Anti-hemojuvelin (HJV) antibodies for the treatment of myelofibrosis

Abstract

Aspects of the present application provide anti-hemojuvelin antibodies and methods of using same in the treatment of myelofibrosis and / or conditions associated with myelofibrosis.

Description

[Technical Field]

[0001] Related applications This application claims the benefits of U.S. Provisional Application No. 63 / 024,416, filed on 13 May 2020, titled "Anti-hemoduvelin (HJV) Antibody for Treating Myelofibrosis," No. 63 / 035,649, filed on 5 June 2020, titled "Anti-hemoduvelin (HJV) Antibody for Treating Myelofibrosis," No. 63 / 047,828, filed on 2 July 2020, and No. 63 / 164,294, filed on 22 March 2021, titled "Anti-hemoduvelin (HJV) Antibody for Treating Myelofibrosis," which are incorporated herein by reference in their entirety under 35 U.S. SC § 119(e). [Background technology]

[0002] Iron is a vital component of oxygen-carrying and storage molecules such as hemoglobin and myoglobin. Iron deficiency leads to anemia, while iron overload leads to tissue damage and fibrosis. Hepcidin is an important peptide hormone regulator of systemic iron homeostasis. It exerts its regulatory function by binding to ferroportin, a transmembrane protein present in hepatocytes, duodenal intestinal cells, macrophages, and adipocytes, which is a cellular iron efflux protein. Hepcidin binding promotes the breakdown of ferroportin, preventing iron efflux from cells and release into the plasma. [Overview of the project] [Problems that the invention aims to solve]

[0003] Aspects of this disclosure provide methods for treating hyperhepcidin disorders such as myelofibrosis, myeloma, Waldenström hypergammaglobulinemia, chronic kidney disease, anemia of chronic disease, or iron-restricted anemia, resulting in functional iron deficiency. In certain embodiments, methods are provided for treating myelofibrosis, which is generally characterized as a myeloproliferative disorder associated with chronic inflammation and progressive myelofibrosis. Anemia is a major clinical challenge in myelofibrosis and is associated with negative outcomes. Such anemia is generally caused by or associated with bone marrow failure, splenomegaly, and / or functional iron deficiency and may contribute to inflammation. Furthermore, in myelofibrosis, pro-inflammatory cytokines that induce hepcidin synthesis, such as IL-6 and oncostatin-M, are typically increased and associated with iron blockade, macrophage iron loading, and myeloproliferation and macrophage activation (see, for example, Figure 11). The resulting increase in hepcidin levels is associated with anemia and negative outcomes.

[0004] A part of this disclosure is beneficially low maximum serum concentration (C max This relates to effective bioavailability achieved by subcutaneous administration of an isolated antibody conjugated to hemoduvelin (HJV) to subjects with myelofibrosis. In some embodiments, subcutaneous administration of an anti-HJV antibody allows for lower maximum concentrations (C) of the anti-HJV antibody compared to intravenous administration of the same antibody. max ) produces equivalent pharmacodynamic effects (e.g., a decrease in circulating hepcidin-25 levels, an increase in TSAT%, and / or an increase in serum iron levels). max This is the maximum (or peak) serum concentration of the drug (e.g., anti-HJV antibody) after administration of the drug / antibody and before administration of the second dose. In some embodiments, low C10 is measured within a short time period after administration of the anti-HJV antibody (e.g., within 12 hours, within 24 hours, etc.). max Achieving this minimizes undesirable increases in the serum iron response and / or the potential for off-target effects of the antibody (e.g., binding to RGMa). In some embodiments, blunting C is achieved by subcutaneous administration of anti-HJV antibody. maxThis avoids an undesirable and sharp increase in the serum iron response. In some embodiments, subcutaneous administration of anti-HJV antibody slows down C max This reduces the off-target effects of antibodies.

[0005] Aspects of the present disclosure also relate to an isolated antibody conjugated to human hemoduvelin (HJV) for use in a method of treating a subject having myelofibrosis, wherein the subject is administered the isolated antibody by subcutaneous injection, and the antibody comprises a heavy chain variable region (VH) containing the amino acid sequence of SEQ ID NO: 38 and a light chain variable region (VL) containing the amino acid sequence of SEQ ID NO: 39.

