) and VL sequences according to SEQ ID NO: 19 (12B3-VL), or (iii) VH sequences according to SEQ ID NO:23 (14B11 -VH) and VL sequences according to SEQ ID NO: 27 (14B11 -VL), or

[0104] (iv) VH sequences according to SEQ ID NO:31 (19C6-VH) and VL sequences according to SEQ ID NO: 35 (19C6-VL).

[0105] Mutated antibodies or antibodies differing from specifically antibodies described herein by amino acid deletion, insertion or substitution, in particular conservative substitution, or in terms of identity exhibit functional properties that are substantially equal or superior to the corresponding functional properties of the herein disclosed antibodies, e.g. antibodies 18A2 and 12B3, HC7LC8 or HC10LC8. By “substantially equal” it is herein intended that said functional variant retains at least about 50%, 60%, 70%, 80%, 90%, 95% or 100% of the corresponding functional property of the herein disclosed antibodies. Such antibodies or fragments may be called functional variants.

[0106] The inventive antibodies or fragments thereof bind to one or more epitopes of IL-11 . In some embodiments, the epitopes to which the present antibodies or antigen binding fragments bind are linear epitopes. Typically, however, the epitopes to which the present antibodies or antigen binding fragments bind are non-linear, conformational epitopes.

[0107] The antibodies of the present disclosure may be produced by any technique known in the art, such as, without limitation, any chemical, biological, genetic or enzymatic technique, either alone or in combination. Typically, knowing the amino acid sequence of the desired sequence, one skilled in the art can readily produce said antibodies, by standard techniques for production of polypeptides. For instance, they can be synthesized using well-known solid phase method, preferably using a commercially available peptide synthesis apparatus (such as that made by Applied Biosystems, Foster City, California) and following the manufacturer's instructions. Alternatively, antibodies of the present disclosure can be synthesized by recombinant DNA techniques well-known in the art. For example, antibodies can be obtained as DNA expression products after incorporation of DNA sequences encoding the antibodies into expression vectors and introduction of such vectors into suitable eukaryotic or prokaryotic hosts that will express the desired antibodies, from which they can be later isolated using well-known techniques.

[0108] Characteristics of the inventive Antibodies

[0109] The inventive anti-IL-11 antibodies are preferably human, chimeric or humanized antibodies, in particular a human or humanized IgG antibodies. Especially preferred are human or humanized lgG1 antibodies comprising a kappa light chain.

[0110] In some embodiments, the antibody of the present disclosure is a chimeric antibody, in particular a chimeric mouse / human antibody.

[0111] The term "chimeric antibody" refers to a monoclonal antibody which comprises a VH domain and a VL domain of an antibody derived from a non-human animal, a CH domain and a CL domain of a human antibody. As non-human animal, any animal such as mouse, rat, hamster, rabbit or the like can be used. In particular, said mouse / human chimeric antibody may comprise the VH and the VL domains of the present reference antibody.

[0112] According to preferred embodiments, the antibody of the present disclosure is a humanized antibody. The inventive antibody is preferably a humanized antibody which comprises a set of 6 CDRs as disclosed herein. In the context of the present disclosure the term "humanized antibody" refers to antibodies in which the framework regions (FRs) have been modified to comprise the FRs from a donor immunoglobulin of different species (for example human species) as compared to that of the parent immunoglobulin (for example CDRs of the rat antibody 18A2).

[0113] According to a preferred embodiment, the inventive anti-IL-11 antibody is a humanized silent antibody. As used herein, the term “silent” antibody refers to an antibody that exhibits no or low antibody-dependent cellular cytotoxicity (ADCC) as measured in an in vitro ADCC activity assay measuring cell lysis of target cells. The term “no or low ADCC activity” means that the silent antibody preferably exhibits an ADCC activity that is at below 50%, for example below 10% of the ADCC activity that is observed with the corresponding wild type (non-silent) antibody for example with a wild type human IgG 1 antibody. Preferably, no detectable ADCC activity is observed in an in vitro ADCC activity assay with a silent antibody as compared to a control Fab antibody. Silenced effector functions can be obtained by mutation in the Fc constant part of the antibodies and have been described in the Art: Strohl 2009 (LALA & N297A); Baudino 2008, D265A (Baudino et al., J. Immunol. 181 (2008): 6664-69, Strohl, CO Biotechnology 20 (2009): 685-91 ). Examples of silent lgG1 antibodies comprise mutations reducing ADCC at positions 234, 235 and / or 331 in the lgG1 Fc amino acid sequence (Ell numbering). Another silent lgG1 antibody comprises the N297A mutation or another mutation such as N297S which leads to aglycosylated or non-glycosylated antibodies.

[0114] The inventive antibody or fragments thereof preferably binds to human IL-11 with a Kd of 5 nM or less, particularly of 1 nM or less as determined by ELISA.

[0115] The inventive antibody or fragments thereof preferably binds to human IL-11 with a Kd of 5 nM or less, particularly of 1 nM or less as determined by Surface Plasmon Resonance.

[0116] The binding behavior in the native IL-11 receptor complex, consisting of IL-11 , the cognate receptor IL-11 RA, and the glycoprotein gp130 in a 2:2:2 configuration, is characterized by a high-affinity binding of IL-11 to the receptor in the sub-nanomolar range (400-800 pM). The orderly sequence of IL-11 binding to the receptor starts with IL-11 binding to IL-11 RA (Karow et al., 1996), which only then enables binding to gp130. In the absence of gp130, IL-11 exhibits significantly reduced affinity for IL-11 RA in the nanomolar range. Therefore, without being bound by theory, it may be advantageous to block this low-affinity binding site on IL-11 for interactions with IL- 11 RA in order to induce strong inhibition. Preferably an inventive anti-IL-11 antibody or binding fragment thereof inhibits binding of IL-11 to IL-11 RA with an IC50 of 10 nM or less, preferably in the absence of gp130.

[0117] It is preferred that an inventive anti-IL-11 antibody or binding thereof inhibits the binding of human IL-11 to human IL-11 RA, and / or to mouse IL-11 RA, and / or to rat IL-11 RA and / or to cyno IL-11 -RA with an IC50 of 10 nM or less, preferably in the absence of gp130.

[0118] It is preferred that an inventive anti-IL-11 antibody or binding fragment thereof inhibits binding of human IL-11 to mouse IL-11 RA with an IC50 of 10 nM or less, preferably in the absence of gp130.

[0119] It is preferred that the inventive anti-IL-11 antibodies or fragments thereof inhibit IL-11 - induced cellular signaling with an IC50 of 5 nM or less, particularly of 1 nM or less (as determined by ELISA).

[0120] It is preferred that the inventive anti-IL-11 antibodies or fragments thereof inhibit the IL- 11 induced or IL-11 dependent cell proliferation, in particular of malignant or cancer cells, in particular of TF-1 erythroleukemia cells, with an IC50 of 5 nM or less preferably with an IC50 of 1 nM or less (as determined by measurement of the cell titer).

[0121] The anti-IL-11 antibodies disclosed herein preferably show no or only minimal binding (cross-specificity) with closely related cytokines, such as human Leukemia Inhibitory Factor (hLIF), human Oncostatin M (hOSM), human lnterleukin-6 (hlL-6), human Interleukin-27 (hlL-27), and mouse Transforming Growth Factor (31 (mTGF-[31 ), (Fig. 2).

[0122] Thus, an inventive anti-IL-11 antibody or fragment thereof may be preferably further characterized by low cross-specificity to closely related cytokines, preferably by low cross-specificity and in particular no cross-specificity against human cytokines closely related by amino acid sequence homology, in particular human Leukemia Factor (hLIF), human Oncostatin M (hOSM), human lnterleukin-6 (hlL-6), human Interleukin- 27 (hlL-27), mouse Transforming Growth Factor [31 (mTGF-[31 ).

[0123] Preferably the antibody or antigen-binding fragment thereof exhibits less binding of the IL-11 binding, preferably less than 10% of the IL-11 binding, to human Leukemia Inhibitory Factor (hLIF) and / or human Interleukin-27 (hlL-27) and / or Transforming Growth Factor (31 (TGF-|31 ), when determined by ELISA.

[0124] Herein, “binding to IL-11” or “antibody affinity” refers to the strength with which the antibody binds to the epitope presented on an antigen, such as IL-11 in the present disclosure, through its antigen-binding site (paratope). The apparent affinity-binding constant value (KD) for IL-11 may be assessed, for example, by methods known to the person skilled in the art or by methods described herein. Suitable methods comprise flow cytometry by measuring the saturation curve of antibody binding and by determination of the EC50 value (Fig 1 ) or by Surface Plasmon Resonance (SPR) (Table 3).

[0125] "Selective binding" typically means that the antibody binds more strongly to a target, such as an epitope, for which it is specific as compared to the binding to another target. Thus, the antibody binds more strongly to a first target as compared to a second target if its affinity for the first target is higher than its affinity for the second target. Typically, an antibody binds more strongly to a first target as compared to a second target if it binds to the first target with a dissociation constant (Kd), or an EC50 as mentioned above, that is lower than the dissociation constant, or the EC50, for the second target. Most specifically the agent does not bind at all to the second target to a relevant extent, or has relatively little detectable reactivity; typically in a binding assay it has no detectable staining, or the staining level is less than a reference value.

