Methods for Treating Anemia in Renal Disease - Patent application

Abstract

Aspects of the present application provide hemojuvelin antagonists and methods of using them in the treatment of anemia of renal disease and conditions associated with these anemias. The methods provided in the present application relate to the treatment of renal disease in subjects with anemia associated with chronic kidney disease and / or subjects with a level of glomerular filtration rate lower than one or more thresholds.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of the filing dates of U.S. Provisional Application No. 63 / 280,594, filed November 17, 2021, and U.S. Provisional Application No. 63 / 419,648, filed October 26, 2022, the entire contents of each of which are incorporated by reference.

[0002] Reference to Electronic Sequence Listing The contents of the electronic sequence listing (D084270006WO00-SEQ-EPG.xml; size: 204,621 bytes; and creation date: November 16, 2022) are incorporated herein by reference in their entirety. [Background technology]

[0003] Kidney disease is a condition characterized by abnormalities in kidney structure or function. Outcomes of kidney disease can include renal failure as well as complications of decreased kidney function and cardiovascular disease. When kidney abnormalities persist for a prolonged period of time, kidney disease can be classified as chronic kidney disease (CKD). Cardiovascular death is the leading cause of death in CKD patients. As of 2017, the estimated prevalence of CKD worldwide is 9.1% of the world's population, and this prevalence is increasing. CKD most commonly manifests in people typically aged 65 years or older. Stages of CKD are defined according to the level of glomerular filtration rate (GFR).

[0004] Many kidney disease patients have or develop anemia. If left untreated, anemia negatively impacts cardiac function, increases the risk of blood transfusion, and seriously impairs quality of life. In some cases, untreated anemia can be fatal, especially in the context of CKD. A new class of drugs, referred to as hypoxia-inducible factor prolyl hydroxylase inhibitor (HIF-PHI) drugs, has emerged for the treatment of anemia in CKD. These drugs are designed to help stimulate the body to make red blood cells. To date, no HIF-PHI has been approved by the FDA. Thus, to date, no HIF-PHI has been demonstrated to be safe and effective for anemia in CKD in US clinical trials. A major efficacy concern with HIF-PHI drugs is overcorrection of serum hemoglobin (Hb) levels, which can lead to adverse cardiovascular outcomes. Thus, there is a need for safe and effective treatment of anemia associated with kidney disease (e.g., CKD). Summary of the Invention

[0005] Aspects of the present disclosure relate to methods of treating a subject with anemia involving the use of a hepcidin antagonist (e.g., a hemojuvelin antagonist). For example, in some embodiments, methods are provided herein that involve administration of a hemojuvelin antagonist to treat anemia in a subject with kidney disease. A further aspect of the present disclosure provides a method of treating anemia in a subject with chronic kidney disease (CKD). In some embodiments, the disclosed methods provide administration of a hemojuvelin antagonist to a subject that has (or exhibits) a certain level of glomerular filtration rate (GFR) as measured or estimated by a physician. In some embodiments, the methods provided herein involve subcutaneous administration of a hemojuvelin antagonist (e.g., an anti-hemojuvelin antagonist antibody) to treat anemia, which is advantageous for administration outside of a hospital or similar health care setting where intravenous and similar administrations or procedures are typically performed. For example, in some embodiments, the methods based on subcutaneous administration provided herein may be performed in a clinic or as self-administration by a subject or patient. Thus, embodiments of the present disclosure are useful for treating anemia in subjects with kidney disease, where the subject is not undergoing therapy involving one or more visits to a hospital or similar health care setting where intravenous therapy can be readily administered (e.g., dialysis therapy, IV iron therapy, and other similar therapies).

[0006] A further aspect of the present disclosure relates to a method for treating 1.73 m of atherosclerosis by administering to a subject an effective amount of a hemojuvelin antagonist. 2The present disclosure relates to treating a subject with anemia, the subject being identified as having a glomerular filtration rate (GFR) level of less than 90 mL / min per day. Certain aspects of the present disclosure relate to treating anemia caused, at least in part, by nutritional iron deficiency, iron deficiency due to blood loss, intrinsic red blood cell damage, hemolysis, inflammation, functional iron deficiency, or any combination thereof. In some embodiments, the subject may have chronic kidney disease. Thus, in some embodiments, the level of glomerular filtration rate in the subject has been sustained for at least 3 months. In some embodiments, the anemia is associated with renal damage in the subject. The renal damage may have been present in the subject for at least 3 months. In some embodiments, the subject has chronic kidney disease, which is a non-dialysis-dependent chronic kidney disease. Similarly, in some embodiments, the subject is not undergoing dialysis therapy.

[0007] Anemia is a common complication in patients with CKD and is associated with multiple adverse outcomes in this population. Increased hepcidin is thought to be a central contributor to the development of anemia, due to both reduced clearance and increased synthesis of hepcidin. Treatment with hepcidin-lowering drugs has been demonstrated to increase iron availability from systemic iron stores and increase hemoglobin (Sheetz et al, Br J Clin Pharmacol. 2019;85:935-948, incorporated herein by reference). It is hypothesized that by downregulating hepcidin, anti-hemojuvelin antibodies (anti-HJV Ab) will have beneficial effects in treating anemia in CKD patients, making iron available for improved erythropoiesis.

[0008] Thus, in some embodiments, the methods provided herein are useful for treating a subject with anemia associated with kidney disease to reduce hepcidin levels or activity. In some embodiments, the subject may experience an improvement in iron intake from the gastrointestinal system (i.e., from the diet). In some embodiments, the subject may experience a partial or complete recovery of iron levels.

[0009] In some embodiments, the methods provided herein are useful for treating subjects with low TSAT levels (e.g., less than 20%, less than 30%, less than 40%, or less than 50%) and / or low ferritin levels (e.g., ferritin of ≦50 ng / mL, ≦100 ng / mL, or ≦300 ng / mL). In some embodiments, the methods provided herein are useful for treating subjects exhibiting low hemoglobin levels (e.g., Hb levels of less than 11 g / dL, 10 g / dL, or 9 g / dL). In some embodiments, the methods provided herein are useful for treating subjects exhibiting normal to relatively high hepcidin levels. In some embodiments, the subject has a 1.73m 2 The patient further exhibits renal impairment, which may be chronic, with a glomerular filtration rate ranging from 15 mL / min to just under 60 (eg, 59) mL / min per day.

[0010] In some embodiments, the hemojuvelin antagonist is an anti-hemojuvelin antibody. In some embodiments, the anti-hemojuvelin antibody preferentially binds to RGMc relative to RGMa and RGMb. In some embodiments, the anti-hemojuvelin antibody has an equilibrium dissociation constant (K D ) binds to RGMc. In some embodiments, the anti-HJV antibody is an anti-HJV antibody in Table 1.

[0011] In some embodiments, the anti-hemojuvelin antibody comprises (a) a heavy chain variable region comprising a CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 3; and / or (b) a light chain variable region comprising a CDR1 comprising the amino acid sequence of SEQ ID NO: 17, a CDR2 comprising the amino acid sequence of SEQ ID NO: 5, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 27. In exemplary embodiments, the antibody comprises a HC CDR1 of SEQ ID NO: 1, a HC CDR2 of SEQ ID NO: 2, a HC CDR3 of SEQ ID NO: 3; a LC CDR1 of SEQ ID NO: 17, a LC CDR2 of SEQ ID NO: 5, and a LC CDR3 of SEQ ID NO: 27. In some embodiments, the anti-hemojuvelin antibody is hHA-008. In some embodiments, the anti-hemojuvelin antibody is hHA-008-QL.

[0012] In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 38, and a VL comprising the amino acid sequence of SEQ ID NO: 39. The antibody may be selected from the group consisting of a full length IgG, a Fab fragment, a F(ab') fragment, a F(ab')2 fragment, a scFv, and an Fv. In some embodiments, the antibody is a full length IgG comprising a heavy chain constant region of isotype IgG1, IgG2, IgG3, or IgG4.

[0013] Thus, in some embodiments, a method is provided for treating a subject having anemia associated with chronic kidney disease by administering an anti-hemojuvelin antibody comprising a VH comprising the amino acid sequence of SEQ ID NO: 38 and a VL comprising the amino acid sequence of SEQ ID NO: 39.

[0014] In some embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 61, and a light chain comprising the amino acid sequence of SEQ ID NO: 62. In other embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 63, and a light chain comprising the amino acid sequence of SEQ ID NO: 62.

[0015] In some embodiments, the anti-hemojuvelin antibody is administered to a subject in need thereof in an amount of 0.1-0.8 mg per kg of subject. In some embodiments, the antibody is administered at a dose of about 7 mg to 56 mg, e.g., 56 mg. Any of these doses may be administered monthly or every two months.

[0016] In some embodiments, the subject is identified prior to treatment as having high hepcidin levels. In some embodiments, the subject is identified as having functional iron deficiency. In some embodiments, the subject is identified as exhibiting inflammation and / or iron-restricted erythropoiesis. In some embodiments, the subject is a human. In some embodiments, the subject has nutritional iron deficiency. In some embodiments, the subject does not have nutritional iron deficiency.

[0017] In some embodiments, the subject has a serum ferritin level above 100 μg / L (100 ng / ml). In other embodiments, the subject has a serum ferritin level below 100 μg / L. In some embodiments, the subject has a reticulocyte hemoglobin content below 26 pg / cell. In some embodiments, the subject has a transferrin saturation level below 50%, 30%, or 25%. In some embodiments, the subject has a liver iron level above 2000 μg / g dry weight. In some embodiments, the subject has a serum iron level in the range below 50 μg / dL. In some embodiments, the subject has a total iron binding capacity in the range below 400 μg / dL. In some embodiments, the subject has a hepcidin level in the range above 55 ng / ml. In some embodiments, the subject has an IL-6 level above 1.8 pg / mL. In some embodiments, the subject has a serum creatinine value above 2 mg / dL.

[0018] In some embodiments, the subject is identified as having a hemoglobin level within a range of 1.5-2.0 g / dL or 2.0-4.0 g / dL or greater below normal hemoglobin levels. In some embodiments, the subject exhibits a serum hemoglobin level below 10 g / dL. In some embodiments, the subject exhibits a serum hemoglobin level below 8 g / dL. In some embodiments, administration of the hepcidin antagonist increases hemoglobin levels by at least 1 g / dL from baseline.

[0019] In some embodiments, the method of treating a subject further comprises administering to the subject one or more additional therapeutic agents, such as a growth differentiation factor (GDF) trap, an erythropoiesis stimulating agent (ESA), oral iron, intravenous iron (IV iron), hypoxia inducible factor prolyl hydroxylase inhibitor (HIF-PHI), or red blood cell transfusion. In some embodiments, a GDF trap, such as sotatercept or raspatercept, is administered. In some embodiments, an ESA, such as erythropoietin (EPO), is administered. In some embodiments, the additional therapeutic agent is oral iron. In certain embodiments, the additional therapeutic agent is oral iron at a dose of about 30 mg twice weekly. In some embodiments, the additional therapeutic agent is HIF-PHI.

[0020] In some embodiments, the methods provided herein for decreasing hepcidin levels or activity may be combined with oral iron therapy to aid in the restoration of iron levels. Thus, in some embodiments, such methods provided herein are useful for treating anemic subjects experiencing intolerance to or an inadequate response to oral iron.

[0021] Thus, combination therapies are provided that include any of the disclosed hemojuvelin antagonists and one or more of growth differentiation factor (GDF) traps, erythropoietin stimulating agents (ESAs), oral iron, IV iron (e.g., MonoFerric®), hypoxia-inducible factor prolyl hydroxylase inhibitors (HIF-PHIs), and red blood cell transfusions.

[0022] In some embodiments, administration of any of the disclosed anti-hemojuvelin antibodies results in or provides an increase in hemoglobin levels in a subject of at least 2, 4, 6, 8, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or 20 g / dL compared to an untreated subject. In some embodiments, administration results in an increase in hemoglobin levels in a subject of about 17 g / dL. Administration may result in an increase in reticulocyte hemoglobin (Ret-HGB) levels in a subject of at least 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.75, 0.8, 0.85, 0.9, 0.95, 1.0, or greater than 1.0 pg compared to an untreated subject. In some embodiments, administration results in an increase in serum iron levels in the subject of about 25, 27.5, 30, 32.5, 35, 38.5, 40, or 45 μmol / L compared to an untreated subject. Administration results in an increase in serum iron levels in the subject of about 1, 2, 2.5, 3, 4, 5, 6, 7, 7.5, 8, 9, or 10×10 compared to an untreated subject. 5 Administration may result in an increase in red blood cell (RBC) count in the subject, in cells / μL. 9 This may result in a reduction in the reticulocyte (Ret) count in the subject, in cells / L.

[0023] The disclosed methods can be particularly useful for subcutaneous, intravenous, or intramuscular administration of a hemojuvelin antagonist (e.g., an anti-hemojuvelin antibody). Thus, the disclosed methods can include the step of administering the antagonist by subcutaneous, intravenous, or intramuscular injection. In exemplary embodiments, the administering step is by subcutaneous injection. In some embodiments, the subcutaneous injection is administered as a self-administration.

[0024] The foregoing and other aspects, implementations, operations, functionality, features, and embodiments of the present teachings may be more fully understood from the following description taken in conjunction with the accompanying drawings.

[0025] The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate certain embodiments and, together with the written description, provide non-limiting examples of certain aspects of the compositions and methods disclosed herein. [Brief description of the drawings]

[0026] [Figure 1] 1A-1G are graphs showing the generation and characterization of anti-HJV antibodies. FIG 1A shows the schematic process of the generation, humanization, and affinity maturation of anti-hemojuvelin antibodies. FIG 1B-1G show the sensorgrams from BIAcore analysis of antibodies HA, hHA-004, hHA-008, hHA-009, and hHA-011. [Diagram 2] 2A-2C are graphs showing the BMP reporter gene assay for anti-HJV antibodies. FIG. 2A shows the general principle of the HJV BMP reporter assay. FIG. 2B shows the effect of anti-HJV antibodies in inhibiting RGMc BMP signaling. FIG. 2C shows the effect of anti-HJV antibodies in inhibiting RGMa BMP signaling. [Diagram 3] 3 is a graph showing non-specific binding of anti-HJV antibodies to HEK293 cells. The bars from left to right are 100 μg / ml, 10 μg / ml, and 1 μg / ml in each group. [Figure 4] Figures 4A-4C are schematic diagrams showing the structure and design of hHA-008 and hHA-008-QL. Figure 4A shows the structure of hHA-008. Figure 4B shows the structure of hHA-008-QL. Figure 4C shows the comparison in antibody structure between hHA-008 and hHA-008-QL. [Diagram 5] 5A-5B are graphs showing CD4+ T cell responses of peripheral blood mononuclear cells (PBMCs) loaded with hHA-008 or hHA-008-QL. [Figure 6]Figures 6A-6C are graphs showing PK / PD analysis of hHA-008 in rats. Figure 6A shows that the maximal effect of hHA-008, as measured by TSAT%, occurred between 4 and 8 days after treatment. Figure 6B shows that hHA-008 reached maximal effect, as measured by TSAT%, approximately 1-4 days after injection in female cynomolgus monkeys. Figure 6C shows that hHA-008 reached maximal effect, as measured by TSAT%, approximately 1-4 days after injection in male cynomolgus monkeys, although one of the males did not respond to hHA-008 treatment. [Figure 7] Figures 7A-7F show PK / PD correlations in cynomolgus monkeys using a single IV dose of 6mpk. Figure 7A shows that the maximum TSAT% increase occurred 1-4 days after injection (Tmax=1-4 days), with one of the animals tested having a dramatic drop in TSAT around day 34. Figure 7B shows that plasma hepcidin-25 concentrations change over time after hHA-008 injection. Figure 7C shows that plasma hHA-008 concentrations change over time after hHA-008 injection. Figures 7D-7F show that hHA-008 had a robust PK / PD correlation of PK (plasma antibody concentration) to TSAT% and plasma hepcidin-25 concentrations. Results for each tested cynomolgus monkey are shown in Figure 7D (cynomolgus monkey 1), Figure 7E (cynomolgus monkey 2), and Figure 7F (cynomolgus monkey 3). [Figure 8] Figures 8A-8C show that hHA-008 antibody modulates TSAT% in a dose-dependent manner. Figure 8A shows TSAT% and hHA008 concentration after animals were treated with 0 (vehicle control) or 0.6 mpk of hHA-008. Figure 8B shows TSAT% and hHA-008 concentration after animals were treated with 0 (vehicle control) or 3 mpk of hHA-008. Figure 8C shows TSAT% and hHA-008 concentration after animals were treated with 0 (vehicle control) or 60 mpk of hHA-008. [Figure 9]Figures 9A-9C are graphs showing a comparison of PK / PD between hHA-008 and hHA-008-QL. Figure 9A shows the TSAT% change over time in cynomolgus monkeys after treatment with hHA-008 or hHA-008-QL. Figure 9B shows the plasma concentration of antibody over time in cynomolgus monkeys after treatment with hHA-008 or hHA-008-QL. Figure 9C shows the time course of decrease in plasma concentration of hHA-008 and hHA-008-QL. [Figure 10] Figures 10A-10D are graphs showing FcRn binding of hHA-008 and hHA-008-QL at pH 6.0 or 7.4. Figures 10A-10B show FcRn binding of hHA-008 and hHA-008-QL at pH 6.0. Figures 10C-10D show FcRn binding of hHA-008 and hHA-008-QL at pH 7.4. X-axis: time. Y-axis: response. [Figure 11] FIG. 11 illustrates the myeloproliferative cycle characteristic of certain high hepcidin disorders. [Figure 12] FIG. 12 shows the hepcidin stimulated pathway and physiological control of iron homeostasis by hepcidin. [Figure 13] FIG. 13 is a graph showing that IL-6 induces hepcidin expression in cynomolgus monkeys and hHA-008 treatment prevents inflammation-induced (IL 6) increases in hepcidin-25 in a dose-dependent manner. [Figure 14] Figure 14 shows that hHA-008 interacts with amino acids 170-183 [SSPMALGANATATR (SEQ ID NO: 121)] on 3720-RG-050. Interactions occur at amino acids 170, 171, 180, 182, and 183 on 3720-RG-050. [Figure 15]Figures 15A-15J show the interaction of 3720-RG-050 and hHA-008. 3720-RG-050 PDB structure was generated by homology using Swiss Model software. 3720-RG-050 amino acids 170-183 [SSPMALGANATATR (SEQ ID NO: 121)] are shown in Figures 15A, 15B, 15C, 15D, 15E: ribbon / surface representation of front view (Figure 15A); back view (Figure 15B), side view 1 (Figure 15C), side view 2 (Figure 15D) and top view (Figure 15E). Figures 15F, 15G, 15H, 15I, 15J: ribbon representation of front view (Figure 15F); back view (Figure 15G), side view 1 (Figure 15H), side view 2 (Figure 15I) and top view (Figure 15J). [Figure 16] Figure 16 shows that hHA-008-QL interacts with amino acids 169-182 [TSSPMALGANATAT (SEQ ID NO: 122)] and 289-300 [SQRLSRSERNRR (SEQ ID NO: 127)] of 3720-RG-050. Interactions occur at amino acids 169, 171, 180, 182; 289, 293, 294, 295, 297, 300 on 3720-RG-050. [Figure 17] Figures 17A-17J show the interaction 3720-RG-050 / hHA-008-QL. The 3720-RG-050 PDB structure was generated by homology using Swiss Model software. 3720-RG-050 amino acids 169-182 [TSSPMALGANATAT (SEQ ID NO: 122)] and 289-291 (SQR) are shown in Figures 17A, 17B, 17C, 17D, 17E: front view (Figure 17A); back view (Figure 17B), side view 1 (Figure 17C), side view 2 (Figure 17D) and top view (Figure 17E) ribbon / surface representation. Figures 17F, 17G, 17H, 17I, 17J: Ribbon representation of front view (Figure 17F); back view (Figure 17G), side view 1 (Figure 17H), side view 2 (Figure 17I) and top view (Figure 17J). [Figure 18] FIG. 18 shows that hHA-008 was effective in preventing IL-6-induced serum iron suppression in a dose-dependent manner in cynomolgus monkeys. [Figure 19]FIG. 19 shows that the decline in PD responses (eg, hepcidin-25 concentrations and TSAT%) coincided with a decrease in hHA-008 serum concentrations following subcutaneous administration of hHA-008 to Sprague-Dawley rats (FIG. 19). [Figure 20] Figures 20A-20D show PK / PD analysis in cynomolgus monkeys following subcutaneous administration of hHA-008. Figure 20A shows that serum concentration-time profiles became indistinguishable between SC and IV injections 4 days after dosing. Figures 20B-20D show that the return of serum iron to baseline levels was consistent with a decrease in hHA-008 serum concentrations following injection of 0.3 mpk, 0.6 mpk, and 1 mpk of hHA-008 by either subcutaneous or intravenous injection. [Figure 21] Figures 21A and 21B show functional iron deficiency in chronic kidney disease (CKD). Figure 21A depicts functional iron deficiency in the context of CKD. Figure 21B shows CKD and inflammatory pathways that lead to anemia. [Figure 22] FIG. 22 depicts the hepcidin stimulated pathway and the physiological regulation of iron homeostasis by hepcidin. [Figure 23] Figures 23A-23G show the role of hepcidin in functional iron deficiency (FID) and an example of regulating hepcidin levels with hepcidin antagonists. Figure 23A depicts the mechanism of functional iron deficiency. Figure 23B shows that functional iron deficiency is a common feature of chronic diseases, including anemia of inflammation and chronic kidney disease (CKD). Figure 23C shows that functional iron deficiency is associated with high iron levels and high hepcidin levels. Figure 23D is a schematic illustration of reducing hepcidin levels to normal by using hepcidin antagonists for the treatment of iron-restricted diseases. Figure 23E depicts the use of anti-HJV antibodies as an example of inhibiting the HJV-induced BMP signaling pathway to reduce hepcidin to normal levels. Figure 23F shows that matriptase-2 negatively regulates hepcidin by cleaving membrane-bound HJV. FIG. 23G depicts examples of possible hepcidin antagonists for modulating hepcidin levels. [Figure 24] FIG. 24 shows serum Fe concentrations following subcutaneous (SC) administration of hHA-008 to male and female rats as a single 6 mg / kg or 30 mg / kg dose. [Diagram 25] FIG. 25 shows serum Fe measured after either 6 mg / kg or 30 mg / kg given as an intravenous (IV) or subcutaneous (SC) dose to rats. [Figure 26] FIG. 26 shows the pharmacokinetics of hHA-008 following IV and SC administration to cynomolgus monkeys at dose levels of 0.3, 0.6, 1.0, and 6.0 mg / kg. [Figure 27] Figures 27A-27D show serum hepcidin-25 concentrations following IV and SC hHA-008 administration at dose levels of 0.3, 0.6, 1.0, and 6.0 mg / kg. [Figure 28] FIG. 28 shows the pharmacokinetics of hHA-008 following IV and SC administration to cynomolgus monkeys at dose levels of 0.3, 0.6, 1.0, and 6.0 mg / kg. [Figure 29] Figures 29A-29D show serum Fe concentrations following IV and SC hHA-008 administration at dose levels of 0.3, 0.6, 1.0, and 6.0 mg / kg. [Diagram 30] Figures 30A-30D show the transferrin saturation % (TSAT%) following IV and SC hHA-008 administration at dose levels of 0.3, 0.6, 1.0, and 6.0 mg / kg. [Diagram 31] FIG. 31 is a schematic showing the in vivo study design to evaluate the effect of treatment with the lead anti-hemojuvelin antibody (anti-HJV Ab) in an anemic rat model of CKD. [Diagram 32] FIG. 32 shows the metabolic effects of adenine diet-induced kidney injury and anemia in a CKD rat model after administration of empty vehicle. [Diagram 33]Figure 33 is a series of line plots showing the variation in HAMP (hepcidin gene) mRNA expression, hepcidin-25 and iron levels in serum of a rat model after anti-HJV Ab administration. This figure shows that anti-HJV Ab reduced hepcidin and increased serum iron levels in CKD rats over the course of treatment. [Diagram 34] FIG. 34 is a series of line plots showing the effect of anti-HJV Abs on reticulocyte hemoglobin, mean corpuscular hemoglobin (MCH), hemoglobin (HGB), reticulocyte count, red blood cell (RBC) count, and mean corpuscular volume (MCV) over the course of treatment. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0027] According to some aspects, the present disclosure provides methods of administering an effective amount of a hemojuvelin antagonist effective for inhibiting hepcidin function and / or reducing hepcidin expression. In some embodiments, these methods also include administering an effective amount of a hemojuvelin antagonist effective for inhibiting hepcidin function and / or reducing hepcidin expression. 2 These methods are particularly useful for treating anemia in subjects who are identified as having a glomerular filtration rate (GFR) level of less than 90mL / min per day. In some embodiments, these methods are particularly useful for treating anemia of kidney disease (e.g. CKD) and / or one or more symptoms or complications thereof. Thus, in some embodiments, these methods can be used to treat subjects with kidney disease, whether the disease is chronic or not. Thus, in related aspects, the present disclosure provides compositions and methods for treating anemia that may be associated with chronic kidney disease.

[0028] A frequent complication of CKD is anemia caused by insufficient erythropoietin (EPO) production by damaged kidneys. As EPO production decreases, the bone marrow responds by decreasing RBC count. Other factors in CKD patients that lead to anemia are iron deficiency, blood loss, and shortened red blood cell (RBC) survival. In some embodiments, the following criteria are generally applied to classify severe CKD patients as anemic or nonanemic: nonanemic [hemoglobin (Hb) of ≥12 / ≥13 g / dL in women / men]; anemia grade 1 (Hb of 10-12 / 13 g / dL in women / men); grade 2 (Hb of 8-10 g / dL); and grade 3+ (Hb of <8 g / dL). See Toft et al., J Nephrol. 2020 Feb; 33(1):147-156, incorporated herein by reference.

[0029] The etiology of anemia associated with CKD is multifactorial and results from any one of the following causes: EPO deficiency, iron malabsorption (functional iron deficiency), inability to utilize stored iron, or shortened red blood cell survival. Subjects suffering from anemia associated with CKD typically exhibit reduced endogenous EPO levels and / or functional iron deficiency. Functional iron deficiency, manifested by low transferrin saturation (TSAT) levels, often leads to EPO resistance. These subjects also typically exhibit reduced hepcidin levels. Hepcidin (-25) is a 25 amino acid protein that is filtered from the blood through the kidneys.

[0030] CKD patients who are not prescribed dialysis are typically managed conservatively. The current standard of care in the treatment of anemia associated with CKD includes erythropoiesis stimulating agents (ESAs), iron supplementation, and RBC transfusions. ESAs are effective and generally exhibit few adverse side effects. The use of ESAs maintains target Hb levels in the majority of patients and reduces the need for transfusions. FDA-approved ESAs include recombinant glycoproteins such as epoetin alfa (Epogen / Procrit), darbepoetin alfa (Aranesp), methoxypolyethylene glycol-epoetin beta (Mircera), epoetin alfa-epbx (Retacrit); and biosimilars of Epogen / Procrit. ESAs are typically administered intravenously; however, they may also be administered subcutaneously. Iron supplementation can be in the form of intravenous (IV) iron or oral iron. In some embodiments, the methods provided herein involve the use of a hepcidin antagonist (eg, a hemojuvelin antagonist) in combination with an erythropoiesis stimulating agent (ESA), iron supplementation, and / or RBC transfusion.

[0031] In some embodiments, CKD patients who are not prescribed dialysis often do not have the habit of frequent visits to the hospital. In some embodiments, because ESAs are often administered intravenously, prescribing ESAs for patients with non-dialysis-dependent CKD (CKD-NDD) often represents a significant burden in that patients must frequently visit the hospital to receive ESAs. Furthermore, patients report experiencing pain during subcutaneous administration of ESAs. Additionally, recent concerns about the safety of ESAs, particularly higher cardiovascular risk, increased cancer progression and mortality, have led to a decline in the use of ESAs worldwide. Also, as with HIF-PHI drugs, a major efficacy concern for ESAs is overcorrection of serum hemoglobin levels, which can lead to adverse cardiovascular outcomes.

[0032] There are several physiological factors and diagnostic criteria that influence a doctor's decision to prescribe dialysis. Exemplary physiological factors include GFR (and then the stage of CKD), ACR, calcium level, and BMI. Patients with GFR values ​​above 15 ml / min are not candidates for dialysis. In contrast, for patients with GFR values ​​below 15 ml / min, dialysis is recommended. Patients with GFR levels below 5 or 6 should immediately undergo dialysis. See Tattersal et al., Nephrol Dial Transplant (2011) 26: 2082-2086, which is incorporated herein by reference. If GFR is above 15, other markers such as ACR, serum calcium level, BMI, and the presence of uremic symptoms such as nausea, anorexia, insomnia, and fatigue are considered in the decision to recommend dialysis. If ACR is below 300, dialysis cannot be recommended. Conversely, dialysis is typically recommended in patients who exhibit uremic symptoms, such as nausea and anorexia. See Wang et al., Renal Failure 2021 43(31): 216-222, incorporated herein by reference.

[0033] Additional non-physiological factors influence the physician's decision to recommend dialysis. Prognosis, expected quality of life (with or without dialysis), treatment burden (if dialysis is performed), support from family members (including assistance with transportation to and from hospital or outpatient facilities), and patient preferences all play a role in the decision. The decision to dialysis is usually made jointly between the physician and the patient in light of the preferences of both parties and the best assessment of these factors and the physiological factors mentioned above. See Murtagh et al., Nephrol Dial Transplant (2007) 22: 1955-1962, incorporated herein by reference.

[0034] There are two types of dialysis: hemodialysis and peritoneal dialysis. In peritoneal dialysis, a cleansing solution is flushed into the abdomen through a tube (catheter). The inner wall of the abdomen (the peritoneum) acts as a filter, removing waste products from the blood. After a set period of time, the fluid containing the filtered waste products is flushed out of the abdomen and discarded. Peritoneal dialysis can be performed by a qualified professional in an outpatient facility or at home. In hemodialysis, blood is removed from the body, filtered through an artificial kidney machine, and then returned to the body. Hemodialysis is typically performed in a hospital. CKD patients undergoing dialysis who suffer from the anemia of CKD are typically given high doses of IV EPO and IV iron supplementation.

[0035] Generally, about 15 percent of anemic CKD patients not undergoing dialysis are treated with oral iron supplementation. (This iron supplementation treatment may be stopped by the physician if dialysis is indicated.) Oral iron may be administered at home or in an outpatient facility. For these patients, treatment with IV treatment (e.g., intravenous antibodies) is burdensome, since outpatient facilities often do not have the facilities for intravenous antibody delivery.

[0036] The present disclosure provides a method of treating anemia of kidney disease with a hepcidin antagonist, such as a hemojuvelin antagonist (e.g., an anti-hemojuvelin antibody). The present disclosure provides an anti-hemojuvelin antibody with a demonstrated safety profile. For example, in some embodiments, a method is provided that involves administration of any of the disclosed antibodies to a subject and does not cause overcorrection of hemoglobin levels in the subject's serum. In some embodiments, these antibodies may be formulated for subcutaneous delivery. Furthermore, in some embodiments, the present disclosure is directed to a method of subcutaneous delivery of an anti-hemojuvelin antibody to treat anemia of CKD. The present disclosure is further directed to a method of subcutaneous delivery of a hemojuvelin antagonist (e.g., an anti-hemojuvelin antibody) at home or in an outpatient facility or clinic, etc., to treat anemia of CKD. Because subcutaneous administration can be performed at home, the disclosed method of subcutaneous administration of a hemojuvelin antagonist may be particularly suitable for CKD patients who are not undergoing dialysis.

[0037] In some embodiments, the methods of the present disclosure may be particularly suitable for subjects who are not undergoing dialysis, such as CKD patients who are not undergoing dialysis.These methods may be particularly suitable for CKD-NDD patients.These methods may be particularly suitable for CKD patients who are undergoing dialysis.These methods may be particularly suitable for the population of anemic CKD patients who are not undergoing dialysis and are candidates for treatment with oral iron supplementation.

[0038] The methods of the present disclosure may be particularly suitable for treating anemia in non-dialysis CKD patients identified as lacking a response to oral iron (non-responsive) (or having CKD refractory to oral iron). The methods of the present disclosure may be particularly suitable for treating anemia in non-dialysis CKD patients identified as non-responsive to IV iron. These methods may be particularly suitable for treating anemia in CKD patients who have not undergone nephrectomy (kidney removal). These methods may be particularly suitable for treating anemia in CKD patients who are not candidates for IV iron (having TSAT less than 20% and serum iron levels less than 20 ng / ml).