[0006] Aspects of the present disclosure also relate to an isolated antibody conjugating to human hemoduvelin (HJV) for use in a method for inhibiting HJV activity in subjects having myelofibrosis, wherein the subject is administered the isolated antibody subcutaneously, and the antibody comprises a heavy chain variable region (VH) containing the amino acid sequence of SEQ ID NO: 38 and a light chain variable region (VL) containing the amino acid sequence of SEQ ID NO: 39. [Means for solving the problem]

[0007] In some embodiments, administration of an anti-HJV antibody (e.g., intravenous or subcutaneous injection) reduces circulating hepcidin-25 (e.g., serum or plasma hepcidin-25), which is the active form of hepcidin.

[0008] In some embodiments, the Disclosure provides a method for reducing hepcidin-25 in a subject having myelofibrosis, comprising administering to the subject an effective amount of an isolated antibody conjugated to human hemoduverin (HJV), wherein the antibody comprises a heavy chain variable region (VH) containing the amino acid sequence of SEQ ID NO: 38 and a light chain variable region (VL) containing the amino acid sequence of SEQ ID NO: 39. In some embodiments, the Disclosure provides an isolated antibody conjugated to hemoduverin (HJV) for use in a method for reducing hepcidin-25 in a subject having myelofibrosis, comprising a heavy chain variable region (VH) containing the amino acid sequence of SEQ ID NO: 38 and a light chain variable region (VL) containing the amino acid sequence of SEQ ID NO: 39. In some embodiments, the subject is administered by subcutaneous injection. In some embodiments, the administration reduces 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 after administration. In some embodiments, the administration reduces hepcidin-25 by at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% of hepcidin levels in the subject before administration.

[0009] Certain embodiments of this disclosure relate to the observation that hemoduvelin (HJV) is a regulatory agent of hepcidin synthesis and that a lack of hemoduvelin function may be associated with iron overload. For example, in some embodiments, homozygous HJV knockdown animals are unable to amplify hepcidin synthesis in response to IL-6 and are unable to initiate an effective hypoirremia response to acute inflammation. Thus, in some embodiments, the method provided herein comprises administering an effective amount of a hepcidin antagonist to a subject in need to treat a hyperhepcidin disorder, which may be a hemoduvelin antagonist. In some embodiments, the hemoduvelin antagonist is an anti-hemoduvelin antibody. In some embodiments, the anti-hemoduvelin antibody binds to RGMc as its primary mode of action (compared to RGMa and RGMb). Thus, in some embodiments, the anti-hemoduvelin antibody preferentially binds to RGMc compared to RGMa and / or RGMb. In some embodiments, the anti-hemoduvelin antibody binds to RGMc at concentrations lower than 100 nanomoles (nM) (K D <100nM) equilibrium dissociation constant (K D ) binds to RGMc. However, in some embodiments, the anti-hemoduvelin antibody binds to RGMc with similar affinity to RGMa and / or RGMb.

[0010] In some aspects, the present disclosure provides a method of treating a subject having myelofibrosis. In some embodiments, the method comprises administering to the subject an effective amount of an isolated antibody that binds to human hemodjuvelin (HJV), the antibody having a heavy chain complementarity determining region 1 (HC CDR1) described as X1YGMN (SEQ ID NO: 105) where X1 can be N or Y; a heavy chain complementarity determining region 2 (HC CDR2) described as MIYYDSSX2KHYADSVKG (SEQ ID NO: 106) where X2 can be E or D; a heavy chain complementarity determining region 3 (HC CDR3) described as GX3TPDX4 (SEQ ID NO: 107) where X3 can be T or S and X4 can be Y, V, or K; and / or a light chain complementarity determining region 1 (LC CDR1) described as 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, 10 RFS (SEQ ID NO: 109) where X9 can be E, D or A, X 10 can be N, S, T, E or H) and a light chain complementarity determining region 2 (LC CDR2) described as X 11 QX 12 TX 13 DPX 14 X 15 (SEQ ID NO: 110) where X 11 can be F or M, X 12 can be V or A, X 13 can be H or Y, X 14 can be M, L or V, X 15 can be T or S) and a light chain complementarity determining region 3 (LC CDR3).