[0126] As used herein, the term "specificity" refers to the ability of an antibody to detectably bind an epitope presented on an antigen, such as IL-11. Within the present invention, it is typically intended to refer to an antibody that binds to human, mouse, rat or cynomolgus monkey IL-11 as described in the examples, while the antibody at the same time has relatively little detectable reactivity with non- IL-11 proteins or structures (such as other proteins having different epitopes). The inventive antibodies have preferably an apparent affinity-binding constant value (KD) (or an EC50) of less than 10 pg / mL, notably less 1 pg / mL, less than 100 ng / mL, less than 10 ng / mL or less than 5 ng / mL as determined in the Examples and Figures. Typically, the apparent affinitybinding constant value (KD) is not more than 0.1 pg / ml, notably 0.03 pg / ml. Generally, apparent affinity binding constant value (KD) (that can be determined as illustrated in the examples) is comprised between 0.1 ng and 10 pg / ml. In some embodiments, it binds to an antigen recombinant polypeptide with a KD of 10nM or less, or 1 nM or less. Typically, the KD is not less than 1 nM notably 0.2 nM. Generally, the KD is comprised between 0.1 pM and 10 nM.

[0127] The term "KD", as used herein, is intended to refer to the equilibrium dissociation constant, which is obtained from the ratio of Kd to Ka (i.e. , Kd / Ka) and is expressed as a molar concentration (M). The KD value relates to the concentration of antibody (the amount of antibody needed for a particular experiment) and so the lower the KD value (lower concentration) and thus the higher the affinity of the antibody. The KD value for an antibody can also be determined using methods well established in the art. For example, methods for determining the KD values of mAbs can be found in Harlow, et al., Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y, 1988), Coligan et al., eds., Current Protocols in Immunology, Greene Publishing Assoc, and Wiley Interscience, N.Y., (1992, 1993), and Muller, Meth. Enzymol. 92:589-601 (1983). A method for determining the KD of an antibody is by using surface plasmon resonance or using a biosensor system such as a Biacore® or Octet® systems. Kd is typically measured using surface plasmon resonance assays using a BIACORE®-2000 or a BIACORE®-3000 (BIAcore, Inc., Piscataway, NJ) or a similar machine.

[0128] An antibody that "competes for binding with IL-11 " with the herein disclosed antibodies e.g. antibodies 18A2 and 12B3, HC7LC8 or HC10LC8 or variants thereof is intended to refer to an antibody that binds IL-11 with an apparent affinity-binding constant value (KD) (or an EC50) equal to or lower than the measured values disclosed in the Examples and Figures, in particular an antibody that binds IL-11 with an apparent affinity-binding constant value of less than 10 pg / mL, notably less 1 pg / mL, less than 100 ng / mL, less than 10 ng / mL or less than 5 ng / mL.

[0129] In a preferred embodiment the antibody according to the present disclosure binds to an epitope on human IL-11 comprising amino acids 105-114 of SEQ ID NO: 60, preferably the binding epitope consist of amino acids 105-114 of SEQ ID NO: 60.

[0130] The inventive anti-IL-11 antibody or fragment thereof may comprise a labeling group and / or an effector group being coupled to the antibody or antigen- binding fragment. Such a labeling group may be a dye, a paramagnetic, radioactive or fluorogenic group that is detectable upon imaging. The effector group may be a therapeutic group, in particular a cytotoxic agent. Cytotoxic agents known to be used in ADCs are particularly preferred.

[0131] It is preferred that an inventive antibody, a fragment or functional variant thereof as described herein retains at least one or more of the properties of the herein disclosed antibodies e.g. antibodies 18A2 and 12B3, HC7LC8 or HC10LC8 preferably

[0132] - high affinity binding to human IL-11

[0133] - binding to a site on IL-11 that overlaps with the binding site of IL-11 RA, in particular to amino acids 105-114 of SEQ ID NO: 60

[0134] - inhibiting the binding of IL-11 to IL-11 RA

[0135] - inhibiting cellular signaling elicited by IL-11 stimulation

[0136] - low cross reactivity with closely related cytokines, in particular low cross reactivity with hLIF and / or hlL-27 and / or mTGF-[31 , preferably the exhibited cross-reactivity is less, preferably less 10%, of its binding to IL-11

[0137] - inhibition of IL-11 induced cell proliferation, in particular of TF-1 erythroleukemia cells In various embodiments, the antibody may exhibit one or two of the desired functional properties above.

[0138] Fc Engineering

[0139] The antibodies as herein disclosed may be characterized by one or more of the functional or structural features described above, or by any combination of selected functional and structural features.

[0140] The antibodies as herein disclosed may be of any suitable isotype.

[0141] The choice of isotype typically will be guided by the desired effector functions, such as ADCC silencing. Exemplary isotypes are lgG1 , lgG2, lgG3, and lgG4. Either of the human light chain constant regions, kappa or lambda, may be used. If desired, the class of an antibody of the present disclosure may be switched by known methods. Typical, class switching techniques may be used to convert one IgG subclass to another, for instance from IgG 1 to lgG2.

[0142] Thus, the effector function of the antibodies of the present disclosure may be changed by isotype switching to, e.g., an lgG1 , lgG2, lgG3, lgG-4, IgD, IgA, IgE, or IgM antibody for various therapeutic uses.

[0143] In some embodiments, the antibody as herein disclosed is a full-length antibody. In some embodiments, the full-length antibody is an lgG1 antibody. In some embodiments, the full-length antibody is an lgG4 antibody.

[0144] In some embodiments, the IL-11 -specific lgG4 antibody is a stabilized lgG4 antibody. Examples of suitable stabilized lgG4 antibodies are antibodies wherein arginine at position 409 in a heavy chain constant region of human lgG4, which is indicated in the Ell index as in Kabat et al. supra, is substituted with lysine, threonine, methionine, or leucine, preferably lysine (described in WO 2006 / 033386) and / or wherein the hinge region comprises a Cys-Pro-Pro-Cys sequence. Other suitable stabilized lgG4 antibodies are disclosed in WO 2008 / 145142.

[0145] In some embodiments, the antibody of the present disclosure does not comprise a Fc portion that induces antibody dependent cellular cytotoxicity (ADCC). The terms "Fc domain," "Fc portion," and "Fc region" refer to a C -terminal fragment of an antibody heavy chain, e.g., from about amino acid (aa) 230 to about aa 450 of human gamma heavy chain or its counterpart sequence in other types of antibody heavy chains (e.g., o, 5, s and p for human antibodies), or a naturally occurring allotype thereof.

[0146] In some embodiments, the antibody of the present disclosure does not comprise an Fc domain capable of substantially binding to a FcgRIIIA (CD16) polypeptide.

[0147] In some embodiments, the antibody of the present disclosure lacks an Fc domain (e.g. lacks a CH2 and / or CHS domain) or comprises an Fc domain of lgG2 or lgG4 isotype.

[0148] In some embodiments, the antibody of the present disclosure consists of or comprises a Fab, Fab', Fab'-SH, F(ah')2, Fv, a diabody, single-chain antibody fragment, or a multispecific antibody comprising multiple different antibody fragments.

[0149] In some embodiments, one or more amino acids can be replaced with a different amino acid residue such that the antibody has altered C2q binding and / or reduced or abolished complement dependent cytotoxicity (CDC). This approach is described in further detail in U.S. Patent Nos. 6,194,551.

[0150] Another possible modification of the inventive antibodies is pegylation. An antibody can be pegylated, for example, to increase the biological (e.g., serum) half-life of the antibody. To pegylate an antibody, the antibody, or fragment thereof, typically is reacted with polyethylene glycol (PEG), such as a reactive ester or aldehyde derivative of PEG, under conditions in which one or more PEG groups become attached to the antibody or antibody fragment. The pegylation can be carried out by an acylation reaction or an alkylation reaction with a reactive PEG molecule (or an analogous reactive water- soluble polymer).

[0151] As used herein, the term "polyethylene glycol" is intended to encompass any of the forms of PEG that have been used to derivatize other proteins, such as mono alkoxy- or aryloxy-poly-ethylene glycol or polyethylene glycol-maleimide. In some embodiments, the antibody to be pegylated is an aglycosylated antibody.

[0152] Another possible modification of the inventive antibodies is a conjugate or a protein fusion of at least the antigen-binding region of the inventive antibody to serum protein, such as human serum albumin or a fragment thereof to increase half-life of the resulting molecule.

[0153] In some embodiments, the disclosure also provides a multispecific antibody. Exemplary formats for the multispecific antibody molecules of the disclosure include, but are not limited to

[0154] (i) two antibodies cross-linked by chemical heteroconjugation, one with a specificity to IL-11 and another with a specificity to a second antigen;

[0155] (ii) a single antibody that comprises two different antigen-binding regions;

[0156] (iii) a single-chain antibody that comprises two different antigen-binding regions, e.g., two scFvs linked in tandem by an extra peptide linker;

[0157] (iv) a dual-variable-domain antibody (DVD-lg), where each light chain and heavy chain contains two variable domains in tandem through a short peptide linkage;

[0158] (v) a chemically-linked bispecific (Fab')2 fragment;

[0159] (vi) a Tandab, which is a fusion of two single chain diabodies resulting in a tetravalent bispecific antibody that has two binding sites for each of the target antigens; (vii) a flexibody, which is a combination of scFvs with a diabody resulting in a multivalent molecule;

[0160] (viii) a so called "dock and lock" molecule, based on the "dimerization and docking domain" in Protein Kinase A, which, when applied to Fabs, can yield a trivalent bispecific binding protein consisting of two identical Fab fragments linked to a different Fab fragment;

[0161] (ix) a so-called Scorpion molecule, comprising, e.g., two scFvs fused to both termini of a human Fab-arm; and

[0162] (x) a diabody.