[0039] These methods may be particularly suitable for CKD patients with i) elevated hepcidin levels, ii) decreased endogenous EPO levels, iii) functional iron deficiency, iv) nutritional iron deficiency (dietary iron intake deficiency), and / or v) decreased hemoglobin levels. In CKD, hepcidin levels are elevated when the kidney begins to fail, and concomitant nutritional iron deficiency and EPO resistance are observed. Therefore, the present disclosure provides a method of administering any of the disclosed hemojuvelin antagonists (e.g., anti-hemojuvelin antibodies) to non-dialysis CKD patients who exhibit low iron levels or low TSAT levels. The disclosed antibodies may be administered to patients who exhibit any one of the following: a) elevated hepcidin levels compared to normal (non-anemic) subjects; b) nutritional iron deficiency (serum iron less than 100 ng / ml); or c) EPO resistance.

[0040] In some embodiments, the disclosed methods are directed to the treatment of anemia in CKD patients with a GFR greater than 15 ml / min. In some embodiments, the disclosed methods are directed to the treatment of anemia in CKD patients with a GFR greater than 15 ml / min but less than 90 ml / min.

[0041] In some embodiments of the disclosed methods, the subject to whom the hemojuvelin antagonist is administered has a blood glucose level of 1.73 m. 2Subjects are identified as having a level of GFR within the range of 15 mL / min to less than 90 mL / min per 1.73 m 2 Subjects may be identified as having a level of GFR within the range of 15 mL / min to less than 60 mL / min per 1.73 m. 2 The subject may have a GFR level within the range of 15 mL / min to less than 30 mL / min per 1.73 m. 2 Less than 30mL / min per 1.73m 2 Less than 15 mL / min per 1.73 m 2 The patient may have a GFR level in the range of less than 7 mL / min per day.

[0042] Thus, provided herein are methods of treating a subject suffering from CKD at stages 1-5, as provided in Table 13 below. In some embodiments, the chronic kidney disease is classified as being at a stage within the range of stages 1-4. In some embodiments, the CKD is classified as being at a stage within the range of stages 2-4. In certain embodiments, the CKD is classified as being at stage 4.

[0043] In some embodiments, the disclosed methods are directed to the treatment of anemia in CKD patients exhibiting albuminuria. Albuminuria (or proteinuria) refers to the presence of albumin in urine and is determined based on the albumin to creatinine (ACR) ratio. An ACR below 30 is categorized as A1 albuminuria, an ACR in the range of 30-300 is categorized as A2, and an ACR above 300 is categorized as A3. In some embodiments, the disclosed methods are directed to the treatment of anemia in CKD patients exhibiting an ACR below 300. In some embodiments, the disclosed methods are directed to the treatment of anemia in CKD patients exhibiting an ACR of 30-300.

[0044] In some embodiments, the disclosed method is directed to the treatment of anemia in CKD patients who exhibit a decrease in the level of endogenous EPO. In some embodiments, the disclosed method is directed to the treatment of anemia in CKD patients who exhibit functional iron deficiency or nutritional iron deficiency. In some embodiments, the disclosed method is directed to the treatment of anemia in CKD patients who exhibit shortened red blood cell survival. Subjects suffering from anemia associated with CKD typically exhibit a decrease in endogenous EPO level and / or functional iron deficiency. In some embodiments, the disclosed method is directed to the treatment of anemia in CKD patients who exhibit a decrease in hepcidin level. In some embodiments, the disclosed method is directed to the treatment of anemia in CKD patients who exhibit non-responsiveness to oral iron or IV iron (e.g., MonoFerric®).

[0045] Further aspects of the disclosure, including descriptions of defined terms, are provided below.

[0046] I. Definition Administration: As used herein, the term "administering" or "administration" means providing a conjugate to a subject in a manner that is physiologically and / or pharmacologically useful (e.g., treating a condition in a subject).

[0047] Anemia of chronic disease: As used herein, the term "anemia of chronic disease" (ACD) refers to a hematological disorder occurring in the context of a disease or condition that induces an active immune / inflammatory response that results in a deficiency in the blood's ability to transport oxygen. A chronic condition (e.g., lasting for 3 months or longer) can result in low levels of iron in the blood, despite normal or even increased levels of iron stores in macrophages and hepatocytes. In this context, inflammation can prevent the use of stored iron to produce sufficient healthy red blood cells, leading to anemia. In some embodiments, ACD is the result of a deficiency of red blood cells, a deficiency of hemoglobin, and / or a deficiency in total blood volume. In some embodiments, ACD is associated with altered iron metabolism and turnover of body iron (e.g., via macrophage sequestration), hemophagocytosis, a decrease in erythropoiesis, and / or a decrease in response to erythropoietin stimulation. In some embodiments, ACD may be associated with or characterized by one or more of the following: failure to produce erythropoietin (EPO), blunted bone marrow erythroid response to EPO, iron-limited erythropoiesis, and a reduced pool of EPO-responsive cells combined with a chronic condition associated with inflammation. Thus, in some embodiments, low serum iron levels can provide a diagnostic indication of the presence of ACD in a subject when observed in the presence of an underlying chronic condition or disease. In some embodiments, ACD is iron-restricted anemia, which can be characterized by functional iron deficiency that may exist in a subject as a result of iron accumulation in tissue macrophages. Conditions associated with ACD include diseases that share the hallmark of immune activation. Examples of conditions associated with ACD include, but are not limited to, chronic kidney disease and kidney disease (e.g., chronic renal failure).

[0048] Antibody: As used herein, the term "antibody" refers to a polypeptide that comprises at least one immunoglobulin variable domain or at least one antigenic determinant, e.g., a paratope, that specifically binds to an antigen. In some embodiments, the antibody is a full-length antibody. In some embodiments, the antibody is a chimeric antibody. In some embodiments, the antibody is a humanized antibody. However, in some embodiments, the antibody is a Fab fragment, a F(ab')2 fragment, an Fv fragment, or an scFv fragment. In some embodiments, the antibody is a nanobody derived from a camelid antibody or a nanobody derived from a shark antibody. In some embodiments, the antibody is a bispecific antibody. In some embodiments, the antibody comprises a framework having a human germline sequence. In another embodiment, the antibody comprises a heavy chain constant domain selected from the group consisting of IgG, IgG1, IgG2, IgG2A, IgG2B, IgG2C, IgG3, IgG4, IgA1, IgA2, IgD, IgM, and IgE constant domains. In some embodiments, the antibody comprises a heavy (H) chain variable region (herein referred to as V H ), and / or a light (L) chain variable region (abbreviated herein as V L In some embodiments, the antibody comprises a constant domain, e.g., an Fc region. Immunoglobulin constant domain refers to a heavy or light chain constant domain. Human IgG heavy and light chain constant domain amino acid sequences and their functional variations are known. With respect to the heavy chain, in some embodiments, the heavy chain of the antibody described herein may be an alpha (α), delta (Δ), epsilon (ε), gamma (γ), or mu (μ) heavy chain. In some embodiments, the heavy chain of the antibody described herein may comprise a human alpha (α), delta (Δ), epsilon (ε), gamma (γ), or mu (μ) heavy chain. In certain embodiments, the antibody described herein comprises a human gamma 1 CH1, CH2, and / or CH3 domain. In some embodiments, the antibody comprises a human gamma 1 CH2, CH3, and / or CH4 domain. HThe amino acid sequence of the domain includes the amino acid sequence of a human gamma (γ) heavy chain constant region, such as any known in the art. Non-limiting examples of human constant region sequences are described in the art, see, e.g., U.S. Patent No. 5,693,780 and Kabat EA et al., (1991), supra. In some embodiments, the V HThe domain comprises an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 98%, or at least 99% identical to any of the variable chain constant regions provided herein. In some embodiments, the antibody is modified, for example, by glycosylation, phosphorylation, sumoylation, and / or methylation. In some embodiments, the antibody is a glycosylated antibody that is conjugated to one or more sugar or carbohydrate molecules. In some embodiments, the one or more sugar or carbohydrate molecules are conjugated to the antibody by N-glycosylation, O-glycosylation, C-glycosylation, glypiation (GPI anchor attachment), and / or phosphoglycosylation. In some embodiments, the one or more sugar or carbohydrate molecules are monosaccharides, disaccharides, oligosaccharides, or glycans. In some embodiments, the one or more sugar or carbohydrate molecules are branched oligosaccharides or branched glycans. In some embodiments, the one or more sugar or carbohydrate molecules comprise a mannose unit, a glucose unit, an N-acetylglucosamine unit, or a phospholipid unit. In some embodiments, the antibody is a construct comprising a linker polypeptide or a polypeptide comprising one or more antigen-binding fragments of the present disclosure linked to an immunoglobulin constant domain. A linker polypeptide comprises two or more amino acid residues joined by peptide bonds and is used to link one or more antigen-binding moieties. Examples of linker polypeptides have been reported [see, e.g., Holliger, P., et al. (1993) Proc. Natl. Acad. Sci. USA 90:6444-6448; Poljak, RJ, et al. (1994) Structure 2:1121-1123]. Additionally, an antibody may be part of a larger immunoadhesion molecule formed by covalent or non-covalent association of the antibody or antibody moiety with one or more other proteins or peptides.Examples of such immunoadhesion molecules include the use of streptavidin core regions to create tetrameric scFv molecules [Kipriyanov, SM, et al. (1995) Human Antibodies and Hybridomas 6:93-101] and the use of cysteine ​​residues, marker peptides and C-terminal polyhistidine tags to create bivalent and biotinylated scFv molecules [Kipriyanov, SM, et al. (1994) Mol. Immunol. 31:1047-1058].

[0049] Affinity matured antibody: "Affinity matured antibody" as used herein refers to an antibody with one or more alterations in one or more CDRs that result in an improvement in the affinity (i.e., KD, kd or ka) of the antibody to the target antigen compared to a parent antibody that does not have the alteration(s). An exemplary affinity matured antibody will have nanomolar or even picomolar affinity to the target antigen. Various procedures for producing affinity matured antibodies are known in the art, including screening combinatorial antibody libraries prepared using biodisplay. For example, Marks et al., BioTechnology, 10: 779-783 (1992) describes affinity maturation by VH and VL domain shuffling. Random mutagenesis of CDR and / or framework residues is described by 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): 3310-3319 (1995);and Hawkins et al., J. Mol. Biol., 226: 889-896 (1992).Selective mutagenesis positions and selective mutations at constant or hypermutation positions with activity-enhancing amino acid residues are described in U.S. Patent No. 6,914,128B1.

[0050] Approximately: As used herein, the term "approximately" or "about" when applied to one or more values ​​of interest refers to a value that is similar to a stated reference value. In certain embodiments, the term "approximately" or "about" refers to a range of values ​​that is within 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or less in either direction (higher or lower) of the stated reference value, unless otherwise stated or otherwise clear from the context (except where such number would exceed 100% of possible values).

[0051] Chronic kidney disease (CKD) is defined by abnormalities in renal structure or function that have been present in a patient for longer than 3 months. In the United States alone, approximately 5.6 million patients have a glomerular filtration rate (GFR) below 45 ml / min. 60 mL / min / 1.73 m 2 A GFR of 0.1 or higher is within the normal range. The standard way to estimate GFR is by assay of creatinine levels in a blood sample. Creatinine is a waste product from the digestion of dietary protein and the normal breakdown of muscle tissue. A substantial decrease in GFR over a sustained period of more than one week suggests some kind of kidney damage. A substantial decrease in GFR for more than three months is usually diagnosed as CKD. GFR is expressed in units of ml / min. Adjusted for body surface area, GFR is ml / min / 1.73m 2 It is expressed in units of 0. Estimated GFR (eGFR) is recommended by clinical practice guidelines for the routine assessment of GFR, while measured GFR is recommended as a confirmatory test when a more precise assessment is required.

[0052] CKD comprises a group of conditions that affect kidney function resulting from damage to kidney structures. There are at least three classes of criteria by which a physician identifies a patient as having CKD. The first is identifying the cause of CKD; the second is assigning a GFR category; and the third is assigning an albuminuria category. Determining the cause of CKD is important to establish whether the patient has a generalized or localized condition based on identifying which kidney structures are damaged. There are three causes of CKD: i) glomerular, ii) tubulointerstitial, and iii) vascular disease. Classification into glomerular, tubulointerstitial, and vascular categories is based on analysis of pathologic-anatomical findings from kidney biopsy or imaging. Nephron loss or fibrosis affecting the glomerulus or tubules is classified as either glomerular or tubulointerstitial disease. Finally, findings from kidney biopsy or imaging that reveal changes to the kidney vasculature are classified as vascular disease.

[0053] There are five stages of CKD: G1, G2, G3, G4 and G5 (also referred to as stage 1, stage 2, stage 3, stage 4, and stage 5). The G3 stage can be further divided into substages 3a and 3b. The GFR rates corresponding to each stage are provided in Table 13 below:

[0054] [Table 1]

[0055] See National Kidney Foundation, K / DOQI clinical practice guidelines for chronic kidney disease: Evaluation, classification, and stratification, Am. J. Kidney Dis. 2002: 39 (Suppl 1): S1-S266, incorporated herein by reference.

[0056] CKD-NDD: CKD patients who are not prescribed dialysis can be classified as being "non-dialysis dependent," or CKD-NDD (or NDD-CKD). These patients are not receiving dialysis (or dialytic) therapy.

[0057] Comorbidity: As used herein, "comorbidity" refers to one or more conditions or disorders that occur together (or co-occur) in an individual with a primary condition (e.g., CKD).

[0058] Albuminuria (or proteinuria) refers to the presence of albumin in the urine. Albuminuria categories are determined based on the albumin-to-creatinine (ACR) ratio. An ACR below 30 is categorized as A1 albuminuria, an ACR between 30-300 is categorized as A2, and an ACR above 300 is categorized as A3.

[0059] CDR: As used herein, the term "CDR" refers to a complementarity determining region in an antibody variable sequence. A typical antibody molecule comprises a heavy chain variable region (VH) and a light chain variable region (VL), which are usually involved in antigen binding. The VH and VL regions can be further subdivided into hypervariable regions, also known as "complementarity determining regions" ("CDRs"), interspersed with more conserved regions known as "framework regions" ("FRs"). Each VH and VL is typically composed of three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The extent of the framework regions and CDRs can be precisely identified using methods known in the art, for example, by the Kabat definition, the IMGT definition, the Chothia definition, the AbM definition, and / or the contact definition, all of which are well known in the art.Furthermore, Kabat, EA, et al. ( 1991 ) Sequences of Proteins of Immunological Interest , Fifth Edition , US Department of Health and Human Services , NIH Publication No . 91-3242;IMGT(Manufacturing), the International ImMunoGeneTics Information System(Manufacturing) (imgt.org), Lefranc, M.-P. http: / / dx.doi.org / 10.1037 / 0021-843X.112.2.213 Ruiz, M. et al., Nucleic Acids Res., 27:209-212 (1999); Nucleic Acids Res., 31:307–310 (2003);Lefranc, M.-P. et al., In Silico Biol., 5, 0006 (2004) [Epub], 5:45-60 (2005);Lefranc, M.-P. et al., Nucleic Acids Res., 33:D593-597 (2005);Lefranc, M.-P. et al., Nucleic Acids Res., 37:D1006-1012 (2009);Lefranc, M.-P. et al., Nucleic Acids Res., 43:D413-422 (2015);Chothia et al., (1989) Nature 342:877; Chothia, C. et al. (1987) J. Mol. Biol. 196:901–917, Al-Lazikani et al (1997) J. Molec. Biol. 273:927-948;and Almagro, J. Mol. Recognize. 17:132-143 (2004) at hgmp.mrc.ac.uk and at bioinf.org.uk / abs.As used herein, CDR can refer to CDRs defined by any method known in the art. Two antibodies with the same CDR means that the two antibodies have the same amino acid sequence of the CDR when determined by the same method, for example, the IMGT definition.

[0060] There are usually three CDRs in each variable region of the heavy and light chains, designated CDR1, CDR2 and CDR3 for each variable region. As used herein, the term "CDR set" refers to a group of three CDRs occurring in a single variable region capable of binding to an antigen. The exact boundaries of these CDRs are defined differently by different systems. The system described by Kabat [Kabat et al., Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, Md. (1987) and (1991))] not only provides a clear residue numbering system that is applicable to any variable region of an antibody, but also provides the exact residue boundaries that define the three CDRs. These CDRs may be referred to as Kabat CDRs. The subportions of the CDRs may be designated as L1, L2 and L3 or H1, H2 and H3, with "L" and "H" indicating the light and heavy chain regions, respectively. These regions may be referred to as Chothia CDRs, and have overlapping boundaries with the Kabat CDRs. Other boundaries defining CDRs that overlap with the Kabat CDRs have been described by Padlan [FASEB J. 9:133-139 (1995)] and MacCallum [J Mol Biol 262(5):732-45 (1996)]. Still other CDR boundary definitions may not strictly follow one of the above systems, but may be shortened or extended to take into account predictions or experimental findings that certain residues or groups of residues or even entire CDRs do not significantly affect antigen binding, despite overlapping with the Kabat CDRs. The methods used herein may utilize CDRs defined according to any of these systems, although exemplary embodiments use CDRs defined by Kabat or Chothia.

[0061] CDR-grafted antibody: The term "CDR-grafted antibody" refers to an antibody that contains heavy and light chain variable region sequences from one species but H and / or V L"CDR" refers to an antibody in which the sequence of one or more CDR regions of the mouse has been replaced with CDR sequence(s) of another species, for example, an antibody having mouse heavy and light chain variable regions in which one or more mouse CDRs (e.g., CDR3) have been replaced with human CDR sequences.

[0062] Chimeric antibody: The term "chimeric antibody" refers to an antibody that contains heavy and light chain variable region sequences from one species and constant region sequences from another species, such as an antibody having murine heavy and light chain variable regions linked to human constant regions.

[0063] Complementarity: As used herein, the term "complementarity" refers to the ability to precisely pair between two nucleotides or two sets of nucleotides. In particular, complementarity is a term that characterizes the degree of hydrogen bond pairing that results in binding between two nucleotides or two sets of nucleotides. For example, if a base at one position of an oligonucleotide can hydrogen bond with a base at the corresponding position of a target nucleic acid (e.g., mRNA), the bases are considered to be complementary to each other at that position. Base pairs can include both classical Watson-Crick base pairs and non-Watson-Crick base pairs (e.g., wobble base pairs and Hoogsteen base pairs). For example, in some embodiments, for complementary base pairing, an adenosine-type base (A) is complementary to a thymidine-type base (T) or a uracil-type base (U), a cytosine-type base (C) is complementary to a guanosine-type base (G), and a universal base, such as 3-nitropyrrole or 5-nitroindole, will hybridize with and be complementary to either A, C, U, or T. Inosine (I) is also considered to be a universal base in the art and is considered to be complementary to either A, C, U, or T.

[0064] Conservative amino acid substitution: As used herein, "conservative amino acid substitution" refers to an amino acid substitution that does not change the relative charge or size characteristics of the protein in which the amino acid substitution is made. Mutants can be prepared according to methods of modifying polypeptide sequences known to those skilled in the art, for example, references that collect such methods, such as Molecular Cloning: A Laboratory Manual, J. Sambrook, et al., eds., Fourth Edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 2012, or Current Protocols in Molecular Biology, FM Ausubel, et al., eds., John Wiley & Sons, Inc., New York. Conservative amino acid substitutions include substitutions made in the amino acids in the following groups: (a) M, I, L, V; (b) F, Y, W; (c) K, R, H; (d) A, G; (e) S, T; (f) Q, N; and (g) E, D.

[0065] Cross-reactivity: As used herein, and in the context of targeting agents (e.g., antibodies), the term "cross-reactivity" refers to the property of an agent that can specifically bind to one or more antigens of a similar type or class (e.g., antigens of various homologs, paralogs, or orthologs) with similar affinity or avidity. For example, in some embodiments, an antibody that is cross-reactive to a similar type or class of human and non-human primate antigens (e.g., human hemojuvelin and non-human primate hemojuvelin) can bind to human antigens and non-human primate antigens with similar affinity or avidity. In some embodiments, the antibody is cross-reactive to a similar type or class of human antigens and rodent antigens. In some embodiments, the antibody is cross-reactive to a similar type or class of rodent antigens and non-human primate antigens. In some embodiments, the antibody is cross-reactive to a similar type or class of human antigens, non-human primate antigens, and rodent antigens.

[0066] Effective amount: As used herein, "effective amount" refers to the amount of each active agent (e.g., hepcidin antagonist, anti-HJV antibody), either alone or in combination with one or more other active agents, required to provide a therapeutic effect to a subject (e.g., in treating anemia associated with CKD, or anemia in a subject with a certain level of GFR). In some embodiments, the therapeutic effect is a decrease in hepcidin levels or activity, an increase in the level of transferrin saturation (TSAT%), a decreased level of circulating transferrin levels, and / or a reduction in a disease state (e.g., a decrease in anemia).

[0067] Framework: As used herein, the term "framework" or "framework sequence" refers to the remaining sequence of the variable region, excluding the CDRs. The exact definition of the CDR sequence may be determined by different systems, so the meaning of the framework sequence assumes correspondingly different interpretations. The six CDRs (CDR-L1, CDR-L2, and CDR-L3 of the light chain and CDR-H1, CDR-H2, and CDR-H3 of the heavy chain) also divide the framework regions of the light and heavy chains into four subregions (FR1, FR2, FR3, and FR4) of each chain, with CDR1 located between FR1 and FR2, CDR2 between FR2 and FR3, and CDR3 between FR3 and FR4. As otherwise mentioned, without specifying a particular subregion as FR1, FR2, FR3, or FR4, the framework region represents the combined FRs in the variable region of a single, naturally occurring immunoglobulin chain. As used herein, FR refers to one of the four subregions, and FRs refers to two or more of the four subregions that make up a framework region. Human heavy and light chain acceptor sequences are known in the art. In one embodiment, acceptor sequences known in the art can be used in the antibodies disclosed herein.

[0068] Glomerular filtration rate: As used herein, "glomerular filtration rate" (GFR) is a measure of kidney function that describes the flow rate of filtered fluid through the kidney. GFR is a measurement of the volume of blood passing through the glomerulus each minute (glomeruli are clusters of capillaries in the nephron that filter waste products from the blood). The standard for calculating the measured GFR is to directly measure the plasma or urinary clearance of exogenous filtration markers (e.g., inulin or iohexol) in a subject. In other embodiments, an exemplary method for calculating the estimated GFR (eGFR) in a patient is calculated from a 24-hour urine collection and is calculated as 1.73 m 2 Normalized to human body surface area of ​​1000 mg / kg. See Tattersal et al., Nephrol Dial Transplant (2011) 26: 2082-2086, incorporated herein by reference. In some embodiments, "glomerular filtration rate" refers to estimated GFR (eGFR). In some embodiments, "glomerular filtration rate" refers to measured GFR.

[0069] Hemojuvelin (HJV): As used herein, the term "hemojuvelin (HJV)" (also known as repulsive guidance molecule C (RGMc) or hemochromatosis type 2 protein (HFE2)) refers to a membrane-bound soluble form protein that controls hepcidin production by the BMP / SMAD signaling pathway. The HFE2 gene encodes two known classes of GPI-anchored and glycosylated HJV molecules, which are targeted to membranes and undergo different fates. HJV exists in multiple isoforms, including two soluble isoforms and two membrane-bound isoforms. In some embodiments, the predominant membrane-bound isoform is a disulfide-linked two-chain form composed of N- and C-terminal fragments. In some embodiments, the full-length single-chain isoform is membrane-bound but is released from the cell surface and accumulates in the extracellular fluid. In some embodiments, the HJV can be of human (NCBI Gene ID148738), non-human primate (e.g., NCBI Gene ID 698805), or rodent (e.g., NCBI Gene ID69585 or NCBI Gene ID310681) origin. In addition to HJV (RGMc), the repulsive guidance molecule family includes repulsive guidance molecule A (RGMa) and repulsive guidance molecule B (RGMb). RGMa and RGMb are expressed in the central nervous system during development and are thought to be involved in regulating axonal patterning and neuronal survival, while HJV is produced in the liver and cardiac and skeletal muscles.

[0070] Hepcidin: As used herein, "hepcidin" refers to an iron-regulating peptide hormone that is mainly produced in the liver and is encoded by the HAMP gene. In some embodiments, hepcidin controls the delivery of iron from enterocytes that absorb iron, from macrophages that recycle red blood cells, and from hepatocytes that store iron to plasma. In some embodiments, hepcidin inhibits iron transport by binding to the iron export channel ferroportin, which is located on the basolateral surface of enterocytes and on the plasma membrane of reticuloendothelial cells (macrophages). In some embodiments, inhibition of ferroportin prevents iron from being exported and iron is sequestered in the cell. In some embodiments, by inhibiting ferroportin, hepcidin prevents enterocytes from entering iron into the hepatic portal system, thereby reducing dietary iron absorption. Hepcidin expression involves multiple aspects, including, for example, the transcription of HAMP gene, the translation of transcribed mRNA, and the post-translational processing of hepcidin precursor into bioactive hepcidin-25 peptide [DTHFPICIFCCGCCHRSKCGMCCKT (SEQ ID NO: 129)].In some embodiments, hepcidin expression is modulated through hemojuvelin-induced BMP signaling pathway.In some embodiments, hepcidin expression is modulated through IL-6-JAK-STAT signaling pathway.

[0071] Hepcidin antagonist: As used herein, "hepcidin antagonist" refers to an agent that reduces (directly or indirectly) hepcidin expression and / or hepcidin activity. In some embodiments, hepcidin antagonists inhibit hepcidin-induced ferroportin degradation. Thus, in some embodiments, hepcidin antagonists indirectly target hepcidin function through hepcidin stimulation pathways and reduce hepcidin expression. In some embodiments, hepcidin antagonists directly target hepcidin function, for example, by binding to hepcidin peptides to capture free hepcidin, or by binding to ferroportin to inhibit hepcidin-ferroportin binding interaction, thereby reducing hepcidin-induced ferroportin degradation. In some embodiments, the hepcidin antagonist is a ferroportin inhibitor that disrupts the ferroportin-hepcidin interaction, e.g., as disclosed in Ross SL, et al., Identification of Antibody and Small Molecule Antagonists of Ferroportin-Hepcidin Interaction. Front Pharmacol. 2017 Nov 21;8:838; Fung E., et al., High-Throughput Screening of Small Molecules Identifies Hepcidin Antagonists. Molecular Pharmacology March 2013, 83 (3) 681-690; and Angeliki Katsarou and Kostas Pantopoulos, Hepcidin Therapeutics. Pharmaceuticals (Basel). 2018 Dec; 11(4): 127, the relevant contents of each of which are incorporated herein by reference.

[0072] Hemojuvelin antagonist: As used herein, the term "hemojuvelin antagonist" refers to a molecule that reduces the expression of hemojuvelin or inhibits hemojuvelin, for example, by binding to hemojuvelin. In some embodiments, the hemojuvelin antagonist is an antisense oligonucleotide (see, for example, U.S. Patent No. 7,534,764; U.S. Patent Application Publication No. 2014 / 127325; and WO 2016 / 180784, which are incorporated herein by reference). In some embodiments, the hemojuvelin antagonist is an antibody. In other embodiments, the hemojuvelin antagonist is a small molecule compound that inhibits hemojuvelin, for example, by competitive binding and / or chemical modification of hemojuvelin.

[0073] HJV-induced BMP signaling: As used herein, the term "HJV-induced BMP signaling" refers to signaling through the BMP receptor induced by hemojuvelin (HJV), a membrane-bound coreceptor of bone morphogenetic protein (BMP) signaling. As described in Xia Y, et al., Hemojuvelin regulates hepcidin expression via a selective subset of BMP ligands and receptors independently of neogenin, Blood. 2008 May 15; 111(10): 5195-5204, in hepatocytes, HJV-induced BMP signaling positively regulates hepcidin mRNA expression. In some embodiments, HJV binds to BMP2, BMP4, BMP5, or BMP6 and induces BMP signaling, e.g., positively regulates hepcidin levels in hepatocytes. In some embodiments, cleavage of HJV by matriptase-2 reduces the amount of cell surface HJV available to participate in BMP signaling. In some embodiments, induction of BMP signaling by HJV is not dependent on neogenin. However, in some embodiments, neogenin promotes induction of BMP signaling by HJV, as described in Zhao et al, Neogenin Facilitates the Induction of Hepcidin Expression by Hemojuvelin in the Liver, J Biol Chem. 2016 Jun 3; 291(23): 12322-12335. In some embodiments, BMP6 is responsible for iron-dependent activation of Smad signaling. In some embodiments, BMP6 is secreted from liver sinusoidal endothelial cells and binds to BMP receptors (BMPR) on hepatocytes, thereby activating the SMAD signaling cascade. In such embodiments, HJV serves as a co-receptor for such BMP6, for example, positively regulating hepcidin levels on hepatocytes. In some embodiments, BMP transmits signals by binding to one or a combination of type I and type II serine / threonine kinase receptors.BMP type II receptors include BMPRII, ActRIIA, and ActRIIB. BMP type I receptors include ALK3, ALK6, and ALK2. In some embodiments, upon ligand binding, constitutively active type II receptor phosphorylates type I receptor, which then phosphorylates intracellular receptor-activated Smads (R-Smads), mainly Smad1, Smad5, and / or Smad8. In such embodiments, activated R-Smads form a complex with common partner Smad4 and translocate to the nucleus to control gene transcription, for example, induction of hepcidin expression.

[0074] Human antibody: The term "human antibody" as used herein is intended to include antibodies having variable and constant regions derived from human germline immunoglobulin sequences. The human antibodies of the present disclosure may include amino acid residues (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo) that are not encoded by human germline immunoglobulin sequences, for example in the CDRs, particularly CDR3. However, the term "human antibody" as used herein is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, have been grafted onto human framework sequences (e.g., CDRs grafted into a xenogeneic framework).

[0075] Humanized antibody: The term "humanized antibody" refers to an antibody that contains heavy and light chain variable region sequences from a non-human species (e.g., mouse), but does not contain any V H and / or V L Humanized antibodies refer to antibodies in which at least a portion of the sequence has been altered to be more "human-like," i.e., more similar to human germline variable sequences. One type of humanized antibody is a CDR-grafted antibody, in which human CDR sequences are grafted onto non-human V sequences. H and V LThe humanized anti-hemojuvelin antibody and antigen-binding portion are introduced into the sequence and replace the corresponding non-human CDR sequence. In one embodiment, a humanized anti-hemojuvelin antibody and antigen-binding portion are provided. Such an antibody can be produced by obtaining a mouse anti-hemojuvelin monoclonal antibody using classical hybridoma technology followed by humanization using in vitro genetic engineering, such as that disclosed in Kasaian et al., International Application PCT2005 / 123126A2.

[0076] Isolated antibody: As used herein, an "isolated antibody" is intended to refer to an antibody that is substantially free of other antibodies having different antigenic specificity (e.g., an isolated antibody that specifically binds to hemojuvelin is substantially free of antibodies that specifically bind to antigens other than hemojuvelin). An isolated antibody that specifically binds to hemojuvelin may, however, have cross-reactivity to other antigens, such as other repulsive guidance molecule (RGM) proteins (e.g., RGMa and / or RGMb). Additionally, an isolated antibody may be substantially free of other cellular material and / or chemicals.

[0077] Kabat numbering: The terms "Kabat numbering", "Kabat definition" and "Kabat labeling" are used interchangeably herein. These terms, as recognized in the art, refer to a system for numbering amino acid residues that are more variable (i.e., hypervariable) than other amino acid residues in the heavy and light chain variable regions of an antibody, or antigen-binding portion thereof [Kabat et al. (1971) Ann. NY Acad, Sci. 190:382-391 and, Kabat, EA, et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, US Department of Health and Human Services, NIH Publication No. 91-3242]. For the heavy chain variable region, the hypervariable region ranges from amino acid 31 to 35 in CDR1, amino acid 50 to 65 in CDR2, and amino acid 95 to 102 in CDR3. For the light chain variable region, the hypervariable region ranges from amino acid positions 24 to 34 in CDR1, amino acid positions 50 to 56 in CDR2, and amino acid positions 89 to 97 in CDR3.

[0078] Renal Damage: As used herein, the term "renal damage" refers to structural or functional abnormalities of the kidney with or without associated reduction in estimated or measured glomerular filtration rate (GFR) that manifest as pathological abnormalities or markers of kidney disease, including abnormalities in blood or urine composition or abnormalities in imaging studies.