[0011] In some embodiments, the antibody comprises a VH comprising an amino acid sequence that is at least 85% identical to SEQ ID NO: 38 and / or a VL comprising an amino acid sequence that is at least 85% identical to SEQ ID NO: 39. 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.

[0012] In some embodiments, the antibody is a humanized antibody. In some embodiments, the humanized antibody includes humanized VH and / or humanized VL.

[0013] In some embodiments, the antibody is selected from the group consisting of full-length IgG, Fab fragment, F(ab') fragment, F(ab')2 fragment, scFv, and Fv.

[0014] In some embodiments, the antibody is full-length IgG. In some embodiments, the antibody includes a heavy chain constant region of isotype IgG1, IgG2, IgG3, or IgG4. In some embodiments, the antibody further includes the heavy chain constant region described in SEQ ID NOs: 46, 48, 112, or 113. In some embodiments, the antibody further includes the light chain constant region described in SEQ ID NOs: 47. In some embodiments, the antibody includes (i) a heavy chain containing an amino acid sequence at least 85% identical to SEQ ID NOs: 61 or 117, and / or a light chain containing an amino acid sequence at least 85% identical to SEQ ID NOs: 62; or (ii) a heavy chain containing an amino acid sequence at least 85% identical to SEQ ID NOs: 63 or 118, and / or a light chain containing an amino acid sequence at least 85% identical to SEQ ID NOs: 62. In some embodiments, the antibody includes a heavy chain containing the amino acid sequence of SEQ ID NOs: 61 or 117, and a light chain containing the amino acid sequence of SEQ ID NOs: 62. In some embodiments, the antibody includes a heavy chain containing the amino acid sequence of SEQ ID NOs: 63 or 118, and a light chain containing the amino acid sequence of SEQ ID NOs: 62.

[0015] In some embodiments, the subject has myelofibrosis-induced mutations in JAK2, MPL, ASXL1, TET2, NFE2, SH2B3, SF3B1, or CALR. In some embodiments, myelofibrosis is associated with elevated levels of pro-inflammatory cytokines (e.g., IL-6, oncostatin-M) in the subject. In some embodiments, the subject exhibits serum hemoglobin levels lower than 10 g / dL. In some embodiments, the subject exhibits serum hemoglobin levels lower than 8 g / dL. In some embodiments, the subject is transfusion-dependent. In some embodiments, the subject is anemic.

[0016] In some embodiments, the subject has previously received an erythropoietin stimulant, a JAK-STAT inhibitor, a growth factor ligand trap, or an antifibrotic agent.

[0017] In some embodiments, the methods described herein further include administering to a subject one or more of the following: an erythropoietin stimulant, a JAK-STAT inhibitor, a growth factor ligand trap, and an antifibrotic agent. In some embodiments, the erythropoietin stimulant is selected from the group consisting of danazol, prednisone, thalidomide, lenalidomide, and pomalidomide. In some embodiments, the JAK-STAT inhibitor is selected from the group consisting of ruxolitinib, fedratinib, momerotinib, pacritinib, INCB039110, AG490, and PpYLKTK. In some embodiments, the growth factor ligand trap is sotatercept or raspatercept. In some embodiments, the antifibrotic agent is PRM-151.

[0018] In some embodiments, the disclosure also provides isolated antibodies conjugated to human hemoduvelin (HJV) and compositions comprising the same. In some embodiments, the antibody comprises a heavy chain complementarity determining region 1 (HC CDR1) described as X1YGMN (SEQ ID NO: 105) (wherein X1 may be N or Y); and / or a heavy chain complementarity determining region 2 (HC CDR2) described as MIYYDSSX2KHYADSVKG (SEQ ID NO: 106) (wherein X2 may be E or D); and / or a heavy chain complementarity determining region 3 (HC CDR3) described as GX3TPDX4 (SEQ ID NO: 107) (wherein X3 may be T or S, and X4 may be Y, V, or K). In some embodiments, the light chain complementarity determination region 1 (LC CDR1) is described as RSSQSLX5X6SDGX7TFLX8 (sequence number 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); and / or X9VSX 10 RFS(Sequence ID 109)(where X9 can be E, D or A, X 10 Light chain complementarity determination region 2 (LC CDR2) (which can be N, S, T, E, or H); and / or X 11 QX 12 TX 13 DPX 14 X 15 (Sequence code 110) (where X 11 It can be F or M, X 12 It can be V or A, and X 13 It can be H or Y, and X 14 It can be M, L, or V, X 15 Light chain complementarity determination region 3 (LC CDR3), which is described as (which can be T or S).