[0163] Another exemplary format for bispecific antibodies are IgG-like molecules with complementary CHS domains to force heterodimerization. Such molecules can be prepared using known technologies, such as, e.g., those known as Triomab / Quadroma (Trion Pharma / Fresenius Biotech), Knob-into-Hole (Genentech), CrossMAb (Roche) and electrostatically-matched (Amgen), LUZ-Y (Genentech), Strand Exchange Engineered Domain body (SEEDbody)(EMD Serono), Biclonic (Merus) and DuoBody (Genmab A / S) technologies. In some embodiments, the bispecific antibody is obtained or obtainable via a controlled Fab-arm exchange, typically using DuoBody technology. In vitro methods for producing bispecific antibodies by controlled Fab-arm exchange have been described in WO 2008 / 119353 and WO 2011 / 131746 (both by Genmab A / S). In one exemplary method, described in WO 2008 / 119353, a bispecific antibody is formed by "Fab-arm" or "half- molecule" exchange (swapping of a heavy chain and attached light chain) between two monospecific antibodies, both comprising lgG4-like CHS regions, upon incubation under reducing conditions. The resulting product is a bispecific antibody having two Fab arms which may comprise different sequences. In another exemplary method of providing bispecific antibodies is described in WO 2011 / 131746.

[0164] Antibody-Drug Conjugates

[0165] In some embodiments, the antibody of the present disclosure is conjugated to a therapeutic moiety, i.e., a drug. The therapeutic moiety can be, e.g., a cytotoxin, a chemotherapeutic agent, a cytokine, an immunosuppressant, an immune stimulator, a lytic peptide, or a radioisotope. Such conjugates are referred to herein as an "antibodydrug conjugates" or "ADCs".

[0166] Pharmaceutical Compositions

[0167] According to another aspect of the invention, a composition is provided, in particular a pharmaceutical composition, containing at least one antibody as disclosed herein, formulated together with a pharmaceutically acceptable carrier. Such compositions may include one or a combination of (e.g., two or more different) antibodies as described above.

[0168] Pharmaceutical compositions disclosed herein also can be administered in combination therapy, i.e., combined with other agents. For example, an antibody of the present disclosure may typically be combined with at least one antiviral, antiinflammatory or another antiproliferative agent. Examples of therapeutic agents that can be used in combination therapy are described in greater detail below in the section on uses of the antibodies of the disclosure.

[0169] The compositions of the present disclosure may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted reservoir. Correspondingly administration includes subcutaneous, intravenous, intramuscular, intra-articular, intra-synovial, intrastemal, intrathecal, intrahepatic, intralesional and intracranial injection or infusion techniques.

[0170] Depending on the route of administration, the active compound, i.e., the antibody, may be coated in a material to protect the compound from the action of acids and other natural conditions that may inactivate the compound. The form of the pharmaceutical compositions, the route of administration, the dosage and the regimen naturally depend upon the condition to be treated, the seventy of the illness, the age, weight, and sex of the patient, etc. As used herein, "pharmaceutically acceptable carrier" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are physiologically compatible. The carrier should be suitable for intravenous, intramuscular, subcutaneous, parenteral, spinal or epidermal administration (e.g., by injection or infusion).

[0171] In preferred embodiments, the carrier is suitable for subcutaneous or intravenous route.

[0172] Pharmaceutically acceptable carriers that may be used in these compositions include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfide, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene- block polymers, polyethylene glycol and wool fat.

[0173] Sterile injectable forms of the inventive compositions may be aqueous or an oleaginous suspension. These suspensions may be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a nontoxic parenterally acceptable diluent or solvent, for example as a solution in 1 ,3- butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. Fatty acids, such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically-acceptable oils, such as olive oil or castor oil, especially in their poly-oxyethylated versions. These oil solutions or suspensions may also contain a long-chain alcohol diluent or dispersant, such as carboxymethyl cellulose or similar dispersing agents that are commonly used in the formulation of pharmaceutically acceptable dosage forms including emulsions and suspensions.

[0174] The inventive compositions may be orally administered in any orally acceptable dosage form including, but not limited to, capsules, tablets, aqueous suspensions or solutions. In the case of tablets for oral use, carriers commonly used include lactose and com starch. Lubricating agents, such as magnesium stearate, are also typically added. For oral administration in a capsule form, useful diluents include, e.g., lactose. When aqueous suspensions are required for oral use, the active ingredient is combined with emulsifying and suspending agents.

[0175] Diagnostic and therapeutic Utilities of the inventive Antibodies

[0176] The inventive antibodies or fragments thereof provide multiple diagnostic and therapeutic utilities.

[0177] The disclosed antibodies and fragments thereof are particularly suitable for treating, preventing or diagnosing Interleukin-11 -related disorders and in particular for treating, preventing or diagnosing proliferative disorders, autoimmune disorders or inflammatory disorders.

[0178] Thus, an aspect of the invention relates to the antibodies or fragments thereof disclosed herein for use in medicine, particularly for therapeutic or diagnostic applications including in vitro and in vivo diagnostic applications.

[0179] According to preferred embodiments, the antibodies or fragments thereof are for use in human medicine, particularly as a medicament in human medicine.

[0180] As used herein, an “IL-11 -related” or “IL-11 -associated" disorder / disease includes conditions associated with or characterized by aberrant IL-11 expression levels. In some embodiments, an “IL-11 -related disorder” or “IL-11 -associated disorder” includes conditions involving cells expressing IL-11 or cells that respond to the binding of IL-11 to the IL-11 receptor, for example, through IL-11 -dependent signal transduction.

[0181] A preferred embodiment of the invention relates to the antibody or described herein for use in a method of preventing and / or treating a disease which is associated with, accompanied by and / or caused by IL-11 activity, e.g. IL-11 -dependent signal transduction. IL-11 activity can be related to IL-11 expression as herein defined.

[0182] The terms "level of expression" or "expression level" in general are used interchangeably and generally refer to the amount of a polynucleotide, mRNA, or an amino acid product or protein in a biological sample. "Expression" generally refers to the process by which gene-encoded information is converted into the structures present and operating in the cell. Therefore, according to the disclosure "expression" of a gene (e.g., the IL-11 gene) may refer to transcription into a polynucleotide, translation into a protein, or even posttranslational modification of the protein. Fragments of the transcribed polynucleotide, the translated protein, or the post- translationally modified protein shall also be regarded as expressed whether they originate from a transcript generated by alternative splicing or a degraded transcript, or from a post-translational processing of the protein, e.g., by proteolysis.

[0183] In some embodiments, "expression level" refers to amount of a protein (e.g., IL-11 ) in a biological sample as determined using methods known in the art or described herein, including but not limited to immunohistochemistry (IHC), immunoblotting (e.g., Western blotting), immunofluorescence (IF), flow cytometry, for example Fluorescence- Activated Cell Sorting (FACS™), or Enzyme-Linked Immunosorbant Assay (ELISA).

[0184] "Increased expression," "increased expression level," "increased levels," "elevated expression," "elevated expression levels," or "elevated levels" refers to an increased expression or increased levels of a biomarker in an individual, i.e. IL-11 in the context of this invention, relative to a control, such as an individual or individuals who are not suffering from the disease or disorder (e.g., cancer or autoimmune disorder) or an internal control (e.g., a housekeeping biomarker).

[0185] "Decreased expression," "decreased expression level," "decreased levels," "reduced expression," "reduced expression levels," or "reduced levels" refers to a decrease expression or decreased levels of a biomarker in an individual, i.e. IL-11 in the context of this invention, relative to a control, such as an individual or individuals who are not suffering from the disease or disorder or an internal control (e.g., a housekeeping biomarker). In some embodiments, reduced expression is little or no expression.

[0186] The disclosure also pertains to the methods of manufacturing a medicament for use in the prevention or treatment of proliferative disorders, autoimmune disorders or inflammatory disorders, comprising an anti-IL-11 antibody or binding fragment thereof.

[0187] Treatment and / or Prevention of IL-11 associated Disorders

[0188] The antibodies or fragments thereof claimed are particularly suitable for use in a method of preventing and / or treating a disease which is selected from an inflammatory disease, an autoimmune disease, a kidney disease, a liver disease, a cardiovascular disease, a pulmonary disease, a pancreatic disease, a muscle disease, a neurological disease, e.g. a disease of the central nervous system (CNS), a metabolic disease and cancer. The subject to be treated is preferably a human.

[0189] The inventive antibodies and fragments thereof are particularly suitable for the treatment of inflammatory and / or autoimmune related disorders.