[0079] Anemia associated with renal injury, as used herein, is anemia that occurs concomitantly with renal injury or is caused directly or indirectly by renal injury.

[0080] Oligonucleotide: As used herein, the term "oligonucleotide" refers to an oligomeric nucleic acid compound up to 200 nucleotides in length. Examples of oligonucleotides include, but are not limited to, RNAi oligonucleotides (e.g., siRNA, shRNA), microRNA, gapmers, mixmers, phosphorodiamidate morpholinos, peptide nucleic acids, aptamers, guide nucleic acids (e.g., Cas9 guide RNA), and the like. Oligonucleotides can be single-stranded or double-stranded. In some embodiments, oligonucleotides can include one or more modified nucleotides (e.g., 2'-O-methyl sugar modifications, purine or pyrimidine modifications). In some embodiments, oligonucleotides can include one or more modified internucleotide bonds. In some embodiments, oligonucleotides can include one or more phosphorothioate bonds, which can be in Rp or Sp stereochemistry.

[0081] Nephrectomy: As used herein, the term "nephrectomy" refers to the removal of part or all of one or more kidneys.

[0082] Recombinant antibody: As used herein, the term "recombinant human antibody" refers to any human antibody that is prepared, expressed, generated or isolated by recombinant means, such as an antibody expressed using a recombinant expression vector transfected into a host cell (as described in more detail in this disclosure), an antibody isolated from a recombinant, combinatorial human antibody library [Hoogenboom HR, (1997) TIB Tech. 15:62-70; Azzazy H., and Highsmith WE, (2002) Clin. Biochem. 35:425-445; Gavilondo JV, and Larrick JW (2002) BioTechniques 29:128-145; Hoogenboom H., and Chames P. (2000) Immunology Today 21:371-378], or an antibody isolated from an animal (e.g., a mouse) that is transgenic for human immunoglobulin genes [e.g., Taylor, LD, et al. (1992) Nucl. Acids 29:128-145; Hoogenboom H., and Chames P. (2000) Immunology Today 21:371-378]. Res. 20:6287-6295; Kellermann SA., and Green LL (2002) Current Opinion in Biotechnology 13:593-597; Little M. et al (2000) Immunology Today 21:364-370], or any other means including splicing of human immunoglobulin gene sequences to other DNA sequences. Such recombinant human antibodies have variable and constant regions derived from human germline immunoglobulin sequences. However, in certain embodiments, such recombinant human antibodies are subjected to in vitro mutagenesis (or in vivo somatic mutagenesis, when transgenic animals of human Ig sequences are used) and thus the V and constant regions of the recombinant antibodies are modified. H and V L The amino acid sequence of the region is H and V LA sequence derived from or related to a sequence, but which may not naturally occur within the human antibody germline repertoire in vivo. One embodiment of the present disclosure provides fully human antibodies capable of binding to human hemojuvelin, which may be generated using techniques well known in the art, for example, using human Ig phage libraries, such as, but not limited to, those disclosed in International Application PCT2005 / 007699A2 to Jermutus et al.

[0083] Selective: As used herein, the term "selective" or "selectively" refers to the ability of a molecule to produce an effect associated with its target molecule compared to a reference molecule. For example, a molecule that selectively inhibits its target molecule means that the molecule can inhibit the target molecule to a degree that can be distinguished from the reference molecule in an inhibition assay or other inhibition environment. For example, with respect to an inhibitor, the term "selectively inhibit" refers to the ability of the inhibitor to inhibit the target molecule to a degree that can be distinguished from a reference molecule that does not substantially inhibit in an inhibition assay, for example, as described herein, allowing selective inhibition of the target molecule. For example, the half maximal inhibitory concentration (IC50) of the target molecule and / or the reference molecule can be tested in a kinase potency assay (e.g., kinase potency assay by Carna Biosciences) as described in Ashoff, M. et al., Momelotinib inhibits ACVR1 / ALK2, decreases hepcidin production, and ameliorates anemia of chronic disease in rodents. Blood. 2017 Mar 30; 129(13): 1823-1830. In this assay, an inhibitor solution (e.g., a solution containing the selective inhibitor to be tested) / kinase substrate is mixed with a target molecule solution (e.g., ALK2) or a reference molecule solution (e.g., JAK1 or JAK2) and incubated at room temperature for 1 hour. After the reaction is over, the signal generated by the enzyme activity on the substrate can be measured. The half maximal inhibitory concentration of the target molecule and the reference molecule can be calculated. In some embodiments, the molecules described herein selectively bind to the target molecule. In some embodiments, the molecules described herein selectively inhibit the target molecule. In some embodiments, the molecules described herein selectively antagonize a target molecule. In some embodiments, the molecules described herein selectively neutralize a target molecule.

[0084] Specific Binding: As used herein, the term "specific binding" refers to the ability of a molecule to bind to a binding partner with a degree of affinity or avidity that allows the molecule to be used to distinguish the binding partner from an appropriate control in a binding assay or other binding environment. With respect to an antibody, the term "specific binding" refers to the ability of an antibody to bind to a particular antigen with a degree of affinity or avidity that allows the antibody to be used to distinguish the particular antigen from others, to the extent that the antibody allows for selective targeting of certain cells, e.g., muscle cells, via binding to the antigen, as compared to an appropriate reference antigen(s), e.g., as described herein. In some embodiments, an antibody has a binding affinity of at least about 10 -4 M, 10 -5 M, 10 -6 M, 10 -7 M, 10 -8 M, 10 -9 M, 10 -10 M, 10 -11 M, 10 -12 M, 10 -13 K for binding the target at or below M D In some embodiments, the antibody specifically binds to a target if it has the following structure:

[0085] Subject: As used herein, the term "subject" refers to a mammal. In some embodiments, the subject is a non-human primate, or a rodent. In some embodiments, the subject is a human. In some embodiments, the subject is a patient, such as a human patient having or suspected of having a disease. In some embodiments, the subject is a human patient having or suspected of having anemia associated with kidney disease and / or one or more conditions associated with or that may cause anemia associated with kidney disease and / or functional iron deficiency.

[0086] Treatment: As used herein, the term "treating" or "treatment" refers to the application or administration of a composition comprising one or more active agents to a subject having a target disease or disorder (e.g., anemia in CKD), a symptom of a disease / disorder, or a predisposition to a disease / disorder, for the purpose of curing, healing, mitigating, alleviating, altering, curing, ameliorating, improving, or affecting the disorder, a symptom of the disease, or a predisposition to a disease or disorder. Alleviating a target disease / disorder includes delaying or preventing the onset or progression of the disease, or reducing the severity of the disease.

[0087] II. Anti-Hemojuvelin (HJV) Antibody In some embodiments, the hemojuvelin antagonist binds to one or more proteins of the repulsive guidance molecule (RGM) family, including RGMa, RGMb, and RGMc (HJV). In some embodiments, the hemojuvelin antagonist selectively binds to hemojuvelin (RGMc) over RGMa and RGMb. In some embodiments, the hemojuvelin antagonist is an antisense oligonucleotide that reduces the expression of hemojuvelin (see, e.g., U.S. Pat. No. 7,534,764; U.S. Patent Application Publication No. 2014 / 127325; and WO 2016 / 180784; which are incorporated herein by reference). In some embodiments, the hemojuvelin antagonist is a small molecule compound that inhibits hemojuvelin, for example, by competitive binding and / or chemical modification of hemojuvelin.

[0088] In some embodiments, the hemojuvelin antagonist is an antibody (e.g., HA001-HA012) specific for hemojuvelin and / or one or more proteins of the RGM-like protein family (e.g., RGMa, RGMb). Suitable antibodies specific for hemojuvelin and / or one or more RGM-like proteins that may be useful in certain methods provided herein are provided, for example, in U.S. Pat. Nos. 10,118,958; and 8,507,435; U.S. Patent Application Publication No. 2013 / 330343; U.S. Patent Application Publication No. 2015 / 166672; and U.S. Patent Application Publication No. 2017 / 029499; and WO 2015 / 171691; and WO 2018 / 009624, which are incorporated herein by reference.

[0089] In some aspects, provided herein are antibodies that bind to human hemojuvelin (HJV) with high specificity and affinity. In some embodiments, the anti-HJV antibodies described herein specifically bind to any extracellular epitope of HJV or epitope exposed to the antibody. In some embodiments, the anti-HJV antibodies provided herein specifically bind to HJV from humans, non-human primates, mice, rats, etc. In some embodiments, the anti-HJV antibodies provided herein bind to human HJV. In some embodiments, the anti-HJV antibodies described herein bind to amino acid segments of human or non-human primate HJV.

[0090] In some embodiments, the anti-HJV antibodies described herein specifically bind to an epitope on human HJV. Human HJV is a 426 amino acid protein with a predicted N-terminal signal peptide of 31 amino acids and a C-terminal GPI attachment signal of 45 amino acids. An exemplary human HJV amino acid sequence is SEQ ID NO:128: MGEPGQSPSPRSSHGSPPTLSTLTLLLLLCGHAHSQCKILRCNAEYVSSTLSLRGGGSSGALRGGGGGGRGGGVGSGGLCRALRSYALCTRRTARTCRGDLAFHSAVHGIEDLMIQHNCSRQGPTAPPPPRGPALPGAGSGLPAPDPCDYEGRFSRLHGRPPGFLHCASFGDPHVRSFHHHFHTCRVQGAWPLLDNDFLFVQATSSPMALGANATATRKLTIIFKNMQECIDQKVYQAEVDNLPVAFEDGSINGGDRPGGSSLSIQTANPGNHVEIQAAYIGTTIIIRQTAGQLSFSIKVAEDVAMAFSAEQDLQLCVGGCPPSQRLSRSERNRRGAITIDTARRLCKEGLPVEDAYFHSCVFDVLISGDPNFTVAAQAALEDARAFLPDLEKLHLFPSDAGVPLSSATLLAPLLSGLFVLWLCIQ (SEQ ID NO: 128).

[0091] In some embodiments, the anti-HJV antibodies described herein may bind to a fragment of human HJV. The fragment of HJV may be about 5 to about 425 amino acids, about 10 to about 400 amino acids, about 50 to about 350 amino acids, about 100 to about 300 amino acids, about 150 to about 250 amino acids, about 200 to about 300 amino acids, or about 75 to about 150 amino acids in length. The fragment may comprise a contiguous number of amino acids from RGMc. Exemplary amino acids of HJV fragments are those set forth in SEQ ID NO: 123: QCKILRCNAEYVSSTLSLRGGGSSGALRGGGGGGRGGGVGSGGLCRALRSYALCTRRTARTCRGDLAFHSAVHGIEDLMIQHNCSRQGPTAPPPPRGPALPGAGSGLPAPDPCDYEGRFSRLHGRPPGFLHCASFGDPHVRSFHHHFHTCRVQGAWPLLDNDFLFVQATSSPMALGANATATRKLTIIFKNMQECIDQKVYQAEVDNLPVAFEDGSINGGDRPGGSSLSIQTANPGNHVEIQAAYIGTTIIIRQTAGQLSFSIKVAEDVAMAFSAEQDLQLCVGGCPPSQRLSRSERNRRGAITIDTARRLCKEGLPVEDAYFHSCVFDVLISGDPNFTVAAQAALEDARAFLPDLEKLHLFPSD (SEQ ID NO: 123).

[0092] In some embodiments, the anti-HJV antibodies described herein bind to different epitopes within human HJV or a fragment of human HJV.

[0093] In some embodiments, the anti-HJV antibody interacts with an epitope within amino acids 160-190 of SEQ ID NO: 123. In some embodiments, the anti-HJV antibody interacts with an epitope having an amino acid sequence of amino acids 170-183 of SEQ ID NO: 123. In some embodiments, the anti-HJV antibody interacts with an epitope having an amino acid sequence of SSPMALGANATATR (SEQ ID NO: 121). In some embodiments, the anti-HJV antibody interacts with different segments within SSPMALGANATATR (SEQ ID NO: 121). In some embodiments, the anti-HJV antibody interacts with amino acids 170-171, 171-180, 180-182, and 182-183 of SEQ ID NO: 123. In some embodiments, the antibody interacts with amino acids 170(S), 171(S), 180(T), 182(T) and 183 I of SEQ ID NO: 123. In some embodiments, hHA-008 interacts with the epitope SSPMALGANATATR (SEQ ID NO: 121). In some embodiments, hHA-008 interacts with amino acids 170(S), 171(S), 180(T), 182(T) and 183(R) of SEQ ID NO: 123.

[0094] In some embodiments, the anti-HJV antibody interacts with an epitope within amino acids 160-190 of SEQ ID NO: 123 and / or amino acids 280-310 of SEQ ID NO: 123. In some embodiments, the anti-HJV antibody interacts with an epitope within amino acids 169-182 of SEQ ID NO: 123 and / or amino acids 289-300 of SEQ ID NO: 123. In some embodiments, the anti-HJV antibody interacts with an epitope within amino acids 169-182 of SEQ ID NO: 123 and amino acids 289-300 of SEQ ID NO: 123. In some embodiments, the anti-HJV antibody interacts with an epitope having the amino acid sequence of TSSPMALGANATAT (SEQ ID NO: 122) and the amino acid sequence SQRLSRSERNRR (SEQ ID NO: 127). In some embodiments, the anti-HJV antibody interacts with different segments within TSSPMALGANATAT (SEQ ID NO: 122) and SQRLSRSERNRR (SEQ ID NO: 127). In some embodiments, the anti-HJV antibody interacts with amino acids 169-171, amino acids 171-180, and amino acids 180-182 of SEQ ID NO: 123, as well as amino acids 289-293, amino acids 293-294, amino acids 294-295, amino acids 295-297, and amino acids 297-300 of SEQ ID NO: 123. In some embodiments, the antibody interacts with amino acids 169(T), 170(S), 171(S), 180(T), 182(T), 289(S), 293(S), 294(R), 295(S), 297(R), and 300(R) of SEQ ID NO: 123. In some embodiments, hHA-008-QL interacts with different segments within TSSPMALGANATAT (SEQ ID NO: 122) and SQRLSRSERNRR (SEQ ID NO: 127). In some embodiments, hHA-008-QL interacts with amino acids 169(T), 170(S), 171(S), 180(T), 182(T), 289(S), 293(S), 294(R), 295(S), 297(R), and 300(R) of SEQ ID NO:123.

[0095] In some embodiments, the anti-HJV antibodies described herein are affinity matured clones. In some embodiments, the anti-HJV antibodies have a affinity of at least about 10 -4 M, 10-5 M, 10 -6 M, 10 -7 M, 10 -8 M, 10 -9 M, 10 -10 M, 10 -11 M, 10 -12 M, 10 -13 M, or a lower binding affinity (e.g., K D ) specifically binds to HJV (e.g., human or non-human primate HJV). For example, the anti-HJV antibodies of the present disclosure can bind to hemojuvelin protein (e.g., human hemojuvelin) with an affinity of 5 pM to 500 nM, e.g., 50 pM to 100 nM, e.g., 500 pM to 50 nM. The present disclosure also includes antibodies that compete with any of the antibodies described herein for binding to hemojuvelin protein (e.g., human hemojuvelin) and have an affinity of 100 nM or less (e.g., 80 nM or less, 50 nM or less, 20 nM or less, 10 nM or less, 500 pM or less, 50 pM or less, or 5 pM or less). The affinity and binding kinetics of the anti-HJV antibodies can be tested using any suitable method, including, but not limited to, biosensor technology (e.g., OCTET or BIACORE). In some embodiments, the anti-HJV antibodies described herein have a K in the subnanomolar range. D In some embodiments, the anti-HJV antibodies described herein selectively bind to RGMc but not to RGMa or RGMb.

[0096] Binding affinity (or binding specificity) can be determined by a variety of methods, including equilibrium dialysis, equilibrium binding, gel filtration, ELISA, surface plasmon resonance (SPR), fluorescence activated cell sorting (FACS) or spectroscopy (e.g., using a fluorescence assay). Exemplary conditions for evaluating binding affinity are in HBS-P buffer [10 mM HEPES pH 7.4, 150 mM NaCl, 0.005% (v / v) surfactant P20] and PBS buffer (10 mM PO4-3, 137 mM NaCl, and 2.7 mM KCl). These techniques can be used to measure the concentration of bound protein as a function of target protein concentration. The concentration of bound protein ([bound]) is generally calculated according to the following formula: [Bound]=[Free] / (Kd+[Free]) is related to the concentration of free target protein ([Free]) by

[0097] K A It is not necessary to make an exact determination of K, and in some cases it may be sufficient to have a quantitative indication of affinity, determined using methods such as, for example, ELISA or FACS analysis, which is K A A comparison can be used to determine whether a compound is more highly affinitive, e.g., two-fold higher, since the affinity is proportional to the affinity of the compound, e.g., activity in a functional assay, e.g., an in vitro or in vivo assay, provides a qualitative indication of affinity or provides an inference of affinity.

[0098] The heavy chain (HC) and light chain (LC) sequences, heavy chain variable domains (VH) and light chain variable domains (VL), CDR sequences, and heavy and light chain constant region sequences of non-limiting examples of anti-HJV antibodies are provided in Table 1.

[0099] [Table 2-1] [Table 2-2]

Table 2-3

Table 2-4

[0100]

Table 2-5

Table 2-6

Table 2-7

Table 2-8

Table 2-9

[0101]

Table 2-10

Table 2-11

Table 2-12

[0102] In some embodiments, the N-terminus of the heavy chain of the anti-HJV antibody described herein is glutamic acid (E). In some embodiments, glutamic acid can spontaneously cyclize to pyroglutamic acid by post-translational modification. The spontaneous cyclization of glutamic acid to pyroglutamic acid has been previously described, for example, in Chelius et al., Formation of Pyroglutamic Acid From N-terminal Glutamic Acid in Immunoglobulin Gamma Antibodies, Anal Chem. 2006;78(7):2370-2376. In some embodiments, the N-terminus of the heavy chain of the anti-HJV antibody described herein is pyroglutamic acid. In some embodiments, the anti-HJV antibody having an N-terminal pyroglutamic acid is an impurity in a population of anti-HJV antibodies (e.g., less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, less than 0.05%, or less than 0.01%) in a population of anti-HJV antibodies. In some embodiments, a population of anti-HJV antibodies comprises a mixture of anti-HJV antibodies having glutamic acid or pyroglutamic acid at the N-terminus of the heavy chain.

[0103] In some embodiments, an anti-HJV antibody of the present disclosure comprises one or more of the HC CDR (e.g., HC CDR1, HC CDR2, or HC CDR3) amino acid sequences from any one of the anti-HJV antibodies selected from Table 1. In some embodiments, an anti-HJV antibody of the present disclosure comprises the HC CDR1, HC CDR2, and HC CDR3 provided for any one of the antibodies selected from Table 1. In some embodiments, an anti-HJV antibody of the present disclosure comprises one or more of the LC CDR (e.g., LC CDR1, LC CDR2, or LC CDR3) amino acid sequences from any one of the anti-HJV antibodies selected from Table 1. In some embodiments, an anti-HJV antibody of the present disclosure comprises the LC CDR1, LC CDR2, and LC CDR3 provided for any one of the anti-HJV antibodies selected from Table 1.

[0104] In some embodiments, the anti-HJV antibody of the present disclosure comprises HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and LC CDR3 according to any one of the anti-HJV antibodies selected from Table 1. In some embodiments, the antibody heavy and light chain CDR3 domains may play a particularly important role in the binding specificity / affinity of the antibody to the antigen. Thus, the anti-HJV antibody of the present disclosure may comprise at least the heavy and / or light chain CDR3 of any one of the anti-HJV antibodies selected from Table 1.

[0105] In some embodiments, the isolated anti-HJV antibody comprises a heavy chain variable region comprising heavy chain CDR1 (HC CDR1), heavy chain CDR2 (HC CDR2), and heavy chain CDR3 (HC CDR3).

[0106] In some embodiments, HC CDR1 may comprise the amino acid sequence of X1YGMN (SEQ ID NO: 105), where X1 can be N or Y, according to the Kabat definition. Alternatively or additionally, HC CDR2 may comprise the amino acid sequence of MIYYDSSX2KHYADSVKG (SEQ ID NO: 106), where X2 can be E or D. Alternatively or additionally, HC CDR3 may comprise the amino acid sequence of GX3TPDX4 (SEQ ID NO: 107), where X3 can be T or S and X4 can be Y, V, or K.

[0107] In some embodiments, the anti-HJV antibody may comprise a light chain variable region comprising a light chain CDR1 (LC CDR1), a light chain CDR2 (LC CDR2), and a light chain CDR3 (LC CDR3) according to the Kabat definition. In some embodiments, the LC CDR1 may comprise the amino acid sequence of RSSQSLX5X6SDGX7TFLX8 (SEQ ID NO: 108), where X5 can be A or E, X6 can be T, S, E, or D, X7 can be D, Y, or G, and X8 can be E or H. Alternatively or additionally, the LC CDR2 may comprise the amino acid sequence of X9VSX 10 RFS (SEQ ID NO: 109) (wherein X9 can be E, D or A, and X 10Alternatively or additionally, the LC CDR3 may comprise the amino acid sequence of X 11 QX 12 TX 13 DPX 14 X 15 (SEQ ID NO: 110) (wherein X 11 can be F or M, and X 12 can be V or A, and X 13 can be H or Y, and X 14 can be M, L or V, and X 15 may comprise the amino acid sequence of

[0108] Also within the scope of the present disclosure are functional variants of any of the exemplary anti-HJV antibodies disclosed herein. Functional variants have a V that is greater than or equal to 100% of the V of the reference antibody while retaining substantially similar binding and biological activity (e.g., substantially similar binding affinity, binding specificity, inhibitory activity, anti-inflammatory activity, or a combination thereof) to the reference antibody. H and / or V L It may contain one or more amino acid residue variations in, or in one or more of the HC CDRs and / or one or more of the LC CDRs.

[0109] In some embodiments, any anti-HJV antibody of the present disclosure has one or more CDR (e.g., HC CDR or LC CDR) sequences substantially similar to any of the HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and / or LC CDR3 sequences from one of the anti-HJV antibodies selected from Table 1. In some embodiments, the position of one or more CDRs of the VH (e.g., HC CDR1, HC CDR2, or HC CDR3) and / or VL (e.g., LC CDR1, LC CDR2, or LC CDR3) regions of an antibody described herein may be altered by 1, 2, 3, 4, 5, or 6 amino acid positions, so long as immunospecific binding to hemojuvelin (e.g., human hemojuvelin) is maintained (e.g., at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% of the binding of the original antibody from which it is derived is substantially maintained). For example, in some embodiments, the positions defining the CDRs of any antibody described herein may be altered by shifting the N-terminal and / or C-terminal boundaries of the CDR by 1, 2, 3, 4, 5, or 6 amino acids compared to the CDR positions of any one of the antibodies described herein, so long as immunospecific binding to hemojuvelin (e.g., human hemojuvelin) is maintained (e.g., at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% of the binding of the original antibody from which it is derived is substantially maintained). In another embodiment, the length of one or more CDRs of the VH (e.g., HC CDR1, HC CDR2, or HC CDR3) and / or VL (e.g., LC CDR1, LC CDR2, or LC CDR3) regions of an antibody described herein may vary (e.g., shorter or longer) by one, two, three, four, five, or more amino acids, so long as immunospecific binding to hemojuvelin (e.g., human hemojuvelin) is maintained (e.g., at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% of the binding of the original antibody from which it is derived is substantially maintained).

[0110] Thus, in some embodiments, the HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and / or LC CDR3 described herein may be one, two, three, four, five, or more amino acids shorter than one or more CDRs described herein (e.g., a CDR from any of the anti-HJV antibodies selected from Table 1), so long as immunospecific binding to hemojuvelin (e.g., human hemojuvelin) is maintained (e.g., substantially maintained at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% compared to the binding of the original antibody from which it is derived). In some embodiments, the HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and / or LC CDR3 described herein may be one, two, three, four, five, or more amino acids longer than one or more CDRs described herein (e.g., a CDR from any of the anti-HJV antibodies selected from Table 1), so long as immunospecific binding to hemojuvelin (e.g., human hemojuvelin) is maintained (e.g., substantially maintained at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% compared to the binding of the original antibody from which it is derived). In some embodiments, the amino portions of the HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and / or LC CDR3 described herein may be extended by one, two, three, four, five, or more amino acids compared to one or more CDRs described herein (e.g., a CDR from any of the anti-HJV antibodies selected from Table 1), so long as immunospecific binding to hemojuvelin (e.g., human hemojuvelin) is maintained (e.g., substantially maintained at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% compared to the binding of the original antibody from which it is derived).In some embodiments, the carboxy portions of HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and / or LC CDR3 described herein may be extended by one, two, three, four, five, or more amino acids compared to one or more CDRs described herein (e.g., a CDR from any of the anti-HJV antibodies selected from Table 1), so long as immunospecific binding to hemojuvelin (e.g., human hemojuvelin) is maintained (e.g., substantially maintained at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% compared to the binding of the original antibody from which it is derived). In some embodiments, the amino portions of the HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and / or LC CDR3 described herein may be truncated by one, two, three, four, five, or more amino acids compared to one or more CDRs described herein (e.g., a CDR from any of the anti-HJV antibodies selected from Table 1), so long as immunospecific binding to hemojuvelin (e.g., human hemojuvelin) is maintained (e.g., substantially maintained at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% compared to the binding of the original antibody from which it is derived). In some embodiments, the carboxy portions of HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and / or LC CDR3 described herein may be truncated by one, two, three, four, five, or more amino acids compared to one or more CDRs described herein (e.g., a CDR from any of the anti-HJV antibodies selected from Table 1), so long as immunospecific binding to hemojuvelin (e.g., human hemojuvelin) is maintained (e.g., substantially maintained at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% compared to the binding of the original antibody from which it is derived).Any method can be used to determine whether immunospecific binding to hemojuvelin (eg, human hemojuvelin) is maintained, for example, using binding assays and conditions described in the art.

[0111] In some examples, any of the anti-HJV antibodies of the present disclosure have one or more CDR (e.g., HC CDR or LC CDR) sequences substantially similar to any one of the anti-HJV antibodies selected from Table 1. For example, the antibody may comprise one or more CDR sequence(s) from any of the anti-HJV antibodies selected from Table 1 that contain up to 5, 4, 3, 2, or 1 amino acid residue variations compared to the corresponding CDR region of any one of the CDRs provided herein (e.g., a CDR from any of the anti-HJV antibodies selected from Table 1), so long as the immunospecific binding to hemojuvelin (e.g., human hemojuvelin) is maintained (e.g., substantially maintained at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% compared to the binding of the original antibody from which it is derived). In some embodiments, any amino acid variation in any of the CDRs provided herein may be a conservative variation. Conservative variations may be introduced into CDRs at positions where the residues would not be involved in interactions with hemojuvelin proteins (e.g., human hemojuvelin proteins), for example, as determined based on a crystal structure. Some aspects of the present disclosure provide anti-HJV antibodies comprising one or more heavy chain variable (VH) and / or light chain variable (VL) domains provided herein. In some embodiments, any VH domain provided herein comprises one or more HC CDR sequences (e.g., HC CDR1, HC CDR2, and HC CDR3) provided herein, for example, any CDR-H sequence provided in any one of the anti-HJV antibodies selected from Table 1. In some embodiments, any VL domain provided herein comprises one or more CDR-L sequences (e.g., LC CDR1, LC CDR2, and LC CDR3) provided herein, for example, any LC CDR sequence provided in any one of the anti-HJV antibodies selected from Table 1.

[0112] In some embodiments, the anti-HJV antibodies of the disclosure include any antibody that comprises the heavy chain variable domain and / or the light chain variable domain of any one of the anti-HJV antibodies selected from Table 1, as well as variants thereof. In some embodiments, the anti-HJV antibodies of the disclosure include any antibody that comprises the heavy chain variable and light chain variable pair of any of the anti-HJV antibodies selected from Table 1.

[0113] Aspects of the present disclosure provide anti-HJV antibodies having heavy chain variable (VH) and / or light chain variable (VL) domain amino acid sequences homologous to any of those described herein. In some embodiments, the anti-HJV antibody comprises a heavy chain variable sequence or a light chain variable sequence that is at least 75% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to any one of the heavy chain variable and / or light chain variable sequences of the anti-HJV antibodies selected from Table 1. In some embodiments, the homologous heavy chain variable and / or light chain variable amino acid sequences do not vary within any of the CDR sequences provided herein. For example, in some embodiments, a degree of sequence variation (e.g., 75%, 80%, 85%, 90%, 95%, 98%, or 99%) may occur within the heavy chain variable and / or light chain variable sequences excluding any of the CDR sequences provided herein. In some embodiments, any of the anti-HJV antibodies provided herein comprises heavy chain and light chain variable sequences comprising framework sequences that are at least 75%, 80%, 85%, 90%, 95%, 98%, or 99% identical to the framework sequences of any of the anti-HJV antibodies selected from Table 1.

[0114] In some embodiments, an anti-HJV antibody of the disclosure is a humanized antibody (e.g., a humanized variant containing one or more CDRs of Table 1). In some embodiments, an anti-HJV antibody of the disclosure comprises HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and LC CDR3 that are the same as the HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and LC CDR3 shown in Table 1, and comprises a humanized heavy chain variable region and / or a humanized light chain variable region.

[0115] A humanized antibody is a human immunoglobulin (recipient antibody) in which residues from a complementarity determining region (CDR) of the recipient are replaced by residues from a CDR of a non-human species (donor antibody), such as mouse, rat, or rabbit, having the desired specificity, affinity, and capacity. In some embodiments, Fv framework region (FR) residues of the human immunoglobulin are replaced by corresponding non-human residues. Furthermore, humanized antibodies may comprise residues which are found neither in the recipient antibody nor in the imported CDR or framework sequences, but are included to further refine and optimize antibody performance. In general, a humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin and all or substantially all of the FR regions are of a human immunoglobulin consensus sequence. A humanized antibody optimally also comprises at least a portion of an immunoglobulin constant region or domain (Fc), typically of a human immunoglobulin. The antibody may have a modified Fc region as described in WO 99 / 58572. Other forms of humanized antibodies have one or more CDRs (one, two, three, four, five, six) that are altered with respect to the original antibody, also referred to as one or more CDRs derived from one or more CDRs from the original antibody. Humanized antibodies may also be affinity matured.

[0116] In some embodiments, humanization is achieved by grafting CDRs (e.g., those shown in Table 1) into a human variable domain (e.g., IGKV1-NL1*01 and IGHV1-3*01 human variable domains). In some embodiments, the anti-HJV antibodies of the disclosure are humanized variants that include one or more amino acid substitutions (e.g., in the VH framework regions) compared to any one of the VHs listed in Table 1, and / or one or more amino acid substitutions (e.g., in the VL framework regions) compared to any one of the VLs listed in Table 1.

[0117] In some embodiments, the anti-HJV antibodies of the disclosure are humanized antibodies that include a VH that contains 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to the VH of any of the anti-HJV antibodies listed in Table 1. Alternatively or additionally, the anti-HJV antibodies of the disclosure are humanized antibodies that include a VL that contains 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to the VL of any one of the anti-HJV antibodies listed in Table 1.

[0118] In some embodiments, an anti-HJV antibody of the disclosure comprises a HC CDR1, a HC CDR2 and a HC CDR3 of a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 7. Alternatively, or in addition, an anti-HJV antibody of the disclosure comprises a LC CDR1, a LC CDR2 and a LC CDR3 of a light chain variable domain having the amino acid sequence of SEQ ID NO: 8.

[0119] In some embodiments, according to the Kabat definition system, an anti-HJV antibody of the present disclosure comprises an HC CDR1 having the amino acid sequence of SEQ ID NO:1, an HC CDR2 having the amino acid sequence of SEQ ID NO:2, an HC CDR3 having the amino acid sequence of SEQ ID NO:3, an LC CDR1 having the amino acid sequence of SEQ ID NO:4, an LC CDR2 having the amino acid sequence of SEQ ID NO:5, and an LC CDR3 having the amino acid sequence of SEQ ID NO:6.

[0120] In some embodiments, the anti-HJV antibody of the present disclosure comprises HC CDR1, HC CDR2, and HC CDR3 that collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variations) compared to HC CDR1 having the amino acid sequence of SEQ ID NO:1, HC CDR2 having the amino acid sequence of SEQ ID NO:2, and HC CDR3 having the amino acid sequence of SEQ ID NO:3. "Total" as used anywhere in this disclosure means that the total number of amino acid variations in all three heavy chain CDRs is within a defined range. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises LC CDR1, LC CDR2, and LC CDR3 that collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variations) compared to LC CDR1 having the amino acid sequence of SEQ ID NO:4, LC CDR2 having the amino acid sequence of SEQ ID NO:5, and LC CDR3 having the amino acid sequence of SEQ ID NO:6.

