Treatment of kidney damage

Abstract

Disclosed are methods for treating or preventing kidney damage through the inhibition of interleukin-11 (IL-11)-mediated signaling, and agents for use in such methods.

Description

[Technical field]

[0001] This application claims priority to UK Patent Publication No. 1902419.9, filed on 22 February 2019, the contents and material of which are incorporated by reference for all purposes as if set forth as a part of this specification.

[0002] The present invention relates to the diagnosis, treatment and prevention of diseases and conditions associated with kidney injury, in particular, but not limited to, acute kidney injury. [Background technology]

[0003] Acute kidney injury (AKI) refers to rapid damage to the kidneys, specifically the renal tubular epithelial cells. Acute kidney injury is often chemical in origin, for example, medicines, chemicals, contrast dyes, and herbal or nutritional supplements can cause kidney damage or injury. Acute kidney injury can also be caused by ischemia, mechanical factors, or immune factors.

[0004] Acute kidney injury is a common condition, with prevalence rates reaching 10% in hospitalized patients (Silver, SA & Chertow, GM The Economic Consequences of Acute Kidney Injury. Nephron 137, 297-301 (2017)). Mortality associated with acute kidney injury is very high, with mortality rates reaching 50% in patients who require intensive care after the onset of acute kidney injury. Even patients who recover from acute kidney injury remain at long-term risk for progression to chronic kidney disease (CKD), end-stage renal failure, renal replacement therapy, or kidney transplantation (Silver, SA & Chertow, GM The Economic Consequences of Acute Kidney Injury. Nephron 137; Mehta, RL et al. Acute Kidney Injury Network: report of an initiative to improve outcomes in acute kidney injury. Crit. Care 11; Zuk, A. et al. Overcoming Translational Barriers in Acute Kidney Injury: A Report from an NIDDK Workshop. Clin. J. Am. Soc. Nephrol. 13, 1113-1123 (2018)).

[0005] The chemotherapy agent cisplatin (dichlorodiaminoplatinum; (SP-4-2)-diamminedichloroplatinum(II)) is widely used to treat a variety of cancers, including head and neck cancer, breast cancer, lung cancer, testicular cancer, ovarian cancer, brain tumors, and bladder cancer. Cisplatin (50–100 mg / m 2A single dose of cisplatin causes some degree of acute kidney injury in most patients, with up to 30% developing nephrotoxicity (Ozkok, A. & Edelstein, CL Pathophysiology of cisplatin-induced acute kidney injury. Biomed Res. Int. 2014, 967826 (2014)). This nephrotoxicity is diagnosed by a rapid decline in renal function, as indicated by an increase in serum creatinine (Mehta et al., supra). In elderly patients with head and neck cancer, up to 20% of them progress from acute kidney injury to severe renal insufficiency (Yao, X., Panichpisal, K., Kurtzman, N. & Nugent, K. Cisplatin nephrotoxicity: a review. Am. J. Med. Sci. 334, 115-124 (2007)). Cisplatin-induced acute kidney injury is due to its dose-limiting toxicity, and although treatment with cisplatin is often divided into multiple doses over several weeks, nephrotoxicity still occurs. The severity of cisplatin-induced acute kidney injury is more severe in the presence of a pre-existing medical history of diabetes, hypertension, nephrotoxic drugs, and advanced age.

[0006] The pathophysiology of acute kidney injury is primarily defined by injury to renal tubular epithelial cells (TECs) due to toxins, chemicals, drugs, immune factors, mechanical factors, or ischemia. The pathophysiology of acute kidney injury also includes vascular and immune system components. In patients treated with cisplatin, kidney cisplatin concentrations can reach up to five times the plasma concentration. In the kidney, cisplatin accumulates in renal tubular epithelial cells, induces reactive oxygen species in renal tubular epithelial cells, depletes glutathione, causes mitochondrial dysfunction, and leads to cell death via apoptosis or necrosis (Ozkok et al, supra; Wang, S., Wei, Q., Dong, G. & Dong, Z. ERK-mediated suppression of cilia in cisplatin-induced tubular cell apoptosis and acute kidney injury. Biochim. Biophys. Acta 1832, 1582-1590 (2013); Jo, S.-K., Cho, WY, Sung, SA, Kim, HK & Won, NH MEK inhibitor, U0126, attenuates cisplatin-induced renal injury by decreasing inflammation and apoptosis. Kidney Int. 67; Nowak, G. Protein Kinase C-α and ERK1 / 2 Mediate Mitochondrial Dysfunction, Decreases in Active Na + Transport, and Cisplatin-induced Apoptosis in Renal Cells. J. Biol. Chem. 277, 43377-43388 (2002)). In addition, when ischemia occurs, renal tubular epithelial cells, which have a very high oxygen demand, become oxygen-deprived and undergo cell death via apoptosis or necrosis.

[0007] Renal function can be restored after acute kidney injury. It is widely recognized that this recovery is due to the proliferation of remaining renal tubular epithelial cells, which have a very high regenerative capacity (Yang, H.-C., Liu, S.-J. & Fogo, AB Kidney regeneration in mammals. Nephron Exp. Nephrol. 126, 50 (2014); Coelho, S., Cabral, G., Lopes, JA & Jacinto, A. Renal regeneration after acute kidney injury. Nephrology 23, 805-814 (2018); Chang-Panesso, M. & Humphreys, BD Cellular plasticity in kidney injury and repair. Nat. Rev. Nephrol. 13, 39-46 (2017)). Insufficient proliferation of renal tubular epithelial cells can lead to nephrotoxicity and, over time, chronic kidney disease, with fibrosis as a secondary phenomenon.Recent studies have shown that a key determinant of tubular epithelial cell dysfunction following the onset of acute kidney injury is re-expression of the SNAIL gene (also known as SNA; SNAH; SNAIL; SLUGH2; SNAIL1) (Grande, MT et al. Snail1-induced partial epithelial-to-mesenchymal transition drives renal fibrosis in mice and can be targeted to reverse established disease. Nat. Med. 21, 989-997 (2015); Simon-Tillaux, N. & Hertig, A. Snail and kidney fibrosis. Nephrol. Dial. Transplant 32, 224-233 (2017); Lovisa, S. et al. Epithelial-to-mesenchymal transition induces cell cycle arrest and parenchymal damage in renal fibrosis. Nat. Med. 21, 998-1009 (2015)). SNAIL is an important gene in embryogenesis, but is hardly expressed in adults, except in cancers where epithelial-mesenchymal transition (EMT) occurs. SNAIL can be induced in renal tubular epithelial cells via the TGFβ1 gene, and this induction of SNAIL is associated with dysfunction and proliferation of renal tubular epithelial cells (Lovisa, S. et al. Epithelial-to-mesenchymal transition induces cell cycle arrest and parenchymal damage in renal fibrosis. Nat. Med. 21, 998-1009 (2015)).

[0008] Interleukin-11 (IL-11), a type of cytokine, has been reported to have a strong protective effect against acute kidney injury after ischemia-reperfusion injury and to suppress apoptosis, necrosis, and inflammation in acute kidney injury (Lee, HT et al. Interleukin-11 protects against renal ischemia and reperfusion injury. Am. J. Physiol. Renal Physiol. 303, F1216-24 (2012)). It has also been shown that IL-11 is upregulated by isoflurane administration in human renal tubular epithelial cells and mouse kidneys, and this upregulation of IL-11 is associated with the important protective effect of IL-11 against acute ischemic kidney injury (Ham, A. et al. Critical role of interleukin-11 in isoflurane-mediated protection against ischemic acute kidney injury in mice. Anesthesiology 119, 1389-1401 (2013)).

[0009] Furthermore, it has been reported that treatment with IL-11 in rats and mice protects them from acute nephrotoxic nephritis by reducing nephritis and preventing renal damage (Stangou, M. et al. Effect of IL-11 on glomerular expression of TGF-beta and extracellular matrix in nephrotoxic nephritis in Wistar Kyoto rats. J. Nephrol. 24, 106-111 (2011); Lai, PC et al. Interleukin-11 attenuates nephrotoxic nephritis in Wistar Kyoto rats. J. Am. Soc. Nephrol. 12, 2310-2320 (2001); Lai, PC et al. Interleukin-11 reduces renal injury and glomerular NF-kappa B activity in murine experimental glomerulonephritis. Nephron Exp. Nephrol. 101, e146-54 (2005)). Summary of the Invention [Means for solving the problem]

[0010] In contrast to the reported protective effects of IL-11 against renal injury and kidney damage, the present invention relates to the treatment and / or prevention of renal injury and disorders, diseases or conditions associated with renal injury via inhibition of IL-11 signaling.

[0011] In one aspect of the present invention, there is provided an agent capable of inhibiting interleukin-11 (IL-11) mediated signaling for use in a method for the treatment or prevention of renal damage and / or disorders, diseases or conditions associated with renal damage.

[0012] In another aspect of the invention, there is provided the use of an agent capable of inhibiting interleukin-11 (IL-11) mediated signaling in the manufacture of a medicament for use in a method for the treatment or prevention of renal damage and / or disorders, diseases or conditions associated with renal damage.

[0013] In another aspect of the present invention, there is provided a method for treating or preventing renal damage and / or disorders, diseases or conditions associated with renal damage, comprising administering to a subject in need of treatment a therapeutically effective amount of an agent capable of inhibiting interleukin-11 (IL-11)-mediated signaling.

[0014] The present invention also provides agents capable of suppressing interleukin-11 (IL-11)-mediated signaling for use in methods of ameliorating renal damage.

[0015] Further provided is the use of an agent capable of inhibiting interleukin-11 (IL-11) mediated signaling in the manufacture of a medicament for use in a method of ameliorating renal damage.

[0016] Further provided is a method for ameliorating renal damage, comprising administering to a subject in need of treatment a therapeutically effective amount of an agent capable of suppressing interleukin-11 (IL-11)-mediated signaling.

[0017] In some embodiments, the kidney injury is acute kidney injury. In some embodiments, the kidney injury is nephrotoxicity. In some embodiments, the kidney injury is drug-induced kidney injury or ischemia-induced kidney injury. In some embodiments, the kidney injury is cisplatin-induced kidney injury or cisplatin-induced nephrotoxicity. In some embodiments, the kidney injury is characterized by damage to renal tubular epithelial cells (TEC).

[0018] The present invention also provides an agent capable of suppressing interleukin-11 (IL-11)-mediated signaling for use in a method of improving renal function in a subject suffering from renal dysfunction.

[0019] Also provided is the use of an agent capable of inhibiting interleukin-11 (IL-11)-mediated signaling in the manufacture of a medicament for use in a method of improving renal function in a subject suffering from renal dysfunction.

[0020] Further provided is a method for improving renal function in a subject suffering from renal dysfunction, comprising administering to a subject in need of treatment a therapeutically effective amount of an agent capable of inhibiting interleukin-11 (IL-11)-mediated signal transduction.

[0021] The present invention also provides agents capable of suppressing interleukin-11 (IL-11)-mediated signaling for use in methods of promoting proliferation, survival and / or function of renal tubular epithelial cells (TECs), and / or proliferation, maintenance and / or function of renal tissue (e.g., following the onset of renal damage).

[0022] Also provided is the use of an agent capable of inhibiting interleukin-11 (IL-11)-mediated signaling in the manufacture of a medicament for use in a method of promoting proliferation, survival and / or function of renal tubular epithelial cells (TECs), and / or proliferation, maintenance and / or function of renal tissue (e.g., following the onset of renal damage).

[0023] Further provided is a method for promoting the proliferation, survival and / or function of renal tubular epithelial cells (TECs) and / or the proliferation, maintenance and / or function of renal tissue (e.g., after the onset of renal damage), comprising administering to a subject in need of treatment a therapeutically effective amount of an agent capable of inhibiting interleukin-11 (IL-11)-mediated signaling.

[0024] The present invention also provides agents capable of inhibiting interleukin-11 (IL-11)-mediated signaling for use in methods of inhibiting expression of SNAIL (eg, following the onset of renal damage).

[0025] Also provided is the use of an agent capable of inhibiting interleukin-11 (IL-11)-mediated signaling in the manufacture of a medicament for use in a method of inhibiting expression of SNAIL (eg, following the onset of renal damage).

[0026] Further provided is a method for suppressing expression of SNAIL (e.g., after the onset of renal damage), comprising administering to a subject in need of treatment a therapeutically effective amount of an agent capable of suppressing interleukin-11 (IL-11)-mediated signal transduction.

[0027] The present invention also provides agents capable of inhibiting interleukin-11 (IL-11)-mediated signaling for use in methods of inhibiting or reversing the transformation of renal tubular epithelial cells (TECs) to a mesenchymal cell-like phenotype (e.g., following the onset of renal injury).

[0028] Also provided is the use of an agent capable of inhibiting interleukin-11 (IL-11)-mediated signaling in the manufacture of a medicament for use in a method of inhibiting or reversing the transformation of renal tubular epithelial cells (TECs) to a mesenchymal cell-like phenotype (e.g., following the onset of renal damage).

[0029] Further provided is a method for inhibiting or reversing the transformation of renal tubular epithelial cells (TECs) to a mesenchymal cell-like phenotype (e.g., after the onset of renal damage), comprising administering to a subject in need of treatment a therapeutically effective amount of an agent capable of inhibiting interleukin-11 (IL-11)-mediated signaling.

[0030] In some embodiments according to various aspects of the invention, the agent is an agent capable of blocking or reducing binding of interleukin-11 (IL-11) to the interleukin-11 receptor (IL-11R).

[0031] In some embodiments, the agent is an agent capable of binding to interleukin 11 (IL-11) or interleukin 11 receptor (IL-11R). In some embodiments, the agent is selected from the group consisting of an antibody or antigen-binding fragment thereof, a polypeptide, a peptide, a nucleic acid, an oligonucleotide, an aptamer, and a small molecule. The agent may be an antibody or antigen-binding fragment thereof. The agent may be a decoy receptor.

[0032] In some embodiments, the agent is an anti-IL-11 antagonist antibody or an antigen-binding fragment thereof against IL-11-mediated signaling.

[0033] In some embodiments, the antibody or antigen-binding fragment thereof comprises: (i) HC-CDR1 having the amino acid sequence of SEQ ID NO: 34; HC-CDR2 having the amino acid sequence of SEQ ID NO: 35, and HC-CDR3 having the amino acid sequence of SEQ ID NO: 36 A heavy chain variable (VH) region incorporating (ii) LC-CDR1 having the amino acid sequence of SEQ ID NO: 37; LC-CDR2 having the amino acid sequence of SEQ ID NO: 38, and LC-CDR3 having the amino acid sequence of SEQ ID NO: 39 and a light chain variable (VL) region incorporating Includes.

[0034] In some embodiments, the antibody or antigen-binding fragment thereof comprises: (i) HC-CDR1 having the amino acid sequence of SEQ ID NO: 40; HC-CDR2 having the amino acid sequence of SEQ ID NO: 41, and HC-CDR3 having the amino acid sequence of SEQ ID NO: 42 A heavy chain variable (VH) region incorporating (ii) LC-CDR1 having the amino acid sequence of SEQ ID NO: 43; LC-CDR2 having the amino acid sequence of SEQ ID NO: 44, and LC-CDR3 having the amino acid sequence of SEQ ID NO: 45 and a light chain variable (VL) region incorporating Includes.

[0035] In some embodiments, the agent is an anti-IL-11Rα antagonist antibody or an antigen-binding fragment thereof for IL-11 mediated signaling.

[0036] In some embodiments, the antibody or antigen-binding fragment thereof comprises: (i) HC-CDR1 having the amino acid sequence of SEQ ID NO: 46; HC-CDR2 having the amino acid sequence of SEQ ID NO: 47, and HC-CDR3 having the amino acid sequence of SEQ ID NO: 48 A heavy chain variable (VH) region incorporating (ii) LC-CDR1 having the amino acid sequence of SEQ ID NO: 49; LC-CDR2 having the amino acid sequence of SEQ ID NO: 50, and LC-CDR3 having the amino acid sequence of SEQ ID NO:51 and a light chain variable (VL) region incorporating Includes.

[0037] In some embodiments, the agent is a decoy receptor for IL-11. In some embodiments, the decoy receptor for IL-11 comprises (i) an amino acid sequence corresponding to a cytokine binding module of gp130 and (ii) an amino acid sequence corresponding to a cytokine binding module of IL-11Rα.

[0038] In some embodiments, the agent is a mutein of IL-11. In some embodiments, the mutein of IL-11 is a mutein having a W147A substitution.

[0039] In some embodiments, the agent is an agent that can block or reduce the expression of interleukin 11 (IL-11) or interleukin 11 receptor (IL-11R). The agent may be an oligonucleotide or a small molecule.

[0040] In some embodiments, the agent is an antisense oligonucleotide that can inhibit or reduce the expression of IL-11. In some embodiments, the antisense oligonucleotide that can inhibit or reduce the expression of IL-11 is an IL-11 targeting siRNA that comprises the sequence of SEQ ID NO: 12, 13, 14 or 15. In some embodiments, the agent is an antisense oligonucleotide that can inhibit or reduce the expression of IL-11Rα. In some embodiments, the antisense oligonucleotide that can inhibit or reduce the expression of IL-11Rα is an IL-11Rα targeting siRNA that comprises the sequence of SEQ ID NO: 16, 17, 18 or 19.

[0041] In embodiments provided herein, the interleukin-11 receptor may be or include IL-11Rα.

[0042] In embodiments provided herein, the renal injury may be any one of acute kidney injury (AKI), nephrotoxicity, drug-induced kidney injury (DIKI), acute renal failure, acute kidney disease, chronic kidney disease, kidney injury, tubular necrosis, acute tubular necrosis, and autoimmune nephropathy.

[0043] In the embodiments provided herein, the agent may be administered prior to, concurrently with, or after the onset of a cause of kidney damage, for example, prior to, concurrently with, or after administration or ingestion of a nephrotoxic pharmaceutical agent, or prior to, concurrently with, or after exposure to a physical, mechanical, chemical, or environmental cause of kidney damage.

[0044] In some embodiments, the medicaments, uses and methods described herein are provided for the treatment and / or prevention of drug-induced nephropathy (DIKI). Drug-induced nephropathy (DIKI) may be intrinsic nephropathy and / or idiosyncratic nephropathy. In embodiments, the medicaments, uses and methods may be provided for the treatment and / or prevention of cisplatin-induced nephropathy and / or cisplatin-induced nephrotoxicity.

[0045] In some embodiments, said medicaments, uses and methods may be provided for the treatment and / or prevention of ischemia-induced kidney injury (IIKI) or ischemia-induced acute kidney injury.

[0046] In the embodiments provided herein, the disorder, disease or condition associated with renal injury may be a disease, disorder or condition in which renal injury is pathologically implicated. The pathology of these diseases, disorders or conditions may include injury to renal tubular epithelial cells and / or conversion of renal tubular epithelial cells to a mesenchymal cell-like phenotype, which may be proximal tubular epithelial cells and / or distal tubular epithelial cells.

[0047] In either embodiment, the method comprises administering the agent to a subject in which expression of interleukin-11 (IL-11) or IL-11 receptor (IL-11R) may be upregulated.

[0048] In either embodiment, the method may include administering the agent to a subject in which upregulated expression of interleukin-11 (IL-11) or IL-11 receptor (IL-11R) has been identified.

[0049] In some embodiments, the method includes determining whether expression of interleukin-11 (IL-11) or IL-11 receptor (IL-11R) is upregulated in a subject, and administering the agent to a subject in which expression of interleukin-11 (IL-11) or IL-11 receptor (IL-11R) is upregulated.

[0050] Also provided is a method for determining a subject's suitability for treatment or prevention of renal impairment and / or disorders, diseases or conditions associated with renal impairment with an agent capable of inhibiting interleukin-11 (IL-11)-mediated signaling, the method comprising determining (in vitro) whether expression of interleukin-11 (IL-11) or IL-11 receptor (IL-11R) is upregulated in the subject.

[0051] Further provided is a method for selecting a subject for treatment or prevention of renal damage and / or a disorder, disease or condition associated with renal damage with an agent capable of inhibiting interleukin-11 (IL-11)-mediated signaling, the method comprising determining (in vitro) whether expression of interleukin-11 (IL-11) or IL-11 receptor (IL-11R) is upregulated in the subject.

[0052] In one aspect, a method is provided for diagnosing renal impairment and / or renal impairment-associated disorders, diseases or conditions in a subject, or diagnosing the risk of developing renal impairment and / or renal impairment-associated disorders, diseases or conditions in a subject, comprising determining (in vitro) whether expression of interleukin-11 (IL-11) or IL-11 receptor (IL-11R) is upregulated in a sample obtained from the subject. In some embodiments, the diagnostic method is a method for confirming a diagnosis of renal impairment and / or renal impairment-associated disorders, diseases or conditions in a subject suspected of suffering from renal impairment and / or renal impairment-associated disorders, diseases or conditions. In some embodiments, the diagnostic method and / or the method for confirming the diagnosis comprises selecting a subject for treatment with an agent capable of suppressing interleukin-11 (IL-11)-mediated signaling.

[0053] Further provided is a method of providing a prognosis for a subject suffering from or suspected of suffering from renal impairment and / or a disorder, disease or condition associated with renal impairment, comprising determining (in vitro) whether expression of interleukin-11 (IL-11) or IL-11 receptor (IL-11R) is upregulated in a sample obtained from the subject, and providing a prognosis for treating the subject with an agent capable of suppressing interleukin-11 (IL-11)-mediated signaling based on this determination. In some embodiments, the method of providing a prognosis comprises selecting a subject identified as having upregulated expression of interleukin-11 (IL-11) or IL-11 receptor (IL-11R) for treatment with an agent capable of suppressing interleukin-11 (IL-11)-mediated signaling.

[0054] In another aspect, a method is provided for diagnosing renal impairment and / or a renal impairment-related disorder, disease or condition, or diagnosing the risk of developing renal impairment and / or a renal impairment-related disorder, disease or condition, comprising measuring in a subject (in vitro) one or more genetic factors predictive of upregulation of IL-11 or IL-11 receptor expression or upregulation of IL-11-mediated signaling. In some embodiments, the method comprises selecting the subject for treatment with an agent capable of suppressing interleukin-11 (IL-11)-mediated signaling.

[0055] Also provided is a method of providing a prognosis to a subject suffering from, or suspected of suffering from, renal damage and / or a disorder, disease or condition associated with renal damage, comprising measuring (in vitro) in the subject one or more genetic factors predictive of upregulation of IL-11 or IL-11 receptor expression or upregulation of IL-11-mediated signaling. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0056] There remains a need for effective prevention and treatment of kidney injury, particularly acute kidney injury.

[0057] It has been reported that IL-11, a type of cytokine, has a protective effect against acute kidney injury following ischemia-reperfusion injury (Lee et al., supra). It has also been reported that IL-11 is upregulated in human renal tubular epithelial cells and mouse kidneys following chemical injury caused by isoflurane, and that the upregulated IL-11 exerts a protective effect (Ham et al., supra).

[0058] In contrast, the present inventors have found that by suppressing IL-11-mediated signal transduction, renal tubular epithelial cells can be effectively protected, which results in proliferation of renal tubular epithelial cells and recovery of renal injury, which is a common causative factor of acute kidney injury.Without wishing to be bound by a particular theory, the present inventors believe that suppressing IL-11 allows proliferation of renal tubular epithelial cells and prevents or reduces the expression of SNAIL, leading to regeneration and recovery of kidney tissue.

[0059] Interleukin-11 and IL-11 receptor Interleukin-11 (IL-11), also known as adipogenic inhibitory factor, is a pleiotropic cytokine and a member of the IL-6 cytokine family that also includes IL-6, IL-11, IL-27, IL-31, oncostatin, leukemia inhibitory factor (LIF), cardiotrophin-1 (CT-1), cardiotrophin-like cytokine (CLC), ciliary neurotrophic factor (CNTF), and neuropoietin (NP-1).

[0060] Interleukin-11 (IL-11) is expressed in various mesenchymal cells. The IL-11 genomic sequence is mapped to the centromeric region of chromosome 7 and chromosome 19, and it is transcribed with a classical signal peptide that allows efficient secretion of IL-11 from cells. The activator protein complex (cJun / AP-1) located in the promoter sequence of the IL-11 gene is important for basal transcriptional regulation of IL-11 (Du and Williams., Blood 1997, Vol 89: 3897-3908). The human IL-11 precursor is a polypeptide of 199 amino acids, and the mature IL-11 is a protein of 178 amino acid residues (Garbers and Scheller. , Biol. Chem. 2013; 394(9):1145-1161). The amino acid sequence of human IL-11 is available under UniProt accession number P20809 (P20809.1 GI:124294; SEQ ID NO:1). Recombinant human IL-11 (Oprelvekin) is also commercially available. Several IL-11 from other species have also been cloned and sequenced, including IL-11 from mouse, rat, pig, cow, several bony fish, and primates.

[0061] As used herein, "IL-11" refers to IL-11 from any species, including isoforms, fragments, variants, or homologs of IL-11 obtained from any species. In a preferred embodiment, the species is human (Homo sapiens). An isoform, fragment, variant, or homolog of IL-11 may be characterized by having at least 70%, preferably 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity with the amino acid sequence of IL-11 precursor or mature IL-11 from a given species (e.g., human). Isoforms, fragments, variants or homologs of IL-11 may be characterized by their ability to bind to IL-11Rα (preferably from the same species) and thereby stimulate signaling of cells expressing IL-11Rα and gp130 (e.g. as described in Curtis et al. Blood, 1997, 90(11) or Karpovich et al. Mol. Hum. Reprod. 2003 9(2): 75-80). The length of the IL-11 fragment may be any length (amino acid length) but may be at least 25% of the length of mature IL-11 and may be up to 50%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% of the length of mature IL-11. The length of the IL-11 fragment may be a minimum of 10 amino acids long and a maximum of 15, 20, 25, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190 or 195 amino acids long.

[0062] IL-11 signals through the ubiquitously expressed homodimer of glycoprotein 130 (gp130; also known as glycoprotein 130, IL-6ST, IL-6-β or CD130). gp130 is a transmembrane protein and a receptor subunit that associates with the IL-6 receptor family to form the type I cytokine receptor. Specificity is exerted by the individual interleukin-11 receptor α subunit (IL-11Rα). Although the IL-11α receptor does not directly participate in signal transduction, upon cytokine binding to the α receptor it associates with gp130 to form the final complex.

[0063] Human gp130 (including a 22 amino acid signal peptide) is a 918 amino acid protein, the mature form of which is 866 amino acids, containing a 597 amino acid extracellular domain, a 22 amino acid transmembrane domain, and a 277 amino acid intracellular domain. The extracellular domain of human gp130 contains the gp130 cytokine binding module (CBM). The gp130 CBM contains the Ig-like domain D1 and the gp130 fibronectin type III domains D2 and D3. The amino acid sequence of human gp130 is available from UniProt accession number P40189-1 (SEQ ID NO:2).

[0064] Human IL-11Rα is a 422 amino acid polypeptide (UniProt Q14626; SEQ ID NO:3) with approximately 85% nucleotide and amino acid sequence identity to mouse IL-11Rα. Two isoforms of IL-11Rα have been reported, which differ in their intracellular domains (Du and Williams, supra). The IL-11 receptor α chain (IL-11Rα) shares many structural and functional similarities with the IL-6 receptor α chain (IL-6Rα). The extracellular domains of IL-11Rα and IL-6Rα share 24% amino acid identity and contain the characteristic Trp-Ser-X-Trp-Ser (WSXWS) conserved motif. The short intracellular domains (34 amino acids long) of IL-11Rα and IL-6α lack the Box1 and Box2 regions required for activation of the JAK / STAT signaling pathway.

[0065] The receptor binding site of mouse IL-11 has been mapped and three sites (site I, site II, and site III) have been identified. Substitutions in the site II region and substitutions in the site III region reduce binding to gp130. Site III mutations have no detectable agonist activity and exhibit antagonist activity against IL-11Rα (Cytokine Inhibitors Chapter 8; co-edited by Gennaro Ciliberto and Rocco Savino, Marcel Dekker, Inc. 2001).

[0066] As used herein, IL-11 receptor (IL-11R) refers to a polypeptide or polypeptide complex capable of binding to IL-11. In some embodiments, the IL-11 receptor is capable of binding to IL-11 and inducing signal transduction in cells expressing the IL-11 receptor.

[0067] The IL-11 receptor may be from any species, including isoforms, fragments, variants or homologs of the IL-11 receptor from any species, in a preferred embodiment, the species is human (Homo sapiens).

[0068] In some embodiments, the IL-11 receptor may be IL-11Rα. In some embodiments, the IL-11 receptor may be a polypeptide complex comprising IL-11Rα. In some embodiments, the IL-11 receptor may be a polypeptide complex comprising IL-11Rα and gp130. In some embodiments, the IL-11 receptor may be gp130 or a gp130-containing complex to which IL-11 binds.

[0069] Isoforms, fragments, variants or homologs of IL-11Rα may be characterized by having at least 70%, preferably 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity with the amino acid sequence of IL-11Rα from a given species (e.g. human). Isoforms, fragments, variants or homologs of IL-11Rα may be characterized by the ability to bind IL-11 (preferably from the same species) and thereby stimulate signaling of cells expressing IL-11Rα and gp130 (e.g. as described in Curtis et al. Blood, 1997, 90(11) or Karpovich et al. Mol. Hum. Reprod. 2003 9(2): 75-80). The length of the IL-11 receptor fragment may be any length (in amino acids), but may be at least 25% of the length of mature IL-11Rα and may be up to 50%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% of the length of mature IL-11Rα. The length of a fragment of the IL-11 receptor may be a minimum of 10 amino acids long and a maximum of 15, 20, 25, 30, 40, 50, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 250, 300, 400 or 415 amino acids long.

[0070] IL-11 Signaling IL-11 binds to IL-11Rα with low affinity (Kd = approx. 10 nmol / L) and is not capable of transmitting biological signals solely through the interaction between these binding partners. Co-expression of IL-11Rα and gp130 is required to form a receptor that can bind with high affinity (Kd = approx. 400-800 pmol / L) and transmit signals (Curtis et al Blood 1997; 90 (11):4403-12; Hilton et al., EMBO J 13:4765, 1994; Nandurkar et al., Oncogene 12:585, 1996). Binding of IL-11 to cell surface IL-11Rα induces heterodimerization, tyrosine phosphorylation, activation of gp130 and downstream signaling primarily via the mitogen-activated protein kinase (MAPK) cascade and the Janus kinase / signal transducer and activator of transcription (Jak / STAT) pathway (Garbers and Scheller, supra).