[0019] In some embodiments, the Disclosure also provides an isolated antibody that binds to human hemoduvelin (HJV), comprising (i) a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 7, a heavy chain complementarity determination region 1 (HC CDR1), a heavy chain complementarity determination region 2 (HC CDR2), and a heavy chain complementarity determination region 3 (HC CDR3); and / or (ii) a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO: 8, a light chain complementarity determination region 1 (LC CDR1), a light chain complementarity determination region 2 (LC CDR2), and a light chain complementarity determination region 3 (LC CDR3). In some embodiments, the antibody comprises HC CDR1 of SEQ ID NO: 1, HC CDR2 of SEQ ID NO: 2, HC CDR3 of SEQ ID NO: 3, and / or LC CDR1 of SEQ ID NO: 4, LC CDR2 of SEQ ID NO: 5, and LC CDR3 of SEQ ID NO: 6. In some embodiments, the antibody comprises VH, which has an amino acid sequence at least 85% identical to SEQ ID NO: 7, and / or VL, which has an amino acid sequence at least 85% identical to SEQ ID NO: 8.

[0020] In some embodiments, the Disclosure provides an isolated antibody that binds to human hemoduvelin (HJV), comprising (i) a heavy chain variable region (VH) containing the amino acid sequence of SEQ ID NO: 7, a heavy chain complementarity determination region 1 (HC CDR1), a heavy chain complementarity determination region 2 (HC CDR2), and a heavy chain complementarity determination region 3 (HC CDR3); and / or (ii) a light chain variable region (VL) containing the amino acid sequence of SEQ ID NO: 30, a light chain complementarity determination region 1 (LC CDR1), a light chain complementarity determination region 2 (LC CDR2), and a light chain complementarity determination region 3 (LC CDR3). In some embodiments, the antibody comprises HC CDR1 of SEQ ID NO: 1, HC CDR2 of SEQ ID NO: 2, HC CDR3 of SEQ ID NO: 3, and / or LC CDR1 of SEQ ID NO: 4, LC CDR2 of SEQ ID NO: 49, and LC CDR3 of SEQ ID NO: 24. In some embodiments, the antibody comprises VH, which has an amino acid sequence at least 85% identical to SEQ ID NO: 7, and / or VL, which has an amino acid sequence at least 85% identical to SEQ ID NO: 30.

[0021] In some embodiments, the Disclosure provides an isolated antibody that binds to hemoduvelin (HJV), comprising (i) a heavy chain variable region (VH) containing the amino acid sequence of SEQ ID NO: 7, a heavy chain complementarity determination region 1 (HC CDR1), a heavy chain complementarity determination region 2 (HC CDR2), and a heavy chain complementarity determination region 3 (HC CDR3); and (ii) a light chain variable region (VL) containing the amino acid sequence of SEQ ID NO: 31, and / or a light chain variable region (VL) containing the amino acid sequence of SEQ ID NO: 31. In some embodiments, the antibody comprises HC CDR1 of SEQ ID NO: 1, HC CDR2 of SEQ ID NO: 2, HC CDR3 of SEQ ID NO: 3, and / or LC CDR1 of SEQ ID NO: 4, LC CDR2 of SEQ ID NO: 18, and LC CDR3 of SEQ ID NO: 25. In some embodiments, the antibody comprises VH, which has an amino acid sequence at least 85% identical to SEQ ID NO: 7, and / or VL, which has an amino acid sequence at least 85% identical to SEQ ID NO: 8.