[0121] In some embodiments, the anti-HJV antibodies of the present disclosure comprise HC CDR1, HC CDR2, and HC CDR3 that are at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to HC CDR1 having the amino acid sequence of SEQ ID NO:1, HC CDR2 having the amino acid sequence of SEQ ID NO:2, and HC CDR3 having the amino acid sequence of SEQ ID NO:3. Alternatively or additionally, the anti-HJV antibodies of the present disclosure comprise LC CDR1, LC CDR2, and LC CDR3 that are at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to LC CDR1 having the amino acid sequence of SEQ ID NO:4, LC CDR2 having the amino acid sequence of SEQ ID NO:5, and LC CDR3 having the amino acid sequence of SEQ ID NO:6.

[0122] In some embodiments, the anti-HJV antibodies of the present disclosure comprise an HC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an HC CDR1 having the amino acid sequence of SEQ ID NO:1; an HC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an HC CDR2 having the amino acid sequence of SEQ ID NO:2; and / or an HC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an HC CDR3 having the amino acid sequence of SEQ ID NO:3. Alternatively or additionally, the anti-HJV antibodies of the present disclosure include an LC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an LC CDR1 having the amino acid sequence of SEQ ID NO:4; an LC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an LC CDR2 having the amino acid sequence of SEQ ID NO:5; and / or an LC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an LC CDR3 having the amino acid sequence of SEQ ID NO:6.

[0123] In some embodiments, an anti-HJV antibody of the present disclosure comprises a VH comprising the amino acid sequence of SEQ ID NO: 7. Alternatively or additionally, an anti-HJV antibody of the present disclosure comprises a VL comprising the amino acid sequence of SEQ ID NO: 8.

[0124] In some embodiments, the anti-HJV antibodies of the present disclosure include a VH that contains 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to the VH set forth in SEQ ID NO: 7. Alternatively, or in addition, the anti-HJV antibodies of the present disclosure include a VL that contains 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to the VL set forth in SEQ ID NO: 8.

[0125] In some embodiments, an anti-HJV antibody of the present disclosure comprises a VH comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to the VH set forth in SEQ ID NO: 7. Alternatively, or in addition, an anti-HJV antibody of the present disclosure comprises a VL comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to the VL set forth in SEQ ID NO: 8.

[0126] In some embodiments, an anti-HJV antibody of the disclosure comprises a HC CDR1, a HC CDR2, and a HC CDR3 of a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 7. Alternatively, or in addition, an anti-HJV antibody of the disclosure comprises a LC CDR1, a LC CDR2, and a LC CDR3 of a light chain variable domain having the amino acid sequence of SEQ ID NO: 30.

[0127] In some embodiments, according to the Kabat definition system, an anti-HJV antibody of the present disclosure comprises an HC CDR1 having the amino acid sequence of SEQ ID NO:1, an HC CDR2 having the amino acid sequence of SEQ ID NO:2, an HC CDR3 having the amino acid sequence of SEQ ID NO:3, an LC CDR1 having the amino acid sequence of SEQ ID NO:4, an LC CDR2 having the amino acid sequence of SEQ ID NO:49, and an LC CDR3 having the amino acid sequence of SEQ ID NO:24.

[0128] In some embodiments, the anti-HJV antibody of the present disclosure comprises HC CDR1, HC CDR2, and HC CDR3 that collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variations) compared to HC CDR1 having the amino acid sequence of SEQ ID NO:1, HC CDR2 having the amino acid sequence of SEQ ID NO:2, and HC CDR3 having the amino acid sequence of SEQ ID NO:3. "Total" as used anywhere in this disclosure means that the total number of amino acid variations in all three heavy chain CDRs is within a defined range. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises LC CDR1, LC CDR2, and LC CDR3 that collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variations) compared to LC CDR1 having the amino acid sequence of SEQ ID NO:4, LC CDR2 having the amino acid sequence of SEQ ID NO:49, and LC CDR3 having the amino acid sequence of SEQ ID NO:24.

[0129] In some embodiments, the anti-HJV antibodies of the present disclosure comprise HC CDR1, HC CDR2, and HC CDR3 that are at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to HC CDR1 having the amino acid sequence of SEQ ID NO: 1, HC CDR2 having the amino acid sequence of SEQ ID NO: 2, and HC CDR3 having the amino acid sequence of SEQ ID NO: 3. Alternatively or additionally, the anti-HJV antibodies of the present disclosure comprise LC CDR1, LC CDR2, and LC CDR3 that are at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to LC CDR1 having the amino acid sequence of SEQ ID NO: 4, LC CDR2 having the amino acid sequence of SEQ ID NO: 49, and LC CDR3 having the amino acid sequence of SEQ ID NO: 24.

[0130] In some embodiments, the anti-HJV antibodies of the present disclosure comprise an HC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an HC CDR1 having the amino acid sequence of SEQ ID NO:1; an HC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an HC CDR2 having the amino acid sequence of SEQ ID NO:2; and / or an HC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an HC CDR3 having the amino acid sequence of SEQ ID NO:3. Alternatively or additionally, the anti-HJV antibodies of the present disclosure include an LC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an LC CDR1 having the amino acid sequence of SEQ ID NO:4; an LC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an LC CDR2 having the amino acid sequence of SEQ ID NO:49; and / or an LC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an LC CDR3 having the amino acid sequence of SEQ ID NO:24.

[0131] In some embodiments, an anti-HJV antibody of the present disclosure comprises a VH comprising the amino acid sequence of SEQ ID NO: 7. Alternatively, or additionally, an anti-HJV antibody of the present disclosure comprises a VL comprising the amino acid sequence of SEQ ID NO: 30.

[0132] In some embodiments, the anti-HJV antibodies of the present disclosure include a VH that contains 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to the VH set forth in SEQ ID NO: 7. Alternatively, or in addition, the anti-HJV antibodies of the present disclosure include a VL that contains 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to the VL set forth in SEQ ID NO: 30.

[0133] In some embodiments, an anti-HJV antibody of the disclosure comprises a VH comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to the VH set forth in SEQ ID NO: 7. Alternatively or additionally, an anti-HJV antibody of the disclosure comprises a VL comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to the VL set forth in SEQ ID NO: 30.

[0134] In some embodiments, an anti-HJV antibody of the disclosure comprises a HC CDR1, a HC CDR2, and a HC CDR3 of a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 7. Alternatively, or in addition, an anti-HJV antibody of the disclosure comprises a LC CDR1, a LC CDR2, and a LC CDR3 of a light chain variable domain having the amino acid sequence of SEQ ID NO: 31.

[0135] In some embodiments, according to the Kabat definition system, an anti-HJV antibody of the present disclosure comprises an HC CDR1 having the amino acid sequence of SEQ ID NO:1, an HC CDR2 having the amino acid sequence of SEQ ID NO:2, an HC CDR3 having the amino acid sequence of SEQ ID NO:3, an LC CDR1 having the amino acid sequence of SEQ ID NO:4, an LC CDR2 having the amino acid sequence of SEQ ID NO:18, and an LC CDR3 having the amino acid sequence of SEQ ID NO:25.

[0136] In some embodiments, the anti-HJV antibody of the present disclosure comprises HC CDR1, HC CDR2, and HC CDR3 that collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variations) compared to HC CDR1 having the amino acid sequence of SEQ ID NO:1, HC CDR2 having the amino acid sequence of SEQ ID NO:2, and HC CDR3 having the amino acid sequence of SEQ ID NO:3. "Total" as used anywhere in this disclosure means that the total number of amino acid variations in all three heavy chain CDRs is within a defined range. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises LC CDR1, LC CDR2, and LC CDR3 that collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variations) compared to LC CDR1 having the amino acid sequence of SEQ ID NO:4, LC CDR2 having the amino acid sequence of SEQ ID NO:18, and LC CDR3 having the amino acid sequence of SEQ ID NO:25.

[0137] In some embodiments, the anti-HJV antibodies of the present disclosure comprise HC CDR1, HC CDR2, and HC CDR3 that are at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical, together, to HC CDR1 having the amino acid sequence of SEQ ID NO: 1, HC CDR2 having the amino acid sequence of SEQ ID NO: 2, and HC CDR3 having the amino acid sequence of SEQ ID NO: 3. Alternatively or additionally, the anti-HJV antibodies of the present disclosure comprise LC CDR1, LC CDR2, and LC CDR3 that are at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical, together, to LC CDR1 having the amino acid sequence of SEQ ID NO: 4, LC CDR2 having the amino acid sequence of SEQ ID NO: 18, and LC CDR3 having the amino acid sequence of SEQ ID NO: 25.

[0138] In some embodiments, the anti-HJV antibodies of the present disclosure comprise an HC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an HC CDR1 having the amino acid sequence of SEQ ID NO:1; an HC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an HC CDR2 having the amino acid sequence of SEQ ID NO:2; and / or an HC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an HC CDR3 having the amino acid sequence of SEQ ID NO:3. Alternatively or additionally, the anti-HJV antibodies of the present disclosure include an LC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an LC CDR1 having the amino acid sequence of SEQ ID NO:4; an LC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an LC CDR2 having the amino acid sequence of SEQ ID NO:18; and / or an LC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an LC CDR3 having the amino acid sequence of SEQ ID NO:25.

[0139] In some embodiments, an anti-HJV antibody of the present disclosure comprises a VH comprising the amino acid sequence of SEQ ID NO: 7. Alternatively, or additionally, an anti-HJV antibody of the present disclosure comprises a VL comprising the amino acid sequence of SEQ ID NO: 31.

[0140] In some embodiments, the anti-HJV antibodies of the present disclosure include a VH that contains 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to the VH set forth in SEQ ID NO: 7. Alternatively, or in addition, the anti-HJV antibodies of the present disclosure include a VL that contains 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to the VL set forth in SEQ ID NO: 31.

[0141] In some embodiments, an anti-HJV antibody of the present disclosure comprises a VH comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to a VH set forth in SEQ ID NO: 7. Alternatively, or in addition, an anti-HJV antibody of the present disclosure comprises a VL comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to a VL set forth in SEQ ID NO: 31.

[0142] In some embodiments, an anti-HJV antibody of the disclosure comprises a HC CDR1, a HC CDR2 and a HC CDR3 of a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 7. Alternatively or additionally, an anti-HJV antibody of the disclosure comprises a LC CDR1, a LC CDR2 and a LC CDR3 of a light chain variable domain having the amino acid sequence of SEQ ID NO: 32.

[0143] In some embodiments, according to the Kabat definition system, an anti-HJV antibody of the present disclosure comprises an HC CDR1 having the amino acid sequence of SEQ ID NO:1, an HC CDR2 having the amino acid sequence of SEQ ID NO:2, an HC CDR3 having the amino acid sequence of SEQ ID NO:3, an LC CDR1 having the amino acid sequence of SEQ ID NO:14, an LC CDR2 having the amino acid sequence of SEQ ID NO:19, and an LC CDR3 having the amino acid sequence of SEQ ID NO:25.

[0144] In some embodiments, the anti-HJV antibody of the present disclosure comprises HC CDR1, HC CDR2, and HC CDR3 that collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variations) compared to HC CDR1 having the amino acid sequence of SEQ ID NO:1, HC CDR2 having the amino acid sequence of SEQ ID NO:2, and HC CDR3 having the amino acid sequence of SEQ ID NO:3. "Total" as used anywhere in this disclosure means that the total number of amino acid variations in all three heavy chain CDRs is within a defined range. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises LC CDR1, LC CDR2, and LC CDR3 that collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variations) compared to LC CDR1 having the amino acid sequence of SEQ ID NO:14, LC CDR2 having the amino acid sequence of SEQ ID NO:19, and LC CDR3 having the amino acid sequence of SEQ ID NO:25.

[0145] In some embodiments, the anti-HJV antibodies of the present disclosure comprise HC CDR1, HC CDR2, and HC CDR3 that are at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to HC CDR1 having the amino acid sequence of SEQ ID NO: 1, HC CDR2 having the amino acid sequence of SEQ ID NO: 2, and HC CDR3 having the amino acid sequence of SEQ ID NO: 3. Alternatively or additionally, the anti-HJV antibodies of the present disclosure comprise LC CDR1, LC CDR2, and LC CDR3 that are at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to LC CDR1 having the amino acid sequence of SEQ ID NO: 14, LC CDR2 having the amino acid sequence of SEQ ID NO: 19, and LC CDR3 having the amino acid sequence of SEQ ID NO: 25.

[0146] In some embodiments, the anti-HJV antibodies of the present disclosure comprise an HC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an HC CDR1 having the amino acid sequence of SEQ ID NO:1; an HC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an HC CDR2 having the amino acid sequence of SEQ ID NO:2; and / or an HC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an HC CDR3 having the amino acid sequence of SEQ ID NO:3. Alternatively or additionally, the anti-HJV antibodies of the present disclosure include an LC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an LC CDR1 having the amino acid sequence of SEQ ID NO: 14; an LC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an LC CDR2 having the amino acid sequence of SEQ ID NO: 19; and / or an LC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an LC CDR3 having the amino acid sequence of SEQ ID NO: 25.

[0147] In some embodiments, an anti-HJV antibody of the disclosure comprises a VH comprising the amino acid sequence of SEQ ID NO: 7. Alternatively, or additionally, an anti-HJV antibody of the disclosure comprises a VL comprising the amino acid sequence of SEQ ID NO: 32.

[0148] In some embodiments, the anti-HJV antibodies of the present disclosure include a VH that contains 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to the VH set forth in SEQ ID NO: 7. Alternatively or additionally, the anti-HJV antibodies of the present disclosure include a VL that contains 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to the VL set forth in SEQ ID NO: 32.

[0149] In some embodiments, an anti-HJV antibody of the present disclosure comprises a VH comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to the VH set forth in SEQ ID NO: 7. Alternatively or additionally, an anti-HJV antibody of the present disclosure comprises a VL comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to the VL set forth in SEQ ID NO: 32.

[0150] In some embodiments, an anti-HJV antibody of the disclosure comprises a HC CDR1, a HC CDR2 and a HC CDR3 of a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 7. Alternatively or additionally, an anti-HJV antibody of the disclosure comprises a LC CDR1, a LC CDR2 and a LC CDR3 of a light chain variable domain having the amino acid sequence of SEQ ID NO: 33.

[0151] In some embodiments, according to the Kabat definition system, an anti-HJV antibody of the present disclosure comprises an HC CDR1 having the amino acid sequence of SEQ ID NO:1, an HC CDR2 having the amino acid sequence of SEQ ID NO:2, an HC CDR3 having the amino acid sequence of SEQ ID NO:3, an LC CDR1 having the amino acid sequence of SEQ ID NO:15, an LC CDR2 having the amino acid sequence of SEQ ID NO:20, and an LC CDR3 having the amino acid sequence of SEQ ID NO:26.

[0152] In some embodiments, the anti-HJV antibody of the present disclosure comprises HC CDR1, HC CDR2, and HC CDR3 that collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variations) compared to HC CDR1 having the amino acid sequence of SEQ ID NO:1, HC CDR2 having the amino acid sequence of SEQ ID NO:2, and HC CDR3 having the amino acid sequence of SEQ ID NO:3. "Total" as used anywhere in this disclosure means that the total number of amino acid variations in all three heavy chain CDRs is within a defined range. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises LC CDR1, LC CDR2, and LC CDR3 that collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variations) compared to LC CDR1 having the amino acid sequence of SEQ ID NO:15, LC CDR2 having the amino acid sequence of SEQ ID NO:20, and LC CDR3 having the amino acid sequence of SEQ ID NO:26.

[0153] In some embodiments, the anti-HJV antibodies of the present disclosure comprise HC CDR1, HC CDR2, and HC CDR3 that are at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to HC CDR1 having the amino acid sequence of SEQ ID NO: 1, HC CDR2 having the amino acid sequence of SEQ ID NO: 2, and HC CDR3 having the amino acid sequence of SEQ ID NO: 3. Alternatively or additionally, the anti-HJV antibodies of the present disclosure comprise LC CDR1, LC CDR2, and LC CDR3 that are at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to LC CDR1 having the amino acid sequence of SEQ ID NO: 15, LC CDR2 having the amino acid sequence of SEQ ID NO: 20, and LC CDR3 having the amino acid sequence of SEQ ID NO: 26.

[0154] In some embodiments, the anti-HJV antibodies of the present disclosure comprise an HC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an HC CDR1 having the amino acid sequence of SEQ ID NO:1; an HC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an HC CDR2 having the amino acid sequence of SEQ ID NO:2; and / or an HC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an HC CDR3 having the amino acid sequence of SEQ ID NO:3. Alternatively or additionally, the anti-HJV antibodies of the present disclosure include an LC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an LC CDR1 having the amino acid sequence of SEQ ID NO: 15; an LC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an LC CDR2 having the amino acid sequence of SEQ ID NO: 20; and / or an LC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an LC CDR3 having the amino acid sequence of SEQ ID NO: 26.

[0155] In some embodiments, an anti-HJV antibody of the disclosure comprises a VH comprising the amino acid sequence of SEQ ID NO: 7. Alternatively, or additionally, an anti-HJV antibody of the disclosure comprises a VL comprising the amino acid sequence of SEQ ID NO: 33.

[0156] In some embodiments, the anti-HJV antibodies of the present disclosure include a VH that contains 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to the VH set forth in SEQ ID NO: 7. Alternatively or additionally, the anti-HJV antibodies of the present disclosure include a VL that contains 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to the VL set forth in SEQ ID NO: 33.

[0157] In some embodiments, an anti-HJV antibody of the present disclosure comprises a VH comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to the VH set forth in SEQ ID NO: 7. Alternatively or additionally, an anti-HJV antibody of the present disclosure comprises a VL comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to the VL set forth in SEQ ID NO: 33.

[0158] In some embodiments, an anti-HJV antibody of the disclosure comprises a HC CDR1, a HC CDR2 and a HC CDR3 of a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 34. Alternatively or additionally, an anti-HJV antibody of the disclosure comprises a LC CDR1, a LC CDR2 and a LC CDR3 of a light chain variable domain having the amino acid sequence of SEQ ID NO: 35.

[0159] In some embodiments, according to the Kabat definition system, an anti-HJV antibody of the present disclosure comprises an HC CDR1 having the amino acid sequence of SEQ ID NO:9, an HC CDR2 having the amino acid sequence of SEQ ID NO:2, an HC CDR3 having the amino acid sequence of SEQ ID NO:3, an LC CDR1 having the amino acid sequence of SEQ ID NO:16, an LC CDR2 having the amino acid sequence of SEQ ID NO:21, and an LC CDR3 having the amino acid sequence of SEQ ID NO:27.

[0160] In some embodiments, the anti-HJV antibody of the present disclosure comprises HC CDR1, HC CDR2, and HC CDR3 that collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variations) compared to HC CDR1 having the amino acid sequence of SEQ ID NO:9, HC CDR2 having the amino acid sequence of SEQ ID NO:2, and HC CDR3 having the amino acid sequence of SEQ ID NO:3. "Total" as used anywhere in this disclosure means that the total number of amino acid variations in all three heavy chain CDRs is within a defined range. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises LC CDR1, LC CDR2, and LC CDR3 that collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variations) compared to LC CDR1 having the amino acid sequence of SEQ ID NO:16, LC CDR2 having the amino acid sequence of SEQ ID NO:21, and LC CDR3 having the amino acid sequence of SEQ ID NO:27.

[0161] In some embodiments, the anti-HJV antibodies of the present disclosure comprise HC CDR1, HC CDR2, and HC CDR3 that are at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to HC CDR1 having the amino acid sequence of SEQ ID NO:9, HC CDR2 having the amino acid sequence of SEQ ID NO:2, and HC CDR3 having the amino acid sequence of SEQ ID NO:3. Alternatively or additionally, the anti-HJV antibodies of the present disclosure comprise LC CDR1, LC CDR2, and LC CDR3 that are at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to LC CDR1 having the amino acid sequence of SEQ ID NO:16, LC CDR2 having the amino acid sequence of SEQ ID NO:21, and LC CDR3 having the amino acid sequence of SEQ ID NO:27.

[0162] In some embodiments, the anti-HJV antibodies of the present disclosure include an HC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an HC CDR1 having the amino acid sequence of SEQ ID NO:9; an HC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an HC CDR2 having the amino acid sequence of SEQ ID NO:2; and / or an HC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an HC CDR3 having the amino acid sequence of SEQ ID NO:3. Alternatively or additionally, the anti-HJV antibodies of the present disclosure include an LC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an LC CDR1 having the amino acid sequence of SEQ ID NO: 16; an LC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an LC CDR2 having the amino acid sequence of SEQ ID NO: 21; and / or an LC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an LC CDR3 having the amino acid sequence of SEQ ID NO: 27.

[0163] In some embodiments, an anti-HJV antibody of the disclosure comprises a VH comprising the amino acid sequence of SEQ ID NO: 34. Alternatively or additionally, an anti-HJV antibody of the disclosure comprises a VL comprising the amino acid sequence of SEQ ID NO: 35.

[0164] In some embodiments, the anti-HJV antibodies of the present disclosure include a VH that contains 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to the VH set forth in SEQ ID NO: 34. Alternatively or additionally, the anti-HJV antibodies of the present disclosure include a VL that contains 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to the VL set forth in SEQ ID NO: 35.

[0165] In some embodiments, an anti-HJV antibody of the disclosure comprises a VH comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to the VH set forth in SEQ ID NO: 34. Alternatively or additionally, an anti-HJV antibody of the disclosure comprises a VL comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to the VL set forth in SEQ ID NO: 35.

[0166] In some embodiments, an anti-HJV antibody of the disclosure comprises a HC CDR1, a HC CDR2 and a HC CDR3 of a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 36. Alternatively or additionally, an anti-HJV antibody of the disclosure comprises a LC CDR1, a LC CDR2 and a LC CDR3 of a light chain variable domain having the amino acid sequence of SEQ ID NO: 37.

[0167] In some embodiments, according to the Kabat definition system, an anti-HJV antibody of the present disclosure comprises an HC CDR1 having the amino acid sequence of SEQ ID NO:1, an HC CDR2 having the amino acid sequence of SEQ ID NO:10, an HC CDR3 having the amino acid sequence of SEQ ID NO:11, an LC CDR1 having the amino acid sequence of SEQ ID NO:17, an LC CDR2 having the amino acid sequence of SEQ ID NO:18, and an LC CDR3 having the amino acid sequence of SEQ ID NO:28.

[0168] In some embodiments, the anti-HJV antibody of the present disclosure comprises HC CDR1, HC CDR2, and HC CDR3 that collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variations) compared to HC CDR1 having the amino acid sequence of SEQ ID NO: 1, HC CDR2 having the amino acid sequence of SEQ ID NO: 10, and HC CDR3 having the amino acid sequence of SEQ ID NO: 11. "Total" as used anywhere in this disclosure means that the total number of amino acid variations in all three heavy chain CDRs is within a defined range. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises LC CDR1, LC CDR2, and LC CDR3 that collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variations) compared to LC CDR1 having the amino acid sequence of SEQ ID NO: 17, LC CDR2 having the amino acid sequence of SEQ ID NO: 18, and LC CDR3 having the amino acid sequence of SEQ ID NO: 28.

[0169] In some embodiments, the anti-HJV antibodies of the present disclosure comprise HC CDR1, HC CDR2, and HC CDR3 that are at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to HC CDR1 having the amino acid sequence of SEQ ID NO: 1, HC CDR2 having the amino acid sequence of SEQ ID NO: 10, and HC CDR3 having the amino acid sequence of SEQ ID NO: 11. Alternatively or additionally, the anti-HJV antibodies of the present disclosure comprise LC CDR1, LC CDR2, and LC CDR3 that are at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to LC CDR1 having the amino acid sequence of SEQ ID NO: 17, LC CDR2 having the amino acid sequence of SEQ ID NO: 18, and LC CDR3 having the amino acid sequence of SEQ ID NO: 28.

[0170] In some embodiments, the anti-HJV antibodies of the present disclosure include an HC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an HC CDR1 having the amino acid sequence of SEQ ID NO:1; an HC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an HC CDR2 having the amino acid sequence of SEQ ID NO:10; and / or an HC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an HC CDR3 having the amino acid sequence of SEQ ID NO:11. Alternatively or additionally, the anti-HJV antibodies of the present disclosure include an LC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an LC CDR1 having the amino acid sequence of SEQ ID NO: 17; an LC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an LC CDR2 having the amino acid sequence of SEQ ID NO: 18; and / or an LC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an LC CDR3 having the amino acid sequence of SEQ ID NO: 28.

[0171] In some embodiments, an anti-HJV antibody of the present disclosure comprises a VH comprising the amino acid sequence of SEQ ID NO: 36. Alternatively or additionally, an anti-HJV antibody of the present disclosure comprises a VL comprising the amino acid sequence of SEQ ID NO: 37.

[0172] In some embodiments, the anti-HJV antibodies of the present disclosure include a VH that contains 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to the VH set forth in SEQ ID NO: 36. Alternatively or additionally, the anti-HJV antibodies of the present disclosure include a VL that contains 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to the VL set forth in SEQ ID NO: 37.

[0173] In some embodiments, an anti-HJV antibody of the present disclosure comprises a VH comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to the VH set forth in SEQ ID NO: 36. Alternatively or additionally, an anti-HJV antibody of the present disclosure comprises a VL comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to the VL set forth in SEQ ID NO: 37.

[0174] In some embodiments, an anti-HJV antibody of the disclosure comprises a HC CDR1, a HC CDR2 and a HC CDR3 of a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 38. Alternatively or additionally, an anti-HJV antibody of the disclosure comprises a LC CDR1, a LC CDR2 and a LC CDR3 of a light chain variable domain having the amino acid sequence of SEQ ID NO: 39.

[0175] In some embodiments, according to the Kabat definition system, an anti-HJV antibody of the present disclosure comprises an HC CDR1 having the amino acid sequence of SEQ ID NO:1, an HC CDR2 having the amino acid sequence of SEQ ID NO:2, an HC CDR3 having the amino acid sequence of SEQ ID NO:3, an LC CDR1 having the amino acid sequence of SEQ ID NO:17, an LC CDR2 having the amino acid sequence of SEQ ID NO:5, and an LC CDR3 having the amino acid sequence of SEQ ID NO:27.

[0176] In some embodiments, the anti-HJV antibody of the present disclosure comprises HC CDR1, HC CDR2, and HC CDR3 that collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variations) compared to HC CDR1 having the amino acid sequence of SEQ ID NO:1, HC CDR2 having the amino acid sequence of SEQ ID NO:2, and HC CDR3 having the amino acid sequence of SEQ ID NO:3. "Total" as used anywhere in this disclosure means that the total number of amino acid variations in all three heavy chain CDRs is within a defined range. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises LC CDR1, LC CDR2, and LC CDR3 that collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variations) compared to LC CDR1 having the amino acid sequence of SEQ ID NO:17, LC CDR2 having the amino acid sequence of SEQ ID NO:5, and LC CDR3 having the amino acid sequence of SEQ ID NO:27.

[0177] In some embodiments, an anti-HJV antibody of the present disclosure comprises an HC CDR1, an HC CDR2, and an HC CDR3 that are at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to an HC CDR1 having the amino acid sequence of SEQ ID NO: 1, an HC CDR2 having the amino acid sequence of SEQ ID NO: 2, and an HC CDR3 having the amino acid sequence of SEQ ID NO: 3. Alternatively or additionally, an anti-HJV antibody of the present disclosure comprises an LC CDR1, an LC CDR2, and an LC CDR3 that are at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to an LC CDR1 having the amino acid sequence of SEQ ID NO: 17, an LC CDR2 having the amino acid sequence of SEQ ID NO: 5, and an LC CDR3 having the amino acid sequence of SEQ ID NO: 27.

[0178] In some embodiments, the anti-HJV antibodies of the present disclosure comprise an HC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an HC CDR1 having the amino acid sequence of SEQ ID NO:1; an HC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an HC CDR2 having the amino acid sequence of SEQ ID NO:2; and / or an HC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an HC CDR3 having the amino acid sequence of SEQ ID NO:3. Alternatively or additionally, the anti-HJV antibodies of the present disclosure include an LC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an LC CDR1 having the amino acid sequence of SEQ ID NO: 17; an LC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an LC CDR2 having the amino acid sequence of SEQ ID NO: 5; and / or an LC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an LC CDR3 having the amino acid sequence of SEQ ID NO: 27.

[0179] In some embodiments, an anti-HJV antibody of the present disclosure comprises a VH comprising the amino acid sequence of SEQ ID NO: 38. Alternatively, or additionally, an anti-HJV antibody of the present disclosure comprises a VL comprising the amino acid sequence of SEQ ID NO: 39.

[0180] In some embodiments, the anti-HJV antibodies of the present disclosure include a VH that contains 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to the VH set forth in SEQ ID NO: 38. Alternatively or additionally, the anti-HJV antibodies of the present disclosure include a VL that contains 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to the VL set forth in SEQ ID NO: 39.

[0181] In some embodiments, an anti-HJV antibody of the present disclosure comprises a VH comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to the VH set forth in SEQ ID NO: 38. Alternatively or additionally, an anti-HJV antibody of the present disclosure comprises a VL comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to the VL set forth in SEQ ID NO: 39.

[0182] In some embodiments, an anti-HJV antibody of the disclosure comprises a HC CDR1, a HC CDR2 and a HC CDR3 of a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 38. Alternatively or additionally, an anti-HJV antibody of the disclosure comprises a LC CDR1, a LC CDR2 and a LC CDR3 of a light chain variable domain having the amino acid sequence of SEQ ID NO: 41.

[0183] In some embodiments, according to the Kabat definition system, an anti-HJV antibody of the present disclosure comprises an HC CDR1 having the amino acid sequence of SEQ ID NO:1, an HC CDR2 having the amino acid sequence of SEQ ID NO:2, an HC CDR3 having the amino acid sequence of SEQ ID NO:3, an LC CDR1 having the amino acid sequence of SEQ ID NO:50, an LC CDR2 having the amino acid sequence of SEQ ID NO:22, and an LC CDR3 having the amino acid sequence of SEQ ID NO:28.

[0184] In some embodiments, the anti-HJV antibody of the present disclosure comprises HC CDR1, HC CDR2, and HC CDR3 that collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variations) compared to HC CDR1 having the amino acid sequence of SEQ ID NO:1, HC CDR2 having the amino acid sequence of SEQ ID NO:2, and HC CDR3 having the amino acid sequence of SEQ ID NO:3. "Total" as used anywhere in this disclosure means that the total number of amino acid variations in all three heavy chain CDRs is within a defined range. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises LC CDR1, LC CDR2, and LC CDR3 that collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variations) compared to LC CDR1 having the amino acid sequence of SEQ ID NO:50, LC CDR2 having the amino acid sequence of SEQ ID NO:22, and LC CDR3 having the amino acid sequence of SEQ ID NO:28.

[0185] In some embodiments, an anti-HJV antibody of the present disclosure comprises an HC CDR1, an HC CDR2, and an HC CDR3 that are at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical, together, to an HC CDR1 having the amino acid sequence of SEQ ID NO: 1, an HC CDR2 having the amino acid sequence of SEQ ID NO: 2, and an HC CDR3 having the amino acid sequence of SEQ ID NO: 3. Alternatively or additionally, an anti-HJV antibody of the present disclosure comprises an LC CDR1, an LC CDR2, and an LC CDR3 that are at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical, together, to an LC CDR1 having the amino acid sequence of SEQ ID NO: 50, an LC CDR2 having the amino acid sequence of SEQ ID NO: 22, and an LC CDR3 having the amino acid sequence of SEQ ID NO: 28.

[0186] In some embodiments, the anti-HJV antibodies of the present disclosure comprise an HC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an HC CDR1 having the amino acid sequence of SEQ ID NO:1; an HC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an HC CDR2 having the amino acid sequence of SEQ ID NO:2; and / or an HC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an HC CDR3 having the amino acid sequence of SEQ ID NO:3. Alternatively or additionally, the anti-HJV antibodies of the present disclosure include an LC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an LC CDR1 having the amino acid sequence of SEQ ID NO:50; an LC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an LC CDR2 having the amino acid sequence of SEQ ID NO:22; and / or an LC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an LC CDR3 having the amino acid sequence of SEQ ID NO:28.

[0187] In some embodiments, an anti-HJV antibody of the disclosure comprises a VH comprising the amino acid sequence of SEQ ID NO: 38. Alternatively or additionally, an anti-HJV antibody of the disclosure comprises a VL comprising the amino acid sequence of SEQ ID NO: 41.