[0071] Furthermore, since in principle soluble IL-11Rα can bind IL-11 to form a biologically active soluble complex (Pflanz et al., 1999 FEBS Lett, 450, 117-122), it is conceivable that IL-11, like IL-6, may bind to soluble IL-11Rα before binding to gp130 on the cell surface (Garbers and Scheller, supra). Curtis et al. (Blood 1997 Dec 1; 90 (11):4403-12) reported the expression of a soluble mouse IL-11 receptor α chain (sIL-11R) and the investigation of signal transduction in gp130-expressing cells. In this study, it was reported that in the presence of gp130 but in the absence of transmembrane IL-11R, soluble IL-11R induced IL-11-dependent differentiation of M1 leukemia cells and proliferation of Ba / F3 cells, as well as early intracellular events (such as phosphorylation of gp130, STAT3, and SHP2), similar to transmembrane IL-11R-mediated signaling. Activation of membrane-bound gp130-mediated signaling by IL-11 bound to soluble IL-11Rα has been demonstrated in recent years (Lokau et al., 2016 Cell Reports 14, 1761-1773). This so-called IL-11 trans-signaling may be important in disease pathogenesis, but its role in human disease awaits further investigation. In a preferred embodiment, inhibition of IL-11-mediated signaling is achieved by disrupting IL-11-mediated cis-signaling.

[0072] As used herein, "IL-11 trans-signaling" refers to signaling initiated by the further binding of IL-11 bound to IL-11Rα to gp130. IL-11 can bind to IL-11Rα through non-covalent binding to form a complex. gp130 is membrane-bound and expressed by cells, and signaling occurs when the IL-11:IL-11Rα complex binds to gp130. In some embodiments, IL-11Rα may be soluble IL-11Rα. In some embodiments, soluble IL-11Rα is a soluble (secreted) isoform of IL-11Rα (e.g., lacking a transmembrane domain). In some embodiments, soluble IL-11Rα is a product liberated by proteolysis of the extracellular domain of membrane-bound IL-11Rα. In some embodiments, IL-11Rα may be membrane-bound, and signal transduction via gp130 may be initiated by IL-11 bound to membrane-bound IL-11Rα further binding to gp130. This is referred to as "cis-signal transduction of IL-11."

[0073] IL-11-mediated signaling has been shown to stimulate hematopoiesis and thrombopoiesis, stimulate osteoclast activity, stimulate neurogenesis, inhibit adipogenesis, reduce the expression of pro-inflammatory cytokines, regulate extracellular matrix (ECM) metabolism, and mediate normal growth control of gastrointestinal epithelial cells (Du and Williams, supra).

[0074] The physiological role of interleukin 11 (IL-11) remains unclear. IL-11 has been most strongly associated with hematopoietic cell activation and platelet production. IL-11 has also been shown to confer protective effects against graft-versus-host disease, inflammatory arthritis, and inflammatory bowel disease, which suggests that IL-11 is an anti-inflammatory cytokine (Putoczki and Ernst, J Leukoc Biol 2010, 88(6):1109-1117). On the other hand, IL-11 has been suggested to be both pro- and anti-inflammatory, pro-angiogenic, and important in tumorigenesis. Recent studies have shown that IL-11 is readily detectable in virus-induced inflammation in mouse arthritis models and cancer, suggesting that IL-11 expression is induced by pathological stimuli. Furthermore, IL-11 has been reported to be associated with Stat3-dependent activation of tumor-promoting target genes in neoplastic gastrointestinal epithelium (Putoczki and Ernst, supra).

[0075] As used herein, "IL-11 signal transduction" and "IL-11-mediated signal transduction" refer to signal transduction via binding of IL-11 to the IL-11 receptor, or via binding of an IL-11 fragment having the function of a mature IL-11 molecule to the IL-11 receptor. It will also be appreciated that "IL-11 signal transduction" and "IL-11-mediated signal transduction" refer to signal transduction initiated by IL-11 or a functional fragment thereof, such as via binding to the IL-11 receptor. Thus, "signal transduction" refers to signal transduction and other cellular processes that control cellular activity.

[0076] Kidney damage Aspects of the present invention relate to the diagnosis, treatment and prevention of renal injury, particularly acute kidney injury (AKI) (also known as acute renal failure) and / or renal injury (eg, acute kidney injury).

[0077] As used herein, "renal injury" refers to damage to the kidney, renal tissue, and / or one or more renal cells. The damage to the cells / tissues / organs may be due to injury to these cells / tissues / organs, e.g., chemical or physical treatment / experience. In some embodiments, the renal injury may be due to chemical injury, e.g., drug-induced renal injury (e.g., cisplatin-induced renal injury). In some embodiments, the renal injury may be due to physical injury, e.g., renal injury caused by crush injury, or due to surgical injury to the renal tissue, e.g., during surgery for treating disease and / or kidney transplantation (e.g., renal injury may be iatrogenic). In some embodiments, the renal injury may be due to hypoxia, e.g., ischemia, or reperfusion. In some embodiments, the renal injury may be due to infection, an immune response to infection, cancer, and / or autoimmunity. The injury may be reversible or irreversible.

[0078] In some embodiments, renal injury includes damage to one or both kidneys. In some embodiments, renal injury includes damage to one or more of renal capsule, renal cortex, renal medulla, renal papilla, renal pyramid, renal trabecula, renal calyx, minor calyx, major calyx, renal hilus, renal pelvis, ureter, renal artery and renal vein. In some embodiments, renal injury includes damage to one or more of renal epithelial cells, renal tubular epithelial cells, proximal tubular epithelial cells, distal tubular epithelial cells, renal parietal cells, podocytes, cells of thin part of Henle's loop, cells of thick ascending limb, principal collecting duct cells, interstitial collecting duct cells and renal interstitial cells. In some embodiments, renal injury includes damage to renal epithelial cells, such as tubular epithelial cells, such as proximal tubular epithelial cells.

[0079] The cell / tissue / organ damage may be characterized by changes in the structure of the cell / tissue / organ and / or its function. For example, the cell / tissue / organ damage may be characterized by a decrease in the level of a correlate of normal function of the cell / tissue / organ and / or an increase in a correlate of dysfunction of the cell / tissue / organ. For example, kidney / renal tissue / renal cell damage may be characterized by a decrease in urine volume in a subject suffering from renal damage and / or an increase in serum creatine concentration in a subject suffering from renal damage. The cell / tissue / organ damage may also be characterized by cell death, e.g., death of cells of the damaged organ / tissue. Such cell death may be due to apoptosis (i.e., programmed cell death) or necrosis (premature cell death due to injury).

[0080] As used herein, "acute kidney injury" generally refers to a sudden decrease in kidney function. Acute kidney injury may be characterized by a sudden increase in serum creatinine and / or a sudden decrease in urine output.

[0081] Acute kidney injury may be defined and staged according to the Summary of Recommendation Statements of the Kidney Disease Improving Global Outcomes (KDIGO) group, Kidney International Supplements (2012) 2, 8-12, which is incorporated herein by reference in its entirety.

[0082] In some embodiments, acute kidney injury is defined as (i) an increase in serum creatine of 0.3 mg / dl or more (26.5 μmol / l or more) within 48 hours, (ii) an increase in serum creatine of 1.5 times baseline (which indicates or is indicative of an onset within the past 7 days), or (iii) a 6-hour urine output of less than 0.5 ml / kg / hr.

[0083] In some embodiments, the acute kidney injury may be stage 1, stage 2, or stage 3 acute kidney injury. Stage 1 acute kidney injury may be defined as (i) an increase in serum creatine 1.5-1.9 times above baseline, (ii) an increase in serum creatine 0.3 mg / dl or more (26.5 μmol / l or more), or (iii) a urine output of less than 0.5 ml / kg / hr for 6-12 hours. Stage 2 acute kidney injury may be defined as (i) an increase in serum creatine 2.0-2.9 times above baseline, or (ii) a urine output of less than 0.5 ml / kg / hr for 12 hours or more. Stage 3 acute kidney injury is defined as (i) an increase in serum creatine ≥3.0 times baseline, (ii) an increase in serum creatine ≥4.0 mg / dL (≥353.6 μmol / L), (iii) initiation of renal replacement therapy, or (iv) an estimated glomerular filtration rate (eGFR) of ≥1.73 m in patients <18 years of age. 2 (v) a urinary output of less than 0.3 ml / kg / hour for at least 24 hours; or (vi) anuria for at least 12 hours.

[0084] Renal injury may be characterized by injury to renal tubular epithelial cells (TECs) and / or conversion of renal tubular epithelial cells to an epithelial-mesenchymal transition cell-like phenotype (i.e., epithelial-mesenchymal transition (EMT)). Conversion of renal tubular epithelial cells to a mesenchymal cell-like phenotype may be characterized, for example, by decreased expression of E-cadherin, increased expression of SNAIL, and / or increased expression of ACTA2.

[0085] The cause of kidney damage may be any, for example, mechanical damage or injury (i.e., physical damage or injury), chemical damage or injury, ischemia, or kidney damage caused by genetic predisposition. The cause of kidney damage or kidney injury usually causes kidney dysfunction, and may even lead to kidney failure.

[0086] The mechanical injury or damage may include physical injury to the subject, kidney, tubular epithelial cells, or podocytes, and may include tubular obstruction / blockage, such as, for example, the urinary tract.

[0087] The chemical damage or injury may be caused by a drug, medicine, toxin, herbal supplement, nutritional supplement, or other chemical agent administered to the subject, or absorbed or ingested by the subject. In some embodiments, the chemical damage is a side effect of such an agent administered to treat a disease or condition occurring in a tissue other than the kidney, or a side effect of such an agent administered to treat a disease or condition occurring in the kidney and one or more other tissues. In some embodiments, the chemical damage is a side effect of the administration of a chemotherapeutic agent administered to the subject for the prevention or treatment of cancer. In some embodiments, the kidney injury is drug-induced kidney injury or drug-induced acute kidney injury. In some embodiments, the renal tubule obstruction / blockage, such as, for example, the urinary tract, may be caused by the administration of certain chemical agents, such as, for example, sulfonamides, methotrexate, acyclovir, diethylene glycol, triamterene, etc.

[0088] Ischemic injury may be caused by the reduction of renal blood flow, which may be caused by various factors, such as hypotension due to sepsis, blood loss, surgery, etc.; or the effect of chemical agents (e.g., medicines or drugs) administered to subjects for the purpose of treating another disease, disorder or condition. Renal injury caused by ischemia may be ischemia-induced renal injury or ischemia-induced acute renal injury. Renal injury caused by crush injury may be ischemia-induced renal injury accompanied by vasoconstriction, or may be renal injury caused by mechanical factors due to renal tubule casts, or may be renal injury caused by the toxic effects of circulating factors (e.g., myoglobin).

[0089] In some embodiments, the kidney injury may be acute kidney injury (AKI), characterized by injury to renal tubular epithelial cells (TECs) (i.e., renal tubular epithelial cells), and in some cases, the kidney injury may include or result in the death of the kidney tubular epithelial cells. The kidney tubular epithelial cells may be proximal or distal tubular epithelial cells, both of which may be injured in acute kidney injury, and glomerular podocytes may also be injured in acute kidney injury. The injury to the kidney tubular epithelial cells may be any type of injury, damage, or injury, such as mechanical, chemical, or ischemic injury, as described above. The injury to the kidney tubular epithelial cells is often seen as a causative factor of kidney injury, especially acute kidney injury. While the proliferation of the kidney tubular epithelial cells provides a mechanism for recovery and repair of kidney function, the failure of the proliferation of the kidney tubular epithelial cells may lead to the development and progression of diseases, such as chronic kidney disease and renal failure. Proliferation of podocyte precursor cells can also restore glomerular function, but this is less well described than proliferation of tubular epithelial cells.

[0090] In some embodiments, the renal damage is nephrotoxicity or is characterized by nephrotoxicity. As used herein, "nephrotoxicity" refers to toxicity to the kidney. Also, "toxicity" refers to damage, for example, as described herein. Nephrotoxicity may result from the toxic effect of certain substances on renal function, and may therefore be considered a result of chemical damage or injury. Similar to chemical damage or injury, nephrotoxicity is a side effect of a drug administered to treat a disease or condition occurring in tissues other than the kidney, or a side effect of a drug administered to treat a disease or condition occurring in the kidney and one or more other tissues. In some embodiments, nephrotoxicity is a side effect of the administration of a chemotherapeutic agent administered to a subject for the purpose of preventing or treating cancer. Thus, nephrotoxicity may be a form of drug-induced nephropathy or drug-induced acute kidney injury.

[0091] As mentioned above, drug-induced nephropathy, drug-induced acute kidney injury, or drug-induced nephrotoxicity may occur as a side effect of a drug or medicine administered to treat a disease, disorder, or condition occurring in tissues other than the kidney or tissues within the kidney, or both. Such side effects are known to occur with many drugs, and may depend on the condition being treated, the subject being treated, the dosage, dosing schedule, or method of administration, and the severity of partial renal failure that the subject exhibited before treatment. For example, drugs that have been reported to be capable of inducing nephropathy include diuretics, beta-blockers, vasodilators, ACE inhibitors, aminoglycoside antibiotics (e.g., gentamicin), amphotericin B, cisplatin, NSAIDs (e.g., aspirin, ibuprofen, diclofenac), cyclosporine, lithium salts, cyclophosphamide, sulfonamides, methotrexate, acyclovir, diethylene glycol, triamterene, beta-lactam antibiotics, vancomycin, rifampicin, ciprofloxacin, ranitidine, cimetidine, furosemide, thiazides, and phenytoin. Drug-induced nephropathy does not have to be folic acid-induced nephropathy.

[0092] In some embodiments, drug-induced nephropathy, drug-induced acute kidney injury or drug-induced nephrotoxicity may occur as a side effect of a chemotherapeutic agent administered to treat or prevent cancer in a subject. Exemplary chemotherapeutic agents include alkylating agents such as cisplatin, carboplatin, mechlorethamine, cyclophosphamide, chlorambucil, ifosfamide; purine or pyrimidine antimetabolites such as azathioprine and mercaptopurine; alkaloids and terpenoids such as vinca alkaloids (e.g., vincristine, vinblastine, vinorelbine, vindesine), podophyllotoxin, etoposide, teniposide, taxanes (e.g., paclitaxel (Taxol™), docetaxel); topoisomerase inhibitors such as type I topoisomerase inhibitors (camptothecin, irinotecan, and topotecan) and type II topoisomerase inhibitors (amsacrine, etoposide, etoposide phosphate, teniposide); dactinomycin, doxorubicin (Adriamycin™), epirubicin, ribavir ... antitumor antibiotics (e.g., anthracycline antibiotics) such as cyclosporine, bleomycin, and rapamycin; anti-VEGF antibodies, anti-TNFα antibodies, anti-IL-2 antibodies, anti-GpIIb / IIIa antibodies, anti-CD52 antibodies, anti-CD20 antibodies, anti-RSV antibodies, anti-HER2 / neu (erbB2) antibodies, anti-TNF receptor antibodies, anti-EGFR antibodies, monoclonal antibodies or antibody fragments (specific examples include cetuximab, panitumumab, and the like); antibody-based agents such as rituximab, infliximab, basiliximab, bevacizumab (Avastin®), abciximab, daclizumab, gemtuzumab, alemtuzumab, rituximab (Mabthera®), palivizumab, trastuzumab, etanercept, adalimumab, nimotuzumab); as well as EGFR inhibitors such as erlotinib, cetuximab, and gefitinib.

[0093] In some preferred embodiments, the present invention relates to the diagnosis, treatment and / or prevention of cisplatin-induced nephropathy. Cisplatin-induced nephropathy includes cisplatin-induced acute kidney injury or cisplatin-induced nephrotoxicity. Cisplatin (dichlorodiaminoplatinum; (SP-4-2)-diamminedichloroplatinum(II)) is a chemotherapeutic agent that is widely used in the treatment of various cancers, such as head and neck cancer, breast cancer, lung cancer, testicular cancer, ovarian cancer, brain tumor, bladder cancer, etc., and is widely recognized to cause nephropathy and renal dysfunction accompanied by tubular damage and tubular necrosis (see, for example, Oh et al., Electrolyte Blood Press 2014 Dec; 12(2): 55-65; PA Arunkumar et al., Asian Pac J Trop Biomed 2012 Aug 2(8): 640-644). Other platinum-based chemotherapeutic agents also cause kidney damage.

[0094] It is recognized that subjects with renal injury may develop renal fibrosis, which may occur as a condition with a distinct etiology from renal injury or as a secondary effect of renal injury, but in some embodiments, the renal injury diagnosed, treated, or prevented is not renal fibrosis (e.g., renal fibrosis). In some embodiments, the subject does not suffer from fibrosis. In some embodiments, the damage to tubular epithelial cells occurs in the absence of fibrosis. In some embodiments, fibrosis separately (e.g., secondarily) leads to acute kidney injury, for example, due to incomplete regeneration of tubular epithelial cells. In some embodiments, the damaged tubular epithelial cells in the subject are not profibrotic tubular epithelial cells. In some embodiments, fibrosis does not develop.

[0095] Drugs that can suppress the action of IL-11 An embodiment of the present invention involves the inhibition of IL-11 mediated signaling.

[0096] As used herein, "inhibition" refers to a decrease, lowering or reduction compared to a control condition. For example, the inhibition of the action of IL-11 by an agent capable of inhibiting IL-11-mediated signal transduction refers to a decrease, lowering or reduction in the intensity / degree of IL-11-mediated signal transduction in the absence of the agent and / or in the presence of a suitable control agent.

[0097] In addition, as used herein, "suppression" may refer to neutralization or antagonism. That is, an agent capable of suppressing IL-11-mediated signal transduction (e.g., interaction, signal transduction or other activity via IL-11 or IL-11-containing complexes) may be said to be a "neutralizing" agent or an "antagonizing" agent for the relevant function or process. For example, an agent capable of suppressing IL-11-mediated signal transduction may be referred to as an agent capable of neutralizing IL-11-mediated signal transduction, or as an antagonist for IL-11-mediated signal transduction.

[0098] There are multiple routes in the IL-11 signal transduction pathway that can inhibit IL-11 signal transduction. For example, a drug capable of inhibiting IL-11-mediated signal transduction may inhibit IL-11 signal transduction by inhibiting the action of one or more factors involved in IL-11 receptor-mediated signal transduction or the action of one or more factors required for IL-11 receptor-mediated signal transduction.

[0099] For example, inhibition of IL-11 signaling may be achieved by disrupting the interaction between IL-11 (or an IL-11-containing complex, e.g., a complex consisting of IL-11 and IL-11Rα) and an IL-11 receptor (e.g., IL-11Rα, a receptor complex containing IL-11Rα, gp130, or a receptor complex containing IL-11Rα and gp130). In some embodiments, inhibition of IL-11-mediated signaling is achieved by inhibiting gene or protein expression of, for example, one or more of IL-11, IL-11Rα, and gp130.

[0100] In another embodiment, the inhibition of IL-11-mediated signaling is achieved by disrupting IL-11-mediated cis-signaling without disrupting IL-11-mediated trans-signaling, e.g., the inhibition of IL-11-mediated signaling is achieved by inhibiting gp130-mediated cis-complexes containing membrane-bound IL-11Rα. In another embodiment, the inhibition of IL-11-mediated signaling is achieved by disrupting IL-11-mediated trans-signaling without disrupting IL-11-mediated cis-signaling, i.e., the inhibition of IL-11-mediated signaling is achieved by inhibiting gp130-mediated trans-signaling complexes, such as IL-11 bound to soluble IL-11Rα and IL-6 bound to soluble IL-6R. In another embodiment, the inhibition of IL-11-mediated signaling is achieved by disrupting IL-11-mediated cis-signaling and IL-11-mediated trans-signaling. Any of the agents described herein may be used to inhibit IL-11-mediated cis-signaling and / or IL-11-mediated trans-signaling.

[0101] In another example, inhibition of IL-11 signaling may be achieved by disrupting a signaling pathway downstream of IL-11 / IL-11Rα / gp130. That is, in some embodiments, the inhibition / antagonism of IL-11-mediated signaling includes inhibition of a signaling pathway / signaling process / signaling factor downstream of signaling via a complex consisting of IL-11 and the IL-11 receptor.

[0102] In some embodiments, the inhibition / antagonism of IL-11-mediated signaling comprises the inhibition of signaling through an intracellular signaling pathway activated by a complex of IL-11 and the IL-11 receptor. In some embodiments, the inhibition / antagonism of IL-11-mediated signaling comprises the inhibition of one or more factors whose expression / activity is upregulated by signaling through a complex of IL-11 and the IL-11 receptor.

[0103] In some embodiments, the method of the present invention uses an agent capable of inhibiting JAK / STAT signaling. In some embodiments, the agent capable of inhibiting JAK / STAT signaling can inhibit the action of JAK1, JAK2, JAK3, TYK2, STAT1, STAT2, STAT3, STAT4, STAT5A, STAT5B and / or STAT6. For example, the agent capable of inhibiting JAK / STAT signaling can inhibit the activation of JAK / STAT proteins, inhibit the interaction between JAK or STAT proteins and cell surface receptors (e.g., IL-11Rα or gp130), inhibit the phosphorylation of JAK proteins, inhibit the interaction between JAK proteins and STAT proteins, inhibit the phosphorylation of STAT proteins, inhibit the dimerization of STAT proteins, inhibit the transport of STAT proteins to the cell nucleus, inhibit the binding of STAT proteins to DNA, and / or promote the degradation of JAK proteins and / or STAT proteins. In some embodiments, the JAK / STAT inhibitor is selected from the group consisting of ruxolitinib (Jakafi / Jakavi; Incyte), tofacitinib (Xeljanz / Jakvinus; NIH / Pfizer), oclacitinib (Apoquel), baricitinib (Olumient; Incyte / Eli Lilly), filgotinib (G-146034 / GLPG-0634; Galapagos), and rituximab (G-146034 / GLPG-0634; Galapagos). NV), gandotinib (LY-2784544; Eli Lilly), lestaurtinib (CEP-701; Teva), momelotinib (GS-0387 / CYT-387; Gilead Sciences), pacritinib (SB1518; CTI), PF-04965842 (Pfizer), upadacitinib (ABT-494; AbbVie), peficitinib (ASP015K / JNJ-54781532; Astellas), fedratinib (SAR302503; Celgene), cucurbitacin I (JSI-124), and CHZ868.

[0104] In some embodiments, the methods of the present invention use agents capable of inhibiting MAPK / ERK signaling. In some embodiments, the agents capable of inhibiting MAPK / ERK signaling can inhibit the action of GRB2, inhibit the action of RAF kinase, inhibit the action of MEK protein, inhibit the activation of MAP3K / MAP2K / MAPK and / or Myc, and / or inhibit the phosphorylation of STAT protein. In some embodiments, the agents capable of inhibiting ERK signaling can inhibit ERK p42 / 44. In some embodiments, the ERK inhibitor is selected from SCH772984, SC1, VX-11e, DEL-22379, sorafenib (Nexavar; Bayer / Onyx), SB590885, PLX4720, XL281, RAF265 (Novartis), encorafenib (LGX818 / Biraftovi; Array BioPharma), dabrafenib (Tafinlar; GSK), vemurafenib (Zelboraf; Roche), cobimetinib (Cotellic; Roche), CI-1040, PD0325901, binimetinib (MEK162 / Mektovi; Array BioPharma), selumetinib (AZD6244; Array / AstraZeneca), and trametinib (GSK1120212 / Mekinist; Novartis). In some embodiments, the method of the present invention uses an agent capable of inhibiting c-Jun N-terminal kinase (JNK) signaling / activity. In some embodiments, the agent capable of inhibiting JNK signaling / activity can inhibit the action and / or phosphorylation of JNK (e.g., JNK1, JNK2, etc.). In some embodiments, the JNK inhibitor is selected from SP600125, CEP 1347, TCS JNK 6o, c-JUN peptide, SU3327, AEG 3482, TCS JNK 5a, BI78D3, IQ3, SR3576, IQ1S, JIP-1(153-163) and CC401 dihydrochloride.

[0105] Widjaja et al., bioRxiv (2019) 830018, shows that signaling through IL-11 / IL-11Rα / gp130 upregulates NOX4 expression and activity. NOX4 is an NADPH oxidase and a source of reactive oxygen species (ROS). Nox4 expression is upregulated in transgenic mice expressing IL-11 specifically in hepatocytes, and stimulation of human primary hepatocytes with IL-11 also upregulates NOX4 expression.

[0106] In some embodiments, the present invention uses a drug that can suppress the expression (gene or protein expression) or its function. In some embodiments, the present invention uses a drug that can suppress the upregulation of NOX4 expression / function via IL-11. The drug that can suppress the expression or function of NOX4 may be referred to as NOX4 inhibitor herein. For example, NOX4 inhibitor can reduce the expression (e.g., gene and / or protein expression) of NOX4, can reduce the expression amount of RNA encoding NOX4, can reduce the expression amount of NOX4 protein, and / or can reduce the activity level of NOX4 (e.g., can reduce the activity of NADPH oxidase via NOX4, and / or can reduce the production of ROS via NOX4).

[0107] NOX4 inhibitors include molecules that bind to NOX4 and molecules that can reduce the expression of NOX4. Inhibitors that bind to NOX4 include peptide / nucleic acid aptamers, antibodies (and antibody fragments), and fragments of interaction partners of NOX4 that behave as antagonists to the function of NOX4, as well as small molecule inhibitors of NOX4. Molecules that can reduce the expression of NOX4 include antisense RNA of NOX4 (e.g., siRNA, shRNA, etc.). In some embodiments, the NOX4 inhibitor is selected from the NOX4 inhibitors described in Altenhofer et al., Antioxid Redox Signal. (2015) 23(5): 406-427 or Augsburder et al., Redox Biol. (2019) 26: 101272, such as GKT137831.

[0108] Binding Agents In some embodiments, the agent capable of suppressing IL-11-mediated signaling may bind to IL-11. In some embodiments, the agent capable of suppressing IL-11-mediated signaling may bind to an IL-11 receptor (e.g., IL-11Rα, gp130, or a complex including IL-11Rα and / or gp130). When such an agent binds to IL-11 or the IL-11 receptor, the binding ability of IL-11 to the IL-11 receptor may be reduced / inhibited, thereby suppressing IL-11-mediated signaling, thereby suppressing downstream signaling. Also, when such an agent binds to IL-11 or the IL-11 receptor, the binding ability of IL-11 to the IL-11 receptor (e.g., IL-11Rα and / or gp130) may be reduced / inhibited, thereby suppressing IL-11-mediated cis signaling and / or IL-11-mediated trans signaling, thereby suppressing downstream signaling. The agent may bind to a trans-signaling complex, such as a complex consisting of IL-11 and soluble IL-11Rα, and inhibit gp130-mediated signaling.

[0109] The agent capable of binding to IL-11 or an IL-11-containing complex or IL-11 receptor may be of any type, but in some embodiments the agent may be an antibody, an antigen-binding fragment thereof, a polypeptide, a peptide, a nucleic acid, an oligonucleotide, an aptamer, or a small molecule. The agent may be provided in an isolated or purified form and may be formulated as a pharmaceutical composition or medicament.

[0110] Antibodies and antigen-binding fragments In some embodiments, the agent capable of binding to IL-11 or IL-11-containing complexes or IL-11 receptor is an antibody or an antigen-binding fragment thereof.In some embodiments, the agent capable of binding to IL-11 or IL-11-containing complexes or IL-11 receptor is a polypeptide, such as a decoy receptor molecule.In some embodiments, the agent capable of binding to IL-11 or IL-11-containing complexes or IL-11 receptor may be an aptamer.

[0111] In some embodiments, the agent capable of binding to IL-11 or an IL-11-containing complex or an IL-11 receptor is an antibody or an antigen-binding fragment thereof. As used herein, the term "antibody" is used in the broadest sense and includes monoclonal antibodies, polyclonal antibodies, monospecific antibodies, multispecific antibodies (e.g., bispecific antibodies) and antibody fragments that exhibit binding to relevant target molecules.

[0112] Modern techniques in monoclonal antibody technology allow the production of antibodies against most antigens. The antigen-binding portion may be a portion of an antibody (e.g., a Fab fragment) or a synthetic antibody fragment (e.g., a single chain Fv fragment [ SThe monoclonal antibody may be a monoclonal antibody (e.g., a monoclonal antibody fragment (cFv)) against a selected antigen. Monoclonal antibodies against a selected antigen may be produced by known techniques, such as those described in "Monoclonal Antibodies: A manual of techniques", H Zola (CRC Press, 1988) and "Monoclonal Hybridoma Antibodies: Techniques and Applications", JGR Hurrell (CRC Press, 1982). Chimeric antibodies have also been reported by Neuberger et al. (1988, 8th International Biotechnology Symposium Part 2, 792-799). Monoclonal antibodies (mAbs) are particularly useful in the methods of the present invention. Monoclonal antibodies (mAbs) are homogeneous antibody populations that specifically target a single epitope on an antigen.

[0113] Polyclonal antibodies are also useful in the methods of the invention. Monospecific polyclonal antibodies are preferred. Suitable polyclonal antibodies can be produced using methods well known in the art.

[0114] Antigen-binding fragments of antibodies, such as Fab and Fab2 fragments, may be used / provided, as well as recombinant antibodies and recombinant antibody fragments. H ) region and the light chain variable (V L ) region is known to be involved in antigen recognition, as first discovered by early protease digestion experiments and confirmed by experiments in "humanizing" rodent antibodies. Variable regions of rodent origin can be fused to constant regions of human origin to produce antibodies that retain the antigen specificity of the parent rodent antibody (Morrison et al (1984) Proc. Natl. Acad. Sd. USA 81, 6851-6855).

[0115] The antibodies and antigen-binding fragments according to the present disclosure comprise complementarity determining regions (CDRs) of an antibody capable of binding to the relevant target molecule (ie, IL-11 / IL-11-containing complex / IL-11 receptor).

[0116] Examples of antibodies capable of binding to IL-11 include the monoclonal mouse anti-human IL-11 antibody clone #22626; catalogue no. MAB218 (R&D Systems, Minnesota, USA), clone 6D9A (Abbiotec), clone KT8 (Abbiotec), clone M3103F11 (BioLegend), clone 1F1 (Abnova Corporation), clone 3C6 (Abnova Corporation), clone GF1 (LifeSpan Biosciences), clone 13455 (Source BioScience), 11h3 / 19.6.1 (Hermann et al., Arthritis Rheum. (1998) 41(8):1388-97), AB-218-NA (R&D Systems), X203 (Ng et al., Sci Transl Med. (2019) 11(511) pii: eaaw1237, and also reported as Ng, et al., "IL-11 is a therapeutic target in idiopathic pulmonary fibrosis." bioRxiv 336537; doi: https: / / doi.org / 10.1101 / 336537), as well as the anti-IL-11 antibodies disclosed in U.S. Patent Publication No. 2009 / 0202533(A1), WO99 / 59608(A2), WO2018 / 109174(A2), and WO 2019 / 238882(A1).

[0117] In particular, the anti-IL-11 antibody clone 22626 (also known as MAB218) has been shown to be an antagonist to IL-11-mediated signaling, for example, in Schaefer et al., Nature (2017) 552(7683):110-115. Hermann et al., Arthritis Rheum. (1998) 41(8):1388-97 discloses that monoclonal antibody 11h3 / 19.6.1 is an anti-IL-11 IgG1 neutralizing antibody. R&D Systems' AB-218-NA, used, for example, in McCoy et al., BMC Cancer (2013) 13:16, is another example of an anti-IL-11 neutralizing antibody. WO 2018 / 109174(A2) and WO 2019 / 238882(A1) disclose further examples of anti-IL-11 antagonist antibodies against IL-11-mediated signaling. Also, Ng, et al., "IL-11 is a therapeutic target in idiopathic pulmonary fibrosis." bioRxiv 336537; doi: https: / / doi.org / 10.1101 / 336537 and X203 (also referred to as Enx203) disclosed in WO 2019 / 238882 (A1) is an anti-IL11 antagonist antibody against IL-11-mediated signal transduction, and includes a VH region as shown in SEQ ID NO: 92 in WO 2019 / 238882(A1) (SEQ ID NO: 22 in the present disclosure) and a VL region as shown in SEQ ID NO: 94 in WO 2019 / 238882(A1) (SEQ ID NO: 23 in the present disclosure). Humanized X203 is described in WO 2019 / 238882(A1), including hEnx203, which comprises a VH region set forth in SEQ ID NO: 117 in WO 2019 / 238882(A1) (SEQ ID NO: 30 in this disclosure) and a VL region set forth in SEQ ID NO: 122 in WO 2019 / 238882(A1) (SEQ ID NO: 31 in this disclosure).Enx108A is yet another example of an anti-IL-11 antagonist antibody against IL-11-mediated signaling, and comprises a VH region set forth in SEQ ID NO:8 in WO 2019 / 238882(A1) (SEQ ID NO:26 in the present disclosure) and a VL region set forth in SEQ ID NO:20 in WO 2019 / 238882(A1) (SEQ ID NO:27 in the present disclosure).

[0118] Examples of antibodies capable of binding to IL-11Rα include monoclonal antibody clone 025 (Sino Biological), clone EPR5446 (Abcam), clone 473143 (R&D Systems), clones 8E2, 8D10 and 8E4 described in U.S. Patent Publication No. 2014 / 0219919(A1), and affinity matured variants of 8E2, monoclonal antibodies described in Blanc et al. (J. Immunol Methods. 2000 Jul 31;241(1-2);43-59), X209 (reported in Widjaja et al., Gastroenterology (2019) 157(3):777-792, and described in Widjaja, et al., "IL-11 neutralising therapies target hepatic stellate cell-induced liver inflammation and fibrosis in NASH." bioRxiv 470062; doi: https: / / doi.org / 10.1101 / 470062), WO2014121325(A1) and U.S. Patent Publication No. 2013 / 0302277(A1), as well as the anti-IL-11Rα antibodies disclosed in U.S. Patent Publication No. 2009 / 0202533(A1), WO99 / 59608(A2), WO2018 / 109170(A2) and WO 2019 / 238884(A1).

[0119] In particular, anti-IL-11Rα antibody clone 473143 (also known as MAB1977) has been shown to be an antagonist to IL-11-mediated signaling, for example, in Schaefer et al., Nature (2017) 552(7683):110-115. US Patent Publication No. 2014 / 0219919(A1) provides sequences of anti-human IL-11Rα antibody clones 8E2, 8D10 and 8E4, and discloses that these anti-human IL-11Rα antibody clones have antagonistic effects on IL-11-mediated signaling (see, for example, paragraphs

[0489] to

[0490] of US Patent Publication No. 2014 / 0219919(A1)). Furthermore, US Patent Publication No. 2014 / 0219919(A1) also provides sequence information for 62 affinity matured variants of clone 8E2, 61 of which are disclosed to have antagonistic activity against IL-11-mediated signaling (see Table 3 in US Patent Publication No. 2014 / 0219919(A1)). WO 2018 / 109170(A2) and WO 2019 / 238884(A1) disclose further examples of anti-IL-11Rα antagonist antibodies against IL-11-mediated signaling. Also, Widjaja, et al., "IL-11 neutralising therapies target hepatic stellate cell-induced liver inflammation and fibrosis in NASH." bioRxiv 470062; doi: https: / / doi.org / 10.1101 / 470062 and X209 (also referred to as Enx209), disclosed in WO 2019 / 238884(A1), is an anti-IL-11Rα antagonist antibody against IL-11-mediated signal transduction, and comprises a VH region as shown in SEQ ID NO: 7 in WO 2019 / 238884(A1) (SEQ ID NO: 24 in this disclosure) and a VL region as shown in SEQ ID NO: 14 in WO 2019 / 238884(A1) (SEQ ID NO: 25 in this disclosure).Humanized X209 is described in WO 2019 / 238884(A1), including hEnx209, which comprises a VH region set forth in SEQ ID NO: 11 in WO 2019 / 238884(A1) (SEQ ID NO: 32 in the present disclosure) and a VL region set forth in SEQ ID NO: 17 in WO 2019 / 238884(A1) (SEQ ID NO: 33 in the present disclosure).

[0120] Those of skill in the art will be familiar with techniques for generating antibodies suitable for therapeutic use in a given species / subject. For example, procedures for generating antibodies suitable for therapeutic use in humans are described in Park and Smolen Advances in Protein Chemistry (2001) 56: 369-421, which is incorporated herein by reference in its entirety.

[0121] Antibodies against a given target protein (e.g., IL-11 or IL-11Rα) can be generated in model species (e.g., rodents or rabbits) and then genetically modified to improve suitability for therapeutic use in a given species / subject. For example, one or more amino acids of a monoclonal antibody generated by immunization of a model species can be substituted to obtain an antibody sequence that is closer to the sequence of human germline immunoglobulins (thus reducing the likelihood of an immune response due to anti-xenoantibodies in human subjects treated with the antibody). Modifications of antibody variable domains can be focused on the framework regions to preserve the paratope of the antibody. Antibody humanization is routine in the field of antibody technology and has been reviewed, for example, in Almagro and Fransson, Frontiers in Bioscience (2008) 13:1619-1633, Safdari et al., Biotechnology and Genetic Engineering Reviews (2013) 29(2): 175-186, and Lo et al., Microbiology Spectrum (2014) 2(1), all of which are incorporated by reference in their entireties. Antibody humanization can be omitted by generating antibodies against a given target protein (e.g., IL-11 or IL-11Rα) in a transgenic model species expressing human immunoglobulin genes to obtain a fully human antibody (e.g., as described in Bruggemann et al., Arch Immunol Ther Exp (Warsz) (2015) 63(2):101-108, which is incorporated herein by reference in its entirety).

[0122] Phage display technology may also be used to identify antibodies against a given target protein (e.g., IL-11 or IL-11Rα), and this method is well known to those skilled in the art. The identification of fully human antibodies against human target proteins using phage display is reviewed, for example, in Hoogenboom, Nat. Biotechnol. (2005) 23, 1105-1116 and Chan et al., International Immunology (2014) 26(12): 649-657, which are incorporated herein by reference in their entirety.

[0123] The antibody / fragment may be an antagonist antibody / fragment that inhibits or reduces the biological activity of IL-11. The antibody / fragment may be a neutralizing antibody that neutralizes the biological effect of IL-11, for example, neutralizing the ability of IL-11 to stimulate protein synthesis signaling through the IL-11 receptor. Neutralizing activity may be measured by assessing the neutralizing ability against IL-11-induced proliferation in T11 mouse plasmacytoma cell line (Nordan, RP et al. (1987) J. Immunol. 139:813).

[0124] Antibodies that bind IL-11 or that bind IL-11Rα can be assessed for antagonistic effects on IL-11-mediated signaling using assays such as those described in U.S. Patent Publication No. 2014 / 0219919(A1) or Blanc et al. (J. Immunol Methods. 2000 Jul 31;241(1-2);43-59). Briefly, antibodies that bind IL-11 and antibodies that bind IL-11Rα can be assessed in vitro for their ability to inhibit proliferation of Ba / F3 cells expressing IL-11Rα and gp130 from an appropriate species in response to stimulation with IL-11 from the appropriate species. In another method, antibodies that bind IL-11 and antibodies that bind IL-11Rα can be analyzed in vitro for their ability to inhibit fibroblast to myofibroblast conversion by assessing αSMA expression after stimulation of fibroblasts with TGFβ1 (e.g., as described in WO 2018 / 109174(A2) (Example 6) and WO 2018 / 109170(A2) (Example 6) and Ng et al., Sci Transl Med. (2019) 11(511) pii: eaaw1237 and Widjaja et al., Gastroenterology (2019) 157(3):777-792).

[0125] An antibody typically contains six CDRs: three CDRs (LC-CDR1, LC-CDR2 and LC-CDR3) in the light chain variable region (VL) and three CDRs (HC-CDR1, HC-CDR2 and HC-CDR3) in the heavy chain variable region (VH). Together, these six CDRs define the paratope of the antibody, which refers to the part of the antibody that binds to the target molecule. The VH and VL regions contain framework regions (FR) on either side of each CDR, which provide a scaffold for the CDRs. The structure of the VH region comprises, from the N-terminus to the C-terminus, N-terminus-[HC-FR1]-[HC-CDR1]-[HC-FR2]-[HC-CDR2]-[HC-FR3]-[HC-CDR3]-[HC-FR4]-C-terminus, and the structure of the VL region comprises, from the N-terminus to the C-terminus, N-terminus-[LC-FR1]-[LC-CDR1]-[LC-FR2]-[LC-CDR2]-[LC-FR3]-[LC-CDR3]-[LC-FR4]-C-terminus.

[0126] There are several conventional methods for defining the CDRs and FRs of an antibody, including, for example, the methods described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD (1991) and Chothia et al., J. Mol. Biol. 196:901-917 (1987), and VBASE2 described in Retter et al., Nucl. Acids Res. (2005) 33 (suppl 1): D671-D674. The CDRs and FRs of the VH and VL regions of the antibodies described herein are defined according to the Kabat system.

[0127] In some embodiments, an antibody or antigen-binding fragment thereof according to the present disclosure is derived from an antibody that specifically binds to IL-11 (e.g., Enx108A, Enx203, or hEnx203). In some embodiments, an antibody or antigen-binding fragment thereof according to the present disclosure is derived from an antibody that specifically binds to IL-11Rα (e.g., Enx209 or hEnx209).

[0128] Antibodies and antigen-binding fragments according to the present disclosure preferably inhibit IL-11 mediated signaling. Such antibodies / antigen-binding fragments may be described as being antagonists to IL-11 mediated signaling and / or as having neutralizing capacity to IL-11 mediated signaling.

[0129] In some embodiments, the antibody / antigen-binding fragment comprises the CDRs of an antibody that binds IL-11. In some embodiments, the antibody / antigen-binding fragment comprises the CDRs of or is derived from the CDRs of an antibody that binds IL-11 described herein (e.g., Enx108A, Enx203, or hEnx203).

[0130] In some embodiments, the antibody / antigen-binding fragment comprises: (1) HC-CDR1 having the amino acid sequence of SEQ ID NO: 34, HC-CDR2 having the amino acid sequence of SEQ ID NO: 35, and HC-CDR3 having the amino acid sequence of SEQ ID NO: 36, or A variant in which one, two or three amino acids are replaced with another amino acid in one or more of HC-CDR1, HC-CDR2 or HC-CDR3. It contains a VH region incorporating

[0131] In some embodiments, the antibody / antigen-binding fragment comprises: (2) LC-CDR1 having the amino acid sequence of SEQ ID NO: 37, LC-CDR2 having the amino acid sequence of SEQ ID NO: 38, and LC-CDR3 having the amino acid sequence of SEQ ID NO: 39, or A variant in which one, two or three amino acids are replaced with another amino acid in one or more of LC-CDR1, LC-CDR2 or LC-CDR3. It contains a VL region incorporating

[0132] In some embodiments, the antibody / antigen-binding fragment comprises: (3) HC-CDR1 having the amino acid sequence of SEQ ID NO: 40, HC-CDR2 having the amino acid sequence of SEQ ID NO: 41, and HC-CDR3 having the amino acid sequence of SEQ ID NO: 42, or A variant in which one, two or three amino acids are replaced with another amino acid in one or more of HC-CDR1, HC-CDR2 or HC-CDR3. It contains a VH region incorporating

[0133] In some embodiments, the antibody / antigen-binding fragment comprises: (4) LC-CDR1 having the amino acid sequence of SEQ ID NO: 43, LC-CDR2 having the amino acid sequence of SEQ ID NO: 44, and LC-CDR3 having the amino acid sequence of SEQ ID NO: 45, or A variant in which one, two or three amino acids are replaced with another amino acid in one or more of LC-CDR1, LC-CDR2 or LC-CDR3. It contains a VL region incorporating

[0134] In some embodiments, the antibody / antigen-binding fragment comprises a VH region incorporating the CDRs described in (1) and a VL region incorporating the CDRs described in (2). In some embodiments, the antibody / antigen-binding fragment comprises a VH region incorporating the CDRs described in (3) and a VL region incorporating the CDRs described in (4).

[0135] In some embodiments, the antibody / antigen-binding fragment comprises the VH and VL regions of an antibody that binds IL-11. In some embodiments, the antibody / antigen-binding fragment comprises the VH and VL regions of, or derived from, the VH and VL regions of an antibody that binds IL-11 described herein (e.g., Enx108A, Enx203, or hEnx203).

[0136] In some embodiments, the antibody / antigen-binding fragment comprises a VH region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 100% sequence identity to the amino acid sequence of SEQ ID NO:26. In some embodiments, the antibody / antigen-binding fragment comprises a VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 100% sequence identity to the amino acid sequence of SEQ ID NO:27. In some embodiments, the antibody / antigen-binding fragment has an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 100% sequence identity to the amino acid sequence of SEQ ID NO:26. and a VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 100% sequence identity, to the amino acid sequence of SEQ ID NO:27.

[0137] In some embodiments, the antibody / antigen-binding fragment comprises a VH region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 100% sequence identity to the amino acid sequence of SEQ ID NO:22. In some embodiments, the antibody / antigen-binding fragment comprises a VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 100% sequence identity to the amino acid sequence of SEQ ID NO:23. In some embodiments, the antibody / antigen-binding fragment has an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 100% sequence identity to the amino acid sequence of SEQ ID NO:22. and a VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 100% sequence identity, to the amino acid sequence of SEQ ID NO:23.

[0138] In some embodiments, the antibody / antigen-binding fragment comprises a VH region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 100% sequence identity to the amino acid sequence of SEQ ID NO:30. In some embodiments, the antibody / antigen-binding fragment comprises a VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 100% sequence identity to the amino acid sequence of SEQ ID NO:31. In some embodiments, the antibody / antigen-binding fragment has an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 100% sequence identity to the amino acid sequence of SEQ ID NO:30. and a VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 100% sequence identity, to the amino acid sequence of SEQ ID NO:31.

[0139] In some embodiments, the antibody / antigen-binding fragment comprises the CDRs of an antibody that binds to IL-11Rα. In some embodiments, the antibody / antigen-binding fragment comprises the CDRs of an antibody that binds to IL-11Rα described herein (e.g., Enx209 or hEnx209) or comprises CDRs derived from the CDRs of these antibodies.

[0140] In some embodiments, the antibody / antigen-binding fragment comprises: (5) HC-CDR1 having the amino acid sequence of SEQ ID NO: 46, HC-CDR2 having the amino acid sequence of SEQ ID NO: 47, and HC-CDR3 having the amino acid sequence of SEQ ID NO: 48, or A variant in which one, two or three amino acids are replaced with another amino acid in one or more of HC-CDR1, HC-CDR2 or HC-CDR3. It contains a VH region incorporating

[0141] In some embodiments, the antibody / antigen-binding fragment comprises: (6) LC-CDR1 having the amino acid sequence of SEQ ID NO: 49, LC-CDR2 having the amino acid sequence of SEQ ID NO: 50, and LC-CDR3 having the amino acid sequence of SEQ ID NO: 51, or A variant in which one, two or three amino acids are replaced with another amino acid in one or more of LC-CDR1, LC-CDR2 or LC-CDR3. It contains a VL region incorporating

[0142] In some embodiments, the antibody / antigen-binding fragment comprises a VH region incorporating the CDRs described in (5) and a VL region incorporating the CDRs described in (6).

[0143] In some embodiments, the antibody / antigen-binding fragment comprises the VH and VL regions of an antibody that binds IL-11Rα. In some embodiments, the antibody / antigen-binding fragment comprises the VH and VL regions of, or derived from, the VH and VL regions of an antibody that binds IL-11Rα described herein (e.g., Enx209 or hEnx209).

[0144] In embodiments of the invention in which one or more amino acids of a reference amino acid sequence (e.g., a CDR sequence, a VH region sequence or a VL region sequence described herein) are replaced with another amino acid, the replacement may be, for example, a conservative replacement according to the following table: In some embodiments, the replacement is between amino acids in the same box in the middle column. In some embodiments, the replacement is between amino acids in the same box in the right column. [Table 1]

[0145] In some embodiments, the substitutions may be functionally conservative substitutions, i.e., in some embodiments, the substitutions may not affect (or may not substantially affect) one or more functional properties (e.g., binding to a target) of the antibody / fragment containing the substitution when compared to the same molecule without the substitution.

[0146] In some embodiments, substitutions to a reference VH or VL region sequence may be made with focus on one or more specific regions of the VH or VL region sequence, for example, changes from the reference VH or VL region sequence may be made with focus on one or more framework regions (FR1, FR2, FR3 and / or FR4).

[0147] Antibodies and antigen-binding fragments according to the present disclosure may be designed and prepared using the sequence of a monoclonal antibody (mAb) capable of binding to the relevant target molecule. Antigen-binding regions of antibodies may also be used / provided, such as single chain variable fragments (scFv), Fab fragments, Fab2 fragments, etc. An "antigen-binding region" or "antigen-binding fragment" is an antibody fragment capable of binding to a target for which the original antibody is specific.

[0148] In some embodiments, the antibody / fragment comprises a VL region and a VH region of an antibody capable of binding to IL-11, an IL-11-containing complex or an IL-11 receptor. The VL region and the VH region in the antigen-binding region of the antibody together constitute an Fv region. In some embodiments, the antibody / fragment comprises or consists of an Fv region of an antibody capable of binding to IL-11, an IL-11-containing complex or an IL-11 receptor. The Fv region may be expressed as a single chain comprising the VH and VL regions covalently linked, for example, by a flexible oligopeptide. Thus, the antibody / fragment may comprise or consist of an scFv comprising the VL and VH regions of an antibody capable of binding to IL-11, an IL-11-containing complex or an IL-11 receptor.

[0149] The light chain variable region (VL) and light chain constant region (CL) and the heavy chain variable region (VH) and heavy chain constant region 1 (CH1) in the antigen-binding region of the antibody together constitute a Fab region. In some embodiments, the antibody / fragment comprises or consists of a Fab region of an antibody capable of binding to IL-11, an IL-11-containing complex or an IL-11 receptor.

[0150] In some embodiments, the antibody / fragment comprises or consists of a full-length antibody capable of binding to IL-11, IL-11-containing complexes or IL-11 receptor. "Full-length antibody" refers to an antibody having a structure substantially similar to that of an immunoglobulin (Ig). For example, Schroeder and Cavacini J Allergy Clin Immunol. (2010) 125(202): S41-S52, which is incorporated herein by reference in its entirety, describes various types of immunoglobulins and their structures. Immunoglobulin G (i.e., IgG) is a glycoprotein of approximately 150 kDa that comprises two heavy chains and two light chains. The heavy chain comprises, from the N-terminus to the C-terminus, a heavy chain variable region (VH) followed by a heavy chain constant region that comprises three constant regions (CH1, CH2 and CH3), and similarly, the light chain comprises a light chain variable region (VL) followed by a light chain constant region (CL). Depending on the type of heavy chain, immunoglobulins may be classified as IgG (e.g., IgG1, IgG2, IgG3, IgG4), IgA (e.g., IgA1, IgA2), IgD, IgE, or IgM. The light chain may be a κ chain or a λ chain.

[0151] In some embodiments, the antibody / antigen-binding fragment of the present disclosure comprises a sequence of an immunoglobulin heavy chain constant region. In some embodiments, the sequence of the immunoglobulin heavy chain constant region may be a sequence of a human immunoglobulin heavy chain constant region. In some embodiments, the sequence of the immunoglobulin heavy chain constant region may be a sequence of a heavy chain constant region of IgG (e.g., IgG1, IgG2, IgG3, IgG4), IgA (e.g., IgA1, IgA2), IgD, IgE or IgM, such as a sequence of a heavy chain constant region of human IgG (e.g., hIgG1, hIgG2, hIgG3, hIgG4), hIgA (e.g., hIgA1, hIgA2), hIgD, hIgE or hIgM, or may be a sequence derived from the sequence of the heavy chain constant region of these antibodies. In some embodiments, the sequence of the immunoglobulin heavy chain constant region is the sequence of a heavy chain constant region of a human IgG1 allotype (e.g., G1m1, G1m2, G1m3 or G1m17) or is a sequence derived from the sequence of a heavy chain constant region of a human IgG1 allotype.

[0152] In some embodiments, the sequence of the immunoglobulin heavy chain constant region is the constant region sequence of the human immunoglobulin G1 constant region (IGHG1; UniProt: P01857-1, v1) or is a sequence derived from the constant region sequence of the human immunoglobulin G1 constant region. In some embodiments, the sequence of the immunoglobulin heavy chain constant region is the constant region sequence of the human immunoglobulin G1 constant region (i.e., allotype G1m3) (IGHG1; UniProt: P01857-1, v1) containing the K214R, D356E and L358M substitutions or is a sequence derived from this human immunoglobulin G1 constant region. In some embodiments, the antibody / antigen-binding fragment comprises an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 100% sequence identity to the amino acid sequence of SEQ ID NO:52.

[0153] In some embodiments, the sequence of the immunoglobulin heavy chain constant region is the constant region sequence of the human immunoglobulin G4 constant region (IGHG4; UniProt: P01861, v1) or is a sequence derived from the constant region sequence of the human immunoglobulin G4 constant region. In some embodiments, the sequence of the immunoglobulin heavy chain constant region is the constant region sequence of the human immunoglobulin G4 constant region (IGHG4; UniProt: P01861, v1) containing the S241P and / or L248E substitutions or is a sequence derived from this human immunoglobulin G4 constant region sequence. The S241P mutation stabilizes the hinge, and the L248E mutation further reduces the already low ADCC effector function of IgG4 (Davies and Sutton, Immunol Rev. 2015 Nov; 268(1):139-159; Angal et al Mol Immunol. 1993 Jan;30(1):105-8). In some embodiments, the antibody / antigen-binding fragment comprises an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 100% sequence identity to the amino acid sequence of SEQ ID NO:53.

[0154] In some embodiments, the antibody / antigen-binding fragment of the present disclosure comprises an immunoglobulin light chain constant region sequence. In some embodiments, the immunoglobulin light chain constant region sequence may be a human immunoglobulin light chain constant region sequence. In some embodiments, the immunoglobulin light chain constant region sequence may be a kappa or lambda light chain sequence or may be a sequence derived from a kappa or lambda light chain. Examples of kappa or lambda light chains include the human immunoglobulin constant region kappa chain (IGKC; Cκ; UniProt: P01834-1, v2) or the human immunoglobulin constant region lambda chain (IGLC; Cλ), such as IGLC1 (UniProt: P0CG04-1, v1), IGLC2 (UniProt: P0DOY2-1, v1), IGLC3 (UniProt: P0DOY3-1, v1), IGLC6 (UniProt: P0CF74-1, v1) or IGLC7 (UniProt: A0M8Q6-1, v3).

[0155] In some embodiments, the antibody / antigen-binding fragment comprises an immunoglobulin light chain constant region sequence. In some embodiments, the immunoglobulin light chain constant region sequence is a human immunoglobulin constant region kappa chain (IGKC; Cκ; UniProt: P01834-1, v2; SEQ ID NO: 90) sequence or a sequence derived from the human immunoglobulin constant region kappa chain. In some embodiments, the immunoglobulin light chain constant region sequence is a human immunoglobulin constant region lambda chain (IGLC; Cλ) sequence, such as IGLC1, IGLC2, IGLC3, IGLC6 or IGLC7. In some embodiments, the antibody / antigen-binding fragment comprises an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 100% sequence identity to the amino acid sequence of SEQ ID NO:54. In some embodiments, the antibody / antigen-binding fragment comprises an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 100% sequence identity to the amino acid sequence of SEQ ID NO:55.

[0156] In some embodiments, the antibody / antigen-binding fragment comprises: (i) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity, preferably 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity, to the amino acid sequence of SEQ ID NO:28; and (ii) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity, preferably 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity, to the amino acid sequence of SEQ ID NO:29.

[0157] In some embodiments, the antibody / antigen-binding fragment comprises: (i) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity, preferably 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity, to the amino acid sequence of SEQ ID NO:56; and (ii) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity, preferably 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity, to the amino acid sequence of SEQ ID NO:57.

[0158] In some embodiments, the antibody / antigen-binding fragment comprises: (i) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity, preferably 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity, to the amino acid sequence of SEQ ID NO:58; and (ii) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity, preferably 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity, to the amino acid sequence of SEQ ID NO:59.

[0159] Fab, Fv, scFv and dAb antibody fragments can all be expressed in E. coli and secreted, and therefore can be easily mass-produced.

[0160] Full length antibodies and F(ab')2 fragments are "bivalent". By "bivalent" we mean that full length antibodies and F(ab')2 fragments have two antigen binding sites. In contrast, Fab, Fv, scFv and dAb fragments are monovalent since they only have one antigen binding site. Synthetic antibodies capable of binding to IL-11, IL-11-containing complexes or IL-11 receptor can also be generated using phage display techniques well known in the art.

[0161] Antibodies may be produced by affinity maturation techniques to produce modified antibodies with improved affinity for antigens compared to the unmodified parent antibody. Affinity matured antibodies may be produced by techniques known in the art, and techniques for producing affinity matured antibodies are described, for example, in Marks et al., Rio / Technology 10:779-783 (1992); Barbas et al. Proc Nat. Acad. Sci. USA 91:3809-3813 (1994); Schier et al. Gene 169:147-155 (1995); Yelton et al. J. Immunol. 155:1994-2004 (1995); Jackson et al., J. Immunol. 154(7):331 0-15 9 (1995); and Hawkins et al, J. Mol. Biol. 226:889-896 (1992).

[0162] Antibodies / fragments include bispecific antibodies, which are composed of two fragments, e.g., from each of two antibodies, and can thereby bind to two antigens. Bispecific antibodies include antibodies / fragments as described herein that can bind to IL-11, IL-11-containing complexes, or IL-11 receptors. Bispecific antibodies may also include another fragment with affinity for a second antigen, which may be any antigen desired. Techniques for producing bispecific antibodies are well known in the art, see, for example, Mueller, D et al. (2010 Biodrugs 24 (2): 89-98), Wozniak-Knopp G et al. (2010 Protein Eng Des 23 (4): 289-297.), and Baeuerle, PA et al. (2009 Cancer Res 69 (12): 4941-4944). Bispecific antibodies and bispecific antigen-binding fragments may be provided in any suitable form, for example, as described in Kontermann MAbs 2012, 4(2): 182-197, which is incorporated herein by reference in its entirety. For example, bispecific antibodies or bispecific antigen-binding fragments may be provided as bispecific antibody complexes (e.g., IgG2, F(ab')2 or CovX-Body), bispecific IgG or IgG-like molecules (e.g., IgG, scFv4-Ig, IgG-scFv, scFv-IgG, DVD-Ig, IgG-sVD, sVD-IgG, 2 in 1-IgG, mAb 2 , or Tandemab with a shared light chain (LC), asymmetric bispecific IgG or IgG-like molecules (e.g. kih IgG, kih IgG with a shared light chain (LC), CrossMab, kih IgG-scFab, mAb-Fv, charge-paired IgG, or SEED-body), small bispecific antibody molecules (e.g. Diabody (Db), dsDb, DART, scDb, tandAb, tandem scFv (taFv), tandem dAb / VHH, triple body, triple head, Fab-scFv, or F(ab')2-scFv2), bispecific Fc-C H3 Fusion proteins (e.g. taFv-Fc, Di-diabody, scDb-C H 3. scFv-Fc-scFv, HCAb-VHH, scFv-kih-Fc or scFv-kih-C H 3), or bispecific fusion proteins (e.g., scFv2-albumin, scDb-albumin, taFv-toxin, DNL-Fab3, DNL-Fab4-IgG, DNL-Fab4-IgG-cytokine2). For details, see Figure 2 in Kontermann MAbs 2012, 4(2): 182-19.

[0163] Methods for producing bispecific antibodies include chemically crosslinking antibodies or antibody fragments, for example, via reducible disulfide bonds or non-reducible thioether bonds, as described, for example, in Segal and Bast, 2001. Production of Bispecific Antibodies. Current Protocols in Immunology. 14:IV:2.13:2.13.1-2.13.16, which is incorporated herein by reference in its entirety. For example, N-succinimidyl-3-(2-pyridyldithio)-propionate (SPDP) can be used to chemically crosslink, for example, Fab fragments, via SH-groups in the hinge region to generate disulfide-linked bispecific F(ab)2 heterodimers.

[0164] Another method for producing bispecific antibodies includes fusing antibody-producing hybridomas, for example with polyethylene glycol, to produce quadroma cells capable of secreting bispecific antibodies, as described, for example, in DM and Bast, BJ 2001. Production of Bispecific Antibodies. Current Protocols in Immunology. 14:IV:2.13:2.13.1-2.13.16.

[0165] Bispecific antibodies and bispecific antigen-binding fragments can also be produced by recombinant techniques, e.g., bispecific antibodies and bispecific antigen-binding fragments can be expressed from nucleic acid constructs encoding the polypeptides of the antigen-binding molecules, e.g., as described in Antibody Engineering: Methods and Protocols, Second Edition (Humana Press, 2012), Chapter 40: Production of Bispecific Antibodies: Diabodies and Tandem scFv (Hornig and Farber-Schwarz), or French, How to make bispecific antibodies, Methods Mol. Med. 2000; 40:333-339.

[0166] For example, a DNA construct can be generated by molecular cloning techniques that contains sequences encoding the light and heavy chain variable regions of two antigen-binding regions (i.e., an antigen-binding region capable of binding to IL-11, an IL-11-containing complex, or an IL-11 receptor, and an antigen-binding region capable of binding to another target protein), as well as a suitable linker or dimerization region linking these antigen-binding regions. This DNA construct can then be expressed (e.g., in vitro) in a suitable host cell (e.g., a mammalian host cell) to produce a recombinant bispecific antibody, and the expressed recombinant bispecific antibody can be purified as needed.

[0167] Decoy Receptor Peptide- or polypeptide-based agents capable of binding to IL-11 or IL-11-containing complexes may be based on the IL-11 receptor, for example based on an IL-11-binding fragment of the IL-11 receptor.

[0168] In some embodiments, the binding agent may comprise an IL-11-binding fragment of the IL-11Rα chain, preferably soluble, and / or may not comprise one or more transmembrane domains, or may not comprise any transmembrane domain at all. In some embodiments, the binding agent may comprise an IL-11-binding fragment of gp130, preferably soluble, and / or may not comprise one or more transmembrane domains, or may not comprise any transmembrane domain at all. Such molecules may be referred to as decoy receptors. Binding of such agents to IL-11 may reduce / inhibit the ability of IL-11 to bind to an IL-11 receptor (e.g., IL-11Rα or gp130), thereby inhibiting IL-11-mediated cis- and / or trans-signaling, and thus inhibiting downstream signaling.

[0169] Curtis et al. (Blood 1997 Dec 1; 90 (11):4403-12) reported that the soluble murine IL-11 receptor α chain (sIL-11R) was able to antagonize the activity of IL-11 when tested on cells expressing the transmembrane IL-11R and gp130. It is proposed that the antagonism of sIL-11R against IL-11 observed in this study depends on the number of gp130 molecules available on cells already expressing the transmembrane IL-11R.

[0170] The use of soluble decoy receptors for inhibition of signaling and therapeutic intervention has also been reported for other signaling molecule and receptor pairs, e.g., VEGF and VEGF receptors (De-Chao Yu et al., Molecular Therapy (2012); 20 5, 938-947; Konner and Dupont Clin Colorectal Cancer 2004 Oct;4 Suppl 2:S81-5).

[0171] Thus, in some embodiments, the binding agent may be a decoy receptor, for example, a soluble receptor for IL-11 and / or IL-11-containing complexes. It has been reported that the decoy receptor competes for IL-11 and / or IL-11-containing complexes, thereby exerting an antagonistic effect on IL-11 (Curtis et al., supra). The decoy receptor for IL-11 is also described in WO 2017 / 103108(A1) and WO 2018 / 109168(A1), which are incorporated herein by reference in their entirety.

[0172] The decoy receptor for IL-11 can preferably inhibit the binding of IL-11 and / or IL-11-containing complexes to gp130, IL-11Rα and / or gp130:IL-11Rα receptors by binding to IL-11 and / or IL-11-containing complexes. In this way, the decoy receptor for IL-11 acts as a "decoy" receptor for IL-11 and IL-11-containing complexes in a manner very similar to that of etanercept, which acts as a decoy receptor for TNFα. IL-11-mediated signaling is reduced compared to signaling in the absence of the decoy receptor.

[0173] The decoy receptor for IL-11 preferably binds to IL-11 via one or more cytokine binding modules (CBM). The CBM is the CBM of a natural IL-11 receptor molecule, is derived from the CBM of a natural IL-11 receptor molecule, or is homologous to the CBM of a natural IL-11 receptor molecule. For example, the decoy receptor for IL-11 may include one or more CBMs of gp130 and / or IL-11Rα, consist of one or more CBMs of gp130 and / or IL-11Rα, include one or more CBMs derived from the CBM of gp130 and / or IL-11Rα, consist of one or more CBMs derived from the CBM of gp130 and / or IL-11Rα, include one or more CBMs homologous to the CBM of gp130 and / or IL-11Rα, or consist of one or more CBMs homologous to the CBM of gp130 and / or IL-11Rα.

[0174] In some embodiments, the decoy receptor for IL-11 may comprise an amino acid sequence corresponding to the cytokine binding module of gp130, or may consist of an amino acid sequence corresponding to the cytokine binding module of gp130. In some embodiments, the decoy receptor for IL-11 may comprise an amino acid sequence corresponding to the cytokine binding module of IL-11Rα. As used herein, an amino acid sequence "corresponding" to a reference region or sequence of a given peptide / polypeptide has at least 60% sequence identity with the amino acid sequence of this reference region / reference sequence, e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity with the amino acid sequence of this reference region / reference sequence.

[0175] In some embodiments, the decoy receptor may be capable of binding to IL-11 with, for example, a binding affinity of at least 100 μM or less, and may bind to IL-11 with a binding affinity of 10 μM or less, 1 μM or less, 100 nM or less, or about 1 to 100 nM. In some embodiments, the decoy receptor may include the entire IL-11 binding domain or a part thereof, or may lack the entire transmembrane domain or a part thereof. The decoy receptor may be fused to a constant region of an immunoglobulin (e.g., an IgG Fc region).

[0176] Inhibitors The present invention contemplates the use of inhibitor molecules capable of binding to one or more of IL-11, IL-11-containing complexes, IL-11Rα, gp130, or complexes containing IL-11Rα and / or gp130, and inhibiting IL-11-mediated signal transduction.

[0177] In some embodiments, the agent is a peptide- or polypeptide-based binding agent based on IL-11, e.g., a mutant, variant, or binding fragment of IL-11. Suitable peptide- or polypeptide-based agents may inhibit initiation of signal transduction or result in insufficient signal transduction by binding to an IL-11 receptor (e.g., IL-11Rα, gp130, or a complex containing IL-11Rα and / or gp130). These types of IL-11 mutants may act as competitive inhibitors of endogenous IL-11.

[0178] For example, W147A is an IL-11 antagonist in which the so-called "site III" of IL-11 is destroyed by mutating the 147th amino acid from tryptophan to alanine. This mutant can bind to IL-11Rα, but does not associate with gp130 homodimers, resulting in an efficient blockade of IL-11 signal transduction (Underhill-Day et al., 2003; Endocrinology 2003 Aug;144(8):3406-14). Also, Lee et al. (Am J respire Cell Mol Biol. 2008 Dec;39(6):739-746) have reported the generation of IL-11 antagonist mutants ("muteins") that can specifically inhibit the binding of IL-11 to IL-11Rα. IL-11 muteins are also described in WO 2009 / 052588(A1).

[0179] Menkhorst et al. (Biology of Reproduction May 1, 2009 vol.80 no.5 920-927) report a pegylated IL-11 antagonist PEGIL11A (CSL Limited, Parkville, Victoria, Australia) that can effectively suppress the action of IL-11 in female mice.

[0180] In addition, Pasqualini et al. (Cancer (2015) 121(14):2411-2421) reported a ligand-targeted peptidomimetic, bone metastasis-targeting peptidomimetic-11 (BMTP-11), that can bind to IL-11Rα.

[0181] In some embodiments, the binding agent capable of binding to the IL-11 receptor may be provided in the form of a small molecule inhibitor of IL-11Rα, gp130, or a complex containing IL-11Rα and / or gp130. In some embodiments, the binding agent may be provided in the form of a small molecule inhibitor of IL-11 or an IL-11-containing complex, such as an IL-11 inhibitor as described in Lay et al., Int. J. Oncol. (2012); 41(2): 759-764, which is incorporated herein by reference in its entirety.

[0182] Aptamers In some embodiments, the agent capable of binding to IL-11 or IL-11-containing complexes or IL-11 receptors (e.g., IL-11Rα, gp130, or complexes containing IL-11Rα and / or gp130) is an aptamer. Aptamers, also called nucleic acid / peptide ligands, are nucleic acid or peptide molecules that are characterized by their ability to bind to target molecules with high specificity and high affinity. Most of the aptamers identified to date are non-natural molecules.

[0183] Aptamers that bind to a given target (e.g., IL-11, IL-11-containing complexes, or IL-11 receptors) are identified using the Systematic Evolution of Ligands by EXponential enrichment (SELEX) method. TMAptamers may be identified and / or created by the SELEX method, or by constructing SOMAmers (slow off-rate modified aptamers) (Gold L et al. (2010) PLoS ONE 5(12):e15004). Aptamers and the SELEX method have been reported by Tuerk and Gold (Science (1990) 249(4968):505-10) and are also described in WO91 / 19813. In the SELEX method and SOMAmer technology, for example, functional groups that mimic amino acid side chains are added to expand the chemical diversity of aptamers. As a result, aptamers with high affinity for the target can be enriched and identified.

[0184] The aptamer may be a DNA molecule, an RNA molecule, and may be single-stranded or double-stranded. The aptamer may also include chemically modified nucleic acids, for example, nucleic acids with chemical modifications at the sugar, phosphate, and / or base. Such modifications may improve the stability of the aptamer, or may confer resistance to degradation to the aptamer, and may include modifications at the 2' position of the ribose.

[0185] Aptamers may be synthesized by methods well known to those skilled in the art. For example, aptamers may be chemically synthesized, for example, on solid support. Solid-phase synthesis using the phosphoramidite method may be used. Briefly, the immobilized nucleotide is detritylated and then coupled with an appropriately activated nucleoside phosphoramidite to form a phosphite triester bond. Capping can then be performed and the phosphite triester can be oxidized with an oxidizing agent, usually iodine. This cycle can be repeated to construct the aptamer (see, for example, Sinha, ND; Biernat, J.; McManus, J.; Koster, H. Nucleic Acids Res. 1984, 12, 4539; and Beaucage, SL; Lyer, RP (1992). Tetrahedron 48 (12): 2223).

[0186] The lower limit of the length of a suitable nucleic acid aptamer may be any of 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 nucleotides. Preferred upper length limits for nucleic acid aptamers are 20 nucleotides, 21 nucleotides, 22 nucleotides, 23 nucleotides, 24 nucleotides, 25 nucleotides, 26 nucleotides, 27 nucleotides, 28 nucleotides, 29 nucleotides, 30 nucleotides, 31 nucleotides, 32 nucleotides, 33 nucleotides, 34 nucleotides, 35 nucleotides, 36 nucleotides, 37 nucleotides, 38 nucleotides, 39 nucleotides, 40 nucleotides, 41 nucleotides, 42 nucleotides, 43 nucleotides, 44 nucleotides, 45 nucleotides, 46 nucleotides, 47 nucleotides, 48 ​​nucleotides, 49 nucleotides, 50 nucleotides, 51 nucleotides, 52 nucleotides, 53 nucleotides, 54 nucleotides, 55 nucleotides, 56 nucleotides, 57 nucleotides, 58 nucleotides, 59 nucleotides, 60 nucleotides, 61 nucleotides, 62 nucleotides, 63 nucleotides, 64 nucleotides, 65 nucleotides, 66 nucleotides, 67 nucleotides, 68 nucleotides, 69 nucleotides, 70 nucleotides, 71 nucleotides, 72 nucleotides, 73 nucleotides, 74 nucleotides, 75 nucleotides, 76 nucleotides, 77 nucleotides, 78 nucleotides, 79 nucleotides, 80 nucleotides, 81 nucleotides, 82 nucleotides, 83 nucleotides, 84 nucleotides, 85 nucleotides, 86 nucleotides, 87 nucleotides, 88 nucleotides, 89 nucleotides, 90 nucleotides, 9 The length may be any of 0 nucleotides, 51 nucleotides, 52 nucleotides, 53 nucleotides, 54 nucleotides, 55 nucleotides, 56 nucleotides, 57 nucleotides, 58 nucleotides, 59 nucleotides, 60 nucleotides, 61 nucleotides, 62 nucleotides, 63 nucleotides, 64 nucleotides, 65 nucleotides, 66 nucleotides, 67 nucleotides, 68 nucleotides, 69 nucleotides, 70 nucleotides, 71 nucleotides, 72 nucleotides, 73 nucleotides, 74 nucleotides, 75 nucleotides, 76 nucleotides, 77 nucleotides, 78 nucleotides, 79 nucleotides, or 80 nucleotides.Preferred lengths of nucleic acid aptamers are 10 nucleotides, 11 nucleotides, 12 nucleotides, 13 nucleotides, 14 nucleotides, 15 nucleotides, 16 nucleotides, 17 nucleotides, 18 nucleotides, 19 nucleotides, 20 nucleotides, 21 nucleotides, 22 nucleotides, 23 nucleotides, 24 nucleotides, 25 nucleotides, 26 nucleotides, 27 nucleotides, 28 nucleotides, 29 nucleotides, 30 nucleotides, 31 nucleotides, 32 nucleotides, 33 nucleotides, 34 nucleotides, 35 nucleotides, 36 nucleotides, 37 nucleotides, 38 nucleotides, 39 nucleotides, 40 nucleotides, 41 nucleotides, 42 nucleotides, 43 nucleotides, 44 nucleotides, 45 nucleotides, 46 nucleotides, 47 nucleotides, 48 ​​nucleotides, 49 nucleotides, 50 nucleotides, 51 nucleotides, 52 nucleotides, 53 nucleotides, 54 nucleotides, 55 nucleotides, 56 nucleotides, 57 nucleotides, 58 nucleotides, 59 nucleotides, 60 nucleotides, 61 nucleotides, 62 nucleotides, 63 nucleotides, 64 nucleotides, 65 nucleotides, 66 nucleotides, 67 nucleotides, 68 nucleotides, 69 nucleotides, 70 nucleotides, 71 nucleotides, 72 nucleotides, 73 nucleotides, 74 nucleotides, 75 nucleotides, 76 nucleotides, 77 nucleotides, 78 nucleotides, 79 nucleotides, 80 nucleotides, 81 The length may be any of 46 nucleotides, 47 nucleotides, 48 ​​nucleotides, 49 nucleotides, 50 nucleotides, 51 nucleotides, 52 nucleotides, 53 nucleotides, 54 nucleotides, 55 nucleotides, 56 nucleotides, 57 nucleotides, 58 nucleotides, 59 nucleotides, 60 nucleotides, 61 nucleotides, 62 nucleotides, 63 nucleotides, 64 nucleotides, 65 nucleotides, 66 nucleotides, 67 nucleotides, 68 nucleotides, 69 nucleotides, 70 nucleotides, 71 nucleotides, 72 nucleotides, 73 nucleotides, 74 nucleotides, 75 nucleotides, 76 nucleotides, 77 nucleotides, 78 nucleotides, 79 nucleotides, and 80 nucleotides.

[0187] Aptamers can be peptides that are selected or constructed to bind to specific target molecules. Peptide aptamers and their methods of making and identifying are reviewed in Reverdatto et al., Curr Top Med Chem. (2015) 15(12):1082-101, which is incorporated herein by reference in its entirety. The lower limit of the length of peptide aptamers can be 2 amino acids, 3 amino acids, 4 amino acids, 5 amino acids, 6 amino acids, 7 amino acids, 8 amino acids, 9 amino acids, or 10 amino acids. The upper limit of the length of the peptide aptamer may be any of 15 amino acids, 16 amino acids, 17 amino acids, 18 amino acids, 19 amino acids, 20 amino acids, 21 amino acids, 22 amino acids, 23 amino acids, 24 amino acids, 25 amino acids, 26 amino acids, 27 amino acids, 28 amino acids, 29 amino acids, 30 amino acids, 31 amino acids, 32 amino acids, 33 amino acids, 34 amino acids, 35 amino acids, 36 amino acids, 37 amino acids, 38 amino acids, 39 amino acids, 40 amino acids, 41 amino acids, 42 amino acids, 43 amino acids, 44 amino acids, 45 amino acids, 46 amino acids, 47 amino acids, 48 ​​amino acids, 49 amino acids, and 50 amino acids. The length of a peptide aptamer may be any of 2 to 30 amino acids, 2 to 25 amino acids, 2 to 20 amino acids, 5 to 30 amino acids, 5 to 25 amino acids, and 5 to 20 amino acids.

[0188] Aptamers have nM or pM K d and K d may be, for example, less than 500 nM, less than 100 nM, less than 50 nM, less than 10 nM, less than 1 nM, less than 500 pM, or less than 100 pM.

[0189] Properties of IL-11 binding agents An agent of the invention capable of binding to IL-11 or an IL-11 containing complex or to the IL-11 receptor may exhibit any one or more of the following properties. Specific binding to IL-11 or IL-11-containing complexes or the IL-11 receptor K below 10μM D , preferably 5 μM or less, 1 μM or less, 500 nM or less, 100 nM or less, 10 nM or less, 1 nM or less, or 100 pM or less. D Binding to IL-11 or IL-11-containing complexes or IL-11 receptors in ·Inhibition of the interaction between IL-11 and IL-11Rα Inhibition of the interaction between IL-11 and gp130 Inhibition of the interaction between IL-11 and the IL-11Rα:gp130 receptor complex · Inhibition of the interaction between IL-11:IL-11Rα complex and gp130

[0190] These properties can be measured by analyzing the relevant factors in a suitable assay, and optionally by comparing performance to appropriate controls. Those skilled in the art can determine appropriate control conditions for a given assay.

[0191] For example, a suitable negative control for the analysis of the binding ability of a test antibody / antigen-binding fragment to an IL-11 / IL-11-containing complex / IL-11 receptor may be an antibody / antigen-binding fragment against a non-target protein (i.e., an antibody / antigen-binding fragment that is not specific for IL-11 / IL-11-containing complex / IL-11 receptor). A suitable positive control may be a validated (e.g., commercially available) known antibody that binds to IL-11 or a known antibody that binds to the IL-11 receptor. The control may be of the same isotype, e.g., have the same constant region, as the antibody / antigen-binding fragment that is putatively bound to an IL-11 / IL-11-containing complex or IL-11 receptor to be analyzed.

[0192] In some embodiments, the agent may be capable of specifically binding to IL-11, an IL-11-containing complex, or an IL-11 receptor (e.g., IL-11Rα, gp130, or a complex comprising IL-11Rα and / or gp130). An agent that specifically binds to a given target molecule preferably binds to the target molecule with higher affinity and / or longer duration than it binds to other non-target molecules.

[0193] In some embodiments, the agent may bind to IL-11 or an IL-11-containing complex with greater affinity than it binds to one or more of the other members of the IL-6 cytokine family (e.g., IL-6, leukemia inhibitory factor (LIF), oncostatin M (OSM), cardiotrophin-1 (CT-1), ciliary neurotrophic factor (CNTF), and cardiotrophin-like cytokine (CLC)). In some embodiments, the agent may bind to an IL-11 receptor (e.g., IL-11Rα, gp130, or a complex containing IL-11Rα and / or gp130) with greater affinity than it binds to one or more of the other members of the IL-6 receptor family. In some embodiments, the agent may bind to IL-11Rα with greater affinity than it binds to one or more of IL-6Rα, leukemia inhibitory factor receptor (LIFR), oncostatin M receptor (OSMR), ciliary neurotrophic factor receptor alpha (CNTFRα), and cytokine receptor-like factor 1 (CRLF1).

[0194] In some embodiments, the extent of binding of a binding agent to a non-target molecule is less than about 10% of the extent of binding of the binding agent to a target molecule, e.g., as measured by ELISA, SPR, biolayer interferometry (BLI), microscale thermophoresis (MST) or radioimmunoassay (RIA). Alternatively, binding specificity may be reflected as binding affinity, where a binding agent has a K D at least 0.1 orders of magnitude (i.e. 0.1 × 10 n (n is an integer representing the number of digits) Small K DThe antibody binds to IL-11, an IL-11-containing complex, or an IL-11 receptor by an order of magnitude of at least 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.5, or 2.0.

[0195] The binding affinity of a given binding agent to a target is determined by its dissociation constant (K D Binding affinity can be measured by methods known in the art, such as, for example, ELISA, surface plasmon resonance (SPR; see, e.g., Hearty et al., Methods Mol Biol (2012) 907:411-442; or Rich et al., Anal Biochem. 2008 Feb 1; 373(1):112-20), biolayer interferometry (see, e.g., Lad et al., (2015) J Biomol Screen 20(4): 498-507; or Concepcion et al., Comb Chem High Throughput Screen. 2009 Sep; 12(8):791-800), microscale thermophoresis (MST) analysis (see, e.g., Jerabek-Willemsen et al., Assay Drug Dev Technol. 2011 Aug; 9(4): 342-353), or radiolabeled antigen binding assay (RIA).

[0196] In some embodiments, the agent has a K of 50 μM or less. D , preferably 10 μM or less, 5 μM or less, 4 μM or less, 3 μM or less, 2 μM or less, 1 μM or less, 500 nM or less, 100 nM or less, 75 nM or less, 50 nM or less, 40 nM or less, 30 nM or less, 20 nM or less, 15 nM or less, 12.5 nM or less, 10 nM or less, 9 nM or less, 8 nM or less, 7 nM or less, 6 nM or less, 5 nM or less, 4 nM or less, 3 nM or less, 2 nM or less, 1 nM or less, 500 pM or less, 400 pM or less, 300 pM or less, 200 pM or less, or 100 pM or less. Dand capable of binding to IL-11, an IL-11-containing complex, or the IL-11 receptor.

[0197] In some embodiments, the agent has an EC 50 is 10,000ng / ml or less, preferably EC 50 or less than 5,000ng / ml, 1000ng / ml, 900ng / ml or less, 800ng / ml or less, 700ng / ml or less, 600ng / ml or less, 500ng / ml or less, 400ng / ml or less, 300ng / ml or less, 200ng / ml or less, 100ng / ml or less, 90ng / ml or less, 80ng / ml or less, 70ng / ml or less, 60ng / ml or less, 50ng / ml or less, 40ng / ml or less, 30ng / ml or less, 20ng / ml or less, 15ng / ml or less, 10ng / ml or less, 7.5ng / ml or less, 5ng / ml or less, 2.5ng / ml or less, or 1ng / ml or less. ELISA can be performed, for example, as described in Antibody Engineering, vol. 1 (2nd Edn), Springer Protocols, Springer (2010), Part V, pp657-665.

[0198] In some embodiments, the agent binds to IL-11 or an IL-11-containing complex in a region important for binding of the IL-11 receptor or IL-11-containing complex to a receptor (e.g., gp130 or IL-11Rα), thereby inhibiting interaction between IL-11 or an IL-11-containing complex and the IL-11 receptor and / or inhibiting signaling through the IL-11 receptor. In some embodiments, the agent binds to the IL-11 receptor in a region important for binding of the IL-11 or IL-11-containing complex, thereby inhibiting interaction between IL-11 or an IL-11-containing complex and the IL-11 receptor and / or inhibiting signaling through the IL-11 receptor.

[0199] The ability of a given binding agent (e.g., an agent capable of binding to IL-11 or an IL-11-containing complex or an IL-11 receptor) to inhibit the interaction between two proteins can be measured, for example, by analyzing the interaction between these interaction partners in the presence of the binding agent or after incubating the binding agent with one or both of the interaction partners. Suitable assays that can determine whether a given binding agent can inhibit the interaction between two interaction partners include competitive ELISA.

[0200] Binding agents capable of inhibiting a given interaction (e.g., the interaction between IL-11 and IL-11Rα, between IL-11 and gp130, between IL-11 and IL-11Rα:gp130, or between IL-11:IL-11Rα and gp130) are identified by a reduced / diminished degree of interaction between these interaction partners in the presence of the binding agent or after incubation of the binding agent with one or both of the interaction partners, compared to the degree of interaction in the absence of the binding agent (or in the presence of a suitable control binding agent). Suitable assays can be performed in vitro, for example, using recombinant interaction partners or cells expressing the interaction partners. Cells expressing an interaction partner may express the interaction partner endogenously or may express the interaction partner from a nucleic acid introduced into the cell. For purposes of carrying out such assays, one or both of the interaction partners and / or the binding agent may be labeled with a detectable substance or used in conjunction with such a label to detect and / or measure the degree of interaction. For example, the binding agent may be labeled with a radioactive atom, a dye molecule, a fluorescent molecule, or any other molecule that can be easily detected. Suitable detectable molecules include fluorescent proteins, luciferase, enzyme substrates, and radioactive labels. The binding agent may be directly or indirectly labeled with a detectable label. For example, the binding agent may be unlabeled and may be detected using another labeled binding agent. Alternatively, the second binding agent may be bound to biotin, and labeled streptavidin may be bound to the biotin to indirectly label the first binding agent.

[0201] The ability of a binding agent to inhibit an interaction between two binding partners can also be identified by analyzing the downstream functional consequences of such an interaction, such as IL-11-mediated signaling. For example, downstream functional consequences of the interaction between IL-11 and IL-11Rα:gp130 or between IL-11:IL-11Rα and gp130 may include, for example, IL-11-mediated processes, or gene / protein expression of, for example, collagen or IL-11.

[0202] Inhibition of the interaction between IL-11 or IL-11-containing complexes and the IL-11 receptor can be achieved by, for example, inhibiting the IL-11 receptor, as described in Curtis et al. Blood, 1997, 90(11) and Karpovich et al. Mol. Hum. Reprod. 2003 9(2): 75-80. 3 H-thymidine incorporation assays and / or Ba / F3 cell proliferation assays can also be used to analyze. Ba / F3 cells co-express IL-11Rα and gp130.

[0203] In some embodiments, the binding agent may be capable of inhibiting the interaction between IL-11 and IL-11Rα to less than 100%, for example 99% or less, 95% or less, 90% or less, 85% or less, 75% or less, 70% or less, 65% or less, 60% or less, 55% or less, 50% or less, 45% or less, 40% or less, 35% or less, 30% or less, 25% or less, 20% or less, 15% or less, 10% or less, 5% or less or 1% or less compared to the extent of interaction between IL-11 and IL-11Rα in the absence of the binding agent (or in the presence of a suitable control binding agent). In some embodiments, the binding agent may be capable of inhibiting the interaction between IL-11 and IL-11Rα to less than 1-fold, for example 0.99-fold or less, 0.95-fold or less, 0.9-fold or less, 0.85-fold or less, 0.8-fold or less, 0.75-fold or less, 0.7-fold or less, 0.65-fold or less, 0.6-fold or less, 0.55-fold or less, 0.5-fold or less, 0.45-fold or less, 0.4-fold or less, 0.35-fold or less, 0.3-fold or less, 0.25-fold or less, 0.2-fold or less, 0.15-fold or less, or 0.1-fold or less, compared to the extent of interaction between IL-11 and IL-11Rα in the absence of the binding agent (or in the presence of a suitable control binding agent).

[0204] In some embodiments, the binding agent may be capable of inhibiting the interaction between IL-11 and gp130 to less than 100%, for example 99% or less, 95% or less, 90% or less, 85% or less, 75% or less, 70% or less, 65% or less, 60% or less, 55% or less, 50% or less, 45% or less, 40% or less, 35% or less, 30% or less, 25% or less, 20% or less, 15% or less, 10% or less, 5% or less or 1% or less compared to the extent of interaction between IL-11 and gp130 in the absence of the binding agent (or in the presence of a suitable control binding agent). In some embodiments, the binding agent may be capable of inhibiting the interaction between IL-11 and gp130 to less than 1-fold, for example 0.99-fold or less, 0.95-fold or less, 0.9-fold or less, 0.85-fold or less, 0.8-fold or less, 0.75-fold or less, 0.7-fold or less, 0.65-fold or less, 0.6-fold or less, 0.55-fold or less, 0.5-fold or less, 0.45-fold or less, 0.4-fold or less, 0.35-fold or less, 0.3-fold or less, 0.25-fold or less, 0.2-fold or less, 0.15-fold or less, or 0.1-fold or less, compared to the extent of interaction between IL-11 and gp130 in the absence of the binding agent (or in the presence of a suitable control binding agent).

[0205] In some embodiments, the binding agent may be capable of inhibiting the interaction between IL-11 and IL-11Rα:gp130 to less than 100%, for example 99% or less, 95% or less, 90% or less, 85% or less, 75% or less, 70% or less, 65% or less, 60% or less, 55% or less, 50% or less, 45% or less, 40% or less, 35% or less, 30% or less, 25% or less, 20% or less, 15% or less, 10% or less, 5% or less, or 1% or less, compared to the extent of interaction between IL-11 and IL-11Rα:gp130 in the absence of the binding agent (or in the presence of a suitable control binding agent). In some embodiments, the binding agent may be capable of inhibiting the interaction between IL-11 and IL-11Rα:gp130 by less than 1-fold, for example 0.99-fold or less, 0.95-fold or less, 0.9-fold or less, 0.85-fold or less, 0.8-fold or less, 0.75-fold or less, 0.7-fold or less, 0.65-fold or less, 0.6-fold or less, 0.55-fold or less, 0.5-fold or less, 0.45-fold or less, 0.4-fold or less, 0.35-fold or less, 0.3-fold or less, 0.25-fold or less, 0.2-fold or less, 0.15-fold or less, or 0.1-fold or less, compared to the extent of interaction between IL-11 and IL-11Rα:gp130 in the absence of the binding agent (or in the presence of a suitable control binding agent).

[0206] In some embodiments, the binding agent may be capable of inhibiting the interaction between IL-11:IL-11Rα complex and gp130 to less than 100%, for example 99% or less, 95% or less, 90% or less, 85% or less, 75% or less, 70% or less, 65% or less, 60% or less, 55% or less, 50% or less, 45% or less, 40% or less, 35% or less, 30% or less, 25% or less, 20% or less, 15% or less, 10% or less, 5% or less, or 1% or less, compared to the extent of interaction between IL-11:IL-11Rα complex and gp130 in the absence of the binding agent (or in the presence of a suitable control binding agent). In some embodiments, the binding agent is capable of inhibiting the interaction between IL-11:IL-11Rα complex and gp130 to less than 1-fold, e.g., 0.99-fold or less, 0.95-fold or less, 0.9-fold or less, 0.85-fold or less, 0.8-fold or less, 0.75-fold or less, 0.7-fold or less, 0.65-fold or less, 0.6-fold or less, 0.55-fold or less, 0.5-fold or less, 0.45-fold or less, 0.4-fold or less, 0.35-fold or less, 0.3-fold or less, 0.25-fold or less, 0.2-fold or less, 0.15-fold or less, or 0.1-fold or less, compared to the extent of interaction between IL-11:IL-11Rα complex and gp130 in the absence of the binding agent.

[0207] Agents capable of reducing the expression of IL-11 or the IL-11 receptor In an embodiment of the present invention, an agent capable of suppressing IL-11 mediated signal transduction may be an agent capable of blocking or reducing the expression of one or more of IL-11, IL-11Rα and gp130.

[0208] The expression may be gene expression or protein expression and may be measured by methods described herein or by methods in the art well known to those of skill in the art. The expression may be expression by a cell / tissue / organ / organ system in a subject.

[0209] Suitable agents may be of any type, but in some embodiments, agents capable of blocking or reducing expression of one or more of IL-11, IL-11Rα and gp130 may be small molecules or oligonucleotides.

[0210] An agent capable of blocking or reducing expression of one or more of IL-11, IL-11Rα, and gp130 may block or reduce expression of one or more of IL-11, IL-11Rα, and gp130, for example, through inhibiting the transcription of a gene encoding IL-11, IL-11Rα, or gp130, inhibiting post-transcriptional processing of RNA encoding IL-11, IL-11Rα, or gp130, reducing the stability of RNA encoding IL-11, IL-11Rα, or gp130, promoting the degradation of RNA encoding IL-11, IL-11Rα, or gp130, inhibiting post-translational processing of an IL-11 polypeptide, IL-11Rα polypeptide, or gp130 polypeptide, reducing the stability of an IL-11 polypeptide, IL-11Rα polypeptide, or gp130 polypeptide, or promoting the degradation of an IL-11 polypeptide, IL-11Rα polypeptide, or gp130 polypeptide.

[0211] Taki et al. (Clin Exp Immunol (1998) Apr; 112(1): 133-138) reported that IL-11 expression was decreased in rheumatoid synovial cells treated with indomethacin, dexamethasone or interferon gamma (IFNγ).

[0212] The present invention further contemplates the use of antisense nucleic acids to block / reduce expression of IL-11, IL-11Rα or gp130. In some embodiments, agents capable of blocking or reducing expression of IL-11, IL-11Rα or gp130 may reduce said expression by RNA interference (RNAi).

[0213] In some embodiments, the agent may be an inhibitory nucleic acid, such as an antisense RNA or a small interfering RNA, including but not limited to an shRNA or an siRNA.

[0214] In some embodiments, the inhibitory nucleic acid is provided in a vector, for example, in some embodiments, the agent may be a lentiviral vector encoding an shRNA against one or more of IL-11, IL-11Rα, and gp130.

[0215] Oligonucleotide molecules (especially RNA) can be used to regulate gene expression.Such oligonucleotide molecules include antisense oligonucleotides; small interfering RNA (siRNA) targeted degradation of mRNA; post-transcriptional gene silencing (PTG); developmentally regulated sequence specific mRNA translation suppression using microRNA (miRNA); and targeted transcriptional gene silencing.

[0216] Antisense oligonucleotide is an oligonucleotide (preferably a single-stranded oligonucleotide) that targets a target oligonucleotide (e.g., mRNA) and binds to it via complementary sequence binding.When the target oligonucleotide is mRNA, the antisense oligonucleotide binds to the mRNA, thereby inhibiting the translation of the mRNA and inhibiting the expression of gene product.Antisense oligonucleotide may be designed to bind to the sense strand of genomic nucleic acid and suppress the transcription of target nucleotide sequence.

[0217] The nucleic acid sequences of known IL-11, IL-11Rα, or gp130 (e.g., accession numbers: BC012506.1 GI:15341754 (human IL-11), BC134354.1 GI:126632002 (mouse IL-11), AF347935.1 GI:13549072 (rat IL-11), NM_001142784.2 GI:391353394 (human IL-11Rα), NM_001163401.1 GI:254281268 (mouse IL-11Rα), NM_139116.1 GI:20806172 (rat IL-11Rα), NM_001190981.1 Oligonucleotides may be designed to inhibit or silence expression of IL-11, IL-11Rα or gp130, taking into account known mRNA sequences available from GenBank at GI:300244534 (human gp130), NM_010560.3 GI:225007624 (mouse gp130), NM_001008725.3 GI:300244570 (rat gp130).

[0218] Such oligonucleotides may be of any length, but are preferably short, for example less than 100 nucleotides in length, for example 10-40 nucleotides in length, or 20-50 nucleotides in length, and may comprise a nucleotide sequence having perfect complementarity or substantial complementarity (e.g., 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% complementarity) to a nucleotide sequence of corresponding length in a target oligonucleotide (e.g., IL-11 mRNA, IL-11Rα mRNA or gp130 mRNA). The complementary region of the nucleotide sequence may be any length, but is preferably at least 5 nucleotides long, and may be up to 50 nucleotides long, for example, 6 nucleotides long, 7 nucleotides long, 8 nucleotides long, 9 nucleotides long, 10 nucleotides long, 11 nucleotides long, 12 nucleotides long, 13 nucleotides long, 14 nucleotides long, 15 nucleotides long, 16 nucleotides long, 17 nucleotides long, 18 nucleotides long, 19 nucleotides long, 20 nucleotides long, 21 nucleotides long, 22 nucleotides long, 23 nucleotides long, 24 nucleotides long, 25 nucleotides long, or less. The length may be any of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 nucleotides.

[0219] By suppressing the expression of IL-11, IL-11Rα or gp130, the amount of IL-11, IL-11Rα or gp130 expressed by a cell / tissue / organ / organ system / subject is preferably reduced. For example, by suppressing IL-11, IL-11Rα or gp130 in a given cell by administration of a suitable nucleic acid, the amount of IL-11, IL-11Rα or gp130 expressed by the cell is reduced compared to untreated cells. The suppression may be partial. The degree of suppression is preferably at least 50%, more preferably at least 60%, 70%, 80%, 85%, or 90%. When the degree of suppression is 90% to 100%, the expression or function is considered to be "silenced".

[0220] The role of the RNAi machinery and small RNAs in targeting heterochromatin complexes and epigenetic gene silencing of specific chromosomal loci has been demonstrated. Double-stranded RNA (dsRNA)-dependent posttranscriptional silencing, also known as RNA interference (RNAi), is a phenomenon in which dsRNA complexes can target and rapidly silence homologous portions of specific genes. RNAi acts as a signal to promote the degradation of mRNAs with sequence identity. 20-nt siRNAs are typically long enough to induce gene-specific silencing and short enough to avoid host responses. Reduction of expression of target gene products can be achieved by using several siRNA molecules, with silencing rates reaching 90%. RNAi-based therapeutics have progressed to phase I, II, and III clinical trials for a variety of indications (Nature 2009 Jan 22; 457(7228):426-433).

[0221] In the art, such RNA sequences are called "short interfering RNA" or "small interfering RNA (siRNA)" or "microRNA (miRNA)" depending on their origin. These RNA sequences can be used to downregulate gene expression by binding to complementary RNA to induce the elimination of mRNA (RNAi) or by inhibiting the translation of mRNA into protein. siRNAs are obtained by processing long double-stranded RNA, and natural siRNAs are usually exogenous. Microinterfering RNA (miRNA) is an endogenously encoded small non-coding RNA that is obtained by processing short hairpin structures. Both siRNA and miRNA can suppress the translation of mRNA with a partially complementary target sequence without cleaving the RNA, and can degrade mRNA with a completely complementary sequence.

[0222] siRNA ligands are typically double-stranded, and to optimize the effectiveness of this RNA in downregulating the function of a target gene, it is preferable to select the length of the siRNA molecule so that it is long enough to be accurately recognized by the RISC complex that mediates recognition of the target mRNA by the siRNA, yet short enough to minimize the host response.

[0223] miRNA ligands are usually single-stranded and have a partially complementary region capable of forming a hairpin structure. miRNAs are RNA genes that are transcribed from DNA but are not translated into proteins. The DNA sequence encoding the miRNA gene is longer than the miRNA and contains the miRNA sequence and a nearly complementary reverse sequence. When this DNA sequence is transcribed into a single-stranded RNA molecule, a partially double-stranded RNA segment is formed from base pairs consisting of the miRNA sequence and its reverse complementary sequence. The design of microRNA sequences is reported in John et al, PLoS Biology, 11(2), 1862-1879, 2004.

[0224] Said RNA ligands mimicking the effect of siRNA or miRNA are usually 10-40 ribonucleotides long (or synthetic analogues thereof), more preferably 17-30 ribonucleotides long, more preferably 19-25 ribonucleotides long, and most preferably 21-23 ribonucleotides long. In some embodiments of the invention using double-stranded siRNA, the double-stranded siRNA molecule may have a symmetric 3'-end overhang, which may be composed of, for example, one or two (ribo)nucleotides, and is usually a 3'-end UU or dTdT overhang. Based on the disclosure herein, a person skilled in the art can easily design suitable siRNA sequences and suitable miRNA sequences, for example using libraries such as Ambion siRNA finder. siRNA sequences and miRNA sequences can be synthetically produced and added exogenously to induce gene downregulation, or can be produced using an expression system (e.g. vector). In a preferred embodiment, the siRNA is synthetically produced.

[0225] Long double-stranded RNA may be processed intracellularly to produce siRNA (see, for example, Myers (2003) Nature Biotechnology 21:324-328). Long dsRNA molecules may have symmetric 3' or 5' overhangs, which may consist of, for example, one or two (ribo)nucleotides, or they may have blunt ends. Long dsRNA molecules may be 25 nucleotides or more in length. Long dsRNA molecules are preferably 25-30 nucleotides in length. Long dsRNA molecules are more preferably 25-27 nucleotides in length. Long dsRNA molecules are most preferably 27 nucleotides in length. dsRNAs of 30 nucleotides or more in length may be expressed using the pDECAP vector (Shinagawa et al., Genes and Dev., 17, 1340-5, 2003).

[0226] In another method, short hairpin RNA molecules (shRNAs) are expressed in cells. shRNAs are more stable than synthetic siRNAs. shRNAs consist of short inverted repeats linked by a short loop sequence. One inverted repeat is complementary to the target gene. In cells, shRNAs are processed by DICER to become siRNAs, which degrade the mRNA of the target gene to suppress its expression. In a preferred embodiment, shRNAs are produced endogenously (intracellularly) by transcription from a vector. shRNAs may be produced intracellularly by transfecting cells with a vector encoding the shRNA sequence under the control of an RNA polymerase III promoter (such as the human H1 promoter or the human 7SK promoter) or an RNA polymerase II promoter. Alternatively, shRNAs may be synthesized exogenously (in vitro) by transcription from a vector. The resulting shRNA may then be directly introduced into cells. The shRNA molecule preferably contains a partial sequence of IL-11, IL-11Rα or gp130. The length of the shRNA sequence is preferably 40 to 100 base pairs, more preferably 40 to 70 base pairs. The length of the stem portion of the hairpin structure is preferably 19 to 30 base pairs. The stem portion may contain GU pairs to stabilize the hairpin structure.

[0227] The siRNA, long dsRNA or miRNA molecule may be produced by recombinant techniques by transcription of a nucleic acid sequence (preferably incorporated into a vector).The siRNA, long dsRNA or miRNA molecule preferably comprises a partial sequence of IL-11, IL-11Rα or gp130.

[0228] In one embodiment, siRNA, long dsRNA or miRNA is produced endogenously (intracellularly) by transcription from vector. Vector can be introduced into cell by any method known in the art. Expression of these RNA sequences can be regulated using tissue-specific (e.g. heart, liver or kidney specific) promoters as needed. In a further embodiment, siRNA, long dsRNA or miRNA is produced exogenously (in vitro) by transcription from vector.

[0229] A suitable vector may be an oligonucleotide vector configured to express an oligonucleotide drug capable of suppressing IL-11, IL-11Rα or gp130. Such vectors may be viral or plasmid vectors. The oligonucleotide therapeutic may be incorporated into the genome of the viral vector and may be operably linked to a regulatory sequence (e.g., a promoter) that induces expression. "Operably linked" may include a situation in which a selected nucleotide sequence and a regulatory nucleotide sequence are covalently linked such that the nucleotide sequence is expressed under the influence or control of the regulatory sequence. Thus, a regulatory sequence is operably linked to a selected nucleotide sequence if the regulatory sequence is capable of inducing transcription of a nucleotide sequence that constitutes the entirety or a portion of the selected nucleotide sequence.

[0230] Viral vectors encoding siRNA sequences whose expression is induced by promoters are known in the art and have the advantage of being able to express oligonucleotide therapeutics for a long period of time.Viral vectors include lentiviruses (Nature 2009 Jan 22; 457(7228):426-433), adenoviruses (Shen et al., FEBS Lett 2003 Mar 27;539(1-3)111-4) and retroviruses (Barton and Medzhitov PNAS November 12, 2002 vol.99, no.23 14943-14945).

[0231] In another embodiment, carriers configured to aid in the delivery of oligonucleotide therapeutics to the site where inhibition of IL-11, IL-11Rα or gp130 expression is required may be used. Such carriers generally include positively charged carriers complexed with oligonucleotides (e.g., cell-permeable cationic peptides, cationic polymers, cationic dendrimers, and cationic lipids); small molecules (e.g., cholesterol, bile acids, and lipids), polymers, antibodies, and RNAs attached to oligonucleotides; or oligonucleotides encapsulated in nanoparticle formulations (Wang et al., AAPS J. 2010 Dec; 12(4): 492-503).

[0232] In one embodiment, the vector may contain nucleic acid sequences in both sense and antisense orientations, such that when the nucleic acid sequences are expressed as RNA, the sense and antisense strand portions associate to form double-stranded RNA.

[0233] Alternatively, siRNA molecules may be synthesized using standard solid-phase or solution-phase synthesis methods known in the art. The linkage between the nucleotides may be a phosphodiester bond or other bond, such as P(O)S (thioate); P(S)S (dithioate); P(O)NR'2; P(O)R'; P(O)OR6; CO; or CONR'2 (wherein R is H (or a salt) or alkyl (C 1~12 ) and R6 is alkyl (C 1~9 ) is linked to the adjacent nucleotide via -O- or -S-.

[0234] In addition to naturally occurring bases, modified nucleotide bases can be used and may impart advantageous properties to the siRNA molecules containing them.

[0235] For example, modified bases may improve the stability of siRNA molecules, thereby reducing the amount of siRNA molecules required for silencing. By adding modified bases, siRNA molecules may be made more or less stable than unmodified siRNAs.

[0236] "Modified nucleotide base" includes nucleotides with modified bases and / or modified sugars covalently attached. For example, modified nucleotides include nucleotides with sugars covalently attached to low molecular weight organic groups other than 3' hydroxyl group and 5' phosphate group. Thus, modified nucleotides further include 2'-substituted sugars such as 2'-O-methyl ribose, 2'-O-alkyl ribose, 2'-O-allyl ribose, 2'-S-alkyl ribose, 2'-S-allyl ribose, 2'-fluoro ribose, 2'-haloribose, 2'-azido ribose; carbocyclic sugar analogs; α-anomeric sugars; epimeric sugars such as arabinose, xylose, lyxose; pyranose sugars, furanose sugars, and sedoheptulose.

[0237] Modified nucleotides are known in the art and include, for example, alkylated purines, alkylated pyrimidines, acylated purines, acylated pyrimidines, and other heterocycles. Such classes of pyrimidines and purines are known in the art and include, for example, pseudoisocytosine, N 4 ,N 4 -Ethanocytosine, 8-hydroxy-N 6 -methyladenine, 4-acetylcytosine, 5-(carboxyhydroxymethyl)uracil, 5-fluorouracil, 5-bromouracil, 5-carboxymethylaminomethyl-2-thiouracil, 5-carboxymethylaminomethyluracil, dihydrouracil, inosine, N 6 -Isopentyl adenine, 1-methyl adenine, 1-methyl pseudouracil, 1-methyl guanine, 2,2-dimethyl guanine, 2-methyl adenine, 2-methyl guanine, 3-methyl cytosine, 5-methyl cytosine, N 6 -Methyladenine, 7-methylguanine, 5-methylaminomethyluracil, 5-methoxyaminomethyl-2-thiouracil, -D-mannosylketosine, 5-methoxycarbonylmethyluracil, 5-methoxyuracil, 2-methylthio-N 6 -isopentenyladenine, uracil-5-oxyacetic acid methyl ester, pseudouracil, 2-thiocytosine, 5-methyl-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil, N-uracil-5-oxyacetic acid methyl ester, uracil-5-oxyacetic acid, keuosine, 2-thiocytosine, 5-propyluracil, 5-propylcytosine, 5-ethyluracil, 5-ethylcytosine, 5-butyluracil, 5-pentyluracil, 5-pentylcytosine, 2,6-diaminopurine, methylpseudouracil, 1-methylguanine and 1-methylcytosine.

[0238] Methods for silencing genes in C. elegans, Drosophila, plants and mammals using RNAi are known in the art (Fire A, et al., 1998 Nature 391:806-811; Fire, A. Trends Genet. 15, 358-363 (1999); Sharp, PA RNA interference 2001. Genes Dev. 15, 485-490 (2001); Hammond, SM, et al., Nature Rev. Genet. 2, 110-1119 (2001); Tuschl, T. Chem. Biochem. 2, 239-245 (2001); Hamilton, A. et al., Science 286, 950-952 (1999); Hammond, SM, et al., Nature 404, 293-296 (2000); Zamore, PD, et al., Cell 101, 25-33 (2000); Bernstein, E., et al., Nature 409, 363-366 (2001); Elbashir, SM, et al., Genes Dev. 15, 188-200 (2001); WO0129058; WO9932619 and Elbashir SM, et al., 2001 Nature 411:494-498).

[0239] Thus, the present invention provides nucleic acids that, when appropriately introduced or expressed in a mammalian cell (e.g., a human cell) that expresses IL-11, IL-11Rα or gp130, are capable of suppressing expression of IL-11, IL-11Rα or gp130 by RNAi techniques.

[0240] IL-11, IL-11Rα or gp130 (e.g., accession numbers: BC012506.1 GI:15341754 (human IL-11), BC134354.1 GI:126632002 (mouse IL-11), AF347935.1 GI:13549072 (rat IL-11), NM_001142784.2 GI:391353394 (human IL-11Rα), NM_001163401.1 GI:254281268 (mouse IL-11Rα), NM_139116.1 GI:20806172 (rat IL-11Rα), NM_001190981.1 Nucleic acid sequences of oligonucleotides to known mRNA sequences available from GenBank at GI:300244534 (human gp130), NM_010560.3 GI:225007624 (mouse gp130), NM_001008725.3 GI:300244570 (rat gp130) may be designed to inhibit or silence expression of IL-11, IL-11Rα or gp130.

[0241] The nucleic acid may have substantial sequence identity to a portion of the mRNA of IL-11, IL-11Rα or gp130, for example, to a portion of the sequence set forth in GenBank Accession No. NM_000641.3 GI:391353405 (IL-11), NM_001142784.2 GI:391353394 (IL-11Rα) or NM_001190981.1 GI:300244534 (gp130), or a sequence complementary to these mRNAs.

[0242] The nucleic acid may be a double-stranded siRNA. (As will be appreciated by those skilled in the art, the siRNA molecule may further comprise a short DNA sequence at the 3' end, which is described in more detail below.)

[0243] Alternatively, the nucleic acid may be DNA (usually double-stranded DNA), which, when transcribed in a mammalian cell, results in an RNA having two complementary portions linked via a spacer, which forms a hairpin structure when the two complementary portions hybridize with each other. In mammalian cells, the hairpin structure portion can be cleaved from the RNA molecule by an enzyme called DICER to obtain a double-stranded RNA in which two different RNA molecules are hybridized.

[0244] In some preferred embodiments, the nucleic acid typically targets a sequence set forth in any of SEQ ID NOs: 4-7 (IL-11) or any of SEQ ID NOs: 8-11 (IL-11Rα).

[0245] Only single-stranded regions of the mRNA transcript (i.e., regions that are not self-hybridized) are expected to be suitable targets for RNAi. Thus, other sequences of the IL-11 or IL-11Rα mRNA transcripts that are highly similar to the sequences set forth by any of SEQ ID NOs: 4-7 and 8-11 are also expected to be suitable targets for RNAi. The length of such target sequences is preferably 17-23 nucleotides long, and preferably overlaps (at one end) at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17 or 18 nucleotides or all 19 nucleotides with any of SEQ ID NOs: 4-7 and 8-11.

[0246] Thus, the present invention provides nucleic acids that, when appropriately introduced or expressed in a mammalian cell expressing IL-11 or IL-11Rα, can suppress the expression of IL-11 or IL-11Rα by RNAi techniques, and these nucleic acids typically target any of the sequences shown in SEQ ID NOs: 4 to 7 and 8 to 11.

[0247] "Regular targeting" refers to the fact that the nucleic acid may target a sequence that overlaps with any of SEQ ID NOs: 4 to 7 and 8 to 11. Specifically, the nucleic acid may target a human IL-11 mRNA sequence or a human IL-11Rα mRNA sequence (preferably 17 to 23 nucleotides in length) that is identical to any of SEQ ID NOs: 4 to 7 and 8 to 11, except that the sequence is slightly longer or slightly shorter than these sequences.

[0248] It is expected that there is a perfect identity / complementarity between the nucleic acid of the present invention and the target sequence, but this is not essential.Thus, the nucleic acid of the present invention may contain a single base mismatch compared to IL-11 mRNA or IL-11Rα mRNA.However, it is preferred that there is no mismatch, since the presence of even a single base mismatch is expected to reduce the efficiency.If there is a 3'-end overhang, the 3'-end overhang may not be taken into account in the number of mismatches.

[0249] "Complementarity" is not limited to base pairing between nucleic acids composed of natural ribonucleotides and / or deoxyribonucleotides, as is commonly seen, but also includes base pairing between the nucleic acids of the present invention and mRNAs containing non-natural nucleotides.

[0250] In one embodiment, the nucleic acid (herein referred to as double-stranded siRNA) comprises the double-stranded RNA sequences shown in SEQ ID NOs: 12 to 15. In another embodiment, the nucleic acid (herein referred to as double-stranded siRNA) comprises the double-stranded RNA sequences shown in SEQ ID NOs: 16 to 19.

[0251] However, slightly shorter or longer sequences targeting the same IL-11 or IL-11Rα mRNA regions are also expected to be effective, specifically double-stranded sequences between 17 and 23 bp in length.

[0252] Each strand constituting the double-stranded RNA may have a short 3'-end overhang of 2 bases, which may be DNA or RNA. A 3'-end DNA overhang has no effect on siRNA activity compared to the use of a 3'-end RNA overhang, but the cost of chemically synthesizing the nucleic acid strand is lower (Elbashir et al., 2001c). For this reason, a 2-base DNA may be preferred.

[0253] If two-base overhangs are present at both 3' ends, these overhangs may be symmetrical to each other, but this is not essential: in fact, the 3' overhang of the sense strand (upper strand) is not relevant for RNAi activity, since it is not involved in mRNA recognition and degradation (Elbashir et al., 2001a, 2001b, 2001c).

[0254] Although RNAi experiments in Drosophila have suggested that the 3' overhang of the antisense strand may be involved in mRNA recognition and targeting (Elbashir et al., 2001c), 3' overhangs are not thought to be required for the RNAi activity of siRNAs in mammalian cells, and therefore misannealing of 3' overhangs is unlikely to have any effect in mammalian cells (Elbashir et al., 2001c; Czauderna et al., 2003).

[0255] Therefore, any two-base overhang may be used in the antisense strand of siRNA. However, the two-base overhang is preferably -UU or -UG (-TT or -TG if the overhang is DNA), and more preferably -UU (or -TT). The two-base overhang consisting of -UU (or -TT) is the most effective and coincides with (i.e. can form part of) the transcription termination signal of RNA polymerase III (the transcription termination signal is TTTTT). Therefore, this two base is most preferred. The two bases AA, CC and GG may be used, but are less effective and therefore less preferred.

[0256] Additionally, the siRNA may not contain any 3' overhangs.

[0257] Furthermore, the present invention provides a single-stranded nucleic acid (referred to as single-stranded siRNA in this specification) that is composed of one of the double-stranded nucleic acids, and this single-stranded siRNA preferably has a 3'-end overhang, but may not have a 3'-end overhang. Furthermore, the present invention provides a kit that includes a pair of such single-stranded nucleic acids, and these single-stranded nucleic acids can hybridize with each other in vitro to form the double-stranded siRNA, and this double-stranded siRNA can then be introduced into a cell.

[0258] Furthermore, the present invention provides DNA that is transcribed in mammalian cells into RNA (also referred to herein as shRNA) in which two complementary portions can self-hybridize to form a double-stranded motif, such as a sequence selected from the group consisting of SEQ ID NOs: 12 to 15 and 16 to 19, or a sequence in which a single base pair has been substituted in any of these sequences.

[0259] The complementary portions are usually linked by a spacer, which has an appropriate length and sequence to allow the two complementary portions to hybridize with each other. The two complementary portions (i.e., the sense strand and the antisense strand) may be linked at the 5' end and the 3' end, or either may be at the 5' end. The spacer may be a short sequence, usually about 4 to 12 nucleotides long, preferably 4 to 9 nucleotides long, more preferably 6 to 9 nucleotides long.

[0260] The 5' end of the spacer (immediately following the 3' end of the upstream complementary portion) preferably consists of the two bases -UU- or -UG-, with -UU- being more preferred (but again, the use of these particular two bases is not essential). A suitable spacer recommended for use with the pSuper system from OligoEngine Inc. (Seattle, Washington, USA) is UUCAAGAGA. When this or other spacers are used, both ends of the spacer hybridize to each other, resulting in a double-stranded motif that is a few base pairs (e.g., one or two base pairs) longer than the exact sequence shown in, for example, SEQ ID NOs: 12-15 or 16-19.

[0261] Similarly, the transcribed RNA preferably comprises a 3' overhang from the downstream complementary portion, again preferably -UU or -UG, more preferably -UU.

[0262] As described above, such shRNA molecules may be cleaved by the DICER enzyme in mammalian cells to form double-stranded siRNAs in which one or both of the single strands constituting the hybridized dsRNA contain a 3'-end overhang.

[0263] It goes without saying that techniques for synthesizing the nucleic acids of the present invention are well known in the art.

[0264] Those skilled in the art can easily construct a transcription vector suitable for the DNA of the present invention using well-known techniques and commercially available materials. Specifically, the DNA of the present invention is ligated to a control sequence such as a promoter and a transcription termination sequence.

[0265] Particularly suitable are the commercially available pSuper and pSuperior systems from OligoEngine (Seattle, WA, USA), which use a polymerase III promoter (H1) and a T5 transcription termination sequence that adds two U residues to the 3' end of the transcript (which is processed by DICER to produce an siRNA with a 3' UU overhang on one RNA strand).

[0266] Another suitable system is described in Shin et al. (RNA, 2009 May; 15(5): 898-910), which uses another polymerase III promoter (U6).

[0267] The double-stranded siRNA of the present invention may be introduced into mammalian cells in vitro or in vivo using known techniques such as those described below to suppress the expression of IL-11 or IL-11 receptor.

[0268] Similarly, a transcription vector containing the DNA of the present invention may be introduced into tumor cells in vitro or in vivo using known techniques such as those described below, to suppress expression of IL-11 or IL-11 receptor by transient or stable expression of RNA.

[0269] Thus, the present invention further provides a method for suppressing expression of IL-11 or IL-11 receptor in mammalian (e.g., human) cells, comprising administering to the cells a double-stranded siRNA of the present invention or a transcription vector of the present invention.

[0270] Similarly, the present invention further provides a method for treating renal damage and / or a disorder, disease or condition associated with renal damage, comprising administering to a subject a double-stranded siRNA of the present invention or a transcription vector of the present invention.

[0271] Furthermore, the present invention provides the double-stranded siRNA of the present invention and the transcription vector of the present invention for use in a method of treatment, preferably a method of treating renal damage and / or a disorder, disease or condition associated with renal damage.

[0272] Furthermore, the present invention provides the use of the double-stranded siRNA of the present invention and the transcription vector of the present invention in the preparation of a medicament for the treatment of renal damage and / or a disorder, disease or condition associated with renal damage.

[0273] Furthermore, the present invention provides a composition comprising the mixture of the double-stranded siRNA of the present invention or the transcription vector of the present invention and one or more pharma- ceutically acceptable carriers.Suitable carriers include lipophilic carriers or vesicles that can improve cell membrane permeability.

[0274] Materials and methods suitable for administration of the double-stranded siRNA and DNA vectors of the present invention are well known in the art, and because of the many potential applications of RNAi technology, improved methods are under development.

[0275] A variety of techniques are generally available for introducing nucleic acids into mammalian cells. The technique used depends on whether the nucleic acid is to be introduced into cultured cells in vitro or into the patient's cells in vivo. Suitable techniques for introducing nucleic acids into mammalian cells in vitro include the use of liposomes, electroporation, microinjection, cell fusion, DEAE-dextran and calcium phosphate precipitation. In vivo gene transfer techniques include transfection using viral vectors (usually retroviral vectors) and transfection using viral coat protein-liposome complexes (Dzau et al. (2003) Trends in Biotechnology 11, 205-210).

[0276] Specifically, suitable techniques for administering the nucleic acid of the present invention to cells in vitro or in vivo are described in the following documents:

[0277] Review: Borkhardt, A. 2002. Blocking oncogenes in malignant cells by RNA interference--new hope for a highly specific cancer treatment? Cancer Cell. 2:167-8. Hannon, G.J. 2002. RNA interference. Nature. 418:244-51. McManus, M.T., and P.A. Sharp. 2002. Gene silencing in mammals by small interfering RNAs. Nat Rev Genet. 3:737-47. Scherr, M., M.A. Morgan, and M. Eder. 2003b. Gene silencing mediated by small interfering RNAs in mammalian cells. Curr Med Chem. 10:245-56. Shuey, D.J., D.E. McCallus, and T. Giordano. 2002. RNAi: gene-silencing in therapeutic intervention. Drug Discov Today. 7:1040-6.

[0278] Editor-in-Chief: Lewis, DL, JE Hagstrom, AG Loomis, JA Wolff, and H. Herweijer. 2002. Efficient delivery of siRNA for inhibition of gene expression in postnatal mice. Night Genet. 32:107-8 Paul, CP, Good PD, Winer I, and English DR. 2002. Effective expression of small interfering RNA in human cells. Nat Biotechnol. 20:505-8 Song , E. , SK Lee , J. Wang , N. Ince , N. Ouyang , J. Min , J. Chen , P. Shankar , and J. Lieberman . 2003. RNA interference targeting Fas protects mice from fulminant hepatitis. Nat Med. 9:347-5 Sorensen , DR , Leirdal M , and M Sioud . 2003. Gene silencing by systemic delivery of synthetic siRNAs in adult mice. J Mol Biol. 327:761-6

[0279] Abbas-Terki, T., W. Blanco-Bose, N. Deglon, W. Pralong, and P. Aebischer. 2002. Lentiviral-mediated RNA interference. Hum Gene Ther. 13:2197–2 Barton , GM , and R. Medzhitov . 2002. Retroviral delivery of small interfering RNA into primary cells. Proc Natl Acad Sci US A. 99:14943–5. Devroe , E. , and PA Silver . 2002. Retrovirus-delivered siRNA. BMC Biotechnol. 2:1 Lori , F. , P. Guallini , L. Galluzzi , and J. Lisziewicz . 2002. Gene therapy approaches to HIV infection. Am J Pharmacogenomics. 2:245-5 Matta , H. , B. Hozayev , R. Tomar , P. Chugh , and PM Chaudhary . 2003. Use of lentiviral vectors for delivery of small interfering RNA. Cancer Biol Ther. 2:206-1 Qin , XF , DS An , IS Chen , and D Baltimore . 2003. Inhibiting HIV-1 infection in human T cells by lentiviral-mediated delivery of small interfering RNA against CCR5. Proc Natl Acad Sci US A. 100:183–8. Scherr , M. , K. Battmer , A. Ganser , and M. Eder . 2003a.Modulation of gene expression by lentiviral-mediated delivery of small interfering RNA. Cell Cycle. 2:251-7. Shen, C., AK Buck, X. Liu, M. Winkler, and SN Reske. 2003. Gene silencing by adenovirus-delivered siRNA. FEBS Lett. 539:111-4.

[0280] Peptide delivery: Morris, MC, L. Chaloin, F. Heitz, and G. Divita. 2000. Translocating peptides and proteins and their use for gene delivery. Curr Opin Biotechnol. 11:461-6. Simeoni, F., MC Morris, F. Heitz, and G. Divita. 2003. Insight into the mechanism of the peptide-based gene delivery system MPG: implications for delivery of siRNA into mammalian cells. Nucleic Acids Res. 31:2717-24. Other techniques that may be suitable for delivering siRNA to target cells include the use of nanoparticles or nanocapsules, such as those described in U.S. Pat. No. 6,649,192(B) and U.S. Pat. No. 5,843,509(B).

[0281] Inhibition of IL-11-mediated signaling In embodiments of the invention, an agent capable of suppressing the action of IL-11 may have one or more of the following functional properties. Inhibition of IL-11-mediated signaling Inhibition of signal transduction via binding of IL-11 to the IL-11Rα:gp130 receptor complex Inhibition of signal transduction via the binding of the IL-11:IL-11Rα complex to gp130 (i.e., trans-signaling of IL-11) Inhibition of IL-11-mediated processes Inhibition of IL-11 and / or IL-11Rα gene / protein expression

[0282] These properties can be measured by assaying the relevant factors in a suitable assay, and may include comparing the performance of the agent to that of an appropriate control. Those skilled in the art can determine appropriate control conditions for a given assay.

[0283] IL-11-mediated signal transduction and / or IL-11-mediated process include signal transduction through IL-11 fragments and signal transduction through polypeptide complexes containing IL-11 or fragments thereof. IL-11-mediated signal transduction may be signal transduction through human IL-11 and / or mouse IL-11. IL-11-mediated signal transduction may be signal transduction caused by binding of IL-11 or IL-11-containing complexes to a receptor to which IL-11 or IL-11-containing complexes bind.

[0284] In some embodiments, an agent of the invention may be capable of inhibiting the biological activity of IL-11 or an IL-11-containing complex.

[0285] In some embodiments, the agents of the invention are antagonists to one or more signaling pathways activated by signaling through receptors comprising IL-11Rα and / or gp130 (e.g., IL-11Rα:gp130). In some embodiments, the agents of the invention can inhibit signaling through one or more immune receptor complexes comprising IL-11Rα and / or gp130 (e.g., IL-11Rα:gp130). In various aspects of the invention, the agents provided herein can inhibit IL-11-mediated cis-signaling and / or trans-signaling. In some embodiments according to various aspects of the invention, the agents provided herein can inhibit IL-11-mediated cis-signaling.

[0286] In some embodiments, an agent of the invention may be capable of inhibiting IL-11-mediated signaling to less than 100%, for example 99% or less, 95% or less, 90% or less, 85% or less, 80% or less, 75% or less, 70% or less, 65% or less, 60% or less, 55% or less, 50% or less, 45% or less, 40% or less, 35% or less, 30% or less, 25% or less, 20% or less, 15% or less, 10% or less, 5% or less, or 1% or less, compared to the amount of IL-11-mediated signaling in the absence of the agent (or in the presence of a suitable control agent). In some embodiments, an agent of the invention can reduce IL-11 mediated signaling to less than 1-fold, for example 0.99-fold or less, 0.95-fold or less, 0.9-fold or less, 0.85-fold or less, 0.8-fold or less, 0.75-fold or less, 0.7-fold or less, 0.65-fold or less, 0.6-fold or less, 0.55-fold or less, 0.5-fold or less, 0.45-fold or less, 0.4-fold or less, 0.35-fold or less, 0.3-fold or less, 0.25-fold or less, 0.2-fold or less, 0.15-fold or less, or 0.1-fold or less, compared to the amount of IL-11 mediated signaling in the absence of the agent (or in the presence of a suitable control agent).

[0287] In some embodiments, IL-11-mediated signaling may be signaling through binding of IL-11 to the IL-11Rα:gp130 receptor. Such signaling can be analyzed, for example, by treating cells expressing IL-11Rα and gp130 with IL-11 or stimulating the production of IL-11 in cells expressing IL-11Rα and gp130.

[0288] IC of inhibition of IL-11-mediated signaling by agents of the invention 50 For example, Ba / F3 cells expressing IL-11Rα and gp130 are cultured in the presence of human IL-11 and the agent of the present invention, and the binding to DNA is 3 In some embodiments, the IC of an agent of the invention in such an assay may be measured by measuring the incorporation of H-thymidine. 50 The value may be 10 μg / ml or less, preferably 5 μg / ml or less, 4 μg / ml or less, 3.5 μg / ml or less, 3 μg / ml or less, 2 μg / ml or less, 1 μg / ml or less, 0.9 μg / ml or less, 0.8 μg / ml or less, 0.7 μg / ml or less, 0.6 μg / ml or less or 0.5 μg / ml or less.

[0289] In some embodiments, IL-11-mediated signaling may be signaling via binding of the IL-11:IL-11Rα complex to gp130. In some embodiments, the IL-11:IL-11Rα complex may be soluble, e.g., a complex of the extracellular domain of IL-11Rα and IL-11, or a complex of a soluble IL-11Rα isoform / fragment and IL-11. In some embodiments, the soluble IL-11Rα is a soluble (secreted) isoform of IL-11Rα or a product liberated by proteolysis of the extracellular domain of membrane-bound IL-11Rα.

[0290] In some embodiments, the IL-11:IL-11Rα complex may be membrane-bound, e.g., a complex consisting of membrane-bound IL-11Rα and IL-11. Signal transduction via binding of the IL-11:IL-11Rα complex to gp130 can be analyzed by treating gp130-expressing cells with the IL-11:IL-11Rα complex, e.g., by treating gp130-expressing cells with a recombinant fusion protein (e.g., hyper IL-11) that includes IL-11 linked to the extracellular domain of IL-11Rα via a peptide linker. Hyper IL-11 was constructed using a fragment of IL-11Rα (amino acid residues 1-317 constituting domains 1-3; UniProtKB: Q14626), IL-11 (amino acid residues 22-199 of UniProtKB: P20809), and a 20 amino acid long linker (SEQ ID NO: 20). The amino acid sequence of hyper IL-11 is shown in SEQ ID NO:21.

[0291] In some embodiments, the agents of the invention may be capable of inhibiting signaling mediated by binding of the IL-11:IL-11Rα complex to gp130 and may also inhibit signaling mediated by binding of IL-11 to the IL-11Rα:gp130 receptor.

[0292] In some embodiments, agents of the invention may be capable of inhibiting IL-11 mediated processes.

[0293] In some embodiments, the agent may be capable of suppressing IL-11 and / or IL-11Rα gene / protein expression. Gene and / or protein expression can be measured by methods described herein or methods well known to those of skill in the art.

[0294] In some embodiments, the agent may be capable of suppressing expression of IL-11 and / or IL-11Rα genes / proteins to less than 100%, for example, 99% or less, 95% or less, 90% or less, 85% or less, 80% or less, 75% or less, 70% or less, 65% or less, 60% or less, 55% or less, 50% or less, 45% or less, 40% or less, 35% or less, 30% or less, 25% or less, 20% or less, 15% or less, 10% or less, 5% or less, or 1% or less, compared to the expression level of IL-11 and / or IL-11Rα genes / proteins in the absence of the agent (or in the presence of an appropriate control agent). In some embodiments, the agent is capable of suppressing expression of the IL-11 and / or IL-11Rα gene / protein to less than 1-fold, for example, 0.99-fold or less, 0.95-fold or less, 0.9-fold or less, 0.85-fold or less, 0.8-fold or less, 0.75-fold or less, 0.7-fold or less, 0.65-fold or less, 0.6-fold or less, 0.55-fold or less, 0.5-fold or less, 0.45-fold or less, 0.4-fold or less, 0.35-fold or less, 0.3-fold or less, 0.25-fold or less, 0.2-fold or less, 0.15-fold or less, or 0.1-fold or less, compared to the expression level of the IL-11 and / or IL-11Rα gene / protein in the absence of the agent (or in the presence of an appropriate control agent).

[0295] Treatment / Prevention of Kidney Damage The present invention provides methods and inventions (medicaments and compositions) for the treatment / prevention of renal damage and disorders, diseases or conditions associated with renal damage, eg, as described herein.

[0296] The treatment is achieved by suppressing IL-11-mediated signaling (i.e., antagonism against IL-11-mediated signaling). That is, the present invention treats / prevents renal damage and disorders, diseases and conditions associated with renal damage through suppression of IL-11-mediated signaling in, for example, cells, tissues / organs / organ systems, subjects, etc. In some embodiments, suppression of IL-11-mediated signaling according to the present disclosure includes suppressing IL-11-mediated signaling in renal cells (e.g., tubular epithelial cells).

[0297] Kind Code: A1 Abstract: Provided are agents capable of inhibiting interleukin-11 (IL-11) mediated signaling for use in methods for the treatment or prevention of renal damage and / or disorders, diseases or conditions associated with renal damage.

[0298] Also provided is the use of an agent capable of inhibiting interleukin-11 (IL-11)-mediated signaling in the manufacture of a medicament for use in a method for the treatment or prevention of renal damage and / or disorders, diseases or conditions associated with renal damage.

[0299] Further provided is a method for treating or preventing renal damage and / or disorders, diseases or conditions associated with renal damage, comprising administering to a subject in need of treatment a therapeutically effective amount of an agent capable of inhibiting interleukin-11 (IL-11)-mediated signal transduction.

[0300] In some embodiments, the present invention treats / prevents the pathology associated with renal injury in disease / condition. That is, the present invention treats / prevents the disease / condition in which renal injury is pathologically involved. The pathology associated with renal injury is described herein. More specifically, the pathology associated with renal injury includes the damage of renal tubule epithelial cells (proximal tubule epithelial cells or distal tubule epithelial cells or both).

[0301] Furthermore, it has been demonstrated herein that agents capable of suppressing interleukin 11 (IL-11)-mediated signal transduction can reverse renal injury, i.e., it has been shown that suppressing IL-11-mediated signal transduction can improve renal function after the onset of renal injury.

[0302] Thus, the present invention contemplates enhancing / improving renal function in subjects who have developed renal dysfunction due to, for example, renal damage, by using antagonists against IL-11-mediated signaling.

[0303] Agents capable of suppressing interleukin-11 (IL-11)-mediated signal transduction are useful for promoting the proliferation of renal tubular epithelial cells (TECs) to form functional renal tissue. Thus, agents capable of suppressing interleukin-11 (IL-11)-mediated signal transduction are provided herein for the purpose of promoting the proliferation, survival and / or function of renal tubular epithelial cells (TECs) and / or the proliferation, maintenance and / or function of renal tissues.

[0304] Provided herein are agents capable of suppressing interleukin-11 (IL-11)-mediated signaling for the purpose of regenerating renal tubular epithelial cells (TECs) and / or renal tissue.

[0305] The transition of tubular epithelial cells from epithelial and / or acta to mesenchymal cell phenotype (also referred to herein as "epithelial-mesenchymal transition (EMT)") is associated with impaired renal function. Epithelial-mesenchymal transition of tubular epithelial cells is induced by soluble factors (such as TGFB1) that are produced in association with tissue injury. It has been shown herein that antagonism against IL-11-mediated signaling maintains / improves renal function by suppressing epithelial-mesenchymal transition of tubular epithelial cells.

[0306] In addition, the expression of SNAIL is involved in the loss of proliferation ability and epithelial-mesenchymal transition (EMT) of renal tubular epithelial cells. It has been demonstrated herein that IL-11-mediated signaling plays a central role in the upregulation of SNAIL expression. Therefore, the present invention envisions the use of an antagonist against IL-11-mediated signaling to suppress the expression of SNAIL, for example, in renal tubular epithelial cells. The antagonism against IL-11-mediated signaling can release the inhibition of renal tubular epithelial cell proliferation mediated by SNAIL.

[0307] Thus, the present invention contemplates the use of antagonists against IL-11-mediated signaling to maintain / increase the number / proportion of kidney cells with a tubular epithelial cell phenotype. The tubular epithelial cell phenotype may be characterized, for example, by the expression of E-cadherin. The present invention also contemplates the use of antagonists against IL-11-mediated signaling to decrease the number / proportion of kidney cells with a mesenchymal cell-like phenotype. The mesenchymal cell-like phenotype may be characterized, for example, by the expression of SNAIL and / or ACTA2, and may be accompanied by a lack of expression of E-cadherin.

[0308] Antagonists against IL-11-mediated signal transduction may be used in methods for maintaining / enhancing the functional level of renal tubular epithelial cells or renal tissue. The function of renal tubular epithelial cells / renal tissue may be evaluated, for example, by evaluating the above-mentioned activity correlates. For example, the function of renal tubular epithelial cells / renal tissue may be evaluated by analyzing serum / blood urea and / or creatine concentrations (e.g., blood urea nitrogen), or by monitoring albumin / creatine ratio (ACR). Antagonists against IL-11-mediated signal transduction may be used in methods for reducing serum / blood creatine and / or urea concentrations, or for reducing albumin / creatine ratio (ACR).

[0309] Those skilled in the art will readily appreciate that the therapeutic and prophylactic benefits of the present invention can be applied to virtually any disease / condition that may benefit from the suppression of renal damage and / or pathology associated with renal damage. The therapeutic and prophylactic benefits of the present invention can be applied to any subject suffering from renal damage and / or a disorder, disease or condition associated with renal damage. The therapeutic and prophylactic benefits of the present invention can also be applied to any subject suffering from a disease in which pathology associated with renal damage exists.

[0310] In some embodiments, the present invention provides for the treatment / prevention of a disease / condition caused by or exacerbated by renal impairment. In some embodiments, the present invention provides for the treatment / prevention of a disease / condition in a subject with renal impairment having a poor prognosis.

[0311] The diagnosis and management of acute kidney injury is described in Rahman M et al., Acute Kidney injury: a guide to diagnosis and management. Am Fam Physician 2012 Oct 1:86(7): 631-9. As stated in this document, acute kidney injury is characterized by a rapid deterioration of renal function, manifested by an increase in serum creatinine concentration, which may be accompanied by a decrease in urinary output.

[0312] In some embodiments, renal impairment and / or renal impairment associated disorders, diseases or conditions treated / prevented by the present invention may be characterized by increased expression of one or both of IL-11 and IL-11Rα in an organ / tissue / subject affected by renal impairment and / or renal impairment associated disorders, diseases or conditions, as compared to, for example, a normal organ / tissue / subject (i.e., an organ / tissue / subject free of renal impairment or renal impairment associated disorders, diseases or conditions).

[0313] In some embodiments, the invention treats / prevents kidney damage in diseases / disorders / conditions associated with kidney damage, such as those described herein. In some embodiments, the invention treats / prevents kidney damage and underlying diseases / disorders / conditions associated with kidney damage. For example, inhibition of IL-11-mediated signaling is useful to antagonize the role of IL-11 in chemotherapy-associated kidney damage, as well as to antagonize the role of IL-11 in cancer.

[0314] The treatment / prevention of renal damage and / or disorders, diseases or conditions associated with renal damage according to the present invention may be treatment / prevention of renal damage and / or disorders, diseases or conditions associated with renal damage associated with upregulation of IL-11, for example treatment / prevention of renal damage and / or disorders, diseases or conditions associated with renal damage associated with upregulation of IL-11 in cells or tissues in which symptoms of said disease / disorder / condition are manifested or in cells or tissues in which symptoms of said disease / disorder / condition may be manifested, or upregulation of extracellular IL-11 or IL-11Rα.

[0315] The disorder, disease or condition associated with renal impairment may affect any tissue, organ or organ system. In some embodiments, the disease / disorder / condition may affect any tissue / organ / organ system. In some embodiments, the disease / disorder / condition affects the kidney.

[0316] In some embodiments, the disorder, disease or condition associated with renal impairment affects one or more of the circulatory system, digestive system, excretory system, respiratory system, renal system, reproductive system, circulatory system, muscular system, endocrine system, exocrine system, lymphatic system, immune system, nervous system and / or skeletal system.

[0317] In some embodiments, the present invention treats / prevents pathology associated with kidney injury in acute kidney injury, nephrotoxicity, drug-induced kidney injury, drug-induced acute kidney injury, drug-induced nephrotoxicity, cisplatin-induced nephroinjury, cisplatin-induced acute kidney injury or cisplatin-induced nephrotoxicity.

[0318] The treatment may be effective in preventing the progression of renal impairment and / or disorders, diseases or conditions associated with renal impairment, e.g., may be effective in reducing / delaying / preventing the deterioration of renal impairment and / or disorders, diseases or conditions associated with renal impairment, or may be effective in reducing / delaying / preventing the onset of renal impairment and / or disorders, diseases or conditions associated with renal impairment. In some embodiments, the treatment may improve symptoms of renal impairment and / or disorders, diseases or conditions associated with renal impairment, e.g., may reduce the severity of symptoms of renal impairment and / or disorders, diseases or conditions associated with renal impairment, and / or may alleviate symptoms of renal impairment and / or disorders, diseases or conditions associated with renal impairment. In some embodiments, the treatment may increase survival rates. In some embodiments, the treatment is effective in alleviating the effects and / or symptoms of renal impairment and / or disorders, diseases or conditions associated with renal impairment.

[0319] "Prevention" may refer to preventing the onset of renal damage and / or disorders, diseases or conditions associated with renal damage, and / or preventing the worsening of renal damage and / or disorders, diseases or conditions associated with renal damage, e.g., preventing the progression of renal damage and / or disorders, diseases or conditions associated with renal damage to a later or chronic stage.

[0320] In some embodiments, the present invention treats / prevents acute kidney injury (AKI), acute renal failure, acute kidney disease, chronic kidney disease, kidney injury, drug-induced kidney injury, tubular necrosis, acute tubular necrosis, autoimmune kidney injury and kidney cancer.

[0321] Acute tubular necrosis is the most common intrinsic acute kidney injury in hospitalized patients (Rahman M et al., supra). Its cause is often ischemic (due to prolonged hypotension) or nephrotoxic (due to drugs that are toxic to tubular cells). Acute kidney injury caused by acute tubular necrosis is often not improved by adequate replacement of intravascular volume and renal blood flow. Both ischemic and nephrotoxic acute tubular necrosis may resolve over time, but temporary renal replacement therapy may be required depending on the severity of renal injury and the presence or absence of pre-existing chronic kidney disease.

[0322] As used herein, "cancer" may refer to any unwanted cell proliferation (or a disease caused by unwanted cell proliferation), neoplasm, or tumor, or an increased risk of or predisposition to unwanted cell proliferation, neoplasm, or tumor. Cancer may be benign or malignant, primary or secondary (metastatic). A neoplasm or tumor may be any abnormal growth or proliferation of cells and may arise in any tissue. Examples of tissues in which a neoplasm or tumor may arise include the adrenal gland, adrenal medulla, anus, appendix, bladder, blood, bone, bone marrow, brain, breast, cecum, central nervous system (including or excluding the brain), cerebellum, cervix, large intestine, duodenum, endometrium, epithelial cells (e.g., kidney epithelium), gallbladder, esophagus, glial cells, heart, ileum, jejunum, kidney, lacrimal gland, larynx, liver, lung, lymph, lymph nodes (abdominal lymph nodes, axillary lymph nodes, etc.) lymph nodes, including lymph nodes, cervical nodes, inguinal nodes, mediastinal nodes, pelvic nodes, and para-aortic nodes), lymphoblasts, maxilla, mediastinal, mesentery, myometrium, nasopharynx, omentum, oral cavity, ovary, pancreas, parotid gland, peripheral nervous system, abdominal cavity, pleura, prostate, salivary gland, sigmoid colon, skin, small intestine, soft tissue, spleen, stomach, testes, thymus, thyroid, tongue, tonsils, trachea, uterus, vulva, and leukocytes.

[0323] The cancer may be a specific type of cancer. Examples of cancer types include astrocytoma, carcinoma (e.g., adenocarcinoma, hepatocellular carcinoma, medullary carcinoma, papillary carcinoma, squamous cell carcinoma), glioma, lymphoma, medulloblastoma, malignant melanoma, myeloma, meningioma, neuroblastoma, and sarcoma (e.g., angiosarcoma, chondrosarcoma, osteosarcoma).

[0324] In this specification, "cancer" includes acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML), adrenal cortical carcinoma, anal cancer, bladder cancer, blood cancer, bone cancer, brain tumor, breast cancer, cancer of the female reproductive system, cancer of the male reproductive system, central nervous system lymphoma, cervical cancer, childhood rhabdomyosarcoma, childhood sarcoma, chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), colorectal cancer, colon cancer, endometrial cancer, endometrial sarcoma, esophageal cancer, eye cancer, gallbladder cancer, stomach cancer, gastrointestinal cancer, hairy cell leukemia, head and neck cancer, hepatocellular carcinoma, Hodgkin's disease, hypopharyngeal cancer, Kaposi's sarcoma, kidney cancer, laryngeal cancer, leukemia, leukemia, liver cancer, lung cancer, The cancer may include one or more of the following: malignant fibrous histiocytoma, malignant thymoma, malignant melanoma, mesothelioma, multiple myeloma, myeloma, nasal cavity and paranasal sinus cancer, nasopharyngeal cancer, nervous system cancer, neuroblastoma, non-Hodgkin's lymphoma, oral cavity cancer, oropharyngeal cancer, osteosarcoma, ovarian cancer, pancreatic cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pituitary tumor, plasmacytoma, primary central nervous system lymphoma, prostate cancer, rectal cancer, respiratory system cancer, retinoblastoma, salivary gland cancer, skin cancer, small intestine cancer, soft tissue sarcoma, gastric cancer, stomach cancer, testicular cancer, thyroid cancer, urinary system cancer, uterine sarcoma, vaginal cancer, vascular system cancer, Waldenstrom's macroglobulinemia, and Wilms' tumor.

[0325] According to various aspects of the invention, the methods of the invention include treating (e.g., in connection with renal impairment) Reducing necrosis of renal tissue and / or renal cells; Reducing fibrosis of the kidney and / or renal tissue; Reducing collagen content in the kidney and / or renal tissue and / or inhibiting collagen deposition in the kidney and / or renal tissue; Increasing / maintaining renal function; Increasing / maintaining urinary output; Reducing the urinary albumin / creatinine ratio; Reducing serum creatinine levels; Reducing serum urea levels; Reducing serum TGF-β1 concentrations; Increase / maintain kidney mass; Increasing / maintaining renal cortical volume; Gaining / maintaining weight; Inhibiting the epithelial-mesenchymal transition of renal tubular epithelial cells; Reducing the number / percentage of ACTA2-positive cells in the kidney; Reducing the expression of SNAIL in renal cells and / or kidney / renal tissue; and Increasing / maintaining E-cadherin expression in renal cells and / or kidney / renal tissue may include one or more of the following:

[0326] Administration Preferably, agents capable of inhibiting IL-11-mediated signaling are administered in a "therapeutically effective" or "prophylactically effective" amount sufficient to benefit the subject.

[0327] In some embodiments, the agent may be administered before, simultaneously with, or after the onset of a cause of kidney damage, for example, before, simultaneously with, or after administration or ingestion of a nephrotoxic pharmaceutical agent, or before, simultaneously with, or after exposure to an environmental cause of kidney damage.

[0328] The actual dosage, administration rate and time course after administration depend on the characteristics and severity of renal impairment and the characteristics of the drug. Prescription of treatment (e.g., dose determination) is under the responsibility of general practitioners and other medical practitioners, and usually takes into account the disease / condition to be treated, the condition of the individual subject, the delivery site, the method of administration, and other factors familiar to medical practitioners. Examples of such methods and protocols are described in Remington's Pharmaceutical Sciences, 20th Edition, 2000, pub. Lippincott, Williams & Wilkins.

[0329] The agents of the invention may be administered multiple times. Another therapeutic agent may be administered in one or more doses or each dose may be administered simultaneously or sequentially.

[0330] The multiple administrations may be spaced apart by a predetermined time interval, which may be 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or 31 days, or 1, 2, 3, 4, 5, or 6 months. As an example, the administration may be once every 7 days, once every 14 days, once every 21 days, or once every 28 days (±3, ±2, or ±1 day).

[0331] In therapeutic applications, agents capable of suppressing IL-11-mediated signal transduction are preferably formulated as pharmaceuticals or pharmaceutical preparations together with one or more other pharma- ceutical acceptable ingredients well known to those skilled in the art, including, but not limited to, pharma- ceutical acceptable carriers, adjuvants, excipients, diluents, fillers, buffers, preservatives, antioxidants, lubricants, stabilizers, solubilizers, surfactants (e.g., wetting agents), masking agents, colorants, flavoring agents, and sweetening agents.

[0332] As used herein, "pharmaceutical acceptable" refers to a compound, ingredient, material, composition, dosage form, etc. that is suitable for use in contact with the tissues of the recipient (e.g., human) without causing excessive toxicity, irritation, allergic response, or other problem or complication, within the scope of sound medical judgment, commensurate with a reasonable benefit-risk ratio. Moreover, each carrier, adjuvant, excipient, etc. must be "acceptable" in terms of compatibility with other ingredients in the formulation.

[0333] Suitable carriers, adjuvants, excipients, etc. can be found in standard pharmaceutical texts, such as, for example, Remington's Pharmaceutical Sciences, 18th edition, Mack Publishing Company, Easton, Pa., 1990; and Handbook of Pharmaceutical Excipients, 2nd edition, 1994.

[0334] The formulation may be prepared by any method well known in the pharmaceutical field. Such a method includes mixing the active compound with a carrier which constitutes one or more accessory ingredients. In general, the formulation is prepared by uniformly and intimately mixing the active compound with a carrier (e.g., liquid carrier, finely divided solid carrier, etc.), and then shaping the resulting mixture as necessary.

[0335] The formulations may be prepared for administration by topical, parenteral, systemic, intravenous, intraarterial, intramuscular, intrathecal, intraocular, intraconjunctival, subcutaneous, oral, or transdermal routes, which may include injection. Injectable formulations may contain the selected agent in a sterile or isotonic solvent. The formulation and method of administration may be selected depending on the agent of the invention and the disease / disorder / condition to be treated.

[0336] In some cases, the invention (e.g., agent / composition) described herein is administered to perform the treatment described herein in combination with another treatment for a disease / disorder / condition associated with renal impairment. Suitable other treatments for diseases / disorders / conditions associated with renal impairment are known in the art. The composition of the invention may be administered alone or in combination with another treatment, either simultaneously or sequentially, depending on the disease / disorder / condition being treated. For example, the invention may be administered before, simultaneously with or after the other treatment. The invention and the other therapeutic agent may be formulated together or separately, for example in the form of a formulation as described above.

[0337] Detection of IL-11 and IL-11 receptor Some aspects and embodiments of the invention relate to detecting expression of IL-11 or an IL-11 receptor (e.g., IL-11Rα, gp130, or a complex comprising IL-11Rα and / or gp130) in a sample obtained from a subject.

[0338] In some aspects and embodiments, the invention relates to the upregulation (overexpression) of expression of IL-11 or IL-11 receptor (either as protein or as oligonucleotides encoding IL-11 or IL-11 receptor) and the detection of upregulation of expression of IL-11 or IL-11 receptor as an indication of suitability for treatment with agents capable of suppressing the action of IL-11 or inhibiting or reducing expression of IL-11 or IL-11 receptor.

[0339] Upregulation of expression includes expression above the normally expected expression level in a given type of cell or tissue. Upregulation may be measured by measuring the expression level of the associated factor in the cell or tissue. The expression level of the associated factor in a cell or tissue sample obtained from a subject may be compared to a reference level of the associated factor (e.g., a value or range of values ​​representing the normal expression level of the associated factor in the same type of cell or tissue or a corresponding cell or tissue). In some embodiments, the reference level may be determined by detecting the expression of IL-11 or IL-11 receptor in a control sample (e.g., a corresponding cell or tissue obtained from a healthy subject or a corresponding cell or tissue obtained from a healthy tissue of the same subject). In some embodiments, the reference level may be obtained from a standard curve or a standard data set.

[0340] The expression level may be quantified by absolute comparison or by relative comparison.

[0341] In some embodiments, IL-11 or an IL-11 receptor (e.g., IL-11Rα, gp130, or a complex containing IL-11Rα and / or gp130) may be considered upregulated if the expression level in the test sample is at least 1.1-fold the reference level. More preferably, the expression level when upregulated may be selected from at least 1.2 times, at least 1.3 times, at least 1.4 times, at least 1.5 times, at least 1.6 times, at least 1.7 times, at least 1.8 times, at least 1.9 times, at least 2.0 times, at least 2.1 times, at least 2.2 times, at least 2.3 times, at least 2.4 times, at least 2.5 times, at least 2.6 times, at least 2.7 times, at least 2.8 times, at least 2.9 times, at least 3.0 times, at least 3.5 times, at least 4.0 times, at least 5.0 times, at least 6.0 times, at least 7.0 times, at least 8.0 times, at least 9.0 times and at least 10.0 times the baseline level.

[0342] The expression level may be measured by any of a variety of known in vitro analytical techniques, such as PCR-based assays, in situ hybridization assays, flow cytometry assays, immunological assays, and immunohistochemical assays.

[0343] As an example, suitable techniques include methods for detecting the amount of IL-11 or IL-11 receptor in a sample by contacting the sample with an agent capable of binding to IL-11 or IL-11 receptor and detecting the formation of a complex consisting of IL-11 or IL-11 receptor and the agent. The agent may be any suitable binding molecule, such as an antibody, a polypeptide, a peptide, an oligonucleotide, an aptamer, a small molecule, etc., and may be labeled so that the complex formed can be detected (e.g., visualized). Suitable labels and methods for detecting such complexes are well known in the art and include, for example, fluorescent labels (e.g., fluorescein, rhodamine, eosin, NDB, green fluorescent protein (GFP); rare earth chelates such as europium (Eu), terbium (Tb), samarium (Sm); tetramethylrhodamine, Texas Red, 4-methylumbelliferone, 7-amino-4-methylcoumarin, Cy3, Cy5), isotopic markers, radioisotopes (e.g., 32 P, 33 P, 35 S), chemiluminescent labels (e.g., acridinium ester, luminol, isoluminol), enzymes (e.g., peroxidase, alkaline phosphatase, glucose oxidase, β-galactosidase, luciferase), antibodies, ligands and receptors. Detection techniques are well known to those skilled in the art and can be selected depending on the labeling agent. Suitable techniques include PCR amplification of oligonucleotide tags, mass spectrometry, detection of fluorescence or color (e.g., produced by enzymatic conversion of a substrate by a reporter protein), or detection of radioactivity.

[0344] The assay may be configured to quantify the amount of IL-11 or IL-11 receptor in a sample. The amount of IL-11 or IL-11 receptor quantified from a test sample may be compared to a reference amount, and such comparison may determine whether the amount of IL-11 or IL-11 receptor contained in the test sample is higher or lower than the reference amount at a selected level of statistical significance.

[0345] The detected IL-11 or IL-11 receptor may be quantified to measure upregulation, downregulation, or amplification of the gene encoding IL-11 or IL-11 receptor. When the test sample contains fibrotic cells, such upregulation, downregulation, or amplification may be compared with a reference amount to determine whether there is a statistically significant difference.

[0346] The sample obtained from the subject may be of any type. The biological sample may be obtained from any tissue or body fluid, for example, a blood sample, a blood-derived sample, a serum sample, a lymph sample, a semen sample, a saliva sample, or a synovial fluid sample. The blood-derived sample may be a selected fraction derived from the patient's blood, for example, a selected cell-containing fraction, a plasma fraction, or a serum fraction. The sample may comprise a tissue sample or a biopsy sample, or cells isolated from the subject. The sample may also be obtained by known techniques, such as biopsy or fine needle aspiration. Furthermore, the sample may be stored until the expression level of IL-11 is measured and / or may be processed before the expression level of IL-11 is measured.

[0347] A sample obtained from a subject may be used to measure upregulation of IL-11 or IL-11 receptor in the subject.

[0348] In some preferred embodiments, the sample may be a tissue sample (e.g., a biopsy sample) obtained from kidney tissue, cardiac tissue, visceral organ tissue, respiratory system organ tissue, or renal-urinary system tissue. The sample may comprise cells.

[0349] Subjects may be selected for treatment / prophylaxis according to the present invention based on the confirmed upregulation of expression of IL-11 or IL-11 receptor (e.g., IL-11Rα, gp130, or a complex containing IL-11Rα and / or gp130) in the subject. Also, upregulation of expression of IL-11 or IL-11 receptor may be used as a marker of renal damage and / or disorders, diseases or conditions associated with renal damage that are suitable for treatment with agents capable of suppressing IL-11-mediated signaling.

[0350] The upregulation may be in a given tissue or in selected cells derived from a given tissue. The preferred tissue may be renal tissue. The upregulation of IL-11 or IL-11 receptor expression may be measured in circulating body fluids (e.g., blood) or in blood-derived samples. The upregulation may be the upregulation of extracellular IL-11 or IL-11Rα. In some embodiments, the expression may be upregulated locally or systemically.

[0351] As described in the following sections, an agent capable of inhibiting IL-11 mediated signaling may be administered to a subject.

[0352] Diagnosis and Prognosis Detection of upregulation of expression of IL-11 or an IL-11 receptor (e.g., IL-11Rα, gp130, or a complex comprising IL-11Rα and / or gp130) may be used in methods of diagnosing renal damage and / or disorders, diseases or conditions associated with renal damage, identifying subjects at risk of developing renal damage and / or disorders, diseases or conditions associated with renal damage, and may be used in methods of prognosticating or predicting a subject's responsiveness to treatment with an agent capable of inhibiting IL-11-mediated signaling.

[0353] The terms "develop" and "development", as well as other forms of the term "develop", may refer to the onset of a disorder / disease, or the continuation or progression of a disorder / disease.

[0354] In some embodiments, a subject may be determined to be suspected of having or suffering from a renal disorder and / or a renal disorder-related disorder, disease or condition, for example, based on the presence or absence of other symptoms indicative of the presence of renal disorder and / or a renal disorder-related disorder, disease or condition in the subject's organism or selected cells / tissues derived therefrom, or may be considered to be at risk for developing a renal disorder and / or a renal disorder-related disorder, disease or condition, for example, due to a genetic predisposition or exposure to environmental conditions known to be risk factors for renal disorder and / or a renal disorder-related disorder, disease or condition. Also, a diagnosis or a suspected diagnosis may be confirmed or a subject may be identified as being at risk for developing a renal disorder and / or a renal disorder-related disorder, disease or condition by measuring the upregulation of IL-11 or IL-11 receptor expression. Also, a renal disorder and / or a renal disorder-related disorder, disease or condition, or a predisposition may be diagnosed as being suitable for treatment with an agent capable of suppressing IL-11-mediated signaling by measuring the upregulation of IL-11 or IL-11 receptor expression.

[0355] Thus, there may be provided a method of providing a prognosis for a subject suffering from renal impairment and / or a disorder, disease or condition associated with renal impairment, or a subject suspected of suffering from renal impairment and / or a disorder, disease or condition associated with renal impairment, comprising determining whether expression of IL-11 or IL-11 receptor is upregulated in a sample obtained from the subject, and based on said determination, providing a prognosis for treatment of the subject with an agent capable of inhibiting IL-11-mediated signaling.

[0356] In some embodiments, a method for prognosticating or predicting a subject's responsiveness to treatment with an agent capable of suppressing IL-11-mediated signal transduction or a diagnostic method may not require the measurement of the expression level of IL-11 or IL-11 receptor, but may be based on measuring genetic factors in the subject that predict upregulation of IL-11 or IL-11 receptor expression or activity. Such genetic factors include measuring genetic mutations, single nucleotide polymorphisms (SNPs) or gene amplifications of IL-11, IL-11Rα and / or gp130 that are correlated with and / or predictive of upregulation of IL-11 or IL-11 receptor expression or activity or upregulation of IL-11-mediated signal transduction. The use of genetic factors to predict predisposition to a disease state or responsiveness to treatment is known in the art, see, e.g., Peter Starkel Gut 2008;57:440-442; Wright et al., Mol. Cell. Biol. March 2010 vol. 30 no. 6 1411-1420.

[0357] The genetic factor may be analyzed by methods known to those skilled in the art, such as PCR-based assays, e.g., quantitative PCR, competitive PCR, etc. For example, by determining the presence or absence of the genetic factor, e.g., in a sample obtained from a subject, a diagnosis may be established, the subject may be classified as at risk for developing renal impairment and / or a disorder, disease or condition associated with renal impairment, and / or the subject may be identified as suitable for treatment with an agent capable of suppressing IL-11-mediated signaling.

[0358] Some methods may include identifying the presence or absence of one or more SNPs associated with renal injury and / or susceptibility to developing disorders, diseases or conditions associated with renal injury or secretion of IL-11. SNPs are usually biallelic and therefore can be easily identified using any of a variety of conventional assays known to those skilled in the art (see, for example, Anthony J. Brookes. The essence of SNPs. Gene Volume 234, Issue 2, 8 July 1999, 177-186; Fan et al., Highly Parallel SNP Genotyping. Cold Spring Harb Symp Quant Biol 2003. 68: 69-78; Matsuzaki et al., Parallel Genotyping of Over 10,000 SNPs using a one-primer assay on a high-density oligonucleotide array. Genome Res. 2004. 14: 414-425).

[0359] The method may include identifying which SNP alleles are present in a sample obtained from the subject. In some embodiments, identifying the presence or absence of a minor allele may identify a susceptibility to developing renal impairment and / or a disorder, disease or condition associated with renal impairment or increased secretion of IL-11.

[0360] Thus, in one aspect of the invention there is provided a method of screening a subject comprising the steps of: obtaining a nucleic acid sample from a subject; and Identifying which alleles are present in the sample at the polymorphic nucleotide positions of one or more SNPs listed in Figure 33, Figure 34 or Figure 35 of WO 2017 / 103108(A1), which is incorporated herein by reference, or determining whether any one of the SNPs listed in these figures is present. 2 Identifying SNPs that exhibit linkage disequilibrium of ≧0.8 The present invention provides a method comprising:

[0361] The step of identifying an allele or SNP may comprise identifying the presence or absence of a minor allele at the selected polymorphic nucleotide position in the sample, and may comprise identifying the presence or absence of zero, one or two minor alleles.

[0362] The screening method may be, or may form part of, a method for identifying a subject's susceptibility to developing renal damage and / or a disorder, disease or condition associated with renal damage, or a diagnostic or prognostic method as described above.

[0363] The method may further comprise identifying the subject as having a susceptibility to developing renal impairment and / or a renal impairment associated disorder, disease or condition, or as being at high risk for developing renal impairment and / or a renal impairment associated disorder, disease or condition, for example if the subject is identified as having a minor allele at the polymorphic nucleotide position. The method may further comprise selecting the subject for treatment with an agent capable of inhibiting IL-11-mediated signaling, and / or administering to the subject an agent capable of inhibiting IL-11-mediated signaling to treat renal impairment and / or a renal impairment associated disorder, disease or condition in the subject, or to prevent the onset or progression of renal impairment and / or a renal impairment associated disorder, disease or condition in the subject.

[0364] In some embodiments, the methods for diagnosing renal damage and / or disorders, diseases or conditions associated with renal damage, identifying subjects at risk for developing renal damage and / or disorders, diseases or conditions associated with renal damage, or prognosing or predicting a subject's responsiveness to treatment with an agent capable of inhibiting IL-11-mediated signaling use indicators that are not for detecting upregulation of IL-11 or IL-11 receptor expression, and that are not genetic factors.

[0365] In some embodiments, the methods of diagnosing renal impairment and / or renal impairment associated disorders, diseases or conditions, identifying subjects at risk of developing renal impairment and / or renal impairment associated disorders, diseases or conditions, or prognosing or predicting a subject's responsiveness to treatment with an agent capable of inhibiting IL-11 mediated signaling are based on detecting, measuring and / or identifying one or more of the following indicators, or performing any one of the following analyses: Elevated serum creatinine, blood urea and / or blood nitrogen -Urinary sodium and creatinine concentrations Decreased urine output Renal ultrasound to identify the presence or absence of ureteral obstruction Kidney biopsy ·Urine testing

[0366] Reference values ​​for laboratory-performed renal function tests are described, for example, in Rahman et al., supra, which is incorporated herein by reference in its entirety.

[0367] The diagnostic or prognostic method may be performed in vitro using a sample obtained from a subject, or may be performed in vitro after processing a sample obtained from a subject. The patient from whom the sample was taken does not need to be present until the in vitro diagnostic or prognostic method is performed, and therefore the methods do not need to be performed on a living human or animal. The sample obtained from the subject may be of any type, as previously described.

[0368] The methods described herein may be used in conjunction with other diagnostic or prognostic tests to improve the accuracy of the diagnosis or prognosis or to confirm the results obtained using the testing methods described herein.

[0369] subject The subject may be an animal or a human. The subject is preferably a mammal, more preferably a human. The subject may be a non-human mammal, more preferably a human. The subject may be male or female. The subject may be a patient. The patient may have a renal disorder and / or a disorder, disease or condition associated with renal disorder as described herein. The subject may be a subject diagnosed as suffering from a renal disorder and / or a disorder, disease or condition associated with renal disorder that requires treatment, a subject suspected of suffering from such a renal disorder and / or a disorder, disease or condition associated with renal disorder, or a subject at risk of developing a renal disorder and / or a disorder, disease or condition associated with renal disorder.

[0370] In an embodiment according to the invention, the subject is preferably a human subject. In an embodiment according to the invention, a subject may be selected for treatment with the method of the invention based on the identification of a particular marker for renal impairment and / or a disorder, disease or condition associated with renal impairment.

[0371] In some embodiments, the subject may be a subject receiving a drug or medicine for the purpose of treating a disease, condition, or disorder that occurs in renal tissue or a disease, condition, or disorder that does not occur in renal tissue. The subject may also be a subject who has developed drug-induced nephropathy due to such treatment (e.g., due to its side effects). Furthermore, the subject may be a subject selected for the treatment or prophylactic therapy according to the present invention based on the development of such drug-induced nephropathy.

[0372] In some embodiments, the subject may be a subject who has been treated with a chemotherapeutic agent, or a subject who is being treated with a chemotherapeutic agent.The subject may also be a subject who has cancer, a subject who is suspected of having cancer, a subject who has recovered from cancer, or a subject who is in remission from cancer, and the administration of a chemotherapeutic agent may be included in part of the treatment of these subjects.Therefore, the subject may be a subject who has chemotherapy-induced nephropathy, chemotherapy-induced acute kidney injury, or chemotherapy-induced nephrotoxicity.

[0373] In some preferred embodiments, the subject may be a subject who has been treated with cisplatin or may be a subject who is being treated with cisplatin. The subject may also be a subject who has cancer, a subject who is suspected of having cancer, a subject who has recovered from cancer, or a subject who is in remission from cancer, and the administration of cisplatin may be included as part of the treatment of these subjects. Thus, the subject may be a subject who has cisplatin-induced nephropathy, cisplatin-induced acute kidney injury, or cisplatin-induced nephrotoxicity.

[0374] The subject may be one undergoing intermittent or regular dialysis.

[0375] Sequence identity Pairwise and multiple sequence alignments to determine percent identity between two or more amino acid or nucleic acid sequences can be performed using various methods known to those skilled in the art, for example, using publicly available computer software such as ClustalOmega software (Soding, J. 2005, Bioinformatics 21, 951-960), T-coffee software (Notredame et al. 2000, J. Mol. Biol. (2000) 302, 205-217), Kalign software (Lassmann and Sonnhammer 2005, BMC Bioinformatics, 6(298)), and MAFFT software (Katoh and Standley 2013, Molecular Biology and Evolution, 30(4) 772-780). When using such software, it is preferable to use default parameters, such as gap penalties and extension penalties.

[0376] [Table 2] TIFF0007638215000003.tif221166TIFF0007638215000004.tif186166TIFF0007638215000005.tif99166

[0377] The present invention includes combinations of the aspects and preferred features described herein except where such combinations are clearly not acceptable or clearly to be avoided.

[0378] The features of the disclosure expressed in the foregoing detailed description, the following claims, or the accompanying drawings as specific forms or means for performing a disclosed function, or methods or processes for conveniently obtaining the results of the disclosure, may be used separately or in any combination to realize the invention in its various forms.

[0379] For the avoidance of doubt, the logical explanations provided herein are provided for the purpose of enhancing the understanding of the reader, and the inventors do not wish to constrain the present invention by these logical explanations.

[0380] All section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described herein.

[0381] Throughout this specification, including the claims which follow, unless otherwise indicated, the terms "comprise" and "include", and their variations "comprises", "comprising" and "including", are understood to include a stated element or step or elements or steps, but not to exclude elements or steps or elements or steps other than those stated.

[0382] As used in this specification and the appended claims, it should be noted that the singular forms "a", "an" and "the" include the plural unless expressly stated otherwise. Numerical ranges are herein expressed as "about" a particular value and / or as a range from "about" a particular value to "about" another particular value. When such a range is stated, another embodiment includes the particular value that is not about, and / or the range from the particular value that is not about, to the other particular value. Similarly, when a particular value is stated as an approximation by use of the antecedent "about", it is understood that the particular value that is not about can constitute another embodiment. The term "about" in connection with a numerical value indicates an arbitrary and average value (e.g., ±10%).

[0383] The methods disclosed herein may be performed in vitro, ex vivo, or in vivo, and the products of the invention may be in vitro, ex vivo, or in vivo products. "In vitro" encompasses experiments using materials, biological matter, cells, and / or tissues in laboratory conditions or culture. In contrast, "in vivo" encompasses experiments and manipulations using living multicellular organisms. "Ex vivo" refers to something that exists or is performed outside of a living organism, e.g., outside of a human or animal body, and may be present or performed in tissues (e.g., whole organs) or cells taken from an organism.

[0384] When a nucleic acid sequence is disclosed herein, the reverse complement thereof is also specifically contemplated.

[0385] For standard molecular biology techniques, see Sambrook, J., Russell, DW Molecular Cloning, A Laboratory Manual. 3 ed. 2001, Cold Spring Harbor, New York: Cold Spring Harbor Laboratory Press.

[0386] Aspects and embodiments of the present invention are described below with reference to the accompanying drawings. Further aspects and embodiments will be readily apparent to those skilled in the art. Any references cited herein are incorporated by reference as part of this specification. The present invention will be described with the following exemplary embodiments, but those skilled in the art will be able to readily understand various equivalent modifications and variations by reference to this disclosure. Therefore, the exemplary embodiments of the present invention described above are considered to be illustrative and not limiting of the present invention. Various modifications may be made to the embodiments described herein without departing from the spirit and scope of the present invention.

[0387] Hereinafter, embodiments and experiments illustrating the principles of the present invention will be described with reference to the accompanying drawings. [Brief description of the drawings]

[0388] [Figure 1] Photomicrographs showing that anti-IL-11 neutralizing antibodies block the progression of acute kidney injury (AKI) to chronic kidney disease (CKD) compared to IgG control antibodies. Mice were subjected to folate-induced kidney injury (a model of acute tubular necrosis (ATN) and secondary fibrosis) and treated with IgG control, Enx203 (anti-IL-11 antibody), or ENx209 (anti-IL-11RA antibody). Kidneys were removed 28 days later and stained with Masson's trichrome stain. Both antibodies were highly effective at blocking collagen deposition.

[0389] [Figure 2A-2B] Figure 2 shows that anti-IL-11 neutralizing antibodies prevent the progression of acute kidney injury. Mice were subjected to folate-induced kidney injury and treated with IgG control, anti-IL-11 (ENx203, AB1) or anti-IL11RA (ENx209, AB2) antibodies. After 28 days, kidneys were harvested for HPA collagen assays, as well as serum and urine (collected in metabolic cages) to assess urinary albumin / creatinine ratios. Markers of kidney injury were comparable between treatment groups at day 2, indicating similar initial kidney damage, although it should be noted that mice treated with anti-IL-11 therapy recovered kidney function to a greater extent, whereas mice treated with IgG recovered kidney function less effectively. (Figure 2A) Renal collagen content and (Figure 2B) urinary albumin / creatinine ratios at day 28 in mice with various treatments are shown.

[0390] [Diagram 3]Graphs and photographs showing the dose-dependent effect of ENx203 and ENx209 on the renal injury response and ERK activation. Mice were injected with each antibody and then treated with folic acid 1 hour later to induce renal injury. After 28 days, collagen content in the kidney was evaluated. Mice were further treated with 10 mg / kg or 20 mg / kg of IgG control antibody twice weekly. No differences were observed between these control groups, so both groups of mice are plotted together. The folic acid-induced renal injury model shows very severe renal injury (see Figure 4). However, even with such a strong chemical stimulus, twice weekly administration of ENx203 at a reduced dose of 5 mg / kg was highly effective (top panel). Each point represents a biological replicate. Data are shown as mean ± SD. Antibody injections twice weekly (biw.) and three times weekly (triw.) are shown. IL-11-mediated ERK activation in fibrotic kidneys was observed by measuring target binding (see figure below), and it was shown that administration of 1 mg / kg ENx203 twice weekly was effective.

[0391] [Figure 4A-4C] Graphs showing the dose-dependent effect of ENx203 or ENx209 on serum creatinine (Figure 4A), urea (Figure 4B) and TGFB1 (Figure 4C). Mice were injected with the respective antibodies and then treated with folic acid 1 h later to induce renal injury. Serum marker concentrations were assessed 28 days later. Mice were further treated with 10 mg / kg or 20 mg / kg of IgG control antibody twice weekly. No differences were observed between these control groups, so both groups of mice are plotted together. Each point represents a biological replicate. Data are shown as mean ± SD. Antibody injections twice weekly (biw.) and three times weekly (triw.) are shown.

[0392] [Figure 5A-5C]Graphs showing the effect of treatment with (FIG. 5A) ENx108A (IgG1), (FIG. 5B) ENx203 or (FIG. 5C) ENx209 on the progression of acute kidney injury (AKI). Mice were treated with the indicated concentrations of each antibody (twice a week) starting the day before folic acid treatment. Collagen content in kidneys was measured 21 days after induction of kidney injury. Data points represent individual mice, and data were obtained in parallel experiments so that each antibody could be compared with each other. Baseline and IgG control mice are also plotted in duplicate on each antibody graph to facilitate data interpretation. Data are means ± SD. Points represent mice in each group. P values ​​corrected by two-tailed Dunnett's test are shown.

[0393] [Figure 6A-6B] 6A-6B show that knocking out Il11ra specifically in fibroblasts in mice protects mice from acute kidney injury (AKI). Mice were treated with tamoxifen to delete Il11ra1 in Col1a1-positive cells, followed by treatment with folic acid to induce kidney injury. After 21 days, (Figure 6A) collagen content was assessed using an HPA assay, and (Figure 6B) kidney function was assessed by serum urea concentration. Mice were protected to a similar extent as antibody treatment, suggesting that IL-11 activity plays a central role in fibroblasts in the pathogenesis of kidney injury and renal dysfunction. P values ​​using Sidak's correction are shown.

[0394] [Figure 7A-7F]7A-7F are schematics, graphs, and photographs of the effect of anti-IL-11 therapy starting 3 days after induction of acute kidney injury. (FIG. 7A) Schematic showing timing of folic acid administration and antibody treatment. (FIG. 7B) Mice treated with anti-IL-11 therapy gain weight immediately after anti-IL-11 therapy. At the end of the experimental period, mice treated with anti-IL-11 therapy had (FIG. 7C) heavier kidney weights, (FIG. 7D) reduced collagen content in the kidneys, (FIG. 7E) normal urine volume, and (FIG. 7F) improved gross morphology than mice treated with IgG.

[0395] [Figure 8A-8F] Schematic diagrams, graphs, and photographs showing that treatment with ENx203 reverses renal injury and renal dysfunction and induces renal regeneration. (Fig. 8A) Schematic diagram showing the timing of folic acid administration and antibody treatment. A folic acid-induced renal injury model was established, and mice were not given any treatment until day 21 after induction of renal injury. This resulted in severe renal function decline and increased collagen content. Next, antibody treatment was started, and renal function was analyzed by evaluating serum urea concentration and collagen content 3, 6, 9, and 12 weeks after the start of treatment. The results revealed that mice treated with ENx203 had significantly decreased collagen content (Fig. 8B) and significantly improved renal function (Fig. 8C) compared with mice before treatment on day 21. Despite the decrease in collagen content in the kidney, kidney weight increased (Fig. 8D), and the kidney surface was less irregular (Fig. 8E) (Fig. 8F) and had a healthy tissue structure (cross-section of kidney, 3 mice). P values ​​are corrected for multiple testing and represent differences between later time points and the FA 21d group (Dunnett's test). Data are presented as mean ± SD.

[0396] [Figure 9]Photographs showing that treatment with ENx203 reduced collagen content and promoted recovery of normal renal parenchymal and cortical volumes. Masson's trichrome staining of the midsection of kidneys from mice at day 21 (D21) after folic acid (FA)-induced renal injury or from control mice, or from mice treated with IgG or anti-IL-11 starting at day 21 after folic acid-induced renal injury and continued for 3, 6, 9 or 12 weeks.

[0397] [Figure 10] 10A-10C are high magnification photographs of the renal cortex from the photograph shown in Figure 9. Shown are Masson's trichrome staining of the renal cortex from mice at day 21 (D21) after induction of renal injury with folic acid (FA) or from control mice, or from mice treated with IgG or anti-IL-11 starting at day 21 after induction of renal injury with folic acid and continued for 3, 6, 9 or 12 weeks.

[0398] [Figure 11A-11B] 11A-11B are graphs showing that treatment with ENx203 after establishment of renal injury increases body weight and restores urinary albumin / creatinine ratio (ACR) by 50%. A folate-induced renal injury model was established and mice were left untreated until 21 days after induction of renal injury. Antibody treatment was then started and mice were continuously evaluated for body weight. (FIG. 11A) Mice treated with ENx203 began to regain body weight immediately after the start of treatment. (FIG. 11B) After 12 weeks of antibody therapy, the urinary albumin / creatinine ratio was restored by approximately 50% compared to the start of treatment 3 weeks after induction of renal injury.

[0399] [Figure 12A-12B]12A-12B are graphs showing that TGFB1-induced partial epithelial-mesenchymal transition of renal tubular epithelial cells is dependent on IL-11. Human primary renal tubular epithelial cells were stimulated with TGFB1, IL-11 (5 ng / ml, 24 h) or a combination of TGFB1 and antibody (2 μg / ml, 24 h). The percentage of ACTA2-positive cells (Figure 12A) and collagen expression (Figure 12B) were monitored using the Operetta high content imaging platform.

[0400] [Figure 13] Photographs of partial epithelial-mesenchymal transition of renal tubular epithelial cells induced by TGFB1. Top panel shows human primary renal tubular epithelial cells stimulated with IL-11 (2.5-20ng / ml, 24 hours). Actin stress fiber formation was visualized with rhodamine phalloidin using Operetta high content imaging. Bottom panel shows collagen expression visualized in the cells.

[0401] [Figure 14A-14B] 14A and 14B are photographs and graphs showing that the expression of essential Snail factors by renal tubular epithelial cells is dependent on IL-11. Human primary renal tubular epithelial cells were stimulated with IL-11 alone or with TGFB in the presence of IgG or anti-IL-11 antibody (ENx203), and the expression of Snail or ACTA2 genes was detected using the Operetta imaging platform. (FIG. 14A) IL-11 and TGFB induce partial epithelial-mesenchymal transition and induce the expression of ACTA2 and Snail. This effect in TGFB-stimulated cells was strongly suppressed in the presence of anti-IL-11 antibody. (FIG. 14B) Quantification of ACTA-positive cells from the micrographs shown in FIG. 14A.

[0402] [Figure 15]Figure 1 shows that the induction of Snail in human primary renal tubular epithelial cells is blocked by anti-IL-11 antibody treatment. Western blots of human primary renal tubular epithelial cells treated with TGFB or IL-11 alone in the presence or absence of anti-IL-11 antibody (ENx203). Top panel shows that induction of SNAIL (a master regulator of epithelial-mesenchymal transition) by TGFB (5ng / ml) is dependent on IL-11 signaling and that SNAIL expression can be induced by IL-11 (5ng / ml) alone. Bottom panel shows that GAPDH control protein confirmed equal protein loading.

[0403] [Figure 16] Photographs showing that anti-IL-11 therapy enhances E-cadherin expression in regenerating kidneys. E-cadherin expression is a standard marker for identifying epithelial cells and is lost when cells undergo epithelial-mesenchymal or partial epithelial-mesenchymal transition. Consistent with the effect of IL-11 in inducing partial epithelial-mesenchymal transition (pEMT) of tubular epithelial cells, kidneys from mice 21 days after folate-induced renal injury and mice treated with IgG for 12 weeks after renal injury showed lower E-cadherin expression than controls when analyzed by Western blot, an effect that was reversed by anti-IL-11 therapy.

[0404] [Figure 17A-17B]17A-B are graphs and bar graphs of the effect of treatment with anti-IL-11 antibody in a unilateral urinary obstruction (UUO)-induced renal injury model. Unilateral urinary obstruction was performed on mice to induce a physically induced renal injury response. Some mice were treated with ENx203 or IgG control antibody (20 mg / kg each) on days 4, 7, and 9 after surgery. (FIG. 17A) Body weights of mice treated with ENx203 and IgG control are shown. (FIG. 17B) Ten days after surgery, collagen content in the kidney was assessed using an HPA assay. Mice treated with ENx203 therapeutic antibody had significantly reduced collagen content compared to mice treated with control antibody. Data are shown as mean ± SD. P values ​​using Sidak's correction method are shown.

[0405] [Fig. 18A-18H] 18A-18H are schematics and graphs showing the effect of treatment with anti-IL-11 antibody in a cisplatin-induced renal injury model. (FIG. 18A) Mice were treated with cisplatin (7 mg / kg cisplatin) once a week for 4 weeks. The control group received saline. Mice were treated with 10 mg / kg X203 or isotype-matched IgG control antibody by intraperitoneal injection twice a week starting from the first week. After 8 weeks, mice were sampled for kidney analysis. (FIG. 18B) Collagen content in kidney assessed by HPA assay. (FIG. 18C-18H) RNA expression levels of (FIG. 18C) Col3, (FIG. 18D) fibronectin, (FIG. 18E) MMP2, (FIG. 18F) TIMP, (FIG. 18G) CCL5, and (FIG. 18H) CCL2 measured by qPCR. Each graph shows the fold change (FC) of expression levels compared to those in saline-treated mice. EXAMPLES

[0406] The following examples demonstrate that anti-IL-11 therapy can attenuate renal injury by broadly regenerating and reversing impaired renal function in models of acute and chronic renal disease.

[0407] Example 1: Materials and Methods 1.1 Primary human renal proximal tubule epithelial cells Culture and stimulation conditions Primary human renal proximal tubule epithelial cells (HRPTEC) (PCS-400-010, ATCC) were grown and maintained in complete growth medium for renal epithelial cells (PCS-400-030 and PCS-400-040, ATCC) at 37 °C and 5% CO2. HRPTEC medium was changed every 2-3 days and cells were passaged at 80% confluence using standard trypsinization techniques. All experiments were performed with cells at passage number P3, where cells were serum starved for 16 h before the respective stimulation in serum-free medium for renal epithelial cells (24 h). Stimulated cells were compared to unstimulated cells grown for the same period under the same conditions (serum-free medium for renal epithelial cells) in the absence of the stimulus.

[0408] Operetta phenotypic assay HRPTECs were cultured at 1 × 10 in a 96-well CellCarrier plate (600550, PerkinElmer). 4Cells were seeded at a density of 1000 cells / well. Cells were stimulated for 24 h with the indicated concentrations of IL-11 (Genscript) or TGF-β1 (PHC 143B, Bio-Rad) in the presence or absence of 2 μg / ml of anti-IL-11 (ENx203) or IgG isotype control antibody. According to the experimental conditions, cells were fixed with 4% paraformaldehyde (PFA) (28908, Thermo Fisher Scientific) and permeabilized with 0.1% Triton X-100 (T8787, Sigma) in PBS. Nonspecific binding sites were blocked with a PBS solution containing 0.5% BSA (A7906, Sigma) and 0.1% Tween-20 (170-6531, Bio-Rad). Cells were incubated overnight (4°C) with primary antibodies (1:500), namely ACTA2 antibody (ab7817, Abcam), collagen I antibody (34710, Abcam), and SNAIL antibody (ab180714, Abcam), and incubated for 1 h (room temperature, in the dark) with the appropriate AlexaFluor 488-conjugated secondary antibodies (goat anti-mouse 488 (ab150113) and goat anti-rabbit 488 (ab150077, Abcam)) (1:1000). Cells were counterstained with blocking solution containing rhodamine phalloidin (R415, Thermo Fisher Scientific) and DAPI (1 μg / ml, D1306, Thermo Fisher Scientific). Plates were scanned and images were acquired with an Operetta high content imaging system (1438, PerkinElmer). At least two wells were used for each condition, and a minimum of seven fields per well were analyzed. ACTA2+ cells were quantified using Harmony software version 3.5.2. Fluorescence intensity of collagen type I per area was measured using Columbus version 2.7.1.

[0409] 1.2 Western blot Western blots were performed on total protein extracts obtained from human kidney proximal tubule epithelial (HRPTE) cells. HRPTE cells were lysed in radioimmunoprecipitation assay (RIPA) buffer containing protease and phosphatase inhibitors (Thermo Scientific) and lysates were clarified by centrifugation. Protein concentrations were measured by Bradford assay (Bio-Rad). Equal amounts of each protein lysate were resolved by SDS-PAGE, transferred to PVDF membranes, and immunoblot analysis was performed for SNAIL (3879, CST) and GAPDH (2118, CST). Proteins were visualized with anti-rabbit HRP antibody (7074, CST) using the ECL detection system (Pierce).

[0410] 1.3 Animal models All animal experiments were approved by the SingHealth Institutional Animal Care and Use Committee (IACUC) and were performed in accordance with the IACUC guidelines. All mice were provided with food and water ad libitum.

[0411] Chemical-induced acute kidney injury mouse model Folic acid (200 mg / kg) dissolved in vehicle (0.3 M NaHCO3) was injected intraperitoneally into 10-week-old male mice to induce kidney damage. Control mice received vehicle only. Mice were sacrificed 28 days after folic acid injection. Mice were also injected intraperitoneally with anti-IL-11 antibody (ENx203), anti-IL-11Rα antibody (ENx209) or IgG isotype control at the same concentration. Treatment duration and dose of antibody therapy are indicated in each figure.

[0412] Surgically-induced acute kidney injury mouse model Twelve-week-old male mice underwent unilateral ureteral obstruction (UUO). Briefly, mice were anesthetized by intraperitoneal injection of ketamine (100 mg / kg) / xylazine (10 mg / kg), and sufficient depth of anesthesia was confirmed by loss of interdigital reflex. The left flank of the mouse was then shaved. A longitudinal incision was made in the skin with a scalpel, and a second incision was made in the peritoneum to expose the kidney. The kidney was brought to the surface of the body using forceps, and the ureter below the kidney was ligated twice with surgical silk. The ligated kidney was gently returned to the anatomically correct position, and sterile saline was added to compensate for fluid loss. The incision was then sutured. Mice were postoperatively treated with the antibiotic enrofloxacin (15 mg / kg, subcutaneous) and the analgesic buprenorphine (0.1 mg / kg, subcutaneous) for three consecutive days.

[0413] 1.4 Colorimetric assays and ELISA Blood urea nitrogen (BUN) concentrations in mouse serum were measured using a urea assay kit (ab83362, Abcam). Urinary albumin concentrations were measured using a mouse albumin ELISA kit (ab108792, Abcam), and urinary creatinine concentrations were measured using a creatinine assay kit (ab204537, Abcam). Serum TGFβ1 concentrations were measured by ELISA. All ELISA and colorimetric assays were performed according to the manufacturer's protocols.

[0414] 1.5 Histological analysis Renal tissues were fixed in 10% neutral buffered formalin (NBF) for 48 hours at room temperature, dehydrated, embedded in paraffin blocks, and sectioned at 4 μm. Sections were stained with Masson's trichrome stain according to standard protocols and observed under a light microscope.

[0415] Example 2: Analysis of the effects of inhibition of IL-11-mediated signaling in a chemically induced renal injury model Renal injury was chemically induced in 10- to 12-week-old littermate mice of similar weight by intraperitoneal (ip) injection of folic acid (180 mg / kg) dissolved in vehicle (0.3 M NaHCO3), and control mice received vehicle alone.

[0416] Enx203 antibody (an antibody capable of binding to mouse IL-11 (and human IL-11) and inhibiting IL-11-mediated signal transduction) or Enx209 antibody (an antibody capable of binding to mouse IL-11 (and human IL-11) and inhibiting IL-11-mediated signal transduction) was administered the day after the folic acid treatment, and then administered three times a week at a dose of 20 mg / kg. Mice were euthanized 28 days after injection.

[0417] Enx203 and Enx209 are antagonists to IL-11-mediated signal transduction. Enx203 is a mouse anti-mouse IL-11 IgG antibody, described, for example, in Ng et al., Sci Transl Med. (2019) 11(511) pii: eaaw1237 (also reported in Ng, et al., "IL-11 is a therapeutic target in idiopathic pulmonary fibrosis." bioRxiv 336537; doi: https: / / doi.org / 10.1101 / 336537). Enx203 is also called "X203". Enx203 comprises a VH region as set forth in SEQ ID NO: 92 in WO 2019 / 238882(A1) (SEQ ID NO: 22 in this disclosure) and a VL region as set forth in SEQ ID NO: 94 in WO 2019 / 238882(A1) (SEQ ID NO: 23 in this disclosure). Enx209 is a mouse anti-mouse IL-11Rα IgG antibody, described, for example, in Widjaja et al., Gastroenterology (2019) 157(3):777-792 (also reported in Widjaja, et al., “IL-11 neutralising therapies target hepatic stellate cell-induced liver inflammation and fibrosis in NASH.” bioRxiv 470062; doi: https: / / doi.org / 10.1101 / 470062). Enx209 is also referred to as “X209”. Enx209 comprises a VH region as set forth in SEQ ID NO: 7 in WO 2019 / 238884(A1) (SEQ ID NO: 24 in this disclosure) and a VL region as set forth in SEQ ID NO: 14 in WO 2019 / 238884(A1) (SEQ ID NO: 25 in this disclosure).

[0418] Mouse plasma urea concentrations were quantified using a urea assay kit (ab83362, Abcam) and creatinine concentrations were quantified using a creatinine assay kit (ab65340, Abcam) according to the manufacturer's instructions. Total collagen content in the kidney was quantified by detecting hydroxyproline by colorimetric quantification using the Quickzyme Total Collagen Quantitation Assay Kit (Quickzyme Biosciences). Both colorimetric quantification methods were performed according to the manufacturer's instructions.

[0419] Tissues were embedded in paraffin, and kidneys were sectioned at a thickness of 3 μm. Paraffin sections were prepared by fixing tissues in 10% neutral buffered formalin (Sigma-Aldrich) for 24 h at room temperature, dehydrating, and embedding in paraffin. Frozen sections were prepared by embedding the organs in Tissue-Tek Optimal Cutting Temperature compound (VWR International) immediately after dissection. The cryomolds were then frozen in a metal beaker containing liquid nitrogen-cooled isopentane, and the sections were stored at -80°C. Total collagen was stained with Masson's Trichrome Staining Kit (HT15, Sigma-Aldrich) according to the manufacturer's instructions. Sections were photographed, and the area stained in blue was measured semiquantitatively using ImageJ software (version 1.49). For immunohistochemical analysis, tissue sections were incubated with anti-ACTA2 antibody (ab5694, Abcam). Primary antibody staining was visualized using the ImmPRESS HRP anti-rabbit IgG polymer detection kit (Vector Laboratories) with ImmPACT DAB peroxidase substrate (Vector Laboratories) as the chromogen. Sections were then counterstained with Mayer's hematoxylin (Merck).

[0420] 1 and 2 show that collagen staining was found to be significantly reduced in mice treated with anti-IL-11 or anti-IL-11Rα antagonist antibodies.

[0421] Furthermore, FIG. 2 shows that treatment with anti-IL-11 or anti-IL-11Rα antibodies significantly reduced the urinary albumin / creatine ratio, indicating a reduction in the extent of renal damage.

[0422] Importantly, serum urea and creatine concentrations were similar between treatment groups 2 days after chemical induction of renal injury (data not shown), but renal function was restored less effectively in mice treated with IL-11 antagonist therapy compared with mice receiving IgG.

[0423] In another experiment, a similar folate-induced nephropathy model was used to evaluate the therapeutic efficacy of antagonists against IL-11-mediated signaling to treat / prevent nephropathy. Mice were administered each antibody and 1 hour later nephropathy was induced by treatment with folic acid as described above. Collagen content in the kidney was evaluated 28 days later. Mice were treated with antibodies 1 mg / kg Enx203 twice weekly, 5 mg / kg Enx203 twice weekly, 5 mg / kg Enx203 three times weekly, 10 mg / kg Enx203 twice weekly, 10 mg / kg Enx203 three times weekly, 20 mg / kg Enx203 three times weekly, 20 mg / kg Enx209 twice weekly, or 10 mg / kg or 20 mg / kg IgG control antibody twice weekly.

[0424] Figure 3 shows that treatment with an antagonistic antibody against IL-11-mediated signaling reduced collagen content associated with folate-induced kidney injury in a dose-dependent manner. Enx203 had an effect on collagen content even when administered at a concentration of 5 mg / kg twice a week.

[0425] IL-11-mediated ERK activation in fibrotic kidneys was monitored by measuring target binding, and ENx203 was shown to be effective when administered at a dose of 1 mg / kg twice weekly.

[0426] In this experiment, serum creatine, urea and TGFβ1 concentrations were also evaluated. Figure 4 shows that treatment with an antagonistic antibody against IL-11-mediated signaling reduced serum levels of these factors, which correlate with renal injury, in a dose-dependent manner. Again, a reduction in serum levels of these factors was observed even when Enx203 was administered at a concentration of 5 mg / kg twice a week.

[0427] In further experiments, a similar folate-induced renal injury model was used to evaluate the therapeutic efficacy of antagonists against IL-11-mediated signaling for the treatment / prevention of renal injury. Mice were administered each antibody and the next day treated with folate to induce renal injury as described above. Collagen content in the kidney was evaluated 21 days later. Mice were treated with antibodies twice weekly with 10 mg / kg IgG control antibody or 0.5 mg / kg, 1 mg / kg, 5 mg / kg or 10 mg / kg Enx203, Enx209 or Enx108A. The results are shown in Figure 5.

[0428] Enx108A is a human anti-human IL-11 IgG antibody that can bind to mouse IL-11 and human IL-11 and inhibit IL-11-mediated signal transduction. Enx108A is described, for example, in WO 2019 / 238882(A1) and includes a VH region as shown in SEQ ID NO:8 (SEQ ID NO:26 in the present disclosure) in WO 2019 / 238882(A1) and a VL region as shown in SEQ ID NO:20 (SEQ ID NO:27 in the present disclosure) in WO 2019 / 238882(A1). In this embodiment, Enx108A is provided in the form of hIgG4(L248E,S241P) containing a lambda light chain (i.e., composed of a heavy chain having the amino acid sequence shown in SEQ ID NO:28 and a light chain having the amino acid sequence shown in SEQ ID NO:29).

[0429] In further experiments, we investigated the effect of knocking out IL11RA1 in this chemical-induced acute kidney injury model. Cre-lox mice, in which IL11RA1 is knocked out specifically in fibroblasts in response to tamoxifen, were treated with tamoxifen to delete IL-11RA1 in col1A2-positive cells, and kidney injury was induced with folic acid (or vehicle treatment) as described above. After 21 days, collagen content was assessed using an HPA assay, and kidney function was assessed by analysis of serum urea concentration. The results are shown in Figure 6. We found that knocking out IL11RA1 in fibroblasts could protect mice from folic acid-induced kidney injury, indicating that IL-11-mediated signaling plays a central role in fibroblasts in the pathology of kidney injury and subsequent kidney dysfunction and secondary fibrosis.

[0430] In another experiment, mice were treated with folic acid as described above and 10 mg / kg Enx203 was administered twice weekly starting on day 3. After 28 days, body weight, kidney weight, collagen content in the kidney, urine volume and gross kidney morphology were assessed. The results are shown in Figure 7. At the end of the experiment, mice treated with anti-IL-11 antibody therapy showed increased kidney weight, decreased collagen content in the kidney, normalized urine volume and improved gross kidney morphology compared to mice administered the IgG control.

[0431] Further experiments were performed in the chemical-induced chronic kidney injury model. Briefly, mice were treated with folic acid to induce kidney injury as described above, and then left untreated for 21 days. 21 days after acute kidney injury was induced by treatment with folic acid, the blood urea nitrogen concentration increased by about 2.5 times (Figure 8B) and the urinary albumin / creatine ratio (ACR) increased by about 2.9 times (Figure 11, lower panel), confirming the development of chronic kidney disease. Kidney weight was reduced by about 33% compared to the initial weight (Figure 8C), and collagen content increased by about 2.8 times (Figure 8A).

[0432] Starting on day 21, mice were treated twice weekly with 10 mg / kg Enx203 or IgG control. Mice were euthanized on the experimental days indicated and assessed for kidney collagen content, serum urea concentration, kidney weight, and gross kidney morphology. Results are shown in Figure 8. Enx203 treatment reduced kidney collagen content (Figure 8A) and serum urea concentration, with a 51% recovery of blood urea nitrogen (BUN) across groups by week 12 of antibody therapy (P<0.001 compared to IgG; Figure 8B).

[0433] Importantly, kidney weight increased over time in Enx203-treated mice, but collagen content in the kidney decreased over time (Fig. 8C and 8A), and serum urea concentration also decreased over time (Fig. 8B). These results indicate that Enx203 treatment suppressed the folate-induced renal tissue injury response and, moreover, potently promoted the regeneration of functional renal tissue and reversed the renal injury phenotype. Kidneys removed from Enx203-treated mice on days 84 and 105 showed characteristics more similar to healthy kidneys than those of IgG control mice (Fig. 8D and 8E).

[0434] Histological analysis of kidney sections prepared from mice with folic acid-induced nephropathy revealed that treatment with Enx203 restored normal renal parenchyma (54% reversal of parenchymal loss by week 12 (P<0.001)) as well as renal cortical thickness, volume, and morphology (Figures 9 and 10).

[0435] In addition, in this experiment, the body weight of the mice was monitored over the course of the experiment, and it was found that treatment with Enx203 increased the body weight of the folic acid-treated mice to the same extent as the untreated control mice (Figure 11, top). The urinary albumin / creatinine ratio (ACR) of the mice was also monitored on days 21, 84, and 105 of the experiment. The results are shown in Figure 11 (bottom). It was found that treatment with Enx203 significantly reduced the urinary albumin / creatinine ratio of the folic acid-treated mice (by about 50% within 12 weeks).

[0436] Taken together, our results demonstrate that antagonism of IL-11-mediated signaling can reverse renal fibrosis and achieve extensive regeneration of the renal parenchyma, which is responsible for the remarkable recovery of renal dysfunction.

[0437] Example 3: Molecular level investigation of suppression / reversal of renal injury by antagonists of IL-11-mediated signaling Further investigations were performed to evaluate the role of IL-11-mediated signaling in renal tissue function and renal injury responses.

[0438] Because the transformation of renal tubular epithelial cells into a mesenchymal cell-like phenotype is involved in renal parenchymal damage and renal dysfunction associated with renal injury (see, e.g., Lovisa et al. Nat. Med. (2015) 21, 998-1009), we investigated whether IL-11-mediated signaling plays a role in the epithelial-mesenchymal transition of renal tubular epithelial cells.

[0439] In vitro, human primary renal tubular epithelial cells (TECs) were stimulated with TGFB1, IL-11 (5 ng / ml, 24 h) or TGFB1+Enx203 (2 μg / ml, 24 h) and the percentage of ACTA2-positive cells and collagen expression were assessed using the Operetta high-content imaging platform.

[0440] The results are shown in Figure 12. We found that TGFB1 and IL-11 increased the proportion of renal tubular epithelial cells expressing ACTA2 and increased the expression of collagen I by these cells. We also found that treatment with Enx203 reduced the expression of ACTA2 and collagen I in renal tubular epithelial cells in response to TGFB1 or IL-11.

[0441] In a separate experiment, human primary renal tubular epithelial cells (TECs) were stimulated in vitro with various concentrations of IL-11 (2.5 ng / ml, 5 ng / ml, 10 ng / ml), TGFB1 (5 ng / ml), or TGFB1 (5 ng / ml) + Enx203 (2 μg / ml) for 24 h, and actin stress fiber formation was visualized with rhodamine phalloidin (Figure 13, top panel) and collagen was also visualized (Figure 13, bottom panel) using the Operetta high-content imaging system.

[0442] As mentioned previously, SNAIL has recently been shown to be a key determinant of renal tubular epithelial cell dysfunction following the onset of acute kidney injury. Expression of SNAIL in renal tubular epithelial cells is associated with renal tubular epithelial cell dysfunction and proliferation failure. Human primary renal tubular epithelial cells (TECs) were stimulated in vitro with various concentrations of IL-11 (5 ng / ml), TGFB1 (5 ng / ml), TGFB1 (5 ng / ml) + Enx203 (2 μg / ml), or TGFB1 (5 ng / ml) + IgG control (2 μg / ml) for 24 h and analyzed for ACTA2 or SNAIL expression using the Operetta high content imaging system.

[0443] The results are shown in Figure 14. Treatment with IL-11 or TGFB1 was found to induce partial epithelial to mesenchymal transition (EMT) and expression of ACTA2 and SNAIL, whereas the induction of ACTA2 and SNAIL was strongly suppressed in the presence of Enx203.

[0444] Cell lysates were prepared from each cell line and analyzed for SNAIL expression by Western blot. Western blots for GAPDH were performed separately as a protein loading control. The results are shown in Figure 15. It was shown that SNAIL expression was upregulated in response to TGFB1 and IL-11, and TGFB-mediated upregulation of SNAIL was completely blocked in the presence of Enx203.

[0445] Thus, TGFB1-mediated induction of SNAIL was found to be dependent on IL-11.

[0446] In addition, protein lysates were prepared from kidney tissues of mice from the previous experiment, and the expression of E-cadherin, a standard marker for identifying epithelial cells, was examined by Western blot. In this experiment, mice were treated with folic acid as described above to induce kidney injury, and from day 21 onwards, the mice were treated twice weekly with 10 mg / kg Enx203 or IgG control.

[0447] The results are shown in Figure 16. It was found that folic acid-induced renal injury reduced the expression of E-cadherin, and treatment with Enx203 restored the expression of E-cadherin.

[0448] Example 4: Analysis of the effects of inhibiting IL-11-mediated signaling in a physically induced renal injury model A mouse model of acute kidney injury was induced by unilateral ureteral ligation (UUO). Briefly, mice were subjected to sham surgery or ureteral obstruction in one ureter.

[0449] Mice were treated with ENx203 (designated "3C6" in FIG. 17B) or IgG control antibody (20 mg / kg each) on days 4, 7, and 9 after surgery. Mouse weights were monitored throughout the experimental period, and collagen content in the kidney was assessed using an HPA assay 10 days after unilateral ureteral ligation.

[0450] The results are shown in Figure 17. Mice treated with Enx203 had reduced collagen content in the kidney and increased body weight compared to mice treated with the IgG control antibody after unilateral ureteral ligation.

[0451] Example 5: Analysis of the effect of inhibition of IL-11-mediated signaling in a cisplatin-induced renal injury model Renal injury was induced in 10-week-old C57Bl / 6J mice by administering 7 mg / kg cisplatin once a week for 4 consecutive weeks, whereas the control group did not receive cisplatin.

[0452] Mice were administered a 10 mg / kg dose of X203 (an antibody capable of binding to mouse IL-11 (and human IL-11) and inhibiting IL-11-mediated signaling) or an isotype-matched IgG control antibody, or saline (control), by intraperitoneal injection twice a week starting from week 1. Samples were taken from the mice after 8 weeks for analysis.

[0453] Total collagen in the kidney was quantified by colorimetric detection of hydroxyproline using the Quickzyme Total Collagen Quantitation Assay Kit (Quickzyme Biosciences). The results are shown in Figure 18B. Cisplatin treatment increased collagen content in the kidney. Kidneys from mice treated with cisplatin and administered anti-IL-11 neutralizing antibody contained less collagen than kidneys from cisplatin-treated mice administered IgG control antibody. Furthermore, RNA expression levels of Col3, fibronectin, MMP2, TIMP, CCL5, and CCL2 were analyzed by qPCR in kidneys from mice with various treatments.

[0454] Briefly, total RNA was extracted from snap-frozen kidney tissues using Trizol (Invitrogen) and the RNeasy Mini kit (Qiagen). PCR amplification was performed using the iScript cDNA synthesis kit (Bio-Rad). Gene expression analysis was performed in duplicate for 40 cycles by TaqMan (Applied Biosystems) or SYBR Green (Qiagen) methods using StepOnePlus (Applied Biosystems). Expression data were normalized to GAPDH mRNA expression levels, and fold changes were calculated compared to saline-treated control subjects.

[0455] The results are shown in Figures 18C to 18H. Treatment with cisplatin increased the expression of Col3, fibronectin, MMP2, TIMP, CCL5, and CCL2. Kidneys from mice treated with cisplatin and administered anti-IL-11 neutralizing antibody tended to have lower expression of Col3, fibronectin, MMP2, TIMP, CCL5, and CCL2 than kidneys from cisplatin-treated mice administered IgG control antibody.

Claims

1. A pharmaceutical for treating, preventing or recovering from drug-induced nephropathy, comprising as an active ingredient a drug capable of suppressing interleukin-11 (IL-11)-mediated signal transduction, the agent capable of inhibiting interleukin-11 (IL-11)-mediated signaling is (i) an anti-IL-11 antagonist antibody or an antigen-binding fragment thereof against IL-11-mediated signaling, or (ii) an anti-IL-11Rα antagonist antibody or an antigen-binding fragment thereof against IL-11-mediated signaling, The pharmaceutical agent, wherein the drug-induced renal damage is renal damage characterized by transformation of renal tubular epithelial cells into a mesenchymal cell-like phenotype.

2. The pharmaceutical composition of claim 1, wherein the drug-induced renal injury is drug-induced acute renal injury.

3. The pharmaceutical product according to claim 1 or 2, wherein the drug-induced renal damage is drug-induced nephrotoxicity.

4. The pharmaceutical product according to any one of claims 1 to 3, wherein the drug-induced nephropathy is nephropathy induced by a drug selected from the group consisting of diuretics, beta-blockers, vasodilators, ACE inhibitors, aminoglycoside antibiotics, gentamicin, amphotericin B, cisplatin, NSAIDs, aspirin, ibuprofen, diclofenac, cyclosporine, lithium salts, cyclophosphamide, sulfonamides, methotrexate, acyclovir, diethylene glycol, triamterene, beta-lactam antibiotics, vancomycin, rifampicin, ciprofloxacin, ranitidine, cimetidine, furosemide, thiazides and phenytoin.

5. A pharmaceutical agent described in any one of claims 1 to 3, wherein the drug-induced nephropathy is nephropathy induced by a chemotherapy agent.

6. The chemotherapeutic agent is selected from the group consisting of alkylating agents, cisplatin, carboplatin, mechlorethamine, cyclophosphamide, chlorambucil, ifosfamide, purine or pyrimidine metabolic antagonists, azathioprine, mercaptopurine, alkaloids, terpenoids, vinca alkaloids, vincristine, vinblastine, vinorelbine, vindesine, podophyllotoxin, etoposide, teniposide, taxanes, paclitaxel, docetaxel, topoisomerase inhibitors, type I topoisomerase inhibitors, camptothecin, irinotecan, topotecan, type II topoisomerase inhibitors, amsacrine, etoposide, etoposide phosphate, teniposide, antitumor antibiotics, anthracycline antibiotics, dactinomycin 6. The pharmaceutical product of claim 5, wherein the therapeutic agent is selected from the group consisting of rifabutin, doxorubicin, epirubicin, bleomycin, rapamycin, antibody-based drugs, anti-VEGF antibodies, anti-TNFα antibodies, anti-IL-2 antibodies, anti-GpIIb / IIIa antibodies, anti-CD52 antibodies, anti-CD20 antibodies, anti-RSV antibodies, anti-HER2 / neu antibodies, anti-TNF receptor antibodies, anti-EGFR antibodies, monoclonal antibodies or antibody fragments, cetuximab, panitumumab, infliximab, basiliximab, bevacizumab, abciximab, daclizumab, gemtuzumab, alemtuzumab, rituximab, palivizumab, trastuzumab, etanercept, adalimumab, nimotuzumab, EGFR inhibitors, erlotinib, cetuximab, and gefitinib.

7. The pharmaceutical product described in claim 5 or 6, wherein the chemotherapeutic agent is selected from the group consisting of cisplatin, carboplatin, mechlorethamine, cyclophosphamide, chlorambucil, and ifosfamide.

8. The pharmaceutical agent according to any one of claims 5 to 7, wherein the drug-induced nephropathy is cisplatin-induced nephropathy or cisplatin-induced nephrotoxicity.

9. The pharmaceutical agent according to any one of claims 1 to 8, wherein the drug-induced nephropathy is nephropathy characterized by damage to renal tubular epithelial cells (TEC).

10. A pharmaceutical composition for suppressing SNAIL expression in the kidney, comprising an active ingredient that can suppress interleukin-11 (IL-11)-mediated signal transduction, The pharmaceutical agent, wherein the drug capable of inhibiting interleukin-11 (IL-11)-mediated signal transduction is (i) an anti-IL-11 antagonist antibody or an antigen-binding fragment thereof against IL-11-mediated signal transduction, or (ii) an anti-IL-11Rα antagonist antibody or an antigen-binding fragment thereof against IL-11-mediated signal transduction.

11. A pharmaceutical for inhibiting or restoring the transformation of renal tubular epithelial cells (TEC) into a mesenchymal cell-like phenotype, characterized in that the pharmaceutical contains as an active ingredient a drug capable of inhibiting interleukin-11 (IL-11)-mediated signal transduction, The pharmaceutical agent, wherein the drug capable of inhibiting interleukin-11 (IL-11)-mediated signal transduction is (i) an anti-IL-11 antagonist antibody or an antigen-binding fragment thereof against IL-11-mediated signal transduction, or (ii) an anti-IL-11Rα antagonist antibody or an antigen-binding fragment thereof against IL-11-mediated signal transduction.

12. The pharmaceutical product according to any one of claims 1 to 11, wherein the drug is capable of blocking or reducing binding of interleukin-11 (IL-11) to interleukin-11 receptor (IL-11R).

13. The pharmaceutical product of claim 12, wherein the interleukin-11 receptor is or comprises IL-11Rα.

14. The pharmaceutical product according to any one of claims 1 to 13, wherein the pharmaceutical product is administered to a subject in which expression of interleukin-11 (IL-11) or IL-11 receptor (IL-11R) is upregulated.

15. The pharmaceutical product according to any one of claims 1 to 14, wherein the pharmaceutical product is administered to a subject in which upregulation of expression of interleukin-11 (IL-11) or IL-11 receptor (IL-11R) has been confirmed.

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