[0022] In some embodiments, the Disclosure provides an isolated antibody that binds to human hemoduvelin (HJV), comprising (i) a heavy chain complementarity determination region 1 (HC CDR1), a heavy chain complementarity determination region 2 (HC CDR2), and a heavy chain complementarity determination region 3 (HC CDR3) of a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 7; and / or (ii) a light chain complementarity determination region 1 (LC CDR1), a light chain complementarity determination region 2 (LC CDR2), and a light chain complementarity determination region 3 (LC CDR3) of a light chain variable region comprising the amino acid sequence of SEQ ID NO: 32. In some embodiments, the antibody comprises HC CDR1 of SEQ ID NO: 1, HC CDR2 of SEQ ID NO: 2, HC CDR3 of SEQ ID NO: 3, and / or LC CDR1 of SEQ ID NO: 14, LC CDR2 of SEQ ID NO: 19, and LC CDR3 of SEQ ID NO: 25. In some embodiments, the antibody comprises VH, which has an amino acid sequence at least 85% identical to SEQ ID NO: 7, and / or VL, which has an amino acid sequence at least 85% identical to SEQ ID NO: 32.

[0023] In some embodiments, the Disclosure provides an isolated antibody that binds to human hemoduvelin (HJV), comprising (i) a heavy chain complementarity determination region 1 (HC CDR1), a heavy chain complementarity determination region 2 (HC CDR2), and a heavy chain complementarity determination region 3 (HC CDR3) of a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 7; and / or (ii) a light chain complementarity determination region 1 (LC CDR1), a light chain complementarity determination region 2 (LC CDR2), and a light chain complementarity determination region 3 (LC CDR3) of a light chain variable region comprising the amino acid sequence of SEQ ID NO: 33. In some embodiments, the antibody comprises HC CDR1 of SEQ ID NO: 1, HC CDR2 of SEQ ID NO: 2, HC CDR3 of SEQ ID NO: 3, and / or LC CDR1 of SEQ ID NO: 15, LC CDR2 of SEQ ID NO: 20, and LC CDR3 of SEQ ID NO: 26. In some embodiments, the antibody comprises VH having an amino acid sequence at least 85% identical to SEQ ID NO: 7, and / or VL having an amino acid sequence at least 85% identical to SEQ ID NO: 33.

[0024] In some embodiments, the Disclosure provides an isolated antibody that binds to human hemoduvelin (HJV), comprising (i) a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 34, a heavy chain complementarity determination region 1 (HC CDR1), a heavy chain complementarity determination region 2 (HC CDR2), and a heavy chain complementarity determination region 3 (HC CDR3); and / or (ii) a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO: 35, a light chain complementarity determination region 1 (LC CDR1), a light chain complementarity determination region 2 (LC CDR2), and a light chain complementarity determination region 3 (LC CDR3). In some embodiments, the antibody comprises SEQ ID NO: 9, SEQ ID NO: 2, HC CDR2, SEQ ID NO: 3, and / or LC CDR1 of SEQ ID NO: 16, LC CDR2 of SEQ ID NO: 21, and LC CDR3 of SEQ ID NO: 27. In some embodiments, the antibody comprises VH having an amino acid sequence at least 85% identical to SEQ ID NO: 34, and / or VL having an amino acid sequence at least 85% identical to SEQ ID NO: 35.

[0025] In some embodiments, the Disclosure provides an isolated antibody that binds to human hemoduvelin (HJV), comprising (i) a heavy chain variable region (VH) containing the amino acid sequence of SEQ ID NO: 36, comprising heavy chain complementarity determination region 1 (HC CDR1), heavy chain complementarity determination region 2 (HC CDR2), and heavy chain complementarity determination region 3 (HC CDR3); and / or (ii) a light chain variable region (VL) containing the amino acid sequence of SEQ ID NO: 37, comprising light chain complementarity determination region 1 (LC CDR1), light chain complementarity determination region 2 (LC CDR2), and light chain complementarity determination region 3 (LC CDR3). In some embodiments, the antibody comprises HC CDR1 of SEQ ID NO: 1, HC CDR2 of SEQ ID NO: 10, HC CDR3 of SEQ ID NO: 11, and / or LC CDR1 of SEQ ID NO: 17, LC CDR2 of SEQ ID NO: 18, and LC CDR3 of SEQ ID NO: 28. In some embodiments, the antibody comprises VH having an amino acid sequence at least 85% identical to SEQ ID NO: 36, and / or VL having an amino acid sequence at least 85% identical to SEQ ID NO: 37.