[0188] In some embodiments, the anti-HJV antibodies of the present disclosure include a VH that contains 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to the VH set forth in SEQ ID NO: 38. Alternatively or additionally, the anti-HJV antibodies of the present disclosure include a VL that contains 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to the VL set forth in SEQ ID NO: 41.

[0189] In some embodiments, an anti-HJV antibody of the disclosure comprises a VH comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to the VH set forth in SEQ ID NO: 38. Alternatively or additionally, an anti-HJV antibody of the disclosure comprises a VL comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to the VL set forth in SEQ ID NO: 41.

[0190] In some embodiments, an anti-HJV antibody of the disclosure comprises a HC CDR1, a HC CDR2 and a HC CDR3 of a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 42. Alternatively or additionally, an anti-HJV antibody of the disclosure comprises a LC CDR1, a LC CDR2 and a LC CDR3 of a light chain variable domain having the amino acid sequence of SEQ ID NO: 43.

[0191] In some embodiments, according to the Kabat definition system, an anti-HJV antibody of the present disclosure comprises an HC CDR1 having the amino acid sequence of SEQ ID NO:1, an HC CDR2 having the amino acid sequence of SEQ ID NO:2, an HC CDR3 having the amino acid sequence of SEQ ID NO:12, an LC CDR1 having the amino acid sequence of SEQ ID NO:15, an LC CDR2 having the amino acid sequence of SEQ ID NO:23, and an LC CDR3 having the amino acid sequence of SEQ ID NO:27.

[0192] In some embodiments, the anti-HJV antibody of the present disclosure comprises HC CDR1, HC CDR2, and HC CDR3 that collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variations) compared to HC CDR1 having the amino acid sequence of SEQ ID NO:1, HC CDR2 having the amino acid sequence of SEQ ID NO:2, and HC CDR3 having the amino acid sequence of SEQ ID NO:12. "Total" as used anywhere in this disclosure means that the total number of amino acid variations in all three heavy chain CDRs is within a defined range. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises LC CDR1, LC CDR2, and LC CDR3 that collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variations) compared to LC CDR1 having the amino acid sequence of SEQ ID NO:15, LC CDR2 having the amino acid sequence of SEQ ID NO:23, and LC CDR3 having the amino acid sequence of SEQ ID NO:27.

[0193] In some embodiments, the anti-HJV antibodies of the present disclosure comprise HC CDR1, HC CDR2, and HC CDR3 that are at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to HC CDR1 having the amino acid sequence of SEQ ID NO: 1, HC CDR2 having the amino acid sequence of SEQ ID NO: 2, and HC CDR3 having the amino acid sequence of SEQ ID NO: 12. Alternatively or additionally, the anti-HJV antibodies of the present disclosure comprise LC CDR1, LC CDR2, and LC CDR3 that are at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to LC CDR1 having the amino acid sequence of SEQ ID NO: 15, LC CDR2 having the amino acid sequence of SEQ ID NO: 23, and LC CDR3 having the amino acid sequence of SEQ ID NO: 27.

[0194] In some embodiments, the anti-HJV antibodies of the present disclosure include an HC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an HC CDR1 having the amino acid sequence of SEQ ID NO:1; an HC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an HC CDR2 having the amino acid sequence of SEQ ID NO:2; and / or an HC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an HC CDR3 having the amino acid sequence of SEQ ID NO:12. Alternatively or additionally, the anti-HJV antibodies of the present disclosure include an LC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an LC CDR1 having the amino acid sequence of SEQ ID NO: 15; an LC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an LC CDR2 having the amino acid sequence of SEQ ID NO: 23; and / or an LC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an LC CDR3 having the amino acid sequence of SEQ ID NO: 27.

[0195] In some embodiments, an anti-HJV antibody of the present disclosure comprises a VH comprising the amino acid sequence of SEQ ID NO: 42. Alternatively, or additionally, an anti-HJV antibody of the present disclosure comprises a VL comprising the amino acid sequence of SEQ ID NO: 43.

[0196] In some embodiments, the anti-HJV antibodies of the present disclosure include a VH that contains 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to the VH set forth in SEQ ID NO: 42. Alternatively or additionally, the anti-HJV antibodies of the present disclosure include a VL that contains 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to the VL set forth in SEQ ID NO: 43.

[0197] In some embodiments, an anti-HJV antibody of the disclosure comprises a VH comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to the VH set forth in SEQ ID NO: 42. Alternatively or additionally, an anti-HJV antibody of the disclosure comprises a VL comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to the VL set forth in SEQ ID NO: 43.

[0198] In some embodiments, an anti-HJV antibody of the disclosure comprises a HC CDR1, a HC CDR2 and a HC CDR3 of a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 44. Alternatively or additionally, an anti-HJV antibody of the disclosure comprises a LC CDR1, a LC CDR2 and a LC CDR3 of a light chain variable domain having the amino acid sequence of SEQ ID NO: 45.

[0199] In some embodiments, according to the Kabat definition system, an anti-HJV antibody of the present disclosure comprises an HC CDR1 having the amino acid sequence of SEQ ID NO:1, an HC CDR2 having the amino acid sequence of SEQ ID NO:2, an HC CDR3 having the amino acid sequence of SEQ ID NO:13, an LC CDR1 having the amino acid sequence of SEQ ID NO:16, an LC CDR2 having the amino acid sequence of SEQ ID NO:21, and an LC CDR3 having the amino acid sequence of SEQ ID NO:29.

[0200] In some embodiments, the anti-HJV antibody of the present disclosure comprises HC CDR1, HC CDR2, and HC CDR3 that collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variations) compared to HC CDR1 having the amino acid sequence of SEQ ID NO:1, HC CDR2 having the amino acid sequence of SEQ ID NO:2, and HC CDR3 having the amino acid sequence of SEQ ID NO:13. "Total" as used anywhere in this disclosure means that the total number of amino acid variations in all three heavy chain CDRs is within a defined range. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises LC CDR1, LC CDR2, and LC CDR3 that collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variations) compared to LC CDR1 having the amino acid sequence of SEQ ID NO:16, LC CDR2 having the amino acid sequence of SEQ ID NO:21, and LC CDR3 having the amino acid sequence of SEQ ID NO:29.

[0201] In some embodiments, an anti-HJV antibody of the present disclosure comprises an HC CDR1, an HC CDR2, and an HC CDR3 that are at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to an HC CDR1 having the amino acid sequence of SEQ ID NO: 1, an HC CDR2 having the amino acid sequence of SEQ ID NO: 2, and an HC CDR3 having the amino acid sequence of SEQ ID NO: 13. Alternatively or additionally, an anti-HJV antibody of the present disclosure comprises an LC CDR1, an LC CDR2, and an LC CDR3 that are at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to an LC CDR1 having the amino acid sequence of SEQ ID NO: 16, an LC CDR2 having the amino acid sequence of SEQ ID NO: 21, and an LC CDR3 having the amino acid sequence of SEQ ID NO: 29.

[0202] In some embodiments, the anti-HJV antibodies of the present disclosure include an HC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an HC CDR1 having the amino acid sequence of SEQ ID NO:1; an HC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an HC CDR2 having the amino acid sequence of SEQ ID NO:2; and / or an HC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an HC CDR3 having the amino acid sequence of SEQ ID NO:13. Alternatively or additionally, the anti-HJV antibodies of the present disclosure include an LC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an LC CDR1 having the amino acid sequence of SEQ ID NO: 16; an LC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an LC CDR2 having the amino acid sequence of SEQ ID NO: 21; and / or an LC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to an LC CDR3 having the amino acid sequence of SEQ ID NO: 29.

[0203] In some embodiments, an anti-HJV antibody of the disclosure comprises a VH comprising the amino acid sequence of SEQ ID NO: 44. Alternatively, or additionally, an anti-HJV antibody of the disclosure comprises a VL comprising the amino acid sequence of SEQ ID NO: 45.

[0204] In some embodiments, the anti-HJV antibodies of the present disclosure include a VH that contains 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to the VH set forth in SEQ ID NO: 44. Alternatively or additionally, the anti-HJV antibodies of the present disclosure include a VL that contains 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to the VL set forth in SEQ ID NO: 45.

[0205] In some embodiments, an anti-HJV antibody of the disclosure comprises a VH comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to the VH set forth in SEQ ID NO: 44. Alternatively or additionally, an anti-HJV antibody of the disclosure comprises a VL comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to the VL set forth in SEQ ID NO: 45.

[0206] The CDRs of an antibody may have different amino acid sequences when different definition systems are used (e.g., the IMGT definition, the Kabat definition, or the Chothia definition). The definition systems annotate each amino acid of a given antibody sequence (e.g., the VH or VL sequence) by a number, and the numbers corresponding to the heavy and light chain CDRs are provided in Table 2. The CDRs listed in Table 1 are defined by the Kabat definition. One skilled in the art can obtain the CDR sequences of the anti-HJV antibodies provided in Table 2 using different numbering systems.

[0207] [Table 3]

[0208] In some embodiments, the anti-HJV antibody of the present disclosure is a chimeric antibody that may contain heavy and light chain constant regions from a human antibody. Chimeric antibody refers to an antibody that has a variable region or a portion of a variable region from a first species and a constant region from a second species. Typically, in these chimeric antibodies, the variable regions of both the light and heavy chains mimic the variable regions of an antibody from one species of mammal (e.g., non-human mammals, such as mice, rabbits, and rats), and the constant portion is homologous to a sequence in an antibody from another mammal, such as a human. In some embodiments, amino acid modifications may be made in the variable and / or constant regions.

[0209] In some embodiments, the anti-HJV antibody described herein is a chimeric antibody that may contain heavy and light chain constant regions from a human antibody. Chimeric antibody refers to an antibody that has a variable region or a portion of a variable region from a first species and a constant region from a second species. Typically, in these chimeric antibodies, the variable regions of both the light and heavy chains mimic the variable region of an antibody from one species of mammal (e.g., non-human mammals, such as mice, rabbits, and rats), and the constant portion is homologous to a sequence in an antibody from another mammal, such as a human. In some embodiments, amino acid modifications may be made in the variable and / or constant regions.

[0210] In some embodiments, the anti-HJV antibodies of the present disclosure comprise a VL domain and / or a VH domain of any one of the anti-HJV antibodies selected from Table 1, and comprise a constant region comprising the amino acid sequence of the constant region of an IgG, IgE, IgM, IgD, IgA or IgY immunoglobulin molecule, any class of immunoglobulin molecule (e.g., IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2), or any subclass (e.g., IgG2a and IgG2b). Non-limiting examples of human constant regions are described in the art, see, for example, Kabat EA et al., (1991), supra. The following human IgG1 constant region sequences (e.g., SEQ ID NO:48, SEQ ID NO:112, SEQ ID NO:113, and SEQ ID NO:130) are variants of the heavy chain constant region sequence set forth in SEQ ID NO:46 (e.g., having 1-5 amino acids that differ from the heavy chain constant region sequence set forth in SEQ ID NO:46). Examples of human IgG1 constant regions are given below: ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 103)

[0211] In some embodiments, the heavy chain of any of the anti-HJV antibodies described herein comprises a mutant human IgG1 constant region. For example, the introduction of a LALA mutation in the CH2 domain of human IgG1 (a mutant derived from mAb b12 mutated to replace the lower hinge residues Leu234 and Leu235 with Ala234 and Ala235) is known to reduce Fcg receptor binding [Bruhns, P., et al. (2009) and Xu, D. et al. (2000)]. The mutant human IgG1 constant region is provided below (mutations are bonded and underlined): TIFF2024540480000016.tif32150

[0212] In some embodiments, the heavy chain of any of the anti-HJV antibodies described herein further comprises a mutation in the human IgG1 constant region. For example, introduction of T250Q and M248L substitutions. In some embodiments, such substitutions may affect FcRn binding and serum half-life (WO2005047307 and WO2013063110). An exemplary IgG1 constant region comprising LALA and QL mutations is provided below (mutations are bolded and underlined): TIFF2024540480000017.tif32150

[0213] It will be appreciated that in some embodiments, during production of the antibody, particularly using Chinese Hamster Ovary cells (CHO cells), the lysine at the C-terminus of the heavy chain is cleaved. Thus, the human IgG1 constant region in the secreted antibody may be: ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO: 111)

[0214] In some embodiments, a mutant human IgG1 comprising a LALA mutation in the secreted antibody may be: TIFF2024540480000018.tif32150

[0215] In some embodiments, a mutant human IgG1 comprising a LALA mutation and a QL mutation can be: TIFF2024540480000019.tif32150

[0216] In some embodiments, the light chain of any of the anti-HJV antibodies described herein may further comprise a light chain constant region (CL), which may be any CL known in the art. In some examples, the CL is a kappa light chain. In other examples, the CL is a lambda light chain. In some embodiments, the CL is a kappa light chain, the sequence of which is provided below: RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 47)

[0217] Other antibody heavy and light chain constant regions are known in the art, such as those provided in the IMGT database (imgt.org) or vbase2.org / vbstat), both of which are incorporated herein by reference.

[0218] In some embodiments, the anti-HJV antibodies described herein comprise a heavy chain comprising any one of the VHs listed in Table 1 or any variant thereof and a heavy chain constant region at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to SEQ ID NO:46, SEQ ID NO:48, SEQ ID NO:112, SEQ ID NO:113, or SEQ ID NO:130. In some embodiments, the anti-HJV antibodies described herein comprise a heavy chain comprising any one of the VHs listed in Table 1 or any variant thereof and a heavy chain constant region containing 20 or less amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or less amino acid variations) compared to SEQ ID NO:46, SEQ ID NO:48, SEQ ID NO:112, SEQ ID NO:113, or SEQ ID NO:130. In some embodiments, the anti-HJV antibodies described herein comprise a heavy chain comprising any one of the VHs listed in Table 1 or any variant thereof and a heavy chain constant region set forth in SEQ ID NO: 46. In some embodiments, the anti-HJV antibodies described herein comprise a heavy chain comprising any one of the VHs listed in Table 1 or any variant thereof and a heavy chain constant region set forth in SEQ ID NO: 48. In some embodiments, the anti-HJV antibodies described herein comprise a heavy chain comprising any one of the VHs listed in Table 1 or any variant thereof and a heavy chain constant region set forth in SEQ ID NO: 112. In some embodiments, the anti-HJV antibodies described herein comprise a heavy chain comprising any one of the VHs listed in Table 1 or any variant thereof and a heavy chain constant region set forth in SEQ ID NO: 113. In some embodiments, the anti-HJV antibodies described herein comprise a heavy chain comprising any one of the VHs listed in Table 1 or any variant thereof and a heavy chain constant region set forth in SEQ ID NO: 130.

[0219] In some embodiments, the anti-HJV antibodies described herein comprise a light chain comprising any one of the VLs listed in Table 1 or any variant thereof and a light chain constant region at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to SEQ ID NO: 47. In some embodiments, the anti-HJV antibodies described herein comprise a light chain comprising any one of the VLs listed in Table 1 or any variant thereof and a light chain constant region containing 20 or less amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or less amino acid variations) compared to SEQ ID NO: 47. In some embodiments, the anti-HJV antibodies described herein comprise a light chain comprising any one of the VLs listed in Table 1 or any variant thereof and a light chain constant region set forth in SEQ ID NO: 47.

[0220] Examples of IgG heavy and light chain amino acid sequences of the described anti-HJV antibodies are provided in Table 1 above.

[0221] In some embodiments, an anti-HJV antibody of the present disclosure comprises a heavy chain that contains 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to a heavy chain set forth in any one of SEQ ID NOs: 51, 57, 59, 61, 63, 66, 68, 114, 115, 116, 117, 118, 119, or 120. Alternatively or additionally, an anti-HJV antibody of the disclosure comprises a light chain that contains 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to a light chain set forth in any one of SEQ ID NOs: 52, 53, 54, 55, 56, 58, 60, 62, 65, 67 or 69. In some embodiments, the anti-HJV antibodies described herein comprise a heavy chain comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to any one of SEQ ID NOs: 51, 57, 59, 61, 63, 66, 68, 114, 115, 116, 117, 118, 119, or 120. Alternatively or additionally, the anti-HJV antibodies described herein comprise a light chain comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to any one of SEQ ID NOs: 52, 53, 54, 55, 56, 58, 60, 62, 65, 67, or 69. In some embodiments, the anti-HJV antibodies described herein comprise a heavy chain comprising the amino acid sequence of any one of SEQ ID NOs: 51, 57, 59, 61, 63, 66, 68, 114, 115, 116, 117, 118, 119, or 120. Alternatively or additionally, the anti-HJV antibodies described herein comprise a light chain comprising the amino acid sequence of any one of SEQ ID NOs: 52, 53, 54, 55, 56, 58, 60, 62, 65, 67, or 69.

[0222] In some embodiments, the anti-HJV antibodies of the present disclosure comprise a heavy chain that contains no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variations) compared to the heavy chain set forth in any one of SEQ ID NOs: 51 or 114. Alternatively or additionally, the anti-HJV antibodies of the present disclosure comprise a light chain that contains no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variations) compared to the light chain set forth in any one of SEQ ID NOs: 52. In some embodiments, the anti-HJV antibodies described herein comprise a heavy chain comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to any one of SEQ ID NOs: 51 or 114. Alternatively or additionally, the anti-HJV antibodies described herein comprise a light chain comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to any one of SEQ ID NOs: 52. In some embodiments, the anti-HJV antibodies described herein comprise a heavy chain comprising an amino acid sequence of any one of SEQ ID NOs: 51 or 114. Alternatively or additionally, the anti-HJV antibodies described herein comprise a light chain comprising an amino acid sequence of any one of SEQ ID NOs: 52.

[0223] In some embodiments, the anti-HJV antibodies of the present disclosure comprise a heavy chain that contains no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variations) compared to the heavy chain set forth in any one of SEQ ID NOs: 51 or 114. Alternatively or additionally, the anti-HJV antibodies of the present disclosure comprise a light chain that contains no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variations) compared to the light chain set forth in any one of SEQ ID NOs: 53. In some embodiments, the anti-HJV antibodies described herein comprise a heavy chain comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to any one of SEQ ID NOs: 51 or 114. Alternatively or additionally, the anti-HJV antibodies described herein comprise a light chain comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to any one of SEQ ID NOs: 53. In some embodiments, the anti-HJV antibodies described herein comprise a heavy chain comprising an amino acid sequence of any one of SEQ ID NOs: 51 or 114. Alternatively or additionally, the anti-HJV antibodies described herein comprise a light chain comprising an amino acid sequence of any one of SEQ ID NOs: 53.

[0224] In some embodiments, the anti-HJV antibodies of the present disclosure comprise a heavy chain that contains no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variations) compared to the heavy chain set forth in any one of SEQ ID NOs: 51 or 114. Alternatively or additionally, the anti-HJV antibodies of the present disclosure comprise a light chain that contains no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variations) compared to the light chain set forth in any one of SEQ ID NOs: 54. In some embodiments, the anti-HJV antibodies described herein comprise a heavy chain comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to any one of SEQ ID NOs: 51 or 114. Alternatively or additionally, the anti-HJV antibodies described herein comprise a light chain comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to any one of SEQ ID NOs: 54. In some embodiments, the anti-HJV antibodies described herein comprise a heavy chain comprising an amino acid sequence of any one of SEQ ID NOs: 51 or 114. Alternatively or additionally, the anti-HJV antibodies described herein comprise a light chain comprising an amino acid sequence of any one of SEQ ID NOs: 54.

[0225] In some embodiments, the anti-HJV antibodies of the present disclosure comprise a heavy chain that contains 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to the heavy chain set forth in any one of SEQ ID NOs: 51 or 114. Alternatively or additionally, the anti-HJV antibodies of the present disclosure comprise a light chain that contains 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to the light chain set forth in any one of SEQ ID NOs: 55. In some embodiments, the anti-HJV antibodies described herein comprise a heavy chain comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to any one of SEQ ID NOs: 51 or 114. Alternatively or additionally, the anti-HJV antibodies described herein comprise a light chain comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to any one of SEQ ID NOs: 55. In some embodiments, the anti-HJV antibodies described herein comprise a heavy chain comprising an amino acid sequence of any one of SEQ ID NOs: 51 or 114. Alternatively or additionally, the anti-HJV antibodies described herein comprise a light chain comprising an amino acid sequence of any one of SEQ ID NOs: 55.

[0226] In some embodiments, the anti-HJV antibodies of the present disclosure comprise a heavy chain that contains no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variations) compared to the heavy chain set forth in any one of SEQ ID NOs: 51 or 114. Alternatively or additionally, the anti-HJV antibodies of the present disclosure comprise a light chain that contains no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variations) compared to the light chain set forth in any one of SEQ ID NOs: 56. In some embodiments, the anti-HJV antibodies described herein comprise a heavy chain comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to any one of SEQ ID NOs: 51 or 114. Alternatively or additionally, the anti-HJV antibodies described herein comprise a light chain comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to any one of SEQ ID NOs: 56. In some embodiments, the anti-HJV antibodies described herein comprise a heavy chain comprising an amino acid sequence of any one of SEQ ID NOs: 51 or 114. Alternatively or additionally, the anti-HJV antibodies described herein comprise a light chain comprising an amino acid sequence of any one of SEQ ID NOs: 56.

[0227] In some embodiments, the anti-HJV antibodies of the present disclosure comprise a heavy chain that contains no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variations) compared to the heavy chain set forth in any one of SEQ ID NOs: 57 or 115. Alternatively or additionally, the anti-HJV antibodies of the present disclosure comprise a light chain that contains no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variations) compared to the light chain set forth in any one of SEQ ID NOs: 58. In some embodiments, the anti-HJV antibodies described herein comprise a heavy chain comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to any one of SEQ ID NOs: 57 or 115. Alternatively or additionally, the anti-HJV antibodies described herein comprise a light chain comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to any one of SEQ ID NOs: 58. In some embodiments, the anti-HJV antibodies described herein comprise a heavy chain comprising an amino acid sequence of any one of SEQ ID NOs: 57 or 115. Alternatively or additionally, the anti-HJV antibodies described herein comprise a light chain comprising an amino acid sequence of any one of SEQ ID NOs: 58.

[0228] In some embodiments, the anti-HJV antibodies of the present disclosure comprise a heavy chain that contains no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variations) compared to the heavy chain set forth in any one of SEQ ID NOs: 59 or 116. Alternatively or additionally, the anti-HJV antibodies of the present disclosure comprise a light chain that contains no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variations) compared to the light chain set forth in any one of SEQ ID NOs: 60. In some embodiments, the anti-HJV antibodies described herein comprise a heavy chain comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to any one of SEQ ID NOs: 59 or 116. Alternatively or additionally, the anti-HJV antibodies described herein comprise a light chain comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to any one of SEQ ID NOs: 60. In some embodiments, the anti-HJV antibodies described herein comprise a heavy chain comprising an amino acid sequence of any one of SEQ ID NOs: 59 or 116. Alternatively or additionally, the anti-HJV antibodies described herein comprise a light chain comprising an amino acid sequence of any one of SEQ ID NOs: 60.

[0229] In some embodiments, the anti-HJV antibodies of the present disclosure comprise a heavy chain that contains no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variations) compared to the heavy chain set forth in any one of SEQ ID NOs: 61 or 117. Alternatively or additionally, the anti-HJV antibodies of the present disclosure comprise a light chain that contains no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variations) compared to the light chain set forth in any one of SEQ ID NOs: 62. In some embodiments, the anti-HJV antibodies described herein comprise a heavy chain comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to any one of SEQ ID NOs: 61 or 117. Alternatively or additionally, the anti-HJV antibodies described herein comprise a light chain comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to any one of SEQ ID NOs: 62. In some embodiments, the anti-HJV antibodies described herein comprise a heavy chain comprising an amino acid sequence of any one of SEQ ID NOs: 61 or 117. Alternatively or additionally, the anti-HJV antibodies described herein comprise a light chain comprising an amino acid sequence of any one of SEQ ID NOs: 62.

[0230] In some embodiments, the anti-HJV antibodies of the disclosure comprise a heavy chain that contains no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variations) compared to the heavy chain set forth in any one of SEQ ID NOs: 63 or 118. Alternatively or additionally, the anti-HJV antibodies of the disclosure comprise a light chain that contains no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variations) compared to the light chain set forth in any one of SEQ ID NOs: 62. In some embodiments, the anti-HJV antibodies described herein comprise a heavy chain comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to any one of SEQ ID NOs: 63 or 118. Alternatively or additionally, the anti-HJV antibodies described herein comprise a light chain comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to any one of SEQ ID NOs: 62. In some embodiments, the anti-HJV antibodies described herein comprise a heavy chain comprising an amino acid sequence of any one of SEQ ID NOs: 63 or 118. Alternatively or additionally, the anti-HJV antibodies described herein comprise a light chain comprising an amino acid sequence of any one of SEQ ID NOs: 62.

[0231] In some embodiments, the anti-HJV antibodies of the present disclosure comprise a heavy chain that contains no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variations) compared to the heavy chain set forth in any one of SEQ ID NOs: 61 or 117. Alternatively or additionally, the anti-HJV antibodies of the present disclosure comprise a light chain that contains no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variations) compared to the light chain set forth in any one of SEQ ID NOs: 65. In some embodiments, the anti-HJV antibodies described herein comprise a heavy chain comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to any one of SEQ ID NOs: 61 or 117. Alternatively or additionally, the anti-HJV antibodies described herein comprise a light chain comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to any one of SEQ ID NOs: 65. In some embodiments, the anti-HJV antibodies described herein comprise a heavy chain comprising an amino acid sequence of any one of SEQ ID NOs: 61 or 117. Alternatively or additionally, the anti-HJV antibodies described herein comprise a light chain comprising an amino acid sequence of any one of SEQ ID NOs: 65.

[0232] In some embodiments, the anti-HJV antibodies of the present disclosure comprise a heavy chain that contains no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variations) compared to the heavy chain set forth in any one of SEQ ID NOs: 66 or 119. Alternatively or additionally, the anti-HJV antibodies of the present disclosure comprise a light chain that contains no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variations) compared to the light chain set forth in any one of SEQ ID NOs: 67. In some embodiments, the anti-HJV antibodies described herein comprise a heavy chain comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to any one of SEQ ID NOs: 66 or 119. Alternatively or additionally, the anti-HJV antibodies described herein comprise a light chain comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to any one of SEQ ID NOs: 67. In some embodiments, the anti-HJV antibodies described herein comprise a heavy chain comprising an amino acid sequence of any one of SEQ ID NOs: 66 or 119. Alternatively or additionally, the anti-HJV antibodies described herein comprise a light chain comprising an amino acid sequence of any one of SEQ ID NOs: 67.

[0233] In some embodiments, the anti-HJV antibodies of the present disclosure comprise a heavy chain that contains no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variations) compared to the heavy chain set forth in any one of SEQ ID NOs: 68 or 120. Alternatively or additionally, the anti-HJV antibodies of the present disclosure comprise a light chain that contains no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variations) compared to the light chain set forth in any one of SEQ ID NOs: 69. In some embodiments, the anti-HJV antibodies described herein comprise a heavy chain comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to any one of SEQ ID NOs: 68 or 120. Alternatively or additionally, the anti-HJV antibodies described herein comprise a light chain comprising an amino acid sequence at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to any one of SEQ ID NOs: 69. In some embodiments, the anti-HJV antibodies described herein comprise a heavy chain comprising an amino acid sequence of any one of SEQ ID NOs: 68 or 120. Alternatively or additionally, the anti-HJV antibodies described herein comprise a light chain comprising an amino acid sequence of any one of SEQ ID NOs: 69.

[0234] The anti-HJV antibodies described herein may be in any antibody form, including, but not limited to, an intact (i.e., full-length) antibody, an antigen-binding fragment thereof (e.g., Fab, F'ab'), F'ab'2, Fv), a single-chain antibody, a bispecific antibody, or a nanobody. In some embodiments, the anti-HJV antibodies described herein are scFv. In some embodiments, the anti-HJV antibodies described herein are scFv-Fab (e.g., scFv fused to a portion of the constant region).

[0235] In some embodiments, conservative mutations may be introduced into an antibody sequence (e.g., CDR or framework sequences) at positions where the residues are unlikely to be involved in interactions with the target antigen (e.g., hemojuvelin), for example, as determined based on a crystal structure. In some embodiments, one, two or more mutations (e.g., amino acid substitutions) are introduced into the Fc region (e.g., in the CH2 domain (residues 231-340 of human IgG1) and / or CH3 domain (residues 341-447 of human IgG1) and / or hinge region, using numbering according to the Kabat numbering system (e.g., EU index in Kabat)) of an anti-HJV antibody described herein to alter one or more functional properties of the antibody, such as serum half-life, complement fixation, Fc receptor binding and / or antigen-dependent cellular cytotoxicity.

[0236] In some embodiments, one, two or more mutations (e.g., amino acid substitutions) are introduced into the hinge region of the Fc region (CH1 domain) such that the number of cysteine ​​residues in the hinge region is altered (e.g., increased or decreased), as described, for example, in U.S. Patent No. 5,677,425. The number of cysteine ​​residues in the hinge region of the CH1 domain can be altered, for example, to facilitate assembly of the light and heavy chains, or to alter (e.g., increase or decrease) the stability of the antibody, or to facilitate linker conjugation.

[0237] In some embodiments, one, two or more mutations (e.g., amino acid substitutions) are introduced into the Fc region (e.g., in the CH2 domain (residues 231-340 of human IgG1) and / or the CH3 domain (residues 341-447 of human IgG1) and / or the hinge region, using the numbering according to the Kabat numbering system (e.g., EU index in Kabat)) of an anti-HJV antibody (muscle-targeting antibody) described herein to increase or decrease the affinity of the antibody for an Fc receptor (e.g., an activated Fc receptor) on the surface of an effector cell. Mutations in the Fc region of an antibody that decrease or increase the affinity of the antibody for an Fc receptor and techniques for introducing such mutations into an Fc receptor or a fragment thereof are known to those of skill in the art. Examples of mutations in the Fc receptors of an antibody that can be made to alter the affinity of the antibody for the Fc receptor are described, for example, in Smith P et al., (2012) PNAS 109: 6181-6186, U.S. Patent No. 6,737,056, and WO 02 / 060919; WO 98 / 23289; and WO 97 / 34631, which are incorporated herein by reference.

[0238] In some embodiments, one, two or more amino acid mutations (i.e., substitutions, insertions or deletions) are introduced into the IgG constant domain, or its FcRn binding fragment (e.g., Fc or hinge-Fc domain fragment) to change (e.g., decrease or increase) the half-life of the antibody in vivo. For example, see WO 02 / 060919; WO 98 / 23289; and WO 97 / 34631; and U.S. Patent Nos. 5,869,046, 6,121,022, 6,277,375 and 6,165,745 for examples of mutations that change (e.g., decrease or increase) the half-life of the antibody in vivo.

[0239] In some embodiments, one, two or more amino acid mutations (i.e., substitutions, insertions or deletions) are introduced into the IgG constant domain, or FcRn-binding fragment thereof (e.g., Fc or hinge-Fc domain fragment) to decrease the half-life of the anti-HJV antibody in vivo. In some embodiments, one, two or more amino acid mutations (i.e., substitutions, insertions or deletions) are introduced into the IgG constant domain, or FcRn-binding fragment thereof (e.g., Fc or hinge-Fc domain fragment) to increase the half-life of the antibody in vivo. In some embodiments, the antibody can have one or more amino acid mutations (e.g., substitutions) in the second constant (CH2) domain (residues 231-340 of human IgG1) and / or the third constant (CH3) domain (residues 341-447 of human IgG1) using the numbering according to the EU index in Kabat [Kabat EA et al., (1991), supra]. In some embodiments, the IgG1 constant region of the antibody described herein comprises a methionine (M) to tyrosine (Y) substitution at position 252, a serine (S) to threonine (T) substitution at position 254, and a threonine (T) to glutamic acid (E) substitution at position 256, according to the EU index in Kabat numbering. See U.S. Patent No. 7,658,921, which is incorporated herein by reference. This type of mutant IgG, referred to as "YTE mutants", has been shown to exhibit a four-fold increase in half-life compared to the wild-type version of the same antibody [see D'll'Acqua WF et al., (2006) J Biol Chem 281: 23514-24]. In some embodiments, the antibody comprises an IgG constant domain comprising one, two, three or more amino acid substitutions of amino acid residues at positions 251-257, 285-290, 308-314, 385-389, and 428-436, numbered according to the EU index in Kabat.