[0190] The antibodies or fragments thereof claimed are especially suitable for use in a method of preventing and / or treating inflammatory disorders in particular rheumatoid arthritis, multiple sclerosis, asthma, inflammatory bowel disease, systemic sclerosis, periodontitis, chronic obstructive pulmonary disease (COPD), psoriasis or neuroinflammation; and / or kidney and liver disease in particular renal fibrosis, acute kidney injury, chronic kidney disease, diabetic kidney disease, kidney injury (e.g., caused by ischemia, drug toxicity, hypertension), non-alcoholic steatohepatitis (NASH), alcohol-related liver disease (ALD), hepatic fibrosis, drug-induced liver damage (e.g. acetaminophen-induced liver injury) or chronic kidney disease; and / or cardiovascular and pulmonary disorders in particular cardiac fibrosis, atrial fibrosis, idiopathic pulmonary fibrosis, heart failure (e.g., chronic heart failure), atrial fibrillation, lung cancer or emphysema; and / or pancreatic and metabolic disorders, in particular pancreatitis, pancreatic cancer, insulin resistance, glucose intolerance, type 2 diabetes mellitus, adipose tissue dysfunction, obesity, metabolic syndrome or liver metabolism dysfunction; and / or (degenerative) diseases of nervous and muscle system muscle aging, sarcopenia, myogenic differentiation neurodegeneration, cerebral ischemia or reperfusion injury; and / or cancer in particular gastrointestinal cancer, breast cancer and tumor growth and / or progression (e.g., epithelial cancers).

[0191] Since many diseases, such as renal fibrosis, chronic kidney disease, hepatic fibrosis, heart failure, atrial fibrillation, or insulin resistance, often arise directly from inflammatory reactions or involve diseases progression as a consequence of inflammatory mechanisms, the inventive antibodies or fragments thereof are particularly suitable for the treatment and / or prevention of such conditions as well.

[0192] The treatment of fibrosis (e.g. Fibrosis of Skin, Lung-, Liver-, Heart-, Renal-, or Pancreatic Tissue) by the inventive antibodies is particularly preferred. Efficacy has been demonstrated for the preferred antibody HC7LC8 in animal studies using an inflammation induced mouse model of lung fibrosis (see Fig. 7 and Fig. 8). Thus, the use of HC7LC8 or fragments thereof for use in a method for treating inflammation and fibrosis, in particular lung fibrosis, is especially preferred.

[0193] The inventive antibodies are particularly suitable for use in a method of treating colon cancer (Example 3) and erythroleukemia (Example 4). As used herein, the term "treatment" or "treat" refer to both prophylactic or preventive treatment as well as curative or disease modifying treatment. The treatment may be administered to a subject having a medical disorder or who ultimately may acquire the disorder, in order to prevent, cure, delay the onset of, reduce the severity of, or ameliorate one or more symptoms of a disorder or recurring disorder, or in order to prolong the survival of a subject beyond that expected in the absence of such treatment.

[0194] While preventing and / or treating a disorder, the inventive antibodies are to be administered with a therapeutically effective amount. In general, the term "therapeutically effective amount" refers to an amount effective, at dosages and for periods of time necessary, to achieve a desired therapeutic result.

[0195] A therapeutically effective amount of the antibody of the present invention may vary according to factors such as the disease state, age, sex, and weight of the individual, and the ability of the antibody of the present disclosure to elicit a desired response in the individual.

[0196] The efficient dosages and dosage regimens for the inventive antibodies depend on the disease or condition to be treated and may be determined by the persons skilled in the art. A physician having ordinary skill in the art may readily determine and prescribe the effective amount of the pharmaceutical composition required.

[0197] An exemplary, non-limiting range for a therapeutically effective amount of an inventive antibody may be about 0.1 -100 mg / kg, such as about 0.1 -50 mg / kg, for example about 0.1 -20 mg / kg, such as about 0.1 -10 mg / kg, for instance about 0.5, about such as 0.3, about 1 , about 3 mg / kg, about 5 mg / kg or about 8 mg / kg. An exemplary, non-limiting range for a therapeutically effective amount of an inventive antibody is 0.02-100 mg / kg, such as about 0.02-30 mg / kg, such as about 0.05-10 mg / kg or 0.1 -3 mg / kg, for example about 0.5-2 mg / kg An effective dose of an inventive antibody may be administered using a daily, weekly, biweekly or triweekly dosing period.

[0198] If desired, an effective daily dose of a pharmaceutical composition may be administered as two, three, four, five, six or more sub-doses administered separately at appropriate intervals throughout the day, optionally, in unit dosage forms. In some embodiments, the inventive antibodies are administered by slow continuous infusion over a long period, such as more than 24 hours, in order to minimize any unwanted side effects.

[0199] The dosing period may be restricted to, e.g., 8 weeks, 12 weeks or until clinical progression has been established. As nonlimiting examples, treatment according to the present disclosure may be provided as a daily dosage of an inventive antibody in an amount of about 0.1 -100 mg / kg, such as 0.2, 0.5, 0.9, 1.0, 1.1 , 1.5, 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, 40, 45, 50, 60, 70, 80, 90 or 100 mg / kg, per day, on at least one of days 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, 31 , 32, 33, 34, 35, 36, 37, 38, 39, or 40, or alternatively, at least one of weeks 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19 or 20 after initiation of treatment, or any combination thereof, using single or divided doses every 24, 12, 8, 6, 4, or 2 hours, or any combination thereof.

[0200] The present disclosure also provides for therapeutic applications where an inventive antibody is used in combination with at least one further therapeutic agent, in particular a therapeutic agent being relevant for the disease or disorder to be prevented and / or treated, as described above. Exemplary therapeutic agents include other anti-cancer antibodies or ADCs, cytotoxic agents, immunotherapy, chemotherapeutic agents, anti- angiogenic agents, anti-cancer immunogens, cell cycle control / apoptosis regulating agents, hormonal regulating agents, and other agents described herein. Preferably, the therapeutic agent may be a chemotherapeutic agent or an immunotherapeutic agent such as a checkpoint inhibitor, e.g., an anti-PD1 , PD-L1 or CTLA-4 antibody.

[0201] Administration of an inventive antibody in combination with a further therapeutic agent may be simultaneous, separate or sequential. For simultaneous administration, the agents may be administered as one composition or as separate compositions, as appropriate.

[0202] Diagnostic and prognostic Methods

[0203] The anti-IL-11 antibodies or fragments thereof provided herein may be useful for detecting the presence of IL-11 in a biological sample.

[0204] Thus, one aspect of the present invention relates to an IL-11 antibody or fragment thereof as herein disclosed for use in a method of diagnosis and / or detection.

[0205] According to another aspect of the invention, the use of an anti-IL-11 antibody or fragment thereof as herein disclosed in the manufacture of a reagent for use in a method of diagnosis or detection.

[0206] In certain embodiments the present invention relates to a method of detecting the presence or expression level of IL-11 in a biological sample from a subject, wherein the method comprises: contacting the biological sample with an anti-IL-11 antibody as herein disclosed and detecting the presence of the bound antibody. The subject is preferably a human.

[0207] As herein understood a biological sample can be a tissue or a cell sample. The source of the tissue or cell sample may be solid tissue as from a fresh, frozen and / or preserved organ or tissue sample or biopsy or aspirate; blood or any blood constituents; bodily fluids such as cerebral spinal fluid, amniotic fluid, peritoneal fluid, or interstitial fluid; cells from any time in gestation or development of the subject. The biological sample can also be obtained from in vitro tissue or cell culture. The tissue sample may contain compounds which are not naturally intermixed with the tissue in nature such as preservatives, anticoagulants, buffers, fixatives, nutrients, antibiotics, or the like.

[0208] Examples of biological samples herein include, but are not limited to, tumor biopsies, circulating tumor cells, serum or plasma, circulating plasma proteins, ascitic fluid, primary cell cultures or cell lines derived from tumors or exhibiting tumor-like properties, as well as preserved tumor samples, such as formalin-fixed, paraffin- embedded tumor samples or frozen tumor samples. In specific embodiments the biological sample is a tissue sample, notably a fixed tissue sample, and more particularly paraffin-embedded tissue sample.

[0209] A sample as referred to herein, is preferably selected from the group comprising a tissue sample, a blood sample, a serum sample, and a plasma sample. More specifically, the sample is a tissue sample, e.g. a fixed tissue sample.

[0210] In some embodiments, the biological sample is from a subject having, predisposed to, or being tested for a proliferative disorder, e.g. inflammatory disease or cancer. More specifically the tissue is a cancer tissue sample. By “cancer tissue”, it is herein intended that the tissue is obtained from a subject suffering from a cancer and is taken from an organ or from a tissue affected by cancer.

[0211] In some embodiments, the biological sample is from a subject having, predisposed to, or being tested for an autoimmune or inflammatory disorder. More specifically, the tissue is a sample from a tissue affected by an autoimmune or inflammatory disease. By 'tissue affected by an autoimmune or inflammatory disease,' it is intended that the tissue is obtained from a subject suffering from such a disease and is taken from an organ or tissue affected by the autoimmune or inflammatory condition. In one embodiment, for example, a method of detecting the presence of IL-11 in a biological sample, described below, is provided. In certain embodiments, the method comprises contacting the biological sample with an IL-11 antibody as described herein under conditions permissive for binding of the anti-IL-11 antibody to IL-11 and detecting whether a complex is formed between the anti-IL-11 antibody and IL-11 . Such method may be an in vitro or in vivo method. Anti-IL-11 antibodies disclosed herein can be used, for example, in immunoassays, including, for example, immunohistochemistry (IHC), immunofluorescence (IF), immunoblotting (e.g., Western blotting), flow cytometry (e.g., FACS™), and Enzyme-linked Immunosorbent Assay (ELISA). Often the immuno assay is IHC and the biological sample is a fixed tissue, notably FFPE.