[0026] In some embodiments, the Disclosure provides an isolated antibody that binds to human hemoduvelin (HJV), comprising (i) a heavy chain complementarity determination region 1 (HC CDR1), a heavy chain complementarity determination region 2 (HC CDR2), and a heavy chain complementarity determination region 3 (HC CDR3) of a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 38; and / or (ii) a light chain complementarity determination region 1 (LC CDR1), a light chain complementarity determination region 2 (LC CDR2), and a light chain complementarity determination region 3 (LC CDR3) of a light chain variable region comprising the amino acid sequence of SEQ ID NO: 39. In some embodiments, the antibody comprises HC CDR1 of SEQ ID NO: 1, HC CDR2 of SEQ ID NO: 2, HC CDR3 of SEQ ID NO: 3, and / or LC CDR1 of SEQ ID NO: 17, LC CDR2 of SEQ ID NO: 5, and LC CDR3 of SEQ ID NO: 27. In some embodiments, the antibody comprises VH, which has an amino acid sequence at least 85% identical to SEQ ID NO: 38, and / or VL, which has an amino acid sequence at least 85% identical to SEQ ID NO: 39.

[0027] In some embodiments, the Disclosure provides an isolated antibody that binds to human hemoduvelin (HJV), comprising (i) a heavy chain complementarity determination region 1 (HC CDR1), a heavy chain complementarity determination region 2 (HC CDR2), and a heavy chain complementarity determination region 3 (HC CDR3) of a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 38; and / or (ii) a light chain complementarity determination region 1 (LC CDR1), a light chain complementarity determination region 2 (LC CDR2), and a light chain complementarity determination region 3 (LC CDR3) of a light chain variable region comprising the amino acid sequence of SEQ ID NO: 41. In some embodiments, the antibody comprises HC CDR1 of SEQ ID NO: 1, HC CDR2 of SEQ ID NO: 2, HC CDR3 of SEQ ID NO: 3, and / or LC CDR1 of SEQ ID NO: 50, LC CDR2 of SEQ ID NO: 22, and LC CDR3 of SEQ ID NO: 28. In some embodiments, the antibody comprises VH having an amino acid sequence at least 85% identical to SEQ ID NO: 38, and / or VL having an amino acid sequence at least 85% identical to SEQ ID NO: 41.

[0028] In some embodiments, the Disclosure provides an isolated antibody that binds to human hemoduvelin (HJV), comprising (i) a heavy chain variable region (VH) containing the amino acid sequence of SEQ ID NO: 42, a heavy chain complementarity determination region 1 (HC CDR1), a heavy chain complementarity determination region 2 (HC CDR2), and a heavy chain complementarity determination region 3 (HC CDR3); and / or (ii) a light chain variable region (VL) containing the amino acid sequence of SEQ ID NO: 43, a light chain complementarity determination region 1 (LC CDR1), a light chain complementarity determination region 2 (LC CDR2), and a light chain complementarity determination region 3 (LC CDR3). In some embodiments, the antibody comprises HC CDR1 of SEQ ID NO: 1, HC CDR2 of SEQ ID NO: 2, HC CDR3 of SEQ ID NO: 12, and / or LC CDR1 of SEQ ID NO: 15, LC CDR2 of SEQ ID NO: 23, and LC CDR3 of SEQ ID NO: 27. In some embodiments, the antibody comprises VH, which has an amino acid sequence at least 85% identical to SEQ ID NO: 42, and / or VL, which has an amino acid sequence at least 85% identical to SEQ ID NO: 43.

[0029] In some embodiments, the Disclosure provides an isolated antibody that binds to human hemoduvelin (HJV), comprising (i) a heavy chain variable region (VH) containing the amino