[0240] In some embodiments, one, two or more amino acid substitutions are introduced into the IgG constant domain Fc region to modify the effector function of the anti-HJV antibody. The effector ligand for which the affinity is modified can be, for example, an Fc receptor or the C1 component of complement. This approach is described in more detail in U.S. Patent Nos. 5,624,821 and 5,648,260. In some embodiments, deletion or inactivation of the constant region domain (through point mutation or other means) can reduce Fc receptor binding of circulating antibodies, thereby increasing tumor localization. See, for example, U.S. Patent Nos. 5,585,097 and 8,591,886 for a description of mutations that delete or inactivate constant domains, thereby increasing tumor localization. In some embodiments, one or more amino acid substitutions may be introduced in the Fc region of an antibody described herein to remove potential glycosylation sites in the Fc region that may reduce Fc receptor binding [see, e.g., Shields RL et al., (2001) J Biol Chem 276: 6591-604].

[0241] In some embodiments, one or more amino acids in the constant region of the anti-HJV antibodies described herein may be replaced with different amino acid residues such that the antibody has altered Clq binding and / or reduced or eliminated complement-dependent cytotoxicity (CDC). This approach is described in further detail in U.S. Pat. No. 6,194,551 (Idusogie et al.). In some embodiments, one or more amino acid residues in the N-terminal region of the CH2 domain of the antibodies described herein are altered to thereby alter the ability of the antibody to fix complement. This approach is further described in WO 94 / 29351. In some embodiments, the Fc region of the antibodies described herein is modified to increase the ability of the antibody to mediate antibody-dependent cellular cytotoxicity (ADCC) and / or increase the affinity of the antibody for Fcγ receptors. This approach is further described in WO 00 / 42072.

[0242] In some embodiments, the heavy and / or light chain variable domain sequences of the antibodies provided herein can be used to generate, for example, CDR-grafted, chimeric, humanized, or composite human antibodies or antigen-binding fragments as described elsewhere herein. As will be appreciated by one of skill in the art, any mutant, CDR-grafted, chimeric, humanized, or composite antibody derived from any of the antibodies provided herein can be useful in the compositions and methods described herein and retains the ability to specifically bind hemojuvelin, such that the mutant, CDR-grafted, chimeric, humanized, or composite antibody has at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% or higher binding to hemojuvelin compared to the original antibody from which it is derived.

[0243] In some embodiments, the antibodies provided herein contain mutations that confer desirable properties to the antibodies. For example, to avoid potential complications due to Fab-arm exchange, which is known to occur with native IgG4 mAbs, the antibodies provided herein may contain a stabilizing "Adair" mutation [Angal S., et al., "A single amino acid substitution abolishes the heterogeneity of chimeric mouse / human (IgG4) antibody," Mol Immunol 30, 105-108; 1993], in which serine 228 (EU numbering; residue 241, Kabat numbering) is converted to proline to provide an IgG1-like hinge sequence. Thus, any of the antibodies may contain a stabilizing "Adair" mutation.

[0244] In some embodiments, the antibody is modified, for example by glycosylation, phosphorylation, sumoylation, and / or methylation. In some embodiments, the antibody is a glycosylated antibody conjugated to one or more sugar or carbohydrate molecules. In some embodiments, the one or more sugar or carbohydrate molecules are conjugated to the antibody by N-glycosylation, O-glycosylation, C-glycosylation, glypiation (GPI anchor attachment), and / or phosphoglycosylation. In some embodiments, the one or more sugar or carbohydrate molecules are monosaccharides, disaccharides, oligosaccharides, or glycans. In some embodiments, the one or more sugar or carbohydrate molecules are branched oligosaccharides or branched glycans. In some embodiments, the one or more sugar or carbohydrate molecules comprise a mannose unit, a glucose unit, an N-acetylglucosamine unit, an N-acetylgalactosamine unit, a galactose unit, a fucose unit, or a phospholipid unit. In some embodiments, there are about 1-10, about 1-5, about 5-10, about 1-4, about 1-3, or about 2 sugar molecules. In some embodiments, the glycosylated antibody is fully or partially glycosylated. In some embodiments, the antibody is glycosylated by chemical reaction or by enzymatic means. In some embodiments, the antibody is glycosylated in vitro or inside a cell, which may be deficient in an enzyme, e.g., a glycosyltransferase, in the N- or O-glycosylation pathway. In some embodiments, the antibody is functionalized with a sugar or carbohydrate molecule, as described in International Patent Application Publication No. 2014065661, published May 1, 2014, entitled "Modified Antibodies, Antibody-Conjugates and Methods for Preparation Thereof."

[0245] In some embodiments, any one of the anti-HJV antibodies described herein may include a signal peptide (e.g., an N-terminal signal peptide) in the heavy and / or light chain sequence. In some embodiments, the anti-HJV antibodies described herein include any one of the VH and VL sequences, any one of the IgG heavy and light chain sequences, or any one of the F'ab') heavy and light chain sequences described herein, and further include a signal peptide (e.g., an N-terminal signal peptide). In some embodiments, the signal peptide includes the amino acid sequence of MEFGLSWLFLVAILKGVQC (SEQ ID NO: 104).

[0246] III. Preparation of anti-HJV antibodies Antibodies capable of binding to hemojuvelin as described herein may be generated by any method known in the art, see, for example, Harlow and Lane, (1998) Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, New York.

[0247] In some embodiments, antibodies specific to a target antigen (e.g., HJV) can be produced by conventional hybridoma technology. A full-length target antigen or a fragment thereof, optionally linked to a carrier protein, e.g., KLH, can be used to immunize a host animal to generate antibodies that bind to the antigen. The route and schedule of immunization of the host animal is generally performed according to established conventional techniques for antibody stimulation and production, as further described herein. General techniques for the production of mouse, humanized, and human antibodies are known in the art and are described herein. It is envisioned that any mammalian subject, including humans, or antibody-producing cells therefrom can be engineered to serve as the basis for the production of mammalian hybridoma cell lines, including human hybridoma cell lines. Typically, the host animal is inoculated intraperitoneally, intramuscularly, orally, subcutaneously, intraplantarly, and / or intradermally with an amount of immunogen, including those described herein.

[0248] If desired, the antibody (monoclonal or polyclonal) of interest (e.g., produced by a hybridoma) may be sequenced, and the polynucleotide sequence may then be cloned into a vector for expression or propagation. The sequence encoding the antibody of interest may be maintained in a vector in a host cell, which may then be expanded and frozen for future use. Alternatively, the polynucleotide sequence may be used for genetic engineering to "humanize" the antibody or to improve the affinity (affinity maturation) or other characteristics of the antibody. For example, the constant region may be engineered to be more similar to human constant regions to avoid immune responses when the antibody is used in clinical trials and treatments in humans. It may be desirable to genetically engineer the antibody sequence to obtain higher affinity and higher efficacy for the target antigen. It is clear to one of skill in the art that one or more polynucleotide changes can be made to an antibody and still maintain its binding specificity for the target antigen.

[0249] In other embodiments, fully human antibodies can be obtained by using commercially available mice engineered to express unique human immunoglobulin proteins. Transgenic animals designed to generate more desirable (e.g., fully human antibodies) or more robust immune responses can also be used for the generation of humanized or human antibodies. Examples of such technologies are XenomouseR™ from Amgen, Inc. (Fremont, CA) and HuMAb-MouseR™ and TC Mouse™ from Medarex, Inc. (Princeton, NJ) or H2L2 Mouse from Harbour Antibodies BV (Holland). As another alternative, antibodies can be produced recombinantly by phage display or yeast technology. See, e.g., U.S. Patent Nos. 5,565,332; 5,580,717; 5,733,743; and 6,265,150; and Winter et al., (1994) Annu. Rev. Immunol. 12:433-455. Alternatively, phage display technology [McCafferty et al., (1990) Nature 348:552-553] can be used to produce human antibodies and antibody fragments in vitro, from immunoglobulin variable (V) domain gene repertoires from unimmunized donors.

[0250] Antigen-binding fragments of intact antibodies (full-length antibodies) can be prepared through routine methods. For example, F(ab')2 fragments can be produced by pepsin digestion of antibody molecules, and Fab fragments can be generated by reducing disulfide bridges of F(ab')2 fragments. Engineered antibodies, such as humanized antibodies, chimeric antibodies, single-chain antibodies, and bispecific antibodies, can be produced, for example, through conventional recombinant techniques. In one example, DNA encoding a monoclonal antibody specific for a target antigen can be easily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes capable of specifically binding to genes encoding the heavy and light chains of a monoclonal antibody). Hybridoma cells serve as an exemplary source of such DNA. Once isolated, the DNA may be placed into one or more expression vectors, which are then transfected into host cells, such as Escherichia coli (E. coli) cells, Simian COS cells, Chinese hamster ovary (CHO) cells, human HEK293 cells, or myeloma cells that do not otherwise produce immunoglobulin proteins, to obtain the synthesis of monoclonal antibodies in the recombinant host cells. See, for example, PCT Publication WO 87 / 04462. The DNA may then be modified, for example, by substituting coding sequences for human heavy and light chain constant domains in place of the homologous mouse sequences [Morrison et al., (1984) Proc. Nat. Acad. Sci. 81:6851], or by covalently joining all or part of the coding sequence of a non-immunoglobulin polypeptide to the immunoglobulin coding sequence. In that manner, engineered antibodies, such as "chimeric" or "hybrid" antibodies, with the binding specificity of the target antigen may be prepared.

[0251] Single chain antibodies can be prepared via recombinant techniques by linking a nucleotide sequence encoding a heavy chain variable region and a nucleotide sequence encoding a light chain variable region, preferably incorporating a flexible linker between the two variable regions.

[0252] Alternatively, techniques described for the production of single chain antibodies (U.S. Pat. Nos. 4,946,778 and 4,704,692) can be applied to produce phage or yeast scFv libraries, and HJV-specific scFv clones can be identified from the libraries according to routine procedures. Positive clones can be subjected to further screening to identify clones with high HJV binding affinity.

[0253] The antibodies obtained according to the methods known in the art and described herein can be characterized using methods well known in the art. For example, one method is to identify the epitope that an antigen binds, or "epitope mapping". There are many methods known in the art for mapping and characterizing the location of epitopes on proteins, including elucidation of the crystal structure of antibody-antigen complexes, competitive assays, gene fragment expression assays, and synthetic peptide-based assays, as described in, for example, Chapter 11 of Harlow and Lane, Using Antibodies, a Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1999. In one example, epitope mapping can be achieved using H / D-Ex (hydrogen deuterium exchange) coupled with protein hydrolysis and mass spectrometry. In an additional example, epitope mapping can be used to determine the sequence to which an antibody binds. Epitopes can be linear epitopes, i.e., those contained in a single stretch of amino acids, or conformational epitopes formed by three-dimensional interactions of amino acids that may not necessarily be contained in a single stretch (primary structure linear sequence). Peptides of variable length (e.g., at least 4-6 amino acids long) can be isolated or synthesized (e.g., recombinantly) and used for binding assays with antibodies. In another example, the epitope to which an antibody binds can be determined in a systematic screen by using overlapping peptides derived from the target antigen sequence and determining binding by the antibody. According to a gene fragment expression assay, an open reading frame encoding a target antigen is fragmented randomly or by a unique genetic organization, and the reactivity of the expressed fragments of the antigen with the antibody to be tested is determined. Gene fragments may be produced, for example, by PCR, then in vitro transcribed and translated into proteins in the presence of radioactive amino acids. Binding of the antibody to the radiolabeled antigen fragments is then determined by immunoprecipitation and gel electrophoresis.Certain epitopes can also be identified by using large libraries of random peptide sequences displayed on the surface of phage particles (phage libraries). Alternatively, a defined library of overlapping peptide fragments can be tested for binding to a test antibody in a simple binding assay. In an additional example, mutagenesis of antigen-binding domains, domain swapping experiments and alanine scanning mutagenesis can be performed to identify residues required, sufficient and / or necessary for epitope binding. Alternatively, a competition assay can be performed using other antibodies known to bind to the same antigen to determine whether the antibody binds to the same epitope as the other antibody. Competition assays are well known to those skilled in the art.

[0254] In some examples, the anti-HJV antibody is prepared by recombinant technology as exemplified below. The nucleic acids encoding the heavy and light chains of the anti-HJV antibody described herein can be cloned into one expression vector, and each nucleotide sequence is operably linked to a suitable promoter. In one example, each of the nucleotide sequences encoding the heavy and light chains is operably linked to a separate promoter. Alternatively, the nucleotide sequences encoding the heavy and light chains can be in operably linked to a single promoter, so that both the heavy and light chains are expressed from the same promoter. If necessary, an internal ribosome entry site (IRES) can be inserted between the sequence encoding the heavy chain and the sequence encoding the light chain.

[0255] In some examples, the nucleotide sequences encoding the two chains of the antibody are cloned into two vectors, and the two vectors can be introduced into the same or different cells. When the two chains are expressed in different cells, each of them can be isolated from the host cell that expresses it, and the isolated heavy and light chains can be mixed and incubated under suitable conditions to allow the formation of the antibody.

[0256] In general, the nucleic acid sequence encoding one or all chains of the antibody can be cloned into a suitable expression vector operably linked with a suitable promoter using methods known in the art.For example, the nucleotide sequence and the vector can be contacted with a restriction enzyme under suitable conditions, which creates complementary ends on each molecule that can pair with each other and join with ligase.Alternatively, synthetic nucleic acid linkers can be ligated to the ends of the gene.These synthetic linkers contain nucleic acid sequences that correspond to specific restriction sites in the vector.The choice of expression vector / promoter depends on the type of host cell used in producing the antibody.

[0257] A variety of promoters can be used for expression of the antibodies described herein, including, but not limited to, the cytomegalovirus (CMV) intermediate early promoter, viral LTRs such as Rous sarcoma virus LTR, HIV-LTR, HTLV-1 LTR, Simian Virus 40 (SV40) early promoter, E. coli lac UV promoter, and herpes simplex tk virus promoter.

[0258] Regulatable promoters can also be used, including those that use the lac repressor from E. coli as a transcriptional modulator to regulate transcription from mammalian cell promoters carrying the lac operator [Brown, M. et al., Cell, 49:603-612 (1987)], and those that use the tetracycline repressor (tetR) [Gossen, M., and Bujard, H., Proc. Natl. Acad. Sci. USA 89:5547-555115 (1992); Yao, F. et al., Human Gene Therapy, 9:1939-1950 (1998); Shockelt, P., et al., Proc. Natl. Acad. Sci. USA, 92:6522-6526 (1995)]. Other systems include FK506 dimers, VP16 or p65 using astradiol, RU486, diphenol murislerone, or rapamycin. Inducible systems are available from Invitrogen, Clontech and Ariad, among others.

[0259] Regulatable promoters containing repressors with operons can be used. In one embodiment, the lac repressor from E. coli can function as a transcriptional modulator to regulate transcription from lac operator-bearing mammalian cell promoters [M. Brown et al., Cell, 49:603-612 (1987)]; Gossen and Bujard (1992); [M. Gossen et al., Natl. Acad. Sci. USA, 89:5547-5551(1992)], and the tetracycline repressor (tetR) is combined with a transcriptional activator (VP 16) to create a tetR-mammalian cell transcriptional activator fusion protein, tTa (tetR-VP 16), and tetO has a minimal promoter derived from the human cytomegalovirus (hCMV) promoter to create a tetR-tet operator system for controlling gene expression in mammalian cells. In one embodiment, a tetracycline-inducible switch is used. The tetracycline repressor (tetR), but not the tetR-mammalian cell transcription factor fusion derivative, can function alone as a potent transmodulator to regulate gene expression in mammalian cells when the tetracycline operator is appropriately placed downstream of the TATA element of the CMVIE promoter (Yao et al., Human Gene Therapy). One particular advantage of this tetracycline-inducible switch is that it does not require the use of tetracycline repressor-mammalian cell transactivator or repressor fusion proteins, which may be toxic to cells in some cases [Gossen et al., Natl. Acad. Sci. USA, 89:5547-5551 (1992); Shockett et al., Proc. Natl. Acad. Sci. USA, 92:6522-6526 (1995)], to achieve its regulatable effect.

[0260] Additionally, the vector can contain, for example, some or all of the following: a selectable marker gene, such as the neomycin gene for the selection of stable or transient transfectants in mammalian cells; an enhancer / promoter sequence from the immediate early gene of human CMV for high level transcription; a transcription termination and RNA processing signal from SV40 for mRNA stability; an SV40 polyoma origin of replication and ColE1 for proper episomal replication; an internal ribosome binding site (IRES), a versatile multiple cloning site; and T7 and SP6 RNA promoters for in vitro transcription of sense and antisense RNA. Suitable vectors and methods for producing vectors containing transgenes are well known and available in the art. Examples of polyadenylation signals useful for carrying out the methods described herein include, but are not limited to, human collagen I polyadenylation signal, human collagen II polyadenylation signal, and SV40 polyadenylation signal.

[0261] One or more vectors (e.g., expression vectors) containing nucleic acids encoding any of the antibodies (e.g., nucleic acid coding sequences listed in Table 3) may be introduced into a suitable host cell to produce the antibody. Non-limiting examples of host cells include Chinese hamster ovary (CHO) cells, dhfr-CHO cells, human embryonic kidney (HEK)-293 cells, verda reno (VERO) cells, non-secretory null (NS0) cells, human embryonic retina (PER.C6) cells, Sp2 / 0 cells, baby hamster kidney (BHK) cells, Madin-Darby canine kidney (MDCK) cells, Madin-Darby bovine kidney (MDBK) cells, and SV40 transformed monkey kidney CV1 line (COS) cells. In some embodiments, the host cell expressing the anti-HJV antibody is a CHO cell. The host cell may be cultured under suitable conditions for expression of the antibody or any polypeptide chain thereof. Such antibodies or polypeptide chains thereof can be recovered by the cultured cells (e.g., from the cells or culture supernatant) via conventional methods, such as affinity purification. If necessary, the polypeptide chains of the antibodies can be incubated under suitable conditions for a suitable time period to allow for production of the antibodies. In some embodiments, the host cell comprises a nucleic acid encoding a heavy chain of the anti-HJV antibody. In some embodiments, the host cell comprises a nucleic acid encoding a light chain of the anti-HJV antibody. In some embodiments, the host cell comprises a nucleic acid encoding a heavy chain and a nucleic acid encoding a light chain.

[0262] In some embodiments, the method of preparing the antibody described herein involves a recombinant expression vector encoding both the heavy and light chains of the anti-HJV antibody, also as described herein. The recombinant expression vector can be introduced into a suitable host cell (e.g., dhfr-CHO cell) by conventional methods, such as calcium phosphate-mediated transfection. Positive transformant host cells can be selected and cultured under suitable conditions that allow the expression of the two polypeptide chains that form the antibody, and the two chains can be recovered from the cells or culture medium. If necessary, the two chains recovered from the host cell can be incubated under suitable conditions that allow the formation of the antibody.

[0263] In one example, two recombinant expression vectors are provided, one encoding the heavy chain of an anti-HJV antibody and the other encoding the light chain of an anti-HJV antibody. Both of the two recombinant expression vectors can be introduced into a suitable host cell (e.g., dhfr-CHO cell) by conventional methods, such as calcium phosphate-mediated transfection.

[0264] Alternatively, each of the expression vectors can be introduced into a suitable host cell. The positive transformants can be selected and cultured under suitable conditions that allow the expression of the polypeptide chains of the antibody. When the two expression vectors are introduced into the same host cell, the antibody produced in the cell can be recovered from the host cell or culture medium. If necessary, the polypeptide chains can be recovered from the host cell or culture medium and then incubated under suitable conditions that allow the formation of the antibody. When the two expression vectors are introduced into different host cells, each of them can be recovered from the corresponding host cell or the corresponding culture medium. The two polypeptide chains can then be incubated under suitable conditions for the formation of the antibody.

[0265] Standard molecular biology techniques are used to prepare the recombinant expression vector, transfect the host cells, select for transformants, culture the host cells, and recover the antibody from the culture medium. For example, some antibodies can be isolated by affinity chromatography using a Protein A or Protein G-linked matrix.

[0266] Any of the nucleic acids encoding the heavy chain, light chain, or both of the anti-HJV antibodies described herein (e.g., provided in Table 3), vectors containing the same (e.g., expression vectors); and host cells containing the vectors are within the scope of the present disclosure.

[0267] [Table 4-1] [Table 4-2]

Table 4-3

Table 4-4

Table 4-5

[0268]

Table 4-6

Table 4-7

Table 4-8

Table 4-9

Table 4-10

Table 4-11

Table 4-12

[0269] In some embodiments, the anti-HJV antibodies described herein are produced by expressing (i) a nucleic acid that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO:71, and / or (ii) a nucleic acid that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO:73.

[0270] In some embodiments, the anti-HJV antibodies described herein are produced by expressing (i) a nucleic acid that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO:71, and / or (ii) a nucleic acid that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO:75.

[0271] In some embodiments, the anti-HJV antibodies described herein are produced by expressing (i) a nucleic acid that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO:71, and / or (ii) a nucleic acid that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO:77.

[0272] In some embodiments, the anti-HJV antibodies described herein are produced by expressing (i) a nucleic acid that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO:71, and / or (ii) a nucleic acid that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO:79.

[0273] In some embodiments, the anti-HJV antibodies described herein are produced by expressing (i) a nucleic acid that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO:71, and / or (ii) a nucleic acid that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO:81.

[0274] In some embodiments, the anti-HJV antibodies described herein are produced by expressing (i) a nucleic acid that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO:84, and / or (ii) a nucleic acid that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO:85.

[0275] In some embodiments, the anti-HJV antibodies described herein are produced by expressing (i) a nucleic acid that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO:88, and / or (ii) a nucleic acid that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO:89.

[0276] In some embodiments, the anti-HJV antibodies described herein are produced by expressing (i) a nucleic acid that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO:92, and / or (ii) a nucleic acid that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO:93.

[0277] In some embodiments, the anti-HJV antibodies described herein are produced by expressing (i) a nucleic acid that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO:94, and / or (ii) a nucleic acid that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO:93.

[0278] In some embodiments, the anti-HJV antibodies described herein are produced by expressing (i) a nucleic acid that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO:92, and / or (ii) a nucleic acid that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO:96.

[0279] In some embodiments, the anti-HJV antibodies described herein are produced by expressing (i) a nucleic acid that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO: 99, and / or (ii) a nucleic acid that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO: 100.

[0280] In some embodiments, the anti-HJV antibodies described herein are produced by expressing (i) a nucleic acid that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO:40, and / or (ii) a nucleic acid that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO:64.

[0281] In some embodiments, the anti-HJV antibodies described herein can be used to deliver a molecular payload to a target cell or tissue (e.g., a cell or tissue expressing HJV). Thus, the anti-HJV antibodies described herein can be linked to a molecular payload. The conjugates described herein can be used in a variety of applications, for example, diagnostic or therapeutic applications.

[0282] In some embodiments, the complex described herein is used to modulate the activity or function of at least one gene, protein, and / or nucleic acid. In some embodiments, the molecular payload is responsible for the modulation of gene, protein, and / or nucleic acid. The molecular payload may be a small molecule, protein, nucleic acid, oligonucleotide, or any molecular entity that has the ability to modulate the activity or function of gene, protein, and / or nucleic acid in cells. In some embodiments, the molecular payload is an oligonucleotide that targets disease-related repeats in muscle cells.

[0283] IV. Pharmaceutical Compositions In addition to the antibody described herein, the encoding nucleic acid or nucleic acid set, the vector containing it, or the host cell containing the vector can be mixed with a pharmaceutically acceptable carrier (excipient) to form a pharmaceutical composition for use in treating a target disease. "Acceptable" means that the carrier must be compatible with the active ingredient of the composition (and preferably has the ability to stabilize the active ingredient) and not harmful to the subject being treated. Pharmaceutically acceptable excipients (carriers) include buffers well known in the art. See, for example, Remington: The Science and Practice of Pharmacy 20th Ed. (2000) Lippincott Williams and Wilkins, Ed. KE Hoover.

[0284] The anti-HJV antibody-containing pharmaceutical composition disclosed herein may further comprise a suitable buffering agent. A buffering agent is a weak acid or base used to maintain the pH of a solution near a selected value after the addition of another acid or base. In some examples, the buffering agent disclosed herein may be a buffering agent that has the ability to maintain a physiological pH regardless of changes in carbon dioxide concentration (produced by cellular respiration). Exemplary buffering agents include, but are not limited to, HEPES [4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid] buffer, Dulbecco's phosphate buffered saline (DPBS) buffer, or phosphate buffered saline (PBS) buffer. Such buffering agents may include disodium hydrogen phosphate and sodium chloride, or potassium dihydrogen phosphate and potassium chloride.

[0285] In some embodiments, the buffering agent in the pharmaceutical compositions described herein may maintain a pH value of about 5 to 8. For example, the pH of the pharmaceutical composition may be about 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, or 8.0. In other examples, the pharmaceutical composition may have a pH value below 7, for example, about 7, 6.8, 6.5, 6.3, 6, 5.8, 5.5, 5.3, or 5.

[0286] The pharmaceutical compositions described herein include one or more suitable salts. Salts are ionic compounds that can be formed by the neutralization reaction of acids and bases [Skoog, DA; West, DM; Holler, JF; Crouch, SR (2004). "chapters 14-16". Fundamentals of Analytical Chemistry (8th ed.)]. Salts are composed of a related number of cations (positively charged ions) and anions (negative ions), so that the product is electrically neutral (has no net charge).

[0287] In some embodiments, the pharmaceutical composition may comprise a pharma- ceutical acceptable carrier, excipient, or stabilizer in the form of a lyophilized formulation or an aqueous solution. [Remington: The Science and Practice of Pharmacy 20th Ed. (2000) Lippincott Williams and Wilkins, Ed. KE Hoover]. In some embodiments, the pharmaceutical composition may be formulated for intravenous injection. In some embodiments, the pharmaceutical composition may be formulated for subcutaneous injection.

[0288] Pharmaceutical compositions used for in vivo administration must be sterile. This is easily accomplished, for example, by filtration through sterile filtration membranes. Therapeutic antibody compositions are generally placed into a container having a sterile access port, for example, an intravenous or subcutaneous solution bag or vial having a stopper pierceable by a hypodermic injection needle.

[0289] V. Combination Therapy In some embodiments, immunomodulatory or erythropoietin stimulating agents for the treatment of anemia, for example, associated with chronic kidney disease, are provided herein. Such agents include erythropoietin stimulating agents (ESAs). Thus, in some embodiments, erythropoietin (EPO) is administered as an additional therapeutic agent in the methods described herein. In some embodiments, the EPO is selected from epoetin alfa (Epogen / Procrit), darbepoetin alfa (Aranesp), methoxypolyethylene glycol-epoetin beta (Mircera), epoetin alfa-epbx (Retacrit); and biosimilars of Epogen / Procrit.

[0290] In some embodiments, any of the disclosed hemojuvelin antagonists may be administered in combination therapy with a therapeutic agent selected from a growth differentiation factor (GDF) trap, oral iron, IV iron, HIF-PHI, and red blood cell transfusion. Exemplary GDF traps include sotatercept and raspatercept.

[0291] In some embodiments, the additional therapeutic agent is IV iron therapy, such as iron isomaltoside (MonoFerric®). Additional exemplary IV iron therapies include, but are not limited to, iron sucrose (Venofer®), ferric carboxymaltose (Ferrinject® or Injectofer®), ferumoxytol (Ferraheme®), iron dextran (Imferon®), and sodium ferric gluconate in iron sucrose solution (Ferrlecit®).

[0292] In some embodiments, the additional therapeutic agent is oral iron therapy. In some embodiments, the hemojuvelin antagonists provided herein may be combined with oral iron therapy to promote iron level recovery and / or treat anemic subjects experiencing intolerance or inadequate response to oral iron. In some embodiments, a combination therapy of anti-hemojuvelin antibody and oral iron is administered to subjects exhibiting serum ferritin levels below 100ng / ml and TSAT below 30%. Examples of oral iron therapy include, but are not limited to, ferrous sulfate, ferric maltol (Accrufer®), ferrous gluconate, ferrous succinate, iron polymaltose, polysaccharide-iron complex, and ferrous sulfate. In other cases, iron may be administered intramuscularly (e.g., as ferrous sorbitol citrate).

[0293] In some embodiments, the additional therapeutic agent is a HIF-PHI, such as daprodustat, roxadustat, and vadadustat. In some embodiments, the additional therapeutic agent is roxadustat.

[0294] Other immunomodulatory agents that may be used in the combination therapy of the present disclosure include, for example, corticosteroids. In some embodiments, immunomodulatory agents are advantageous in that they have beneficial effects in reducing inflammation and promoting erythropoiesis. For example, danazol is a steroid compound that has hematopoietic stimulating and immunomodulatory effects. For example, in some embodiments, danazol has an antagonist effect on glucocorticoid receptors, resulting in an upregulatory effect on erythropoiesis (see, for example, Chai KY, et al., Danazol: An Effective and Underutilized Treatment Option in Diamond-Blackfan Anaemia. Case Reports in Hematology. Volume 2019, Article ID 4684156.). Other useful immunomodulatory agents include thalidomide and its derivatives or analogs, such as lenalidomide and pomalidomide.

[0295] VI. Treatment Method Embodiments of the present disclosure relate to methods of treating anemia of kidney disease and / or one or more conditions resulting from anemia of kidney disease in a subject. Certain embodiments of the present disclosure relate to methods of treating anemia of CKD. Additional embodiments of the present disclosure relate to methods of treating anemia of CKD. 2 Less than 90mL / min per 1.73m 2 Less than 60mL / min per 1.73m 2 Less than 30mL / min per 1.73m 2 Less than 15 mL / min per 1.73 m 2 In some embodiments, the subject has a GFR level of 15-59 ml / min. Additional aspects relate to methods of treating anemia from iron deficiency associated with or concurrent with CKD.

[0296] In some embodiments, the methods provided herein are useful for treating a subject with anemia associated with kidney disease to decrease hepcidin levels or activity. In some embodiments, the subject may experience an improvement in iron intake from the gastrointestinal system (i.e., from the diet). In some embodiments, the subject may experience partial or complete restoration of iron levels. In some embodiments, the subject may have an inadequate response to oral iron treatment. In some embodiments, the subject may have non-dialysis-dependent CKD (CKD-NDD). In some embodiments, the subject has non-hemodialysis-dependent chronic kidney disease.

[0297] In exemplary embodiments, the subject has a transferrin saturation (TSAT) level of less than 50%, less than 40%, less than 30%, or less than 20%. In some embodiments, the subject is identified as having a hemoglobin level within a range of 1.5-2.0 g / dL or 2.0-4.0 g / dL or greater below normal hemoglobin levels. In some embodiments, the subject exhibits a serum hemoglobin level of less than 11 g / dL, 10 g / dL, 9 g / dL, or 8 g / dL. In some embodiments, the subject has a serum ferritin level of less than 300 ng / mL, 200 ng / mL, or 100 ng / mL. In some embodiments, the subject exhibits a serum ferritin level of less than 100 ng / ml and a TSAT of less than 30%.

[0298] In some embodiments, administration of a hepcidin antagonist (e.g., an anti-HJV antibody) increases hemoglobin (HGB) levels by at least 1 g / dL from baseline. In some embodiments, administration of a hepcidin antagonist increases hemoglobin levels in a subject by at least 1 g / dL compared to an untreated subject. In some embodiments, any of the disclosed methods of administration of a hepcidin antagonist results in an increase in hemoglobin levels in a subject of at least 2, 4, 6, 8, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or greater than 20 g / dL compared to an untreated subject. In certain embodiments, any of the disclosed methods results in an increase in hemoglobin levels of about 17 g / dL (or 170 g / L). In some embodiments, any of the disclosed methods results in an increase in hemoglobin levels of about 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20% compared to untreated subjects. These increased HGB levels may be observed within 10, 20, 30, 40, or more than 40 days of treatment. In particular, these increased HGB levels may be observed after about 42 days of treatment.

[0299] In some embodiments, administration of a hepcidin antagonist (e.g., an anti-HJV antibody) increases reticulocyte hemoglobin (Ret-HGB) levels in a subject by at least 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.75, 0.8, 0.85, 0.9, 0.95, 1.0, or greater than 1.0 pg compared to an untreated subject. Ret-HGB is a measure of cellular hemoglobinization. In certain embodiments, any of the disclosed methods results in an increase in Ret-HGB levels of about 0.85 pg. In some embodiments, any of the disclosed methods results in an increase in hemoglobin levels of about 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, or greater than 5% compared to an untreated subject. These increased Ret-HGB levels may be observed within 10, 20, 30, 40, or more than 40 days of treatment. In particular, these increased Ret-HGB levels may be observed after about 42 days of treatment.