[0212] In one embodiment, an anti-IL-11 antibody is used to select subjects eligible for therapy with an anti-IL-11 antibody (in particular with the same anti-IL-11 antibody), typically, where IL-11 is a biomarker for selection of patients.

[0213] The disclosure further provides for the use of an anti-IL-11 antibody in a method of diagnosing a subject suffering from a disorder (e.g., a cancer, an inflammatory disorder or an autoimmune disease), the method comprising: determining the presence or expression level of IL-11 in a sample obtained from the subject by contacting the sample with an anti-IL-11 antibody as previously herein described and detecting the presence of the bound antibody.

[0214] The present disclosure yet further provides for the use of a IL-11 antibody in the manufacture of a reagent for use in a method of diagnosing a subject suffering from a disorder (e.g., a cancer, an inflammatory disorder or an autoimmune disease), the method comprising: determining the presence or expression level of IL-11 in a sample obtained from the subject by contacting the sample with an anti- IL-11 antibody as herein disclosed and detecting the presence of the bound antibody.

[0215] In another embodiment, the present disclosure provides a method for identifying a subject suffering from a disorder (e.g., cancer, an inflammatory disorder or an autoimmune disease) who is likely to respond to a treatment, the method including: determining the presence or expression level of IL-11 in a sample obtained from the subject by contacting the sample with an anti-IL-11 antibody as herein disclosed and detecting the presence of the bound antibody, wherein the presence or expression level of IL-11 in the sample indicates that the subject is likely to respond to the treatment.

[0216] More specifically in embodiments of the present disclosure as previously described, the presence and / or expression level / amount of IL-11 in a sample may be determined using IHC and staining protocols using an anti-IL-11 antibody of the present disclosure. IHC staining of tissue sections has been shown to be a reliable method of determining or detecting the presence of proteins in a sample.

[0217] In one embodiment, expression level of IL-11 is determined using a method comprising: (a) performing IHC analysis of a sample (such as a tumor sample obtained from a subject) with an anti-IL-11 antibody as herein disclosed; and (b) determining the presence and / or expression level of IL-11 in the sample. In some embodiments, IHC staining intensity is determined relative to a reference as described below. In some embodiments, the reference may be a reference sample (e.g., a control cell line staining sample, a tissue sample from a healthy subject or a reference sample known to have a pre-determined level of IL-11 expression).

[0218] The term "detecting" as used herein encompasses quantitative or qualitative detection. The present disclosure therefore comprises detecting the presence and / or expression level of IL-11 in the sample. The detection may be performed by methods known in the art, e.g. by immunohistochemistry or FACS.

[0219] All diagnosis and / or detection methods described herein may comprise a step of comparing the determined presence and / or expression level of IL-11 in the sample with a reference, such a reference sample, reference tissue or reference value. The result of such a comparison can form the basis for a diagnosis and / or can be correlated with a disorder to be treated. In general, “reference" as used herein, refers to a sample, cell, tissue, standard, value or level that is used for comparison purposes. In one embodiment, a reference is obtained from a healthy and / or non-diseased part of the body (e.g., tissue or cells) of the same subject or individual. For example, the reference may be healthy and / or nondiseased cells or tissue adjacent to the diseased cells or tissue (e.g., cells or tissue adjacent to a tumor or an inflamed tissue). In another embodiment, a reference sample is obtained from an untreated tissue and / or cell of the body of the same subject or individual. In yet another embodiment, a reference sample is obtained from a healthy and / or non-diseased part of the body (e.g., tissues or cells) of an individual who is not the subject or individual. In even another embodiment, a reference sample is obtained from an untreated tissue and / or cell of the body of an individual who is not the subject or individual.

[0220] Further Aspects of the Invention

[0221] A further aspect of the invention relates to a nucleic acid coding for one or more of the antibodies or fragments thereof described herein.

[0222] More particularly the nucleic acid molecule may encode a heavy chain or a light chain of an antibody of the present disclosure. The nucleic acid molecule may comprise a VH or VL coding region having at least 85%, 90%, 95% or 100% of identity to the corresponding nucleic acid encoding heavy chain variable region (VH region) or light chain variable region (VL) disclosed herein.

[0223] Typically, said nucleic acid is a DNA or RNA molecule, which may be included in any suitable vector, such as a plasmid, cosmid, episome, artificial chromosome, phage or a viral vector. As used herein, the terms "vector", "cloning vector" and "expression vector" mean a vehicle by which a DNA or RNA sequence (e.g., a foreign gene) can be introduced into a host cell, so as to transform the host and promote expression (e.g., transcription and translation) of the introduced sequence. Thus, another aspect of the invention relates to a vector comprising an inventive nucleic acid.

[0224] Such vectors may comprise regulatory elements, such as a promoter, enhancer, terminator and the like, to cause or direct expression of said antibody upon administration to a subject. Examples of promoters and enhancers used in the expression vector for animal cell include early promoter and enhancer of SV40, LTR promoter and enhancer of Moloney mouse leukemia virus, promoter and enhancer of immunoglobulin H chain and the like. Any expression vector for animal cell can be used, so long as a gene encoding the human antibody C region can be inserted and expressed. Examples of suitable vectors include pAGE107, pAGE103, pHSG274, pKCR, pSGI beta d2-4 and the like. Other examples of plasmids include replicating plasmids comprising an origin of replication, or integrative plasmids, such as for instance pUC, pcDNA, pBR, and the like. Other examples of viral vector include adenoviral, retroviral, herpes virus and AAV vectors.

[0225] Such recombinant viruses may be produced by techniques known in the art, such as by transfecting packaging cells or by transient transfection with helper plasmids or viruses. Typical examples of virus packaging cells include PA317 cells, PsiCRIP cells, GPenv+ cells, 293 cells, etc.

[0226] The nucleic acids as herein disclosed may be used to produce an antibody of the present disclosure in a suitable expression system. The term "expression system" means a host cell and compatible vector under suitable conditions, e.g., for the expression of a protein coded for by foreign DNA carried by the vector and introduced to the host cell. Common expression systems include E. coli host cells and plasmid vectors, insect host cells and Baculovirus vectors, and mammalian host cells and vectors. Other examples of host cells include, without limitation, prokaryotic cells (such as bacteria) and eukaryotic cells (such as yeast cells, mammalian cells, insect cells, plant cells, etc.). Specific examples include E.coli, Kluyveromyces or Saccharomyces yeasts, mammalian cell lines (e.g., Veto cells, CHO cells, 3T3 cells, COS cells, etc.) as well as primary or established mammalian cell cultures (e.g., produced from lymphoblasts, fibroblasts, embryonic cells, epithelial cells, nervous cells, adipocytes, etc.).

[0227] Yet another aspect of the invention relates to a host cell comprising a vector as herein described. The host cell may be a procaryotic or eukaryotic cell.

[0228] LEGENDS OF THE FIGURES

[0229] Figure 1 : Binding of the antibodies to human and mouse IL-11

[0230] Shown is concentration dependent binding of primary antibodies to (A) human IL-11 and (B) mouse IL-11 . (BG - background).

[0231] Figure 2: Binding of antibodies to related cytokines

[0232] Shown is the binding of primary antibodies to cytokine coated ELISA plates at two different concentrations: (A) 100 pg / ml and (B) 10 pg / ml.

[0233] Figure 3: Inhibition of IL-11 binding to IL-11 receptor

[0234] Shown is the Inhibition of IL-11 binding to IL-11 receptor by various antibodies

[0235] Figure 4: Inhibition of IL-11 -induced cellular signaling

[0236] Antibodies inhibit IL-11 -induced cellular signaling in IL-11 stimulated human DLD-1 colon cancer cells. Shown are controls together with (A) 18A2: rat anti-IL-11 antibody, HC7LC8, HC10LC8; (B) HC7LC8, and comparator antibodies.

[0237] Figure 5: Inhibition of IL-11 induced cell proliferation

[0238] Shown is the concentration-dependent inhibition of IL-11 -induced proliferation of human FT-1 erythroleukemia cells by anti-IL-11 antibodies.

[0239] Figure 6: Antibody epitope determination

[0240] HC7LC8 binds a different epitope from commercial MAB218 antibody. Shown is the binding of (A) HC7LC8 and (B) MAB218 to human IL-11 derived epitope sequences and the localization (arrows) of binding sites in the structure of human IL-11 for (C) HC7LC8 and (D) MAB218.

[0241] Figures 7 - 9: Efficacy of HC7LC8 in a mouse model of inflammatory lung disease

[0242] Figure 7 shows the Outline of Bleomycin lung fibrosis model in mice

[0243] Figure 8 shows (A) body weight development in the treatment and control groups, and (B) the respiratory parameter of enhanced pause (Penh) in each respective group.

[0244] Figure 9 illustrates that Anti-IL-11 HC7LC8 prevents development of collagen disposition in the lung. Shown is the effect of anti-IL-11 HC7LC8 treatment on Ashcroft scores, with (A) average scores per animal, (B) average scores across all individual sections per treatment group, and (C) representative collagen distribution in lung tissue stained with Picrosirius Red.

[0245] Figure 10: Inhibition of colon cancer growth in combination with anti-PD1

[0246] Shown is (A) the tumor volume of MC38 colon cancer cells treated twice weekly with vehicle PBS, anti-IL11 18A2 (20 mg / kg), anti-PD1 RMP1 -14 (10 mg / kg) or the combination of anti-IL11 and anti-PD1 and (B) the body weight of the treated mice.

[0247] Examples

[0248] The following examples serve to further illustrate the invention. They are not intended to limit the invention in any way.

[0249] Example 1 : Specific IL-11 Binding

[0250] The binding affinity of the antibodies listed in Table 1 to orthologous IL-11 cytokines from the species human, rat, mouse, and cynomolgus was determined using ELISA and surface plasmon resonance (SPR). For the ELISA (Fig. 1 ), a direct assay set up was employed where the antigen adhered to the ELISA plate and binding of the candidate antibodies detected via secondary antibodies conjugated to alkaline phosphatase (Kontermann, R., and Dubel, S. (2001 ). Antibody Engineering (Berlin, Heidelberg: Springer Berlin Heidelberg). For the SPR assay, the antibody was bound via its Fc region to protein A / G coated SPR sensors on a MASS-2 instrument (Sierra Sensors / Bruker) and binding of soluble IL-11 recorded when run over the sensor chip surface. As a comparison, antibodies known in the art were recombinantly produced in CHO cells and purified to homogeneity. For this, the antibody sequences were used which were published in the following patents. The antibodies 3C6, Enleo3 and Enleo4 were derived from Seq IDs 91 / 92 (VH / VL), 8 / 83 and 8 / 20, respectively of US 11084874 B2. Antibodies mu18A10 and mu8C8 were derived from WO 2023 / 143590 A1 Seq. IDs 5 / 6 (VH / VL) and 7 / 8, respectively. In addition, a commercially available known blocking antibody, MAB218 (R&D Systems), was tested (Tab. 1 ).

[0251] Table 1 : All antibodies bind to human IL-11.

[0252] Some of the disclosed antibodies (18A2 HC7LC8, HC10LC8 and 12B3) exhibit comparable affinity also for other orthologous cytokines (Tab. 2 and 3), whereas the comparator antibodies, including Enleo3, Enleo4, and 3C6 possess a significantly lower affinity to human and mouse IL-11 IL-11 in the SPR assay.

[0253] Table 2: Binding affinity (Kd) of the antibodies for IL-11 IL-11 orthologs determined by ELISA

[0254] Binding to the IL-11 orthologs was determined as described in Fig. 1. The Kd values were determined by fitting the data to a one site specific binding model using GraphPad Prism 7.03. Kd in nM, Average of 3-4 experiments, NA - Not applicable / no binding, ND - Not determined

[0255] Table 3: Binding affinity of the antibodies for IL-11 IL-11 orthologs as determined by Surface Plasmon Resonance (SPR)

[0256] Shown are Kd-values in nM, Average of 2-3 experiments, ND - Not determined

[0257] The specificity of the antibody binding to IL-11 was interrogated by an ELISA that used cytokines related in amino acid sequence to IL-11 as the antigen coated to the ELISA plate (Fig. 2). ELISA plates were coated with 1 pg / ml cytokine. After blocking with BSA, the plates were incubated with 100 pg / ml (A) or 10 pg / ml antibody (B). Binding of the primary antibodies was detected by means of alkaline phosphatase-coupled secondary antibodies directed against the constant part of rat (18A2), mouse (MAB218) or human IgG (HC7LC8, Enleo3, Enleo4). The following cytokines were probed: hLIF - human Leukemia Inhibitory Factor, hOSM - human Oncostatin M, hlL- 6 - human lnterleukin-6, hlL-27 - human Interleukin-27, hlL-11 - human Interleukin- 11 , mTGF-b1 - mouse Transforming Growth Factor b1. As can be seen in Fig. 2, binding of HC7LC8 and 18A2 was specific for IL-11 whereas comparator antibody MAB21 8 showed also considerable binding to IL27 and comparator antibodies Enleo3, Enleo4 exhibited substantial binding to LIF and TGF-0 and comparator antibody 3C6 did not bind to any of the cytokines, including IL-11 , in this assay set up.

[0258] These experiments show that the 3C6 antibody binds to an IL-11 epitope that is not accessible when the ligand is coated to a solid support but that is free to bind the antibody in a more physiological set up when the ligand is in solution and the antibody attached to a solid support, as in the SPR assay. In contrast, Enleo3 and Enleo4 bind an epitope that appears to be mostly accessible when the ligand is coated but barely when the ligand is in solution as exemplified by the ELISA and SPR set ups and binding data with these antibodies. The remaining antibodies including 18A2 and HC7LC8 do not exhibit this dependency on the molecular orientation of the ligand since the ELISA and SPR methods and set ups do not result in such strikingly different affinities.

[0259] Example 2: Inhibition of IL-11 binding to IL-11 -receptor

[0260] To investigate whether the antibodies inhibited IL-11 binding to its receptor, an in vitro IL-11 / IL-11 Ra binding assay was set up without incorporating gp130. For this, ELISA plates were coated with an anti-human Fc antibody (Fig. 3A) or anti-mouse-Fc antibody (Fig. 3B) and blocked with BSA. Subsequently, a solution containing mouse IL-11 - receptor-human-Fc fusion protein (R&D Systems #490-IR-200) or mouse IL-11 - receptor-mouse-Fc fusion protein (R&D Systems #7405) was added and incubated over night at 4°C. On the next day, human or mouse IL-11 and a concentration range of the candidate and comparator antibodies or isotype controls was added to the plates and incubated at room temperature for 2h. After washing, binding of IL-11 to the IL-11 receptor was revealed by means of a biotinylated antibody against human (R&D Systems #BAF218) or mouse (R&D Systems #BAF418) IL-11 and alkaline phosphatase-coupled streptavidin whose concentration was estimated using the AttoPhos substrate (Promega). For the IC50 values, data were fitted by non-linear regression to an inhibitor- response model using GraphPad Prism 7.03

[0261] As can be seen in Fig. 3 and Tab. 4, several of the disclosed antibodies, in particular 18A2 and HC7LC8, were considerably more potent inhibiting IL-11 binding to the IL-11 receptor than the comparator antibodies. Specifically, the comparator antibodies Enleo3, Enleo4, mu8C8 and mu18A10 did not inhibit binding of IL-11 to IL-11 RA while the comparator antibodies 3C6 and MAB218 were significantly less potent at inhibiting human or mouse IL-11 binding to IL-11 RA than the antibodies 18A2 and HC7LC8.

[0262] Table 4: IC50 for inhibition of IL-11 binding to mouse IL-11Ra

[0263] Example 3: Inhibition of IL-11 -induced cellular signaling

[0264] The antibodies 18A2, HC7LC8 and HC10LC8 disclosed here are potent inhibitors of IL-11 induced cell signaling in human DLD-1 colon cancer cells (Fig. 4), murine NIH3T3 fibroblasts and rat L6 myoblasts. (Tab. 5). The comparator antibodies mu18A10 and 3C6 also show good inhibition of human and mouse IL-11. In contrast, Enleo3 and Enleo4 have no recognizable activity and mu8C8 appears to have only an effect on mouse IL-11 stimulation. To determine the species-dependent inhibition of IL-11 -induced cell signaling, human (DLD-1 colon carcinoma cells), mouse (NIH-3T3 mouse fibroblasts), or rat (L6 rat myoblasts) cells were seeded in 24-well plates on day 1 in growth medium containing 10% FBS (RPMI for DLD-1 , DMEM for NIH-3T3 and L6). On the next day, the cell culture medium was replaced by medium without FBS. On day 3, human, mouse, or rat IL-11 (10 ng / ml human or rat IL-11 , 1 ng / ml mouse IL-11 ) were incubated for 2 hours in medium containing a concentration range of the antibodies. Subsequently, the medium on the cells was replaced with the medium containing IL-11 and the antibodies. Human IL-11 containing medium was used for DLD-1 , whereas mouse and rat IL-11 were used for NIH-3T3 and L6 cells, respectively. After 20 minutes incubation at 37°C, the cells were lysed on ice and the lysates used to measure STAT3 phosphorylation using a commercial pSTAT3 sandwich ELISA (Cell Signaling Technology #7300C). For the calculation of the IC50 values, background pSTAT3 signal without IL-11 was subtracted and the resultant values fitted by non-linear regression to an inhibitorresponse model using GraphPad Prism 7.03.

[0265] Table 5: IC50 values (nM) of the antibodies for inhibition of hlL-11-induced cellular signaling

[0266] NA - Not applicable / no inhibition, ND - Not determined

[0267] Example 4: Inhibition of IL-11 -induced proliferation

[0268] The inhibition of IL-11 -dependent proliferation of cancer cells was demonstrated using human TF-1 erythroleukemia cells as an example. For this, the GM-CSF that is normally used in the RPMI medium alongside 10% FBS was substituted by 10 ng / ml human IL-11. In addition, a concentration range of the antibodies was added in 96-well plates and the cells incubated for 3 days. On the last day, the cell viability was measured using the CellTiter-Glow luminescent assay (Promega). For the calculation of IC50 values for the inhibition of IL-11 -induced cell proliferation, background values without IL-11 stimulation were subtracted and the resulting values normalized to the IL-11 stimulation without antibody thus resulting in % inhibition of IL-11 -induced cell proliferation. The values were subsequently fitted to a 3-parameter inhibitor-response model using non-linear regression in GraphPad Prism 7.03. An example assays is shown in Fig 5 and average values of three independent experiments in Tab. 6.

[0269] Table 6: IC50 for inhibition of IL-11 -induced TF-1 cell proliferation

[0270] NA - Not applicable / no inhibition

[0271] Since the mu18A10 and mu8C8 antibodies inhibit IL-11 -induced effects in cells but not IL-11 binding to IL-11 Ra (Example 2), it follows that the mechanism of action and epitope of these antibodies is different from the other here tested antibodies, including 18A2 and HC7LC8. The Enleo3 and Enleo4 antibodies do not bind to IL-11 in solution explaining their lack of effect in assays interrogating inhibition of ligand action (Examples 2-4). Example 5: Epitope determination of the candidate antibodies

[0272] Epitope mapping was performed by peptide scanning of overlapping human IL-11 - derived peptides bound to an ELISA plate and covering the whole IL-11 sequence. Binding of HC7LC8 (Fig. 6A) and MAB218 (Fig. 6B) was revealed by means of a secondary anti-human-Fc (A) or mouse-Fc (B) antibody coupled to horse raddish peroxidase. HC7LC8 binds primarily to a stretch of amino acids at position 105-114 (aa sequence: LRHVQWLRRA) whereas MAB218 binds to amino acids 130-143 (aa sequence: ARLDRLLRRLQLL). The location of the epitopes in the structure of human IL-11 (PDB 6040) is depicted by arrows and shading in Fig. 6C (HC7LC8) and Fig. 6D (MAB218).

[0273] Example 6: Attenuation of disease pathology in a mouse model of inflammatory lung disease

[0274] Antibody HC7LC8 was also used in a mouse model of inflammatory lung disease which results in collagen disposition obstructing the breathing function in the mice. For this, Bleomycin was dosed once by oropharyngeal aspiration (OP) on Day 1 at 1.30 mg / kg (1696 lU / mg) in a total of 26 C57BL / 6 female mice. A control group of 6 mice received only saline by oropharyngeal aspiration. 13 Bleomycin treated mice each were subsequently injected i.v. with 20 mg / kg HC7LC8 or isotype control antibody on days 2, 6, 10, 13, 16, and 20. Body weight and clinical signs were monitored daily and on days 8 and 19, respiratory changes were estimated by whole body plethysmography (Fig. 7). As shown in Fig. 8A, treatment with HC7LC8 led to a slower transient loss of body weight compared to isotype control on days 7-11 which are typically characterized by lung inflammation. Additionally, the respiratory parameter of enhanced pause (Penh) was reduced in HC7LC8-treated mice suggesting that the anti-IL-11 antibody partially prevented Bleomycin-induced airway resistance by the ongoing inflammation (Fig. 8B). On day 16, 2 mice of each group were sacrificed, the lungs removed and frozen for further gene expression analysis. During the study, one animal of the isotype control group and 2 animals of the HC7LC8 were prematurely sacrificed due to body weight loss that was unrelated to the antibody treatments. On day 21 , the remaining mice were sacrificed and the lungs fixed in neutral buffered formalin. 9 sections were prepared for each lung covering left and right lobes thus obtaining a total of 90 sections for the Isotype control group from 10 animals and 81 sections for the anti-IL-11 (HC7LC8) group from 9 animals. The sections were stained for collagen deposition using Picrosirius Red and evaluated by a board-certified veterinary pathologist who assigned a modified Ashcroft score to each section in a blinded fashion. As can be seen in Fig. 9, treatment with anti-IL-11 HC7LC8 reduced the Ashcroft scores of collagen staining when the scores of the sections were averaged per animal (Fig. 9A) or when all 90 and 81 single section Ashcroft values were averaged per treatment group (Fig. 9B). Taken together, these data suggest that HC7LC8 can reduce Bleomycin-induced lung inflammation, collagen deposition and breathing impairments.

[0275] Example 7: Inhibition of tumor growth in combination with immune checkpoint inhibitor

[0276] The antibody 18A2 was also used in a mouse model of colon cancer to test its effect on tumor growth in combination with immune checkpoint inhibitors. For this, 7- to 9- week-old female C57BL / 6 mice were inoculated subcutaneously in the right lower flank region with 1 x 106MC38 tumor cells in 0.1 mL of PBS. Tumor development was monitored and mice randomized into 4 groups of 10 mice each with average tumor group size of approximately 96 mm3. The treatment was initiated on the same day (day 0) and comprised intraperitoneal twice-weekly injections of the groups with PBS, 20 mg / kg 18A2, 10 mg / kg anti-PD1 (Programmed cell death protein 1 ) or the combination of 18A2 and anti-PD1 at the same doses. Tumor volumes and body weights were measured 3 times per week. Tumor volumes were measured in two dimensions using a caliper, and the volume was expressed in mm3using the formula: V = (L x W x W) / 2, where V was tumor volume, L is tumor length (the longest tumor dimension) and W was tumor width (the longest tumor dimension perpendicular to L). As can be seen in Fig. 10A, treatment with the combination of anti-IL11 and anti-PD1 led to a statistically significant tumor growth inhibition (One-Way ANOVA) compared to the PBS control group whereas the single agent treatments did not result in a significant reduction of tumor growth. Thus, there was a combinatorial effect of anti-IL11 with immune checkpoint blockade suggesting that both treatments impact anti-tumor immunity. There was no adverse effect on body weight by any of antibodies (Fig. 10B). Example 8: Variable heavy- and light-chain Sequences of anti-IL-11 antibodies

[0277] Table 7 shows heavy and light chain variable Sequences of the rat anti-IL-11 antibodies 18A2 (18A2-VH / 18A2-VL) and 12B3 (12B3-VH / 12B3-VI) and of humanized anti-IL- 11 antibodies HC7LC8 (HC7-VH / LC8-VL) HC10LC8 (HC10-VH / LC8-VL), antibody- A (14B11 -VH / 14B11-VL), antibody-B (19C6-VH / 14BII-VL), and antibody-C (19C6- VH / 19C6-VL).

[0278] CDRs of respective antibodies are marked in Table 7:

[0279] - according to Kabat numbering in “bold” and

[0280] - according the IMGT-numbering by “underlining” respective CDR sequences.

[0281] Table 7

[0282] Table 8: Sequence Listing

[0283]

Claims

CLAIMS1 . A monoclonal antibody or antigen-binding fragment thereof which binds to Interleukin-11 (IL-11 ) comprising(a) a variable heavy chain (VH) region comprising complementaritydetermining regions (CDRs) CDR-H1 , CDR-H2 and CDR-H3 (according to Kabat), wherein(i) the CDR-H1 comprises an amino acid sequence according to SEQ ID NO: 1 , 12, 20, or 28, or an amino acid sequence comprising a deletion, insertion and / or substitution, particularly a conservative substitution, of 1 or 2 amino acids,(ii) the CDR-H2 comprises an amino acid sequence according to SEQ ID NO: 2, 13, 21 , or 29, or an amino acid sequence comprising a deletion, insertion and / or substitution, particularly a conservative substitution, of 1 or 2 amino acids,(iii) the CDR-H3 comprises an amino acid sequence according to SEQ ID NO: 3, 14, 22, or 30, or an amino acid sequence comprising a deletion, insertion and / or substitution, particularly a conservative substitution, of 1 or 2 amino acids, optionally(b) a variable light chain (VL) region, particularly a VL region comprising complementarity-determining regions (CDRs) CDR-L1 , CDR-L2 and CDR-L3, wherein(i) the CDR-L1 comprises an amino acid sequence according to SEQ ID NO: 7, 16, 24, or 32,or an amino acid sequence comprising a deletion, insertion and / or substitution, particularly a conservative substitution, of 1 or 2 amino acids,(ii) the CDR-L2 comprises an amino acid sequence according to SEQ ID NO: 8, 17, 25, or 33, or an amino acid sequence comprising a deletion, insertion and / or substitution, particularly a conservative substitution, of 1 or 2 amino acids,(iii) the CDR-L3 comprises an amino acid sequence according to SEQ ID NO: 9, 18, 26, or 34, or an amino acid sequence comprising a deletion, insertion and / or substitution, particularly a conservative substitution, of 1 or 2 amino acids.

2. The antibody or antigen-binding fragment of claim 1 comprising(i) a VH region comprising the CDR-H1 of SEQ ID NO:1 , the CDR- H2 of SEQ ID NO:2, and the CDR-H3 of SEQ ID NO:3, and optionally(ii) a VL region comprising the CDR-L1 of SEQ ID NOT, the CDR-L2 of the SEQ ID NO:8 and the CDR-L3 of the SEQ ID NO:9.

3. The antibody or antigen-binding fragment of any of claims 1 -2 comprising(a) a VH region comprising an amino acid sequence according to SEQ ID NO: 4, 5 or 6 or an amino acid sequence having an identity thereof of at least 85%, at least 90%, at least 95% or at least 99%, and optionally(b) a VL region, particularly a VL region comprising an amino acid sequence according to SEQ ID NO: 10 or 11 or an amino sequence having an identity thereof of at least 85%, at least 90%, at least 95% or at least 99%.

574. The antibody or antigen-binding fragment of any of claims 1 or 2 comprising(a) a VH region comprising an amino acid sequence according to SEQ ID NO: 4 or an amino acid sequence having an identity thereof of at least 85%, at least 90%, at least 95% or at least 99%, and optionally(b) a VL region, particularly a VL region comprising an amino acid sequence according to SEQ ID NO: 10 or an amino sequence having an identity thereof of at least 85%, at least 90%, at least 95% or at least 99%.

5. The antibody or antigen-binding fragment of any of claims 1 -2 comprising(a) a VH region comprising an amino acid sequence according to SEQ ID NO: 5 or an amino acid sequence having an identity thereof of at least 85%, at least 90%, at least 95% or at least 99%, and optionally(b) a VL region, particularly a VL region comprising an amino acid sequence according to SEQ ID NO: 10 or an amino sequence having an identity thereof of at least 85%, at least 90%, at least 95% or at least 99%.

6. The antibody or antigen-binding fragment of claim 1 comprising(i) a VH region comprising the CDR-H1 of SEQ ID NO: 12, the CDR- H2 of SEQ ID NO: 13, and the CDR-H3 of SEQ ID NO: 14, and optionally(ii) a VL region comprising the CDR-L1 of SEQ ID NO: 16, the CDR- L2 of the SEQ ID NO: 17 and the CDR-L3 of the SEQ ID NO: 18.

7. The antibody or antigen-binding fragment of claim 5 comprising(a) a VH region comprising an amino acid sequence according to SEQ ID NO: 15 or an amino acid sequence having an identity thereof of at least 85%, at least 90%, at least 95% or at least 99%, and optionally58(b) a VL region, particularly a VL region comprising an amino acid sequence according to SEQ ID NO: 19 or an amino sequence having an identity thereof of at least 85%, at least 90%, at least 95% or at least 99%.

8. The antibody or antigen-binding fragment of claim 1 comprising(i) a VH region comprising the CDR-H1 of SEQ ID NO: 20, the CDR- H2 of SEQ ID NO: 21 , and the CDR-H3 of SEQ ID NO: 22, and optionally(ii) a VL region comprising the CDR-L1 of SEQ ID NO: 24, the CDR- L2 of the SEQ ID NO: 25 and the CDR-L3 of the SEQ ID NO: 26.

9. The antibody or antigen-binding fragment of claim 8comprising(a) a VH region comprising an amino acid sequence according to SEQ ID NO: 23 or an amino acid sequence having an identity thereof of at least 85%, at least 90%, at least 95% or at least 99%, and optionally(b) a VL region, particularly a VL region comprising an amino acid sequence according to SEQ ID NO: 27 or an amino sequence having an identity thereof of at least 85%, at least 90%, at least 95% or at least 99%.

10. The antibody or antigen-binding fragment of claim 1 comprising(i) a VH region comprising the CDR-H1 of SEQ ID NO: 28, the CDR- H2 of SEQ ID NO: 29, and the CDR-H3 of SEQ ID NO: 30, and optionally(ii) a VL region comprising the CDR-L1 of SEQ ID NO: 32, the CDR- L2 of the SEQ ID NO: 33 and the CDR-L3 of the SEQ ID NO: 34.11 . The antibody or antigen-binding fragment of claim 10 comprising59(a) a VH region comprising an amino acid sequence according to SEQ ID NO: 31 or an amino acid sequence having an identity thereof of at least 85%, at least 90%, at least 95% or at least 99%, and optionally(b) a VL region, particularly a VL region comprising an amino acid sequence according to SEQ ID NO: 35 or an amino sequence having an identity thereof of at least 85%, at least 90%, at least 95% or at least 99%.

12. The antibody or antigen-binding fragment of any of claims 1 to 11 which binds to human IL-11 with a Kd of 5 nM or less, particularly of 1 nM or less (as determined by ELISA).

13. The antibody or antigen-binding fragment of any of claims 1 to 12 which binds to human IL-11 with a Kd of 5 nM or less, particularly of 1 nM or less (as determined by Surface Plasmon Resonance).

14. The antibody or antigen-binding fragment of any of claims 1 to 13 which inhibits binding of IL-11 to IL-11 RA with an IC50 of 10 nM or less (as determined by ELISA).

15. The antibody or antigen-binding fragment of claim 14 wherein the binding of IL-11 to IL-11 RA is inhibited in the absence of gp130.

16. The antibody or antigen-binding fragment of any of claims 1 to 15 which inhibits IL-11 -induced cellular signaling with an IC50 of 5 nM or less, particularly of 1 nM or less (as determined by ELISA).

17. The antibody or antigen-binding fragment of any of claims 1 to 16 which inhibits IL-11 -induced cell proliferation of cancer cells, particularly of TF-601 erythroleukemia cells, with an IC50 of 5 nM or less, particularly of 1 nM or less (as determined by measurement of the cell titer).

18. The antibody or antigen-binding fragment of any of claims 1 to 17 which binds to an epitope on human IL-11 comprising amino acids 105-114 (as determined by peptide scanning).

19. The antibody or antigen-binding fragment of any of claims 1 to 18 which is a human or humanized antibody, particularly a human or humanized IgG antibody, more particularly a human or humanized lgG1 antibody and even more particularly a human or humanized lgG1 K antibody.

20. The antibody or antigen-binding fragment of any of claims 1 to 19 consisting of or comprising a Fab, Fab', Fab'-SH, F(ah')2, Fv, a diabody, single-chain antibody fragment, or a multispecific antibody.

21. The antibody or antigen-binding fragment of any of claims 1 to 20 comprising a labeling group and / or an effector group being coupled to the antibody or antigen- binding fragment.

22. The antibody or antigen-binding fragment of claim 21 wherein the labeling group is a dye, a paramagnetic, radioactive or fluorogenic group that is detectable upon imaging.

23. The antibody or antigen-binding fragment of claim 21 wherein the effector group is a therapeutic group, in particular a cytotoxic agent.

24. The antibody or fragment of any one of claims 1 -23 for use in medicine, particularly for therapeutic or diagnostic applications including in vitro and in vivo diagnostic applications.

25. The antibody or fragment of any one of claims 1 -23 for use in human medicine, particularly as a medicament in human medicine.

26. The antibody or fragment of any one of claims 1 -23 for use according to claim 24 or 25 in a method of preventing and / or treating a disease which is associated with, accompanied by and / or caused by IL-11 activity.

27. The antibody or fragment of any one of claims 1 -23 for use according to any one of claims 24 to 26 in a method of preventing and / or treating a disease which is selected from an inflammatory disease, an autoimmune disease, a kidney disease, a liver disease, a cardiovascular disease, a pulmonary disease, a pancreatic disease, a muscle disease, a neurological disease, e.g. a disease of the central nervous system (CNS), a metabolic disease and cancer.

28. The antibody or fragment of any one of claims 1 -23 for use according to any one of claims 24 to 27 in a method of preventing and / or treating rheumatoid arthritis, multiple sclerosis, asthma, inflammatory bowel disease, systemic sclerosis, periodontitis, chronic obstructive pulmonary disease (COPD), psoriasis, pancreatitis, renal fibrosis, acute kidney injury, chronic kidney disease, diabetic kidney disease, kidney injury, e.g. from various insults such as ischemia, drug toxicity, hypertension and urinary tract obstruction, non-alcoholic steatohepatitis (NASH), alcohol- related liver disease (ALD), hepatic fibrosis, drug-induced liver damage, e.g., acetaminophen-induced liver injury, cardiac fibrosis, heart failure, e.g., chronic heart failure, atrial fibrillation, atrial fibrosis, idiopathic pulmonary fibrosis, lung cancer, emphysema, pancreatitis, pancreaticcancer, muscle aging, sarcopenia, cancer-associated cachexia, myogenic differentiation, neuroinflammation, neurodegeneration, cerebral ischemia, reperfusion injury, insulin resistance, glucose intolerance, type 2 diabetes mellitus, adipose tissue dysfunction, obesity, metabolic syndrome, liver metabolism dysfunction, or tumor growth and / or progression, e.g. in epithelial cancers, e.g., gastrointestinal cancer, and in breast cancer.

29. The antibody or fragment of any one of claims 1 -23 for use according to any one of claims 24 to 28 in combination with a further therapeutic agent.

30. The antibody or fragment of any one of claims 1 -23 for use according to claim 29 wherein the further therapeutic agent is a chemotherapeutic agent or an immunotherapeutic agent such as a checkpoint inhibitor, e.g., an anti-PD1 antibody.31 . A nucleic acid coding for one or more of the antibodies according to any one claims 1 -23, or for at least one VL and / or one VH of any one of the antibodies of claims 1 -23.

32. A vector comprising the nucleic acid according to claim 31 .

33. A host cell comprising the vector of claim 32.

34. A pharmaceutical composition comprising an antibody or antigen-binding fragment thereof according to any of claims 1 -23 and a pharmaceutically acceptable carrier.63