NZ753260A - Methods for reducing proteinuria in a human subject suffering from immunoglobulin a nephropathy - Google Patents

Abstract

In one aspect, the invention provides methods for reducing proteinuria in a human subject suffering, or at risk of developing Immunoglobulin A Nephropathy (IgAN). The methods comprise the step of administering, to a subject in need thereof, an amount of a MASP-2 inhibitory antibody effective to inhibit MASP-2-dependent complement activation.

Description

METHODS FOR REDUCING PROTEINURIA IN A HIHVIAN SUBJECT SUFFERING FROM IM]VIUNOGLOBULIN A NEPHROPATHY STATEMENT ING SEQUENCE LISTING The sequence listing ated with this application is provided in text format in lieu of a paper copy and is hereby incorporated by reference into the specification. The name of the text file containing the sequence listing is MP_1_0269_PCT_Sequence_Listing_20171013_ST25. The text file is 136 KB, was created on October 10, 2017, and is being submitted via EFS-Web with the filing of the specification.

BACKGROUND The complement system provides an early acting mechanism to initiate, amplify and orchestrate the immune response to ial infection and other acute insults (MK. Liszewski and JP. Atkinson, 1993, in Fundamental Immunology, Third n, edited by WE. Paul, Raven Press, Ltd., New York), in humans and other vertebrates.

While complement activation provides a le first-line defense against potential pathogens, the ties of complement that promote a protective immune response can also represent a potential threat to the host (KR. Kalli, et al., Springer Semin. lmmunopalhol.15:417-431, 1994, BR Morgan, Eur. J. Clinical lnveslig. -228, 1994). For example, C3 and C5 proteolytic ts recruit and activate neutrophils.

While indispensable for host defense, ted neutrophils are indiscriminate in their release of destructive enzymes and may cause organ damage. In addition, complement activation may cause the deposition of lytic complement components on nearby host cells as well as on microbial targets, resulting in host cell lysis.

The ment system has also been implicated in the pathogenesis of numerous acute and chronic disease , including: myocardial infarction, stroke, ARDS, reperfusion injury, septic shock, capillary leakage following thermal burns, postcardiopulmonary bypass inflammation, transplant rejection, rheumatoid arthritis, 2017/056386 multiple sclerosis, myasthenia gravis, and Alzheimer's disease. In almost all of these conditions, complement is not the cause but is one of several factors involved in pathogenesis. Nevertheless, complement activation may be a major pathological mechanism and represents an effective point for clinical control in many of these disease states. The growing recognition of the importance of complement-mediated tissue injury in a variety of disease states underscores the need for effective complement inhibitory drugs. To date, Eculizumab (Solaris®), an antibody against C5, is the only complement- targeting drug that has been approved for human use. Yet, C5 is one of several effector molecules d "downstream" in the complement system, and blockade of C5 does not inhibit tion of the complement system. Therefore, an inhibitor of the initiation steps of complement activation would have significant advantages over a "downstream" ment inhibitor.

Currently, it is widely accepted that the complement system can be activated through three distinct pathways: the classical y, the lectin pathway, and the alternative pathway. The classical pathway is usually triggered by a compleX composed of host antibodies bound to a foreign particle (1'.e., an antigen) and thus requires prior exposure to an antigen for the generation of a specific antibody response. Since activation of the classical pathway depends on a prior adaptive immune response by the host, the classical y is part of the ed immune system. In contrast, both the 2O lectin and alternative ys are independent of adaptive ty and are part of the innate immune system.

The activation of the complement system results in the sequential activation of serine protease zymogens. The first step in activation of the classical pathway is the g of a ic recognition le, Clq, to antigen-bound IgG and IgM molecules. Clqis ated with the Clr and Cls serine protease proenzymes as a compleX called Cl. Upon binding of Clq to an immune compleX, autoproteolytic cleavage of the Arg-Ile site of Clr is followed by Clr-mediated cleavage and activation of Cls, which thereby acquires the ability to cleave C4 and C2. C4 is cleaved into two fragments, designated C4a and C4b, and, similarly, C2 is cleaved into C2a and C2b. C4b fragments are able to form nt bonds with adjacent yl or amino groups and generate the C3 convertase (C4b2a) through noncovalent interaction with the C2a fragment of activated C2. C3 convertase (C4b2a) tes C3 by proteolytic cleavage into C3a and C3b subcomponents leading to generation of the C5 convertase (C4b2a3b), 2017/056386 which, by cleaving C5 leads to the formation of the membrane attack complex (C5b combined with C6, C7, C8 and C-9, also ed to as "MAC") that can disrupt cellular membranes leading to cell lysis. The activated forms of C3 and C4 (C3b and C4b) are covalently deposited on the foreign target surfaces, which are recognized by complement receptors on multiple phagocytes.

Independently, the first step in activation of the complement system through the lectin pathway is also the binding of specific recognition les, which is ed by the activation of associated serine protease proenzymes. However, rather than the binding of immune complexes by Clq, the recognition molecules in the lectin pathway comprise a group of carbohydrate-binding proteins (mannan-binding lectin (lVfl3L), H-f1colin, M-ficolin, lin and C-type lectin CL-l l), collectively referred to as lectins. See J. Lu et al., m. Biophys. Acta 1572:387-400, (2002); Holmskov et al., Annu. Rev. Immunol. 21:547-578 (2003); Teh et al., Immunology [01:225—232 (2000)).

See also J. Luet et al., Biochim s Acta 1572:387-400 (2002); Holmskov et al, Annu Rev Immunol 21:547-578 ; Teh et al., Immunology 1012225-232 (2000); Hansen et al, J. l 185(10):6096-6104 (2010).

Ikeda et a1. first demonstrated that, like Clq, MBL could activate the ment system upon binding to yeast mannan-coated erythrocytes in a C4—dependent manner (Ikeda et al., J. Biol. Chem. 262:7451-7454, (1987)). MBL, a member of the collectin n family, is a calcium-dependent lectin that binds carbohydrates with 3- and 4-hydroxy groups oriented in the equatorial plane of the pyranose ring. Prominent ligands for MBL are thus D-mannose and N—acetyl-D-glucosamine, while carbohydrates not fitting this steric requirement have undetectable affinity for MBL (Weis et al., Nature 360:127-134, (1992)). The interaction between MBL and monovalent sugars is extremely weak, with dissociation constants typically in the single-digit millimolar range.

MBL achieves tight, c binding to glycan ligands by avidity, i.e., by interacting aneously with multiple monosaccharide residues located in close proximity to each other (Lee et al., Archiv. Biochem. Biophys. 299:129-136, (1992)). MBL recognizes the carbohydrate patterns that commonly decorate microorganisms such as bacteria, yeast, parasites and certain viruses. In contrast, 1Vfl3L does not recognize D-galactose and sialic acid, the penultimate and ultimate sugars that usually decorate "mature" complex glycoconjugates present on mammalian plasma and cell e glycoproteins. This binding specificity is thought to e recognition of "foreign" surfaces and help protect from "self-activation." However, MBL does bind with high y to clusters of high-mannose "precursor" glycans on N—linked glycoproteins and ipids sequestered in the endoplasmic reticulum and Golgi of mammalian cells (Maynard eta1., J. Biol.

Chem. 257:3788-3794, (1982)), Therefore, damaged cells are potential targets for lectin pathway tion via MBL binding.

The ficolins possess a different type of lectin domain than MBL, called the gen-like domain. Ficolins bind sugar residues in a Ca++-independent manner. In humans, three kinds of ficolins (L-ficolin, M-ficolin and H-ficolin) have been identified.

The two serum s, L-ficolin and H-ficolin, have in common a specificity for N—acetyl-D-glucosamine, however, H-ficolin also binds N—acetyl-D-galactosamine. The ence in sugar specificity of L-ficolin, H-ficolin, CL-ll, and MBL means that the different lectins may be complementary and target different, though overlapping, glycoconjugates. This concept is supported by the recent report that, of the known lectins in the lectin pathway, only L-ficolin binds cally to lipoteichoic acid, a cell wall onjugate found on all Gram-positive bacteria (Lynch et al., J. Immunol. 172:1198-1202, (2004)). The collectins (i.e., lVfl3L) and the ficolins bear no significant similarity in amino acid sequence. However, the two groups of proteins have similar domain organizations and, like Clq, assemble into oligomeric structures, which maximize the possibility of multisite binding. 2O The serum concentrations of MBL are highly variable in healthy tions and this is genetically controlled by polymorphisms/mutations in both the er and coding s of the lVfl3L gene. As an acute phase protein, the expression of MBL is further upregulated during inflammation. L-ficolin is t in serum at concentrations similar to those of lVfl3L. Therefore, the L-ficolin branch of the lectin pathway is potentially comparable to the MBL arm in strength. lVfl3L and ficolins can also function as opsonins, which allow phagocytes to target lVfl3L- and ficolin-decorated surfaces (see Jack et al., J Leukoc Biol., 77(3):328-36 (2004), Matsushita and Fujita, Immunobiolog, 205(4—5):490—7 (2002), Aoyagi et al., J Immunol, l74(l):418—25(2005). This opsonization requires the interaction of these proteins with yte receptors (Kuhlman et al., J. Exp. Med. 169:1733, (1989); Matsushita et al., J. Biol. Chem. 271:2448-54, (1996)), the identity of which has not been ished.

Human MBL forms a specific and ffinity interaction through its collagen-like domain with unique Clr/Cls-like serine proteases, termed MBL-associated serine proteases ). To date, three MASPs have been described. First, a single enzyme "MASP" was identified and characterized as the enzyme responsible for the initiation of the complement cascade (i.e., cleaving C2 and C4) (Matsushita et al., J Exp Med 176(6):1497-1502 (1992); Ji et al., J. Immunol. [50:571-578, (1993)). It was subsequently determined that the MASP activity was, in fact, a mixture of two proteases: MASP—1 and MASP-2 (Thiel et al., Nature 6-510, ). However, it was demonstrated that the MBL-MASP-2 x alone is sufficient for complement activation (Vorup-Jensen et al., J. l. 165:2093-2100, (2000)). Furthermore, only MASP-2 cleaved C2 and C4 at high rates (Ambrus et al., J. Immunol. 170:1374-1382, (2003)). Therefore, MASP-2 is the protease responsible for activating C4 and C2 to generate the C3 convertase, C4b2a. This is a cant difference from the Cl complex of the classical pathway, where the coordinated action of two specific serine ses (Clr and Cls) leads to the activation of the complement system. In on, a third novel protease, , has been isolated (Dahl, M.R., et al., ty 15:127-35, 2001). MASP—l and MASP-3 are alternatively spliced products of the same gene.

MASPs share identical domain organizations with those of Clr and Cls, the enzymatic components of the Cl complex (Sim et al., Biochem. Soc. Trans. 28:545, (2000)). These domains include an N—terminal Clr/Cls/sea urchin VEGF/bone morphogenic protein (CUB) domain, an epidermal growth factor—like domain, a second 2O CUB domain, a tandem of complement control protein s, and a serine protease . As in the Cl proteases, activation of MASP-2 occurs through cleavage of an Arg-Ile bond adjacent to the serine protease domain, which splits the enzyme into disulfide-linked A and B chains, the latter consisting of the serine protease domain. lVfl3L can also associate with an alternatively sliced form of MASP-2, known as MBL-associated protein of 19 kDa (MApl9) or small lVfl3L-associated protein (sMAP), which lacks the catalytic activity of MASP-2. (Stover, J. Immunol. 162:3481-90, (1999), Takahashi et al., Int. Immunol. 11:859-863, ). MAp19 comprises the first two s of MASP-2, followed by an extra sequence of four unique amino acids. The function of MApl9 is unclear (Degn et al., J Immunol. Methods, 2011). The MASP-1 and MASP-2 genes are located on human chromosomes 3 and 1, respectively eble et al., Immunobiology 205:455-466, (2002)).

Several lines of evidence suggest that there are different MBL—MASP complexes and a large fraction of the MASPs in serum is not complexed with MBL (Thiel, et al., J.

Immunol. [65:878-887, (2000)). Both H- and L-flcolin bind to all MASPs and activate the lectin complement pathway, as does MBL (Dahl et al., Immunity 15:127-35, (2001); Matsushita et a1., J. Immunol. 168:3502-3506, (2002)). Both the lectin and classical pathways form a common C3 tase (C4b2a) and the two pathways ge at this step.

The lectin pathway is widely thought to have a major role in host defense against infection in the naive host. Strong evidence for the involvement of MBL in host defense comes from analysis of patients with decreased serum levels of functional MBL (Kilpatrick, Biochim. Biophys. Acta 1572:401-413, (2002)). Such patients display susceptibility to recurrent bacterial and fungal infections. These symptoms are usually evident early in life, during an apparent window of vulnerability as maternally d dy titer wanes, but before a full repertoire of antibody responses develops. This syndrome often results from mutations at several sites in the collagenous portion of lVfl3L, which interfere with proper formation of lVfl3L oligomers. However, since MBL can function as an opsonin independent of complement, it is not known to what extent the increased susceptibility to ion is due to impaired complement activation.

All three pathways (i.e., the classical, lectin and alternative) have been thought to converge at C5, which is cleaved to form products with multiple proinflammatory effects, The converged pathway has been referred to as the al complement pathway. C5a is 2O the most potent anaphylatoxin, ng alterations in smooth muscle and vascular tone, as well as vascular permeability. It is also a ul chemotaxin and activator of both neutrophils and monocytes. C5a-mediated ar activation can significantly amplify inflammatory responses by inducing the release of multiple additional inflammatory mediators, ing cytokines, hydrolytic enzymes, arachidonic acid metabolites, and reactive oxygen s. C5 cleavage leads to the formation of C5b-9, also known as the membrane attack complex (MAC). There is now strong evidence that sublytic MAC deposition may play an important role in inflammation in addition to its role as a lytic pore-forming complex.

In addition to its ial role in immune defense, the complement system contributes to tissue damage in many al conditions. Although there is extensive evidence implicating both the classical and alternative complement ys in the pathogenesis of non-infectious human diseases, the role of the lectin pathway is just ing to be evaluated. Recent studies provide evidence that activation of the lectin pathway can be sible for complement activation and related inflammation in ischemia/reperfusion injury. Collard et al. (2000) reported that cultured endothelial cells ted to oxidative stress bind MBL and show deposition of C3 upon exposure to human serum (Collard et al., Am. J. Pathol. 156:1549-1556, (2000)). In addition, treatment of human sera with blocking anti-MBL monoclonal antibodies inhibited lVfl3L binding and complement activation. These findings were extended to a rat model of myocardial ischemia-reperfusion in which rats treated with a blocking antibody directed against rat MBL showed significantly less myocardial damage upon occlusion of a coronary artery than rats treated with a control dy (Jordan et al., Circulation 104:1413-1418, ). The molecular mechanism of MBL binding to the vascular endothelium after oxidative stress is unclear; a recent study suggests that activation of the lectin pathway after oxidative stress may be mediated by MBL binding to ar elial cytokeratins, and not to glycoconjugates (Collard et al., Am. J. Pathol. 159:1045-1054, (2001)). Other studies have implicated the cal and alternative pathways in the pathogenesis of ia/reperfusion injury and the role of the lectin pathway in this disease remains controversial (Riedermann, N.C., et al, Am. J. Pathol. 162:363-367, 2003).

Fibrosis is the formation of excessive connective tissue in an organ or tissue, ly in response to damage or injury. A hallmark of fibrosis is the production of excessive extracellular matrix ing local trauma. The normal physiological response to injury results in the deposition of connective tissue, but this initially beneficial reparative process may persist and become pathological, altering the architecture and function of the tissue. At the cellular level, epithelial cells and asts proliferate and differentiate into myofibroblasts, resulting in matrix contraction, increased rigidity, microvascular compression, and hypoxia. An influx of inflammatory cells, including macrophages and lymphocytes, results in cytokine release and amplifies the deposition of collagen, fibronectin and other molecular s of fibrosis. Conventional therapeutic approaches have y been targeted towards the inflammatory s of s, using osteroids and immunosuppressive drugs. Unfortunately, these anti- inflammatory agents have had little to no clinical effect. Currently there are no effective treatments or therapeutics for fibrosis, but both animal studies and tal human reports suggest that fibrotic tissue damage may be reversed (Tampe and Zeisberg, Nat Rev Nephrol, Vol 10:226-237, 2014).

The kidney has a limited capacity to recover from injury. Various renal ogies result in local inflammation that causes scarring and fibrosis of renal tissue. The perpetuation of inflammatory stimuli drives tubulointerstitial inflammation and fibrosis and progressive renal functional impairment in chronic kidney disease. Its progression to end-stage renal e is associated with significant morbidity and mortality. Since tubulointerstitial is is the common end point of multiple renal pathologies, it represents a key target for therapies aimed at preventing renal failure. Risk factors (e.g., proteinuria) independent of the primary renal disease contribute to the pment of renal fibrosis and loss of renal ory function by driving local inflammation, which in turn enhances disease progression.

In view of the role of fibrosis in many diseases and disorders, such as, for e, tubulointerstitial fibrosis leading to chronic kidney disease, there is a pressing need to develop therapeutically effective agents for treating es and conditions caused or exacerbated by fibrosis. In further view of the paucity of new and existing treatments targeting inflammatory pro-fibrotic pathways in renal disease, there is a need to p therapeutically effective agents to treat, t, prevent and/or reverse renal fibrosis and thereby prevent progressive chronic kidney disease.

SUMMARY This summary is provided to introduce a selection of concepts in a simplified form that are further described below in the Detailed Description. This summary is not ed to identify key features of the claimed subject matter, nor is it intended to be used as an aid in determining the scope of the claimed subject matter.

In a particular aspect, the present invention provides the use of a MASP-2 tory monoclonal antibody, or antigen-binding fragment thereof that specifically binds to human MASP-2 and inhibits MASPdependent complement activation in an amount effective to improve renal function in the manufacture of a medicament for ng a human subject suffering from steroid-dependent lupus nephritis (LN), wherein the composition is to be administered in an amount ient to improve renal function and decrease the corticosteroid dosage in said subject and wherein the MASP-2 inhibitory antibody or n-binding fragment thereof comprises a heavy chain variable region comprising CDR-H1, CDR-H2 and CDR-H3 of the amino acid sequence set forth as SEQ ID NO:67 and a light chain variable region comprising CDR-L1, CDR-L2 and CDR-L3 of the amino acid sequence set forth as SEQ ID NO:69. (followed by page 8a) In another particular aspect, the present invention provides the use of a MASP-2 inhibitory onal antibody, or antigen-binding fragment thereof, comprising a heavy chain variable region comprising CDR-H1, CDR-H2 and CDR-H3 of the amino acid sequence set forth as SEQ ID NO:67 and a light chain variable region comprising CDR-L1, CDR-L2 and CDR-L3 of the amino acid ce set forth as SEQ ID NO:69 in the manufacture of a medicament for reducing nuria in a human t suffering from steroid-dependent Immunoglobulin A Nephropathy (IgAN) wherein the medicament is adapted for administration according to a dosage regimen as follows: a. the medicament comprising about 4 mg/kg of said antibody is to be administered to a subject suffering from IgAN once weekly intravenously for a treatment period of at least 12 weeks; wherein the medicament reduces proteinuria in said human subject.

In one aspect, the invention provides a method for treating, inhibiting, alleviating or preventing fibrosis in a mammalian subject suffering, or at risk of developing a disease or disorder caused or exacerbated by fibrosis and/or inflammation, comprising administering to the subject an amount of a MASP-2 inhibitory agent ive to t fibrosis. In one embodiment, the MASP-2 inhibitory agent is a MASP-2 dy or fragment thereof. In one embodiment, the MASP-2 inhibitory agent is a MASP-2 monoclonal antibody, or fragment thereof that specifically binds to a portion of SEQ ID NO:6. In one embodiment, the MASP-2 inhibitory agent selectively inhibits lectin pathway complement activation without substantially inhibiting C1q-dependent complement activation. In one embodiment, the subject is suffering from a disease or [FOLLOWED BY PAGE 9] - 8a - disorder caused by or exacerbated by at least one of (i) fibrosis and/or inflammation associated with an ischemia reperfusion injury, (ii) renal fibrosis and/or renal inflammation (e.g., tubulointerstitial fibrosis, chronic kidney disease, chronic renal failure, ular disease (e.g., focal segmental glomerulosclerosis), an immune complex disorder (e.g., IgA nephropathy, membraneous nephropathy), lupus nephritis, nephrotic syndrome, diabetic nephropathy, tubulointerstitial damage and glomerulonepthlitis (e.g., C3 glomerulopathy), (iii) pulmonary fibrosis and/or inflammation (e.g., chronic obstructive pulmonary disease, cystic fibrosis, pulmonary fibrosis associated with derma, iectasis and pulmonary ension), (iv) hepatic fibrosis and/or inflammation (e.g., cirrhosis, oholic fatty liver disease (steatohepatitis)), liver fibrosis secondary to alcohol abuse, liver fibrosis secondary to acute or chronic tis, biliary disease and toxic liver injury (e.g., hepatotoxicity due to drug-induced liver damage induced by acetaminophen or other drug), (v) cardiac fibrosis and/or inflammation (e.g., cardiac fibrosis, myocardial infarction, valvular fibrosis, atrial fibrosis, endomyocardial s arrhythmogenic right ventricular cardiomyopathy (ARVC), (vi) vascular fibrosis (e. g., vascular disease, an atherosclerotic vascular disease, vascular stenosis, restenosis, vasculitis, phlebitis, deep vein osis and abdominal aortic aneurysm), (vii) s of the skin (e.g., excessive wound healing, scleroderma, systemic sclerosis, keloids, connective tissue es, scarring, and hypertrophic scars), (viii) fibrosis of the joints (e.g., arthrofibrosis), (ix) fibrosis of the l nervous system (e.g., stroke, traumatic brain injury and spinal cord injury), (x) fibrosis of the digestive system (e.g., Crohn’s disease, pancreatic fibrosis and ulcerative colitis), (xi) ocular fibrosis (e.g., anterior subcapsular ct, posterior capsule opacification, macular degeneration, and retinal and vitreal retinopathy), (xii) fibrosis of musculoskeletal soft- tissue ures (e.g., adhesive capsulitis, ren’s contracture and myelofibrosis), (xiii) fibrosis of the reproductive organs (e.g., endometriosis and Peyronie’s disease), (xiv) a chronic infectious disease that causes fibrosis and/or inflammation (e.g., alpha virus, Hepatitis A, Hepatitis B, Hepatitis C, ulosis, HIV and influenza), (xv) an autoimmune disease that causes fibrosis and/or ation (e.g., scleroderrna and systemic lupus erythematosus (SLE), (xvi) ng associated with trauma (e.g., wherein the scarring associated with trauma is selected from the group consisting of surgical complications (e,g., surgical adhesions wherein scar tissue can form between internal organs causing cture, pain and can cause infertility), chemotherapeutic drug-induced fibrosis, radiation-induced fibrosis and scarring associated with burns), or (xvii) organ transplant, breast fibrosis, muscle s, retroperitoneal fibrosis, d fibrosis, lymph node fibrosis, bladder fibrosis and pleural s.

In r aspect, the present invention provides a method for treating, inhibiting, alleviating or preventing renal fibrosis in a mammalian subject suffering, or at risk of ping a disease or disorder caused or bated by renal fibrosis and/or inflammation, comprising administering to the subject an amount of a MASP—Z inhibitory agent effective to inhibit renal fibrosis. In one embodiment, the MASP-2 inhibitory agent is a MASP—2 dy or fragment thereof. In one embodiment, the MASP-2 inhibitory agent is a MASP-2 monoclonal antibody, or fragment thereof that specifically binds to a portion of SEQ ID NO:6. In one embodiment, the MASP-Z dy or fragment thereof specifically binds to a polypeptide comprising SEQ ID N06 with an y of at least times greater than it binds to a different antigen in the complement system. In one embodiment, the dy or fragment thereof is selected from the group consisting of a recombinant antibody, an antibody having reduced effector function, a chimeric antibody, a humanized antibody and a human dy. In one embodiment, the MASP-2 inhibitory agent selectively inhibits lectin pathway complement activation without substantially inhibiting Clq-dependent complement activation. In one embodiment, the MASP-Z inhibitory agent is administered subcutaneously, intraperitoneally, intra- muscularly, intra-arterially, intravenously, or as an inhalant. In one embodiment, the MASP-Z inhibitory agent is administered in an amount effective to inhibit tubulointerstitial fibrosis. In one embodiment, the MASP-Z tory agent is administered in an amount effective to , delay or eliminate the need for dialysis in the subject. In one embodiment, the subject is suffering from a renal disease or disorder selected from the group ting of chronic kidney disease, chronic renal failure, glomerular disease (e.g., focal segmental glomerulosclerosis), an immune complex disorder (e.g., IgA nephropathy, membraneous pathy), lupus nephritis, nephrotic syndrome, diabetic nephropathy, tubulointerstitial damage and glomerulonepthritis (e.g., C3 ulopathy). In one embodiment, the subject is suffering from proteinuria and the MASP-2 inhibitory agent is administered in an amount effective to reduce proteinuria in the subject. In one embodiment, the MASP-2 inhibitory agent is administered in an amount and for a time effective to achieve at least a 20 percent reduction (e.g., at least a percent reduction, or at least a 40 percent reduction, or at least a 50 percent reduction) in r urine n excretion as compared to baseline r urine protein excretion in the subject prior to treatment. In one embodiment, the subject is suffering from a renal disease or disorder associated with proteinuria selected from the group consisting of nephrotic syndrome, pre-eclampsia, eclampsia, toxic lesions of kidneys, amyloidosis, en vascular es (e.g., systemic lupus erythematosus), dehydration, glomerular diseases (e.g. membranous glomerulonephritis, focal segmental glomerulonephritis, C3 glomerulopathy, minimal change disease, lipoid sis), strenuous exercise, stress, benign orthostatis (postural) proteinuria, focal segmental glomerulosclerosis, IgA nephropathy (i.e., Berger’s disease), IgM nephropathy, membranoproliferative glomerulonephritis, membranous nephropathy, minimal change disease, sarcoidosis, Alport’s syndrome, diabetes mellitus (diabetic nephropathy), drug- induced ty (e.g., NSAIDS, nicotine, llamine, lithium carbonate, gold and other heavy metals, ACE inhibitors, antibiotics (e. g., adriamycin) or opiates (e.g. heroin) or other toxins); Fabry’s disease, infections (e.g., HIV, is, hepatitis A, B or C, poststreptococcal infection, urinary schistosomiasis), aminoaciduria, Fanconi syndrome, hypertensive nephrosclerosis, interstitial nephritis, sickle cell disease, hemoglobinuria, multiple myeloma, myoglobinuria, organ rejection (e.g., kidney transplant rejection), ebola hemorrhagic fever, Nail patella syndrome, familial mediterranean fever, HELLP syndrome, systemic lupus erythematosus, Wegener’s granulomatosis, Rheumatoid arthritis, Glycogen e disease type 1, Goodpasture’s syndrome, Henoch-Schénlein purpura, urinary tract infection which has spread to the kidneys, SjOgren’s syndrome and post-infections glomerulonepthritis. In one embodiment, the subject is suffering from IgA nephropathy. In one ment, the subject is suffering from membranous nephropathy.

In another aspect, the present invention es a method of preventing or reducing renal damage in a subject suffering from a disease or condition associated with proteinuria comprising administering an amount of a MASP-2 inhibitory agent effective to reduce or prevent proteinurea in the subject. In one ment, the MASP-2 inhibitory agent is a MASP-Z antibody or fragment thereof. In one embodiment, the MASP-2 inhibitory agent is a MASP-Z monoclonal antibody or nt thereof that specifically binds to a n of SEQ ID NO:6. In one embodiment, the MASP-Z inhibitory agent selectively inhibits lectin pathway complement activation Without substantially ting Clq-dependent complement activation. In one embodiment, the disease or condition associated with nuria is selected from the group consisting of nephrotic syndrome, pre-eclampsia, sia, toxic lesions of kidneys, amyloidosis, en vascular diseases (e.g., systemic lupus matosus), dehydration, ular diseases (e.g. membranous ulonephritis, focal segmental glomerulonephritis, C3 glomerulopathy, minimal change e, lipoid nephrosis), strenuous exercise, stress, benign orthostatis (postural) proteinuria, focal segmental glomerulosclerosis, IgA nephropathy (i.e., Berger’s disease), IgM nephropathy, membranoproliferative glomerulonephritis, membranous nephropathy, minimal change disease, dosis, Alport’s me, diabetes mellitus (diabetic nephropathy), drug-induced toxicity (e.g., NSAIDS, nicotine, penicillamine, lithium carbonate, gold and other heavy metals, ACE inhibitors, antibiotics (e.g., adriamycin) or opiates (e.g. heroin)); Fabry’s disease, infections (e.g., HIV, syphilis, hepatitis A, B or C, poststreptococcal infection, urinary schistosomiasis); aminoaciduria, Fanconi syndrome, ensive nephrosclerosis, interstitial nephritis, sickle cell disease, hemoglobinuria, multiple myeloma, myoglobinuria, organ rejection (e.g., kidney transplant rejection), ebola hemorrhagic fever, Nail a syndrome, familial mediterranean fever, HELLP syndrome, systemic lupus erythematosus, Wegener’s granulomatosis, Rheumatoid arthritis, Glycogen storage disease type 1, Goodpasture’s syndrome, Henoch-Schonlein purpura, urinary tract infection which has spread to the kidneys, SjOgren’s syndrome and nfections glomerulonepthlitis. In one embodiment, the MASP-2 inhibitory agent is administered in an amount and for a time effective to achieve at least a 20 t ion (e.g., at least a 30 percent reduction, or at least a 40 percent reduction, or at least a 50 percent reduction) in 24-hour urine protein excretion as compared to baseline r urine protein excretion in the subject prior to treatment.

In another aspect, the present invention provides a method of inhibiting the progression of chronic kidney disease, comprising administering an amount of a MASP-2 inhibitory agent effective to reduce or prevent renal fibrosis, e.g., tubulointerstitial is, in a t in need thereof. In one embodiment, the MASP-2 inhibitory agent is a MASP-2 dy or fragment thereof. In one embodiment, the MASP-2 inhibitory agent is a MASP-2 monoclonal antibody, or fragment thereof that specifically binds to a portion of SEQ ID NO:6. In one embodiment, the MASP-2 inhibitory agent selectively ts lectin pathway complement activation Without substantially inhibiting Clq- dependent complement activation. In one embodiment, the subject in need thereof ts proteinuria prior to administration of the MASP-2 inhibitory agent and stration of the MASP-Z inhibitory agent decreases proteinuria in the subject. In one embodiment, the MASP-Z inhibitory agent is administered in an amount and for a time effective to achieve at least a 20 percent ion (e.g., at least a 30 percent reduction, or at least a 40 percent reduction, or at least a 50 percent reduction) in 24-hour urine protein ion as compared to baseline 24-hour urine protein excretion in the subject prior to treatment. In one embodiment, the MASP-Z inhibitory agent is administered in an amount effective to reduce, delay or eliminate the need for dialysis in the subject.

In another aspect, the invention provides a method of protecting a kidney from renal injury in a subject that has undergone, is undergoing, or will o treatment with one or more nephrotoxic agents, comprising administering an amount of a MASP-2 inhibitory agent effective to prevent or ameliorate drug-induced pathy. In one embodiment, the MASP-2 inhibitory agent is a MASP-2 antibody or fragment thereof. In one embodiment, the MASP-2 inhibitory agent is a MASP-2 monoclonal antibody or fragment thereof that specifically binds to a n of SEQ ID NO:6. In one embodiment, the MASP-Z inhibitory agent selectively inhibits lectin y complement activation without substantially inhibiting Clq—dependent complement activation.

In another aspect, the invention provides a method of ng a human subject ing from Immunoglobulin A Nephropathy (IgAN) comprising administering to the subject a composition comprising an amount of a MASP-2 inhibitory antibody, or antigen-binding nt thereof, effective to inhibit MASP-Z-dependent complement activation. In one embodiment, the subject is ing from steroid-dependent IgAN. In one embodiment, the MASP-2 tory antibody is a monoclonal antibody, or fragment thereof that specifically binds to human . In one embodiment, the antibody or fragment thereof is selected from the group consisting of a recombinant antibody, an antibody having reduced effector on, a chimeric antibody, a humanized antibody, and a human antibody. In one embodiment, the MASP-2 inhibitory antibody does not substantially inhibit the classical pathway. In one embodiment, the MASP-2 inhibitory dy inhibits C3b deposition in 90% human serum with an IC50 of 30 nM or less. In one embodiment, the method further comprises identifying a human subject having steroid—dependent IgAN prior to the step of administering to the subject a composition comprising an amount of a MASP-2 inhibitory antibody, or antigen-binding fragment thereof, effective to improve renal function. In one embodiment, the MASP-2 inhibitory antibody or antigen-binding fragment thereof is administered in an amount effective to improve renal on. In one embodiment, the MASP-2 inhibitory antibody or n-binding fragment thereof is administered in an amount effective and for a time sufficient to achieve at least a 20 percent reduction in r urine protein excretion as compared to baseline 24-hour urine protein excretion in the subject prior to treatment. In one embodiment, the composition is administered in an amount sufficient to improve renal function and se the corticosteroid dosage in said subject. In one embodiment, the MASP-2 tory antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising CDRH1 , CDR-H2 and CDR-H3 of the amino acid sequence set forth as SEQ ID NO:67 and a light chain variable region comprising CDR-L1, CDR-L2 and CDR-L3 of the amino acid sequence set forth as SEQ ID NO:69.

In another aspect, the invention provides a method of treating a human subject suffering from membranous nephropathy (MN) comprising administering to the subject a composition comprising an amount of a MASP-2 inhibitory antibody, or antigen-binding fragment thereof, effective to inhibit MASPdependent complement activation. In one ment, the subject is suffering from steroid-dependent MN. In one embodiment, the MASP-2 inhibitory antibody is a monoclonal antibody, or fragment f that specifically binds to human MASP-2. In one embodiment, the MASP-2 tory antibody or nbinding fragment thereof is administered in an amount effective to improve renal function. In one embodiment, the MASP-2 inhibitory antibody or antigen-binding nt thereof is administered in an amount effective and for a time sufficient to achieve at least a 20 t reduction in 24-hour urine protein excretion as compared to baseline r urine n ion in the subject prior to treatment. In one embodiment, the composition is administered in an amount sufficient to improve renal function and decrease the corticosteroid dosage in said subject. In one embodiment, the MASP-2 inhibitory antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising CDRH1 , CDR-H2 and CDR-H3 of the amino acid sequence set forth as SEQ ID NO:67 and a light chain variable region comprising CDR-L1, CDR-L2 and CDR-L3 of the amino acid sequence set forth as SEQ ID NO:69.

In another aspect, the invention provides a method of treating a human subject suffering from Lupus tis (LN) comprising administering to the t a composition comprising an amount of a MASP-2 inhibitory antibody, or antigen-binding nt f, effective to inhibit MASPdependent complement activation. In one embodiment, the subject is suffering from steroid-dependent LN. In one embodiment, the MASP-2 inhibitory antibody is a monoclonal antibody, or fragment thereof that specifically binds to human MASP-2. In one embodiment, the MASP-2 inhibitory antibody or antigen-binding fragment thereof is administered in an amount effective to improve renal function. In one embodiment, the MASP-2 inhibitory antibody or antigen-binding fragment f is administered in an amount effective and for a time sufficient to achieve at least a 20 percent reduction in 24-hour urine protein excretion as compared to baseline 24-hour urine protein ion in the subject prior to treatment. In one embodiment, the composition is administered in an amount sufficient to improve renal function and decrease the osteroid dosage in said subject. In one embodiment, the MASP-2 inhibitory antibody or antigen-binding fragment f comprises a heavy chain variable region comprising CDR-H1, CDR-H2 and CDR-H3 of the amino acid sequence set forth as SEQ ID NO:67 and a light chain variable region comprising CDR-L1, CDR-L2 and CDR-L3 of the amino acid sequence set forth as SEQ ID NO:69.

In another aspect, the ion provides a method of reducing proteinuria in a human subject suffering from IgAN comprising administering to the subject a MASP-2 inhibitory antibody, or antigen-binding fragment thereof, comprising a heavy chain variable region comprising CDR-H1, CDR-H2 and CDR-H3 of the amino acid sequence set forth as SEQ ID NO:67 and a light chain variable region comprising CDR-L1, CDR-L2 and CDR-L3 of the amino acid sequence set forth as SEQ ID NO:69 according to a dosage n as follows: a. administering about 4 mg/kg (i.e., from 3.6 mg/kg to 4.4 mg/kg) of said antibody to a t ing from IgAN once weekly intravenously for a treatment period of at least 12 weeks; or b. stering from about 180 mg to about 725 mg (i.e., from 162 mg to 797 mg) of said antibody to a subject suffering from IgAN once weekly intravenously for a treatment period of at least 12 weeks, wherein the method reduces proteinuria in said human subject.

W0 2018.107170] DESCRIPTION OF THE DRAWINGS The ing aspects and many of the attendant advantages of this invention will become more readily appreciated as the same become better understood by reference to the following detailed description, when taken in conjunction with the accompanying drawings, wherein: FIGURE 1 is a diagram illustrating the genomic structure of human MASP-2;