[0300] In some aspects, the present disclosure relates to compositions and methods for treating CKD in subjects.In some embodiments, the subject to be treated according to the present disclosure can be identified based on suitable diagnostic methodology, for example, as described in National Kidney Foundation, K / DOQI clinical practice guidelines for chronic kidney disease: Evaluation, classification, and stratification, Am. J. Kidney Dis. 2002: 39 (Suppl 1): S1-S266; Cullis, JO, Diagnosis and management of anaemia of chronic disease: current status British Journal of Haematology, Volume 154, Issue 3, August 2011 pages 289-300; and Madua AJ and Ughasoro MD, Anaemia of Chronic Disease: An In-Depth Review Med Princ Pract. 2017 Jan; 26(1): 1-9, the contents of each of which are incorporated herein by reference. Typically, diagnosis of anemia involves evaluation of the signs and symptoms of the underlying chronic condition combined with evaluation of signs of anemia and / or defects in iron metabolism, including, for example, through analysis of complete blood count (CBC), serum iron, ferritin, transferrin, reticulocyte count, and other markers.

[0301] In some embodiments, a subject in need of treatment according to the present disclosure may be identified based on a decrease in EPO production.For example, under normal physiological conditions, the level of EPO is inversely correlated with hemoglobin levels and tissue oxygenation, but in chronic inflammatory conditions, the EPO response is blunted, leading to insufficient levels of EPO due to the degree of anemia, which is believed to be mediated through inflammatory cytokines such as IL-1 and tumor necrosis factor-α (TNF-α).Thus, in some embodiments, a blunted EPO response may be useful in diagnosing anemia of chronic disease (ACD) in a subject, such as anemia of chronic kidney disease.

[0302] In some embodiments, subjects in need of treatment according to the present disclosure may be identified based on decreased erythroid responsiveness. For example, anemia may be characterized by decreased proliferation and differentiation of erythroid progenitor cells. It has been shown that macrophages from patients with anemia suppress colony formation in vitro due to the inhibitory effect of inflammatory cytokines (e.g., interferon-γ) on the proliferation of erythroid burst-forming units (BFU-E) and erythroid colony-forming units (CFU-E), and that this effect could be overcome by adding high concentrations of EPO to the culture system. It has also been shown that bone marrow cultures from patients with active rheumatoid arthritis showed defective growth when compared with normal controls, and that there was an inverse correlation between colony proliferation and the level of TNF-α in the culture supernatant. Furthermore, these effects were reversed after treatment with infliximab, an antibody against TNF-α, both in vitro and in vivo. Thus, in some embodiments, decreased erythrocyte responsiveness in a subject may be identified using these or similar such assays for assessing erythrocyte responsiveness.

[0303] In some embodiments, subjects in need of treatment according to the present disclosure may be identified based on mild to moderate and / or normochromic and normocytic anemic status (although as the disease progresses, the anemia may become microcytic). In some embodiments, subjects are identified based on low reticulocyte counts. Inflammation in a subject may be inferred from other features of blood counts, such as neutrophilia, monocytosis, or thrombocytosis, and through the measurement of non-specific inflammatory markers, such as C-reactive protein (CRP) or erythrocyte sedimentation rate (ESR).

[0304] In some embodiments, the subject is identified by determining the ratio of serum transferrin receptor (sTFR) to ferritin. Measurement of sTFR, a cleaved fragment of membrane receptor, has been described as an indicator for distinguishing anemia from IDA in subjects. Transferrin receptor is found on virtually all cells in the body, but is present at high levels on erythroid progenitor cells. sTFR levels increase in IDA because iron availability for erythropoiesis is reduced, while in anemia, levels may not differ from steady state because transferrin receptor expression is negatively affected by inflammatory cytokines. The ratio of sTFR to the logarithm of serum ferritin may be used in diagnosing anemia in subjects, and in some cases, may be used to distinguish anemia from IDA. A ratio of less than 1 may indicate anemia, while a ratio of greater than 2 suggests that iron stores are deficient, with or without anemia.

[0305] In some embodiments, subjects in need of treatment according to the present disclosure may be identified using red blood cell indices. For example, reticulocyte hemoglobin content (CHr) and percentage of hypochromic red blood cells (%HYPO) can provide information about iron supply to the red blood cell system and may be useful in guiding the management of anemias [e.g., ACD (e.g., anemia of chronic kidney disease)]. CHr is a measure of hemoglobin in the most recently formed red blood cells, and %HYPO indicates the percentage of cells with a hemoglobin content of <280 g / l. The former gives a relatively acute assessment of recent bone marrow activity (e.g., 48 hours), while the latter gives a time-averaged picture (e.g., 20-120 days). Similar indices may be reported by the Sysmex XE-2100 analyzer (Sysmex, Mundelein, IL, USA), which derives RET-Y (equivalent to CHr) and RBC-Y (equivalent to HYPO%). In addition to detecting early iron deficiency, CHr has been shown to be a useful tool in monitoring early response to iron therapy.

[0306] In some embodiments, the subject has previously received an erythropoietin stimulating agent, hi some embodiments, the erythropoietin stimulating agent is selected from the group consisting of danazol, prednisone, thalidomide, lenalidomide, and pomalidomide.

[0307] Whether the amount of HJV antagonist achieves therapeutic effect is clear to those skilled in the art based on the teachings provided herein. Effective amount will vary according to the specific condition being treated, the severity of the condition, individual patient parameters including age, health, size, sex and weight, duration of treatment, nature of concomitant therapy (if any), specific route of administration and factors within the knowledge and expertise of medical practitioners, as recognized by those skilled in the art. These factors are well known to those skilled in the art and can be addressed by simple routine experimentation. The specific administration regimen, i.e., dosage, timing and frequency, used in the methods described herein will depend on the specific subject and the subject's medical history, as described herein.

[0308] Empirical considerations, such as time to maximum effect, half-life, and / or time above a particular concentration, will usually contribute to the determination of dosage.

[0309] In some embodiments, the dosage of the HJV antagonist described herein can be empirically determined in individuals given one or more doses of the antibody. Individuals are given increasing doses of the antagonist. Disease / disorder indicators can be followed to assess the effectiveness of the antagonist.

[0310] The frequency of administration may vary according to the claimed method. In some embodiments, the composition is administered once. In some embodiments, the treatment is administered on multiple occasions. In some embodiments, the frequency of administration is weekly, every 2 weeks, every 3 weeks, every 4 weeks, every 5 weeks, every 6 weeks, every 7 weeks, every 8 weeks, every 9 weeks, or every 10 weeks; or once a month, every 2 months, or every 3 months, or more. In some embodiments, the composition is administered daily, twice a week, weekly, twice a month, monthly, semi-monthly, quarterly, or at any time interval that provides suitable (e.g., maximum) efficacy while minimizing safety risks to the subject. In some embodiments, the dosing frequency is for a period of about 45 days, 50 days, 55 days, 60 days, or 65 days. In some embodiments, the dosing period is 40-60 days, 40-55 days, 50-60 days, or 50-55 days. In some embodiments, the dosing period is 50-55 days. In general, efficacy and treatment and safety risks may be monitored throughout the course of treatment. In some embodiments, the hemojuvelin antagonist is administered once a month. In some embodiments, the hemojuvelin antagonist is administered once every three months.

[0311] In some embodiments, administration of HJV antagonists results in a decrease in serum hepcidin-25 concentration and / or increases serum TSAT%, and in some embodiments, these effects are sustained over a period of time (e.g., one month or longer). Thus, in some embodiments, the timing and frequency of administration of HJV antagonists can be determined by monitoring one or more biomarkers, such as criteria for assessing iron availability or for determining possible iron overload. For example, in some embodiments, HJV antagonists are administered intermittently or according to the level of a particular biomarker, such as serum hepcidin-25 level, or transferrin saturation percentage (TSAT%). In some embodiments, the biomarker levels described herein can be used to determine whether a subject is a candidate for treatment. However, in some embodiments, biomarkers can be used to determine, for example, whether treatment with HJV antagonists should be continued, or whether treatment should be resumed, or whether treatment should be stopped.

[0312] In some embodiments, administration of an HJV antagonist results in a decrease in urinary hepcidin concentration and / or an increase in serum TSAT%.

[0313] For example, in some embodiments, if the subject's TSAT% is 70% or more, 75% or more, 80% or more, 85% or more, 90% or more, or 95% or more, the subject may not be considered a candidate for treatment. In some cases, if the subject's TSAT% is 70% or more, 75% or more, 80% or more, 85% or more, 90% or more, or 95% or more, ongoing treatment with an HJV antagonist may be stopped or temporarily stopped, for example, to prevent iron overload. In other embodiments, administration of an anti-HJV antibody may be performed if the subject's TSAT% is 95% or less, 90% or less, 80% 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, or 30% or less. Thus, in some embodiments, the subject's TSAT% can be monitored, for example, continuously or periodically, while the patient is undergoing treatment or visiting a doctor who treats, for example, anemia, to prevent iron overload, or to evaluate whether further treatment is appropriate. However, it will be appreciated that other suitable markers (including, for example, ferritin levels, serum iron levels, creatinine levels, etc.) can be monitored to determine dosage and frequency of administration according to the methods provided herein.

[0314] In some embodiments, a subject may be administered a composition provided herein (e.g., an HJV antagonist) at one or more intervals for a period of time. In some cases, at one or more intervals, the period during which the subject is administered the composition may be separated by a period during which the subject is not administered the composition. In some embodiments, the relative duration of each period may depend on the subject's response to the treatment or the severity of the disease or both, and / or may be determined based on the treating physician's judgment. For example, in some embodiments, during the course of a year, a subject may be administered the composition weekly, biweekly, monthly, or bimonthly for two months, and then administration may be stopped for ten months. In some embodiments, during the course of a year, a subject may be administered the composition weekly, biweekly, monthly, or bimonthly for three months, and then administration may be stopped for nine months. In some embodiments, during the course of a year, a subject may be administered the composition weekly, biweekly, monthly, or bimonthly for four months, and then administration may be stopped for eight months. In some embodiments, during the course of a year, the subject may be administered the composition weekly, every two weeks, every month, or every two months for 5 months, and then administration may be stopped for 7 months. In some embodiments, during the course of a year, the subject may be administered the composition weekly, every two weeks, every month, or every two months for 6 months, and then administration may be stopped for 6 months. In some embodiments, during the course of a year, the subject may be administered the composition weekly, every two weeks, every month, or every two months for 7 months, and then administration may be stopped for 5 months. In some embodiments, during the course of a year, the subject may be administered the composition weekly, every two weeks, every month, or every two months for 8 months, and then administration may be stopped for 4 months. In some embodiments, during the course of a year, the subject may be administered the composition weekly, every two weeks, every month, or every two months for 9 months, and then administration may be stopped for 3 months. In some embodiments, over the course of a year, a subject may be administered the composition weekly, every two weeks, monthly or every two months for 10 months, and then administration is discontinued for two months.In some embodiments, during the course of a year, the subject may be administered the composition weekly, every two weeks, monthly or every two months for two months and discontinued for two months; or administered for three months and discontinued for three months; or administered for four months and discontinued for four months.In some embodiments, during the course of a year, the subject may be administered the composition every three months for the entire period of the year (i.e., four times).In some embodiments, during the course of a year, the subject may be administered the composition two or three times, but not four times, every three months.

[0315] Generally, for administration of any of the antibodies described herein, the dosage may be about 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.2 mg / kg, 0.3 mg / kg, 0.4 mg / kg, 0.5 mg / kg, 0.6 mg / kg, 0.8 mg / kg, 1 mg / kg, 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 6 mg / kg, 7 mg / kg, 8 mg / kg, 9 mg / kg, 10 mg / kg, 20 mg / kg, 30 mg / kg, 40 mg / kg, 50 mg / kg, 60 mg / kg, 70 mg / kg, 80 mg / kg, 90 mg / kg or 100 mg / kg.

[0316] In some embodiments, the dosage of the anti-HJV antibody is up to 0.01 mg / kg, up to 0.05 mg / kg, up to 0.1 mg / kg, up to 0.2 mg / kg, up to 0.3 mg / kg, up to 0.4 mg / kg, up to 0.5 mg / kg, up to 0.6 mg / kg, up to 0.8 mg / kg, up to 1 mg / kg, up to 2 mg / kg, up to 3 mg / kg, up to 4 mg / kg, up to 5 mg / kg, up to 6 mg / kg, up to 7 mg / kg, up to 8 mg / kg, up to 9 mg / kg, up to 10 mg / kg, up to 20 mg / kg, up to 30 mg / kg, up to 40 mg / kg, up to 50 mg / kg, up to 60 mg / kg, up to 70 mg / kg, 80 mg / kg, up to 90 mg / kg, up to 100 mg / kg or more.

[0317] However, in some embodiments, the dose of anti-HJV antibody may be in the range of 0.01 mg / kg to 100 mg / kg, 0.01 mg / kg to 10 mg / kg, 0.5 mg / kg to 15 mg / kg, 0.6 mg / kg to 15 mg / kg, 0.8 mg / kg to 15 mg / kg, 1 mg / kg to 15 mg / kg, 5 mg / kg to 25 mg / kg, 10 mg / kg to 30 mg / kg, 20 mg / kg to 40 mg / kg, 30 mg / kg to 50 mg / kg, 40 mg / kg to 60 mg / kg, 50 mg / kg to 75 mg / kg, or 50 mg / kg to 100 mg / kg.

[0318] In some embodiments, the antibodies described herein are administered to a subject in need of treatment in an amount sufficient to inhibit the activity of the target antigen in vivo by at least 20% (e.g., 30%, 40%, 50%, 60%, 70%, 80%, 90% or more) (e.g., an amount sufficient to inhibit HJV-induced BMP signaling). In other embodiments, the antibodies are administered in an amount effective to reduce the activity level of the target antigen by at least 20% (e.g., 30%, 40%, 50%, 60%, 70%, 80%, 90% or more).

[0319] In some embodiments, the antibody can be administered parenterally.For example, the parenterally administered composition can be administered subcutaneously, intradermally, intravenously, intraperitoneally, intratumorally, intramuscularly, intraarticularly, intraarterially, or by infusion techniques.Furthermore, it can be administered to a subject via an injectable reservoir route of administration, such as using 1, 3, or 6 month reservoir injectable or biodegradable materials and methods.

[0320] In some embodiments, the antibody (e.g., an anti-HJV antibody) is administered intravenously. In some embodiments, the antibody (e.g., an anti-HJV antibody) is administered subcutaneously. In some embodiments, subcutaneous administration of an anti-HJV antibody results in similar bioavailability compared to intravenous administration of the same antibody at the same dose.

[0321] In some embodiments, subcutaneous administration of an anti-HJV antibody results in a lower maximum concentration (C) of the anti-HJV antibody compared to intravenous administration of the same antibody. max ) produces comparable pharmacodynamic effects (e.g., a decrease in circulating hepcidin-25 levels, an increase in TSAT%, and / or an increase in serum iron levels). max is the maximum (or peak) serum concentration of the drug (e.g., anti-HJV antibody) after the drug is administered and before administration of the second dose. In some embodiments, a low C within a short time period (e.g., within 12 hours, within 24 hours, etc.) after administration of the anti-HJV antibody max Achieving this minimizes undesirable increases in serum iron response and / or minimizes potential off-target effects of the antibody (e.g., binding to RGMa). In some embodiments, subcutaneous administration of anti-HJV antibodies blunts C. max avoids an undesirable sharp increase in serum iron response. In some embodiments, blunting C by subcutaneous administration of anti-HJV antibodies. max In some embodiments, the C achieved by subcutaneous administration reduces the off-target effects of the antibody. max This is achieved by intravenous administration of anti-HJV antibodies. max at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95% lower.

[0322] In some embodiments, any of the disclosed methods (e.g., any of the disclosed methods of subcutaneous administration) result in an increase in serum iron levels in a subject of about 25, 27.5, 30, 32.5, 35, 38.5, 40, or 45 μmol / L compared to an untreated subject. In particular embodiments, any of the disclosed methods result in an increase in serum iron levels of about 35 μmol / L. In some embodiments, any of the disclosed methods result in an increase in serum iron levels of about 95%, 97.5%, 100%, 102.5%, 105%, 110%, 115%, or 120% compared to an untreated subject. In particular embodiments, any of the disclosed methods result in an increase in serum iron levels of about 100-110%, particularly about 109% or 119% on average. These increased serum levels may be observed within 10, 20, 30, 40, or more than 40 days of treatment. In particular, these increased serum levels may be observed after about 42 days of treatment.

[0323] In some embodiments, any of the disclosed methods (e.g., any of the disclosed methods of subcutaneous administration) reduces or improves the survival rate by about 1, 2, 2.5, 3, 4, 5, 6, 7, 7.5, 8, 9, or 10×10 compared to an untreated subject. 5 In certain embodiments, any of the disclosed methods results in an increase in red blood cell (RBC) count (e.g., mature RBC count) in a subject of about 4 or 5×10 cells / μL. 5 Results in an increase in RBC count of cells / μL. In some embodiments, any of the disclosed methods results in an increase in RBC count of about 5%, 6%, 6.5%, 7%, 7.25%, 7.5%, 7.75%, 8%, 8.5%, 9%, 9.25%, 9.5%, 10%, or greater than 10% compared to untreated subjects. In certain embodiments, any of the disclosed methods results in an increase in RBC count of greater than about 7% (e.g., 7.25%). In some embodiments, the RBC count is elevated between 7% and 9.5%. These increased RBC counts may be observed within 10, 20, 30, 40, or greater than 40 days of treatment. Notably, these RBC counts may be observed about 42 days after treatment.

[0324] In some embodiments, any of the disclosed methods (e.g., any of the disclosed methods of subcutaneous administration) can improve cellular function by about 90, 100, 100, 120, or 125×10 compared to untreated subjects. 9 In certain embodiments, any of the disclosed methods results in a reduction in reticulocyte (Ret) count in a subject of about 100×10 cells / L. 9 Resulting in a reduction in Ret count of cells / μL. In some embodiments, any of the disclosed methods result in a reduction in Ret count of about 45%, 50%, or 55% compared to untreated subjects. These reduced Ret counts may be observed within 10, 20, 30, 40, or more than 40 days of treatment. In particular, these Ret counts may be observed about 42 days after treatment.

[0325] For intravenous injection, water-soluble antibodies can be administered by drip infusion, and a pharmaceutical formulation containing the antibody and a physiologically acceptable excipient is injected. Physiologically acceptable excipients may include, for example, 5% dextrose, 0.9% saline, Ringer's solution, or other suitable excipients. Other injectable compositions may contain various carriers, such as vegetable oils, dimethylactamide, dimethylformamide, ethyl lactate, ethyl carbonate, isopropyl myristate, ethanol, and polyols (such as glycerol, propylene glycol, and liquid polyethylene glycol). In some cases, a suitable preparation of the antibody in a soluble salt form, for example, a sterile formulation, can be dissolved in a pharmaceutical excipient, such as water for injection, 0.9% saline, or 5% glucose solution, and then administered.

[0326] In one embodiment, antibody is administered via site-specific or targeted local delivery technology.Examples of site-specific or targeted local delivery technology include antibody various implantable depot sources or local delivery catheters, such as injection catheters, indwelling catheters, or needle catheters, synthetic grafts, adventitial wraps, shunts and stents or other implantable devices, site-specific carriers, direct injection, or direct application.See, for example, PCT Publication No. WO 2000 / 53211 and U.S. Patent No. 5,981,568.

[0327] In some embodiments, more than one antibody, or a combination of an antibody and another suitable therapeutic agent may be administered to a subject in need of treatment.Antibody may also be used in combination with other agents that serve to enhance and / or complement the effectiveness of the agent.The effectiveness of treatment against target disease / disorder may be evaluated by methods well known in the art.

[0328] The anti-HJV antibodies and treatment methods therewith described in this disclosure may be utilized in combination with other types of therapy for the target disease or disorder disclosed herein. In this context, the antibody composition and the therapeutic agent may be given either simultaneously or sequentially. Examples include chemotherapy, immunotherapy (e.g., therapy with other HJV antagonists), surgery, radiation, gene therapy, etc., or anti-infectious disease therapy. Such therapy may be administered simultaneously or sequentially (in any order) with the treatment according to the present disclosure.

[0329] For example, the combination therapy can include the anti-HJV antibody and pharmaceutical composition described herein, which are formulated and / or co-administered with at least one additional therapeutic agent described herein. Such combination therapy can advantageously utilize lower dosages of the therapeutic agent administered, thus preventing possible toxicity or complications associated with various monotherapies. Furthermore, the additional therapeutic agent disclosed herein can act in a pathway in addition to or separate from the hepcidin / BMP pathway, thus enhancing and / or synergizing with the effect of the anti-HJV antibody.

[0330] In some embodiments, the subject has previously received an erythropoietin stimulating agent, hi some embodiments, the erythropoietin stimulating agent is selected from the group consisting of danazol, prednisone, thalidomide, lenalidomide, and pomalidomide.

[0331] In some embodiments, the subject has previously been administered a JAK-STAT pathway inhibitor. In some embodiments, the JAK-STAT pathway inhibitor is a JAK inhibitor or a STAT inhibitor. In some embodiments, the JAK inhibitor is selective for one or both of the subtypes JAK1 and JAK2 (e.g., a JAK1 / 2 inhibitor). In some embodiments, the STAT inhibitor is a STAT3 inhibitor. In some embodiments, the JAK1 / 2 or STAT3 inhibitor is selected from the group consisting of ruxolitinib, fedratinib, momelotinib, pacritinib, INCB039110, AG490, and PpYLKTK [wherein "pY" represents a phosphorylated tyrosine (Y) residue (PY*LKTK); SEQ ID NO: 131].

[0332] In some embodiments, the subject has previously received a growth factor ligand trap. In some embodiments, the growth factor ligand trap is a transforming growth factor beta (TGF-β) ligand trap. In some embodiments, the TGF-β ligand trap is a GDF trap, such as sotatercept or raspatercept. See U.S. Patent No. 8,216,997, the contents of each of which are incorporated herein by reference. In some embodiments, the subject has previously received an antifibrotic agent. In some embodiments, the antifibrotic agent is PRM-151.

[0333] In some embodiments, the subject in need of treatment according to the present disclosure continues to receive a therapeutic treatment for a blood disorder. The present disclosure thus provides compositions and methods for treating anemia (e.g., ACD (e.g., anemia of chronic kidney disease) and / or one or more conditions resulting from anemia) in some aspects by administering an anti-HJV antibody to a subject in need thereof in combination with one or more therapeutic treatments for a blood disorder.

[0334] In some embodiments, anemia is characterized based on reticulocyte hemoglobin content (RET-He or CHr). Reticulocyte hemoglobin content is a measure of the amount of hemoglobin in reticulocytes. The normal range for CHr is about 28-36 pg / cell. In some embodiments, the subject has a CHr lower than the normal range. In some embodiments, the subject has a CHr of less than 36 pg / cell, less than 35 pg / cell, less than 34 pg / cell, less than 33 pg / cell, less than 32 pg / cell, less than 31 pg / cell, 30 pg / cell, less than 29 pg / cell, less than 28 pg / cell, less than 27 pg / cell, less than 26 pg / cell, less than 25 pg / cell, less than 24 pg / cell, less than 23 pg / cell, less than 21 pg / cell, or less than 20 pg / cell. However, it should be understood that other suitable markers (e.g., TSAT%, serum iron levels, total iron binding capacity (TIBC), ferritin levels, hemoglobin levels, liver iron content, hepcidin levels, IL-6 levels, creatinine levels, etc.) may be assessed to determine whether a subject is suitable for the treatment methods described herein.

[0335] In some embodiments, the anemia is characterized by liver iron levels. In some embodiments, the normal range of liver iron levels is 200-2,400 μg per gram dry weight in men and 400-1,600 μg per gram dry weight in women. In some embodiments, the subject has higher than normal liver iron levels. In some embodiments, the patient has liver iron levels greater than 1000 μg per gram dry weight (e.g., about 1000 μg-1200 μg per gram dry weight, about 1000 μg-1500 μg per gram dry weight, or about 1200 μg-1500 μg per gram dry weight), greater than 1500 μg per gram dry weight (e.g., about 1500 μg-1800 μg per gram dry weight, about 1500 μg-2000 μg per gram dry weight, or about 1800 μg-2000 μg per gram dry weight), or greater than normal liver iron levels (e.g., about 1000 μg-1200 μg per gram dry weight, about 1000 μg-1500 μg per gram dry weight, or about 1200 μg-1500 μg per gram dry weight). have a liver iron level of greater than 2000 μg (e.g., about 2000 μg to 2200 μg per gram dry weight, about 2000 μg to 2500 μg per gram dry weight, or about 2200 μg to 2500 μg per gram dry weight), greater than 2500 μg per gram dry weight (e.g., about 2500 μg to 2800 μg per gram dry weight, about 2500 μg to 3000 μg per gram dry weight, or about 2800 μg to 3000 μg per gram dry weight), or greater than 3000 μg per gram dry weight.

[0336] In some embodiments, the anemia is characterized by low total iron binding capacity (TIBC). In some embodiments, the normal range of TIBC is 250-400 μg / dL. In some embodiments, the subject has a lower than normal TIBC. In some embodiments, the subject has a TIBC of less than 400 μg / dL, less than 350 μg / dL, less than 300 μg / dL, less than 250 μg / dL, less than 200 μg / dL, less than 150 μg / dL, less than 100 μg / dL, less than 90 μg / dL, less than 80 μg / dL, less than 70 μg / dL, less than 60 μg / dL, less than 50 μg / dL, less than 40 μg / dL, less than 30 μg / dL, less than 20 μg / dL, or less than 10 μg / dL.

[0337] In some embodiments, the subject is administered a hemojuvelin antagonist (e.g., an anti-HJV antibody and compositions thereof) in combination with an erythropoietin stimulating agent. In some embodiments, the erythropoietin stimulating agent is EPO. In some embodiments, the erythropoietin stimulating agent is selected from an immunomodulatory agent selected from the group consisting of danazol, prednisone, thalidomide, lenalidomide, and pomalidomide.

[0338] In some embodiments, the subject is administered a hemojuvelin antagonist (e.g., an anti-HJV antibody and compositions thereof) in combination with a JAK-STAT pathway inhibitor. In some embodiments, the JAK-STAT pathway inhibitor is a JAK inhibitor or a STAT inhibitor. In some embodiments, the JAK inhibitor is selective for one or both of the subtypes JAK1 and JAK2 (e.g., a JAK1 / 2 inhibitor). In some embodiments, the STAT inhibitor is a STAT3 inhibitor. In some embodiments, the JAK1 / 2 or STAT3 inhibitor is selected from the group consisting of ruxolitinib, fedratinib, momelotinib, pacritinib, INCB039110, AG490, and PpYLKTK (SEQ ID NO: 131). In some embodiments, the subject is administered a hemojuvelin antagonist (e.g., an anti-HJV antibody and compositions thereof) in combination with ruxolitinib.

[0339] In some embodiments, the hemojuvelin antagonist (e.g., anti-HJV antibody and compositions thereof) reduces the extent to which the subject exhibits an anemic response to a JAK-STAT pathway inhibitor. For example, in some embodiments, a subject treated with a JAK-STAT pathway inhibitor as a monotherapy may be characterized as having a deficiency in the blood's ability to transport oxygen compared to the subject's pre-treatment state, a deficiency in red blood cells compared to the subject's pre-treatment state, a deficiency in hemoglobin compared to the subject's pre-treatment state, and / or a deficiency in total blood volume compared to the subject's pre-treatment state. Thus, in some embodiments, the hemojuvelin antagonist (e.g., anti-HJV antibody) reduces the extent to which the subject exhibits an anemic response to a JAK-STAT pathway inhibitor selected from the group consisting of ruxolitinib, fedratinib, momelotinib, pacritinib, INCB039110, AG490, and PpYLKTK (SEQ ID NO: 131). In some embodiments, the hemojuvelin antagonists (eg, anti-HJV antibodies and compositions thereof) reduce the extent to which a subject exhibits an anemic response to ruxolitinib administration.

[0340] In some embodiments, the subject is administered a hemojuvelin antagonist (e.g., an anti-HJV antibody and compositions thereof) in combination with a growth factor ligand trap. In some embodiments, the growth factor ligand trap is a transforming growth factor beta (TGF-β) ligand trap. In some embodiments, the TGF-β ligand trap is sotatercept or raspatercept. In some embodiments, the subject is administered a hemojuvelin antagonist (e.g., an anti-HJV antibody and compositions thereof) in combination with an anti-fibrotic agent. In some embodiments, the anti-fibrotic agent is PRM-151.

[0341] The success of the subject treatment according to the present disclosure may be determined by methods known in the art or by those skilled in the art.In some embodiments, HJV antagonist (e.g., anti-HJV antibody and composition thereof) treatment is evaluated based on the hepcidin (e.g., circulating hepcidin-25 level) level in the subject.For example, in some embodiments, the baseline hepcidin (e.g., circulating hepcidin-25 level) level in the subject is determined (e.g., before treatment with HJV antagonist (e.g., anti-HJV antibody and composition thereof) or in the absence of HJV antagonist (e.g., anti-HJV antibody and composition thereof) treatment at the time of determination otherwise), and compared with the hepcidin (e.g., circulating hepcidin-25 level) level after treatment in the subject. In some embodiments, the subject is successfully treated and the HJV antagonist (e.g., anti-HJV antibodies and compositions thereof) reduces hepcidin (e.g., circulating hepcidin-25 levels) levels in the subject by about 1 ng / mL to about 300 ng / mL. In some embodiments, the HJV antagonist (e.g., anti-HJV antibodies and compositions thereof) reduces hepcidin (e.g., circulating hepcidin-25 levels) levels in the subject by about 1 ng / mL to about 200 ng / mL, about 1 ng / mL to about 100 ng / mL, about 1 ng / mL to about 50 ng / mL, about 1 ng / mL to about 10 ng / mL, about 10 ng / mL to about 100 ng / mL, or about 10 ng / mL to about 50 ng / mL.

[0342] In some embodiments, the present disclosure provides a method of reducing hepcidin (e.g., circulating hepcidin-25 levels) in a subject with anemia (e.g., ACD (e.g., anemia of chronic kidney disease)). In some embodiments, the disclosed administration methods reduce hepcidin-25 within 4 hours, 6 hours, 8 hours, 12 hours, 28 hours, 24 hours, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, or 2 weeks of administration. In some embodiments, the disclosed administration methods reduce hepcidin (e.g., circulating hepcidin-25 levels) by at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 99%, or 100% compared to hepcidin (e.g., circulating hepcidin-25 levels) levels in the subject prior to administration. In some embodiments, the disclosed administration methods reduce hepcidin levels by about 96%.

[0343] In some embodiments, the disclosure provides methods of reducing hepcidin expression (e.g., Hamp gene expression) in the liver of a subject with anemia (e.g., ACD (e.g., anemia of chronic kidney disease)). In some embodiments, the disclosed administration methods reduce Hamp mRNA expression within 4 hours, 6 hours, 8 hours, 12 hours, 28 hours, 24 hours, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, or 2 weeks of administration. In some embodiments, the disclosed administration methods reduce Hamp mRNA expression within 7, 10, 14, 20-21, 24, 28, 30, 35, 40, or 42 days of administration. In some embodiments, the disclosed administration methods reduce Hamp mRNA expression by at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 99%, or 100% of hepcidin compared to expression levels in the subject prior to administration. In some embodiments, the disclosed administration methods reduce Hamp expression levels in the liver of the subject by about 96%.

[0344] In some embodiments, HJV antagonist (e.g., anti-HJV antibody and composition thereof) treatment is evaluated based on the serum ferritin level in the subject.For example, in some embodiments, the baseline serum ferritin level in the subject is determined (e.g., before treatment with HJV antagonist (e.g., anti-HJV antibody and composition thereof) or in the absence of HJV antagonist (e.g., anti-HJV antibody and composition thereof) treatment at the time of determination otherwise) and compared with the serum ferritin level after treatment in the subject.

[0345] In some embodiments, the subject is successfully treated and the HJV antagonist (e.g., anti-HJV antibodies and compositions thereof) reduces serum ferritin levels in the subject by about 1 ng / mL to about 200 ng / mL. In some embodiments, the HJV antagonist (e.g., anti-HJV antibodies and compositions thereof) reduces serum ferritin levels in the subject by about 1 ng / mL to about 100 ng / mL, about 1 ng / mL to about 50 ng / mL, about 1 ng / mL to about 25 ng / mL, about 1 ng / mL to about 10 ng / mL, about 10 ng / mL to about 100 ng / mL, or about 10 ng / mL to about 50 ng / mL.

[0346] In some embodiments, HJV antagonist (e.g., anti-HJV antibody and composition thereof) treatment is evaluated based on the serum hemoglobin level in the subject. For example, in some embodiments, the baseline serum hemoglobin level in the subject is determined (e.g., before treatment with HJV antagonist (e.g., anti-HJV antibody and composition thereof) or in the absence of HJV antagonist (e.g., anti-HJV antibody and composition thereof) treatment at the time of determination otherwise) and compared with the serum hemoglobin level in the subject after treatment.

[0347] In some embodiments, the subject is successfully treated and the HJV antagonist (eg, anti-HJV antibodies and compositions thereof) increases serum hemoglobin levels in the subject by about 0.01 g / dL to about 5 g / dL.

[0348] In some embodiments, the HJV antagonists (e.g., anti-HJV antibodies and compositions thereof) reduce serum ferritin levels in a subject by about 0.01 g / dL to about 1 g / dL, about 0.1 g / dL to about 5 g / dL, about 1 g / dL to about 5 g / dL, about 0.01 g / dL to about 0.1 g / dL, about 0.5 g / dL to about 2.5 g / dL, or about 0.1 g / dL to about 1 g / dL.

[0349] VII. Comorbidities and Risk Factors The disclosed methods may be particularly suitable for CKD patients who have CKD comorbidities.In particular, these methods may be useful for treating the anemia of CKD in subjects with any of the following comorbidities: diabetes, hypertension, autoimmune disease (e.g., lupus or another autoimmune condition), complications caused by certain medicines, such as nonsteroidal anti-inflammatory drugs (NSAIDs), or congenital heart failure.These and other comorbidities may be a major consideration for physicians when advising patients on whether or not to undergo dialysis.

[0350] These methods may be useful for treating anemia of CKD in subjects who have certain other comorbidities or risk factors, such as congenital defects, fatigue, nausea, insomnia, anorexia, or high BMI; or where renal damage is associated with polycystic kidney disease or glomerular disease, such as acute glomerulonephritis. Several rare diseases and birth defects occur simultaneously with CKD, including diabetes (e.g., type II diabetes), bacterial endocarditis, granulomatosis with polyangiitis, hepatitis B and C, human immunodeficiency virus (HIV) infection, hyperkalemia, light chain deposition disease, mixed connective tissue disease, mixed cryoglobulinemia, hyperplasia, nonalcoholic fatty liver disease (NAFLD), parasitic infections, reflux nephropathy, rheumatoid arthritis, scleroderma, Shiga toxin or Streptococcus pneumoniae-associated HUS, some cancers, systemic lupus erythematosus, syphilis, and thrombotic thrombocytopenic purpura (TTP). In addition, use of the prescription drugs captopril and penicillamine (a chelating agent) has been identified as a risk factor for CKD. Recreational use of heroin is another risk factor for CKD. See Zipfel PF, et al. Front Immunol. 2019; 10:2166; and Alwahaibi NY, et al. Saudi J Kidney Dis Transpl. 2018 Nov-Dec; 29(6):1256-1266, each of which is incorporated by reference herein.

[0351] Thus, provided herein are methods of treating anemia in chronic kidney disease associated with one or more of the following comorbidities: diabetes, hypertension, autoimmune diseases such as lupus, congenital heart defects, complications related to administration of one or more NSAIDs (e.g., Aleve, Tylenol, Advil, or Motrin), rare diseases, and congenital defects.

[0352] Further provided herein is a method for treating anemia in chronic kidney disease associated with polycystic kidney disease. Primary or secondary human glomerular diseases, such as acute glomerulonephritis, Alport syndrome, amyloidosis (hereditary Amyl), anti-GBM (Goodpasture) disease, antineutrophil cytoplasmic antibody-mediated vasculitis (ANCA), atypical hemolytic uremic syndrome (aHUS), C3 glomerulopathy, chronic graft-mediated glomerulopathy, diabetic nephropathy (DN), focal segmental glomerulosclerosis (FSGS), glomerulosclerosis, Henoch-Schönlein purpura (HSP), hypertension (HTN). Further provided herein are methods of treating anemia in chronic kidney disease associated with IgA nephropathy (IgAN), immune complex membranoproliferative glomerulonephritis, renal ischemia-reperfusion injury, lupus nephritis (LN), membranous nephropathy, membranoproliferative glomerulonephritis (MPGN), mesangial proliferative glomerulonephritis (MesGN), membranous glomerulopathy (MG), minimal change disease (MCD), post-streptococcal glomerulonephritis (PSGN), rapidly progressive (crescentic) glomerulonephritis (RPGN).

[0353] These methods may be useful for subjects who have previously been administered medicines for treating certain conditions, such as diabetes.In some embodiments, provided herein is a method for treating anemia in subjects with CKD who have previously been administered or are currently administered type II diabetes therapy.Further provided is a method for treating anemia in subjects with CKD who have previously been administered or are currently administered hypertension medicines, such as ACE inhibitors.

[0354] In some embodiments of the disclosed methods, the subject has not previously undergone a nephrectomy or kidney transplant. EXAMPLES

[0355] [Example 1] Generation and characterization of anti-HJV antibodies From rats immunized with human hemojuvelin, hybridoma clones with the ability to bind to human hemojuvelin were identified. Anti-hemojuvelin monoclonal antibodies (mAbs with rat IgG1 / κ) were humanized by CDR grafting (hHA antibodies with hIgG1 constant regions with L234A, L235A mutations). Affinity matured anti-HJV (e.g., HA-001 to HA-011) in vitro yeast display assay. The general method for the generation of humanized affinity matured anti-HJV antibodies is shown in Figure 1A.

[0356] The binding affinity of anti-HJV antibodies to soluble human RGMa, rat RGMa and human RGMc was measured by BIAcore analysis. Table 14 shows the affinity of anti-hemojuvelin antibodies to human RGMa. Table 15 shows the affinity of anti-hemojuvelin antibodies to rat RGMa. Table 16 shows the affinity of anti-hemojuvelin antibodies to human RGMc.

[0357] [Table 5]

[0358] [Table 6]

[0359] [Table 7]

[0360] Among these antibodies, at least hHA-004, hHA-008, hHA-009 and hHA-011 showed strong binding to human RGMc and were selected for further testing. Sensorgrams from BIAcore analysis of antibodies HA, hHA-004, hHA-008, hHA-009 and hHA-011 are shown in Figure 1B-1G. hHA-008 was further tested for its binding affinity to human RGMa, cynomolgus monkey RGMa, rat RGMa, human RGMc, cynomolgus monkey RGMc, rat RGMc, and human RGMb, and the respective binding affinities are shown in Table 7. hHA-008 showed high affinity binding to human RGMa and RGMc along with strong cross-reactivity to cynomolgus monkey and rodent species.

[0361] [Table 8]

[0362] The RGMc (HJV) BMP reporter gene assay was used to screen anti-HJV antibodies and characterize their potency in blocking membrane-bound RGMc-induced BMP / Smad1 / 5 / 8 signaling. The assay directly relates to the mechanism of action of anti-HJV mAbs in inhibiting the RGMc / BMP / Smad1 / 5 / 8 signaling pathway responsible for the induction of iron hormone hepcidin gene expression. The principle of the HJV BMP reporter assay is shown in Figure 2A.

[0363] The BRE-Luc reporter gene vector, originally described by Korchynskyi and Dijke (J. Biol. Chem. 2002; 277:4883c), was used to transiently transfect the porcine kidney epithelial cell line LLC-PK1 with or without co-transfection of an RGMa expression vector by Babbit et al (J. Biol. Chem. 2005; 280:29820) to determine whether RGMa expression modulates BMP signaling. In BRE-Luc transiently transfected cells, RGMa was demonstrated to enhance BMP signaling through the Smad1 / 5 / 8 signaling pathway, consistent with the role of RGMa as a BMP coreceptor. All RGM members (RGMa, RGMb, and RGMc) act as BMP coreceptors and enhance BMP signaling. The BRE-Luc reporter vector was constructed and established in the RGM BMP reporter gene assay in HEK293 cells. The human RGMc expression vector used in the assay was pcDNA-hRGMc.

[0364] HEK293 cells were cultured in growth medium [basal medium DMEM (Invitrogen catalog #11965-092) containing 10% fetal bovine serum (Gibco #10438-026) and 1% sodium pyruvate (Invitrogen, catalog #11360-070)]. To prepare cells for transfection, 9 × 10 6HEK293 cells were plated in 10 cm dishes and incubated at 37°C, 5% CO2 for 6 hours. Cells were then transfected using 20 μl of PolyJet (SignaGen, Cat# SL100688) with a 2:1 PolyJet (μL):DNA (μg) ratio, specifically 5 μg of pGL[luc2P / BRE / Hygro]DNA (Abbvie) and 5 μg of pcDNA-hRGMc (Abbvie) for 16 hours at 37°C, 5% CO2. Media was replaced with fresh HEK293 growth media for 6 hours. Cells were then trypsinized using TrpLE (ThermoFisher, Cat# 12605010), counted, and plated at 1 × 10 in 96-well white assay plates (Thermo Scientific Nunclon delta F96, Cat# 136102). 5 Cells were plated at 1000 x 1000 cells / well. Cells were treated with anti-RGMc mAb, anti-BMP2 / 4 mAb (R&D Systems, Cat# MAB3552) (as a positive control), or isotype control mAb at 37°C, 5% CO2 for 16 hours. Luciferase activity was detected using One-Glo Luciferase kit (Promega, Cat# E6120) and measurement on a TopCount luminescence counter. The results showed that anti-HJV antibodies dose-dependently inhibited RGMc-enhanced BMP signaling (Figure 2B). The IC50 (nM) of each of the antibodies tested in inhibiting RGMc signaling in the BMP reporter assay is shown in Table 18.

[0365] [Table 9]

[0366] In addition, anti-HJV antibodies were tested for their ability to inhibit RGMa signaling in BMP reporter assays. hHA-004 and hHA-011 showed strong inhibition of membrane-bound human RGMa in RGMa BMP reporter gene assays, while hHA-008 and hHA-009 showed no or minimal inhibition of RGMa activity. hHA antibodies showed no inhibition of membrane-bound RGMa. Rat mAb HA showed inhibition of RGMa, but to a much lesser extent compared to hHA-004 and hHA-011. Figure 2C. The IC50 (nM) of each of the antibodies tested in inhibiting RGMa signaling in BMP reporter assays is shown in Table 9.

[0367] [Table 10]

[0368] The above data showed that the potency of anti-HJV antibodies in neutralizing membrane RGMc in the BMP reporter assay correlated with their binding affinity to soluble RGMc. A summary of the binding affinity of anti-HJV antibodies to soluble human RGMc and their ability to inhibit membrane-bound RGMc and RGMa signaling is shown in Table 20.

[0369] [Table 11]

[0370] Antibodies including hHA, hHA-004, hHA-008, hHA-008-QL, hHA-009, and hHA-011 were tested for non-specific cell binding to HEK293 cells by FACS analysis. The data showed that hHA and its hHA-008, hHA-008-QL, and hHA-011 showed no or minimal non-specific cell binding to HEK293 cells at concentrations up to 100ug / ml. hHA-004 and hHA-009 showed some non-specific cell binding at higher concentrations, but to a much lesser extent compared to the positive control IgG (Figure 3).

[0371] [Example 2] Generation of hHA-008-QL IgG serum half-line (t 1 / 2 hHA-008-QL was developed to prolong the life of IgG. Previous studies have shown that the neonatal Fc receptor (FcRn) protects IgG from catabolism, thereby increasing IgG serum half-life. Thus, the Fc portion of hHA-008-QL was engineered to have T250Q and M428L mutations (QL mutations) to have enhanced binding to FcRn. Figures 4A and 4B illustrate the structures of hHA-008 and hHA-008-QL, respectively. Further examples of hHA-008 and hHA-008-QL are shown in Figure 4C.

[0372] First, hHA-008-QL was tested for its RGMa and RGMc binding ability, and the data showed that hHA-008-QL had binding affinity for RGMa and RGMc comparable to hHA-008 (Table 21).

[0373] [Table 12]

[0374] [Example 3] Immunogenicity study of hHA-008 and hHA-008-QL hHA-008 and hHA-008-QL were tested for their immunogenicity using peripheral blood mononuclear cells (PBMCs) from 50 donors (representing all major HLA-DR and HLA-DQ haplotypes) loaded with hHA-008 or hHA-008-QL in a CD4+ T cell response assay. The results showed that only 4% of the 50 donors (2 / 50) showed a T cell response, suggesting that both antibodies have a low risk of immunogenicity. In this assay, Herceptin was used as a negative control, to which 8% of the donors had a T cell response. Bydureon and keyhole limpet hemocyanin (KLH) were used as positive controls. 32% of the donors showed a T cell response to Bydureon and 100% of the donors had a T cell response to KLH. Figure 5 showed the T cell responses in 50 donors for hHA-008 and hHA-008-QL.

[0375] In addition, FcγR binding was tested for hHA-008 and hHA-008-QL as another parameter of immunogenicity. The control group, wild-type unrelated IgG1, had significant binding to both high and low affinity Fc gamma receptors. As expected, the binding of wild-type unrelated IgG4 was significantly lower than that of wild-type unrelated IgG1. Binding to high and low affinity receptors was significantly reduced for both hHA-008 and hHA-008-QL compared to wild-type IgG1, suggesting low immunogenicity for both antibodies.

[0376] [Example 4] Pharmacokinetic / Pharmacodynamic (PK / PD) Modeling of hHA-008 and hHA-008-QL In an initial study of the PK / PD of hHA-008, male rats were treated with a single dose of hHA-008 at 5 mg / kilogram body weight (mpk) by intravenous injection. Transferrin saturation (TSAT%; determined as the percentage of serum iron to total serum iron binding capacity) was examined over time. Data showed that the maximal effect of increased TSAT occurred between 4 and 8 days after treatment (Figure 6A).

[0377] Additionally, a similar study was performed in the non-human primate cynomolgus monkey. Both female (n=2) and male (n=2) cynomolgus monkeys were given a single dose of hHA-008 at 5 mg / kilogram body weight (mpk) by intravenous injection. Transferrin saturation (TSAT%) was examined over time. Both female cynomolgus monkeys showed a maximal effect of increased TSAT% approximately 1-4 days after injection (Figure 6B). One of the males showed the same effect, while the other did not respond to hHA-008 treatment (Figure 6C). By examining the hematological profile of this male, it was subsequently determined that the lack of response was the result of this cynomolgus monkey having plasma hepcidin-25 levels below the limit of detection (below 2 ng / mL). Furthermore, prior to injection, this male cynomolgus monkey had lower baseline TSAT% and serum iron levels than the other cynomolgus monkeys in the group, with a baseline TSAT% level of 18% and a serum iron level of 77 μg / dL 2 days prior to injection; and a baseline TSAT% level of 14% and a serum iron level of 61 μg / dL 1 day prior to injection. The low baseline TSAT% and serum iron levels, combined with the low hepcidin-25 levels, were indicative that this male cynomolgus monkey had absolute iron deficiency rather than functional iron deficiency.

[0378] Further studies examining the effects of hHA-008 administration in more than 70 cynomolgus monkeys revealed that animals with normal baseline serum iron levels (e.g., within the range of 80 μg / dL-180 μg / dL) and hepcidin-25 levels greater than 2 ng / mL were responsive to hHA-008 treatment in that they exhibited a significant increase in TSAT% levels immediately after injection (e.g., within 1-2 days after treatment) in response to a decrease in hepcidin-25 expression.

[0379] Additional experiments were performed in cynomolgus monkeys. A single dose of 6 mpk of hHA-008 was administered via intravenous injection to three cynomolgus monkeys, and TSAT% (determined as serum iron relative to total iron binding capacity), plasma hepcidin-25 concentration, and plasma hHA-008 concentration were examined for each animal and time point. Similarly, the data showed that the maximal effect of increased TSAT% occurred 1-4 days after injection (T max = 1-4 days), which was consistent with an increase in hHA-008 concentrations. One of the animals had a dramatic drop in TSAT% around day 34 (Figure 7A), which was consistent with a drop in plasma hHA-008 concentration around that time point. Plasma hepcidin-25 concentrations were inversely correlated with the concentration of hHA-008; hepcidin-25 was undetectable after antibody injection, and for the animal that had a dramatic drop in TSAT%, hepcidin-25 levels increased around the same time point (Figure 7B). In this cynomolgus monkey, hHA-008 increased plasma t 1 / 2 The animals showed a decrease in TSAT% and an increase in hepcidin-25 levels at about day 34, which correlated well with the decrease in hHA-008 from plasma (Figure 7C). 1 / 2 supports a dosing frequency of at least once per month in humans.

[0380] Additionally, hHA-008 demonstrated robust PK / PD correlations of PK (plasma antibody concentration) versus TSAT% and plasma hepcidin-25 concentration. Results for each tested cynomolgus monkey are shown in Figure 7D (cynomolgus monkey 1), Figure 7E (cynomolgus monkey 2), and Figure 7F (cynomolgus monkey 3). hHA-008 demonstrated a t of 10.3 days in cynomolgus monkey 1. 1 / 2 , t of 8.8 days in cynomolgus monkeys 1 / 2 and a t of 5.1 days in cynomolgus monkeys3. 1 / 2 Hepcidin-25 levels were reduced to undetectable levels (<2ng / ml) after hHA-008 treatment. Interestingly, the return of hepcidin-25 levels to circulation was consistent with the t 1 / 2 (Figure 7F).

[0381] hHA-008 antibody modulates TSAT% in a dose-dependent manner. A multiple dose study was performed in cynomolgus monkeys. Cynomolgus monkeys (n=4 per dose level, 2 males and 2 females) were treated with either 0 (vehicle control), 0.6mpk hHA-008, 3mpk hHA-008, or 60mpk hHA-008. The resulting concentrations of hHA-008 and the corresponding TSAT% responses are presented in Figure 8A, Figure 8B, and Figure 8C for treatment with 0.6, 3, and 60mpk, respectively, all plotted against vehicle control. Cynomolgus monkeys were dosed every 14 days. The dotted lines represent the days of dosing. TSAT% increased after dosing, and the modulation percentage was consistent with dose level; at 0.6mpk, TSAT reached about 60%, and at 3mpk and 60mpk, TSAT% was saturated, indicating that hHA-008 modulates TSAT% in a dose-dependent manner.Furthermore, after the first dose at 0.6mpk, TSAT% level was maintained at about 60%, and at higher dose levels, TSAT% reached 100%, suggesting that TSAT can be modulated by selecting appropriate dose levels / regimen (Figures 8A-8C).

[0382] [Example 5] hHA-008-QL confers a longer serum half-life compared to hHA-008, and takes longer to reach maximum efficacy This study was designed to administer hHA-008 or hHA-008-QL at 6mpk intravenously to cynomolgus monkeys (n=3). Samples were collected from each animal 48 and 24 hours prior to treatment, and 0, 0.04, 0.08, 0.167, 0.333, 0.5, 1, 2, 4, 8, 14, 21, 28, 35, and 42 days after treatment. TSAT% and plasma hepcidin-25 levels were measured in each sample. The data showed that hHA-008 had a maximal effect (T max ) (Figure 9A). It is noted that there were non-responders in animals treated with hHA-008-QL. As discussed above, non-responders had low hepcidin-25 levels and low serum iron at baseline before antibody treatment. On the other hand, hHA-008-QL had a longer serum half-life (T1 / 2=12.1 days) compared to hHA-008 (T1 / 2=6.76 days) (Figure 9B). Plasma concentrations of both antibodies were measured over time, as shown in Figure 9C. Peak plasma concentrations were determined. A summary of the data between hHA-008 and hHA-008-QL is shown in Table 22.

[0383] [Table 13]

[0384] To investigate whether the longer serum lifetime for hHA-008-QL is attributable to a higher affinity for FcRn, both antibodies were analyzed for binding to FcRn at pH 6.0 and pH 7.4 using BIAcore. Non-specific IgG1 and IgG4 were used as controls. The dissociation constants (KD) of each of the antibodies and controls for FcRn at pH 6.0 and 7.4 are shown in Table 23, and the response curves are shown in Figures 10A-10B. No binding was observed for either antibody at pH 7.4 (Figure 10B).

[0385] [Table 14]

[0386] Mutations L247A and L248A (Kabat) (L234A and L235A, EU numbering) do not appear to significantly affect FcRn binding. The overall affinity and response (RMax) for FcRn binding at pH 6.0 is increased for hHA-008-QL compared to hHA-008, suggesting that the QL mutation confers a longer t1 / 2 through binding to the receptor.

[0387] [Example 6] hHA-008 reduces IL-6-induced hepcidin-25 expression in non-human primates As depicted in Figure 11, in iron sequestration anemia, pro-inflammatory cytokines such as IL-6 and oncostatin-M, which induce hepcidin synthesis, are typically increased and are associated with iron sequestration, macrophage iron loading, as well as bone marrow proliferation and macrophage activation. To test whether IL-6 indeed increases hepcidin expression and whether anti-HJV antibodies can inhibit IL-6-induced hepcidin expression in non-human primates, sinuses were loaded with IL-6 on day 1 and divided into three groups. On day 4, sinuses in group 1 received vehicle control, sinuses in group 2 received hHA-008 antibody at 0.6 mg / kg, and sinuses in group 3 received hHA-008 antibody at 6.0 mg / kg. On day 11, sinuses in all three groups were loaded with IL-6 again, and plasma hepcidin-25 in all sinuses was measured. As shown in FIG. 13, IL-6 loading increased plasma hepcidin-25 concentrations on day 1 compared to pre-loading baseline (BL) in sinuses of all three groups. After a second IL-6 loading on day 11, sinuses o...

Claims

1. A pharmaceutical composition comprising an antibody that binds to human hemojuvelin (HJV) for use in treating anemia in a subject, wherein the subject has an anemia of 1.73 m ２ The pharmaceutical composition has been identified as having a glomerular filtration rate (GFR) level of less than 90 mL / min per day. A method comprising:

2. 10. The pharmaceutical composition of claim 1, wherein the anemia is caused, at least in part, by nutritional iron deficiency, iron deficiency due to blood loss, intrinsic red blood cell damage, hemolysis, inflammation, functional iron deficiency, or any combination thereof.

3. The target is 1.73 m ２ 3. The pharmaceutical composition of claim 1 or 2, wherein the composition is identified as having a GFR level within the range of 15 mL / min to less than 90 mL / min per minute.

4. The target is 1.73 m ２ 10. The pharmaceutical composition of claim 1, wherein the composition is identified as having a GFR level within the range of 15 mL / min to less than 60 mL / min per minute.

5. The target is 1.73 m ２ 10. The pharmaceutical composition of claim 1, wherein the composition is identified as having a GFR level within the range of 15 mL / min to less than 30 mL / min per minute.

6. The target is 1.73 m ２ 10. The pharmaceutical composition of claim 1, wherein the composition is identified as having a GFR level within the range of less than 30 mL / min per minute.

7. The target is 1.73 m ２ 10. The pharmaceutical composition of claim 1, wherein the composition is identified as having a GFR level of less than 15 mL / min per minute.

8. The target is 1.73 m ２ 10. The pharmaceutical composition of claim 1, wherein the composition is identified as having a GFR level of less than 7 mL / min per minute.

9. 2. The pharmaceutical composition of claim 1, wherein the level of glomerular filtration rate is sustained for at least 3 months.

10. 10. The pharmaceutical composition of claim 1, wherein the anemia is associated with kidney damage in the subject.

11. 11. The pharmaceutical composition of claim 10, wherein the renal damage has been present for at least 3 months.

12. 12. The pharmaceutical composition of claim 10 or 11, wherein the kidney damage is associated with polycystic kidney disease or a glomerular disease, such as acute glomerulonephritis.

13. 10. The pharmaceutical composition of claim 1, wherein the subject is identified as having chronic kidney disease.

14. The pharmaceutical composition according to claim 13, wherein the chronic kidney disease is non-dialysis-dependent chronic kidney disease.

15. The pharmaceutical composition of claim 1 , wherein the subject is not undergoing dialysis therapy.

16. 16. The pharmaceutical composition of any one of claims 13 to 15, wherein the chronic kidney disease is associated with one or more of the following comorbidities: diabetes, hypertension, an autoimmune disease, such as lupus or other autoimmune conditions, congenital heart defects, complications related to the administration of one or more nonsteroidal anti-inflammatory drugs, rare diseases, and congenital defects.

17. 16. The pharmaceutical composition of any one of claims 13 to 15, wherein the chronic kidney disease is classified as being at a stage within the range of stages 1 to 4.

18. 16. The pharmaceutical composition of any one of claims 13 to 15, wherein the chronic kidney disease is classified as being at a stage within the range of stages 2 to 4.

19. The pharmaceutical composition of any one of claims 13 to 15, wherein the chronic kidney disease is classified as being at stage 4.

20. The pharmaceutical composition of claim 1 , wherein the subject has not undergone a nephrectomy or kidney transplant.

21. The pharmaceutical composition of claim 1, for administration by subcutaneous, intravenous, or intramuscular injection.

22. The pharmaceutical composition of claim 1, for administration by subcutaneous injection.

23. 23. The pharmaceutical composition of claim 22, for subcutaneous self-administration.

24. The pharmaceutical composition of claim 1, wherein the antibody preferentially binds to RGMc compared to RGMa and RGMb.

25. The antibody according to claim 25, wherein the antibody has an equilibrium dissociation constant (K Ｄ 25. The pharmaceutical composition of claim 24, which binds to RGMc at 26. The pharmaceutical composition of claim 25, wherein the antibody is an anti-hemojuvelin antibody listed in Table 1.

27. 27. The pharmaceutical composition of any one of claims 24 to 26, wherein the antibody comprises a HC CDR1 of SEQ ID NO: 1, a HC CDR2 of SEQ ID NO: 2, a HC CDR3 of SEQ ID NO: 3; a LC CDR1 of SEQ ID NO: 17, a LC CDR2 of SEQ ID NO: 5, and a LC CDR3 of SEQ ID NO:

27.

28. The pharmaceutical composition of any one of claims 24 to 26, wherein the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 38 and a VL comprising the amino acid sequence of SEQ ID NO:

39.

29. The pharmaceutical composition of any one of claims 24 to 26, wherein the antibody is selected from the group consisting of a full-length IgG, a Fab fragment, a F(ab') fragment, a F(ab')2 fragment, a scFv, and an Fv.

30. The pharmaceutical composition of any one of claims 24 to 26, wherein the antibody is a full-length IgG comprising a heavy chain constant region of isotype IgG1, IgG2, IgG3, or IgG4.

31. The pharmaceutical composition of any one of claims 24 to 26, wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 61 and a light chain comprising the amino acid sequence of SEQ ID NO:

62.

32. The pharmaceutical composition of any one of claims 24 to 26, wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 63 and a light chain comprising the amino acid sequence of SEQ ID NO:

62.

33. A pharmaceutical composition comprising an antibody that binds to human hemojuvelin (HJV) for use in treating anemia associated with chronic kidney disease in a subject, wherein the antibody comprises a VH having the amino acid sequence of SEQ ID NO: 38 and a VL having the amino acid sequence of SEQ ID NO:

39.

34. 34. The pharmaceutical composition of claim 13 or 33, wherein the chronic kidney disease is refractory to treatment with intravenous iron or blood transfusions.

35. 34. The pharmaceutical composition of claim 13 or 33, wherein the chronic kidney disease is refractory to treatment with oral iron.

36. 34. The pharmaceutical composition of claim 1 or 33, wherein the subject is identified as having functional iron deficiency.

37. 34. The pharmaceutical composition of claim 1 or 33, wherein the subject is identified as exhibiting inflammation and / or iron-restricted erythropoiesis.

38. 34. The pharmaceutical composition of claim 1 or 33, wherein the subject has anemia associated with a secondary cause.

39. 34. The pharmaceutical composition of claim 1 or 33, wherein the subject is a mammal.

40. 34. The pharmaceutical composition of claim 1 or 33, wherein the subject is a human.

41. 34. The pharmaceutical composition of claim 1 or 33, wherein the anemia is not caused by nutritional iron deficiency, iron deficiency due to blood loss, intrinsic red blood cell disorder, hemolysis, inflammation, or functional iron deficiency.

42. 34. The pharmaceutical composition of claim 1 or 33, wherein the subject has previously undergone treatment with intravenous iron or red blood cell transfusion.

43. 34. The pharmaceutical composition of claim 1 or 33, wherein the subject has not previously undergone treatment with intravenous iron or red blood cell transfusions.

44. 34. The pharmaceutical composition of claim 1 or 33, wherein the subject has a serum ferritin level greater than 100 μg / L.

45. 34. The pharmaceutical composition of claim 1 or 33, wherein the subject has a reticulocyte hemoglobin content of less than 26 pg / cell.

46. 34. The pharmaceutical composition of claim 1 or 33, wherein the subject has a transferrin saturation (TSAT) level of less than 30%, 40%, or 50%.

47. 34. The pharmaceutical composition of claim 1 or 33, wherein the subject has a liver iron level greater than 2000 μg / g dry weight.

48. 34. The pharmaceutical composition of claim 1 or 33, wherein the subject has a serum iron level in the range of less than 50 μg / dL.

49. 34. The pharmaceutical composition of claim 1 or 33, wherein the subject has a total iron binding capacity in the range of less than 400 μg / dL.

50. 34. The pharmaceutical composition of claim 1 or 33, wherein the subject has elevated hepcidin levels compared to a subject without anemia.

51. 34. The pharmaceutical composition of claim 1 or 33, wherein the subject has a hepcidin level in the range of greater than 55 ng / ml.

52. 34. The pharmaceutical composition of claim 1 or 33, wherein the subject has endogenous erythropoietin (EPO) levels that are decreased compared to a suitable control subject.

53. 53. The pharmaceutical composition of claim 52, wherein a suitable control subject is a healthy subject who does not have anemia and / or does not have renal impairment.

54. 34. The pharmaceutical composition of claim 33, wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 61 and a light chain comprising the amino acid sequence of SEQ ID NO:

62.

55. 34. The pharmaceutical composition of claim 33, wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 63 and a light chain comprising the amino acid sequence of SEQ ID NO:

62.

56. 34. The pharmaceutical composition of claim 33, wherein the chronic kidney disease is non-dialysis-dependent chronic kidney disease.

57. 34. The pharmaceutical composition of claim 33, wherein the chronic kidney disease is associated with one or more of the following comorbidities: diabetes, hypertension, an autoimmune disease, such as lupus or other autoimmune conditions, congenital heart defects, complications related to the administration of one or more nonsteroidal anti-inflammatory drugs, rare diseases, and birth defects.

58. 34. The pharmaceutical composition of claim 33, wherein the chronic kidney disease is classified as being at a stage within the range of stages 1 to 4.

59. 34. The pharmaceutical composition of claim 33, wherein the chronic kidney disease is classified as being at a stage within the range of stages 2 to 4.

60. The pharmaceutical composition of claim 33, for administration by subcutaneous, intravenous, or intramuscular injection.

61. The pharmaceutical composition of claim 33, for administration by subcutaneous injection.

62. 62. The pharmaceutical composition of claim 61, for subcutaneous self-administration.

63. The pharmaceutical composition of claim 33, comprising the antibody at an average dose of about 1-10 mg / day, 10-20 mg / day, 20-30 mg / day, 30-45 mg / day, 45-60 mg / day, 60-75 mg / day, or 75-100 mg / day.

64. The pharmaceutical composition of claim 1 or 33, comprising the antibody in an amount of 0.1 to 10.0 mg per kg of subject.

65. The pharmaceutical composition of claim 1 or 33, comprising the antibody in an amount of 0.1 to 0.6 mg per kg of subject.

66. The pharmaceutical composition of claim 1 or 33, for administration to a subject once per month or once every three months.

67. A pharmaceutical composition described in claim 1 or 33, administered together with one or more additional therapeutic agents.

68. The pharmaceutical composition of claim 67, wherein the one or more additional therapeutic agents are selected from a growth differentiation factor (GDF) trap, an erythropoiesis stimulating agent (ESA), oral iron, intravenous iron, hypoxia-inducible factor prolyl hydroxylase inhibitor (HIF-PHI), or red blood cell transfusion.

69. 69. The pharmaceutical composition of claim 68, wherein the GDF trap is sotatercept or laspatercept.

70. 69. The pharmaceutical composition of claim 68, wherein the ESA is erythropoietin (EPO).

71. The pharmaceutical composition of claim 67, wherein the one or more additional therapeutic agents is oral iron.

72. The pharmaceutical composition of claim 67, wherein the one or more additional therapeutic agents is HIF-PHI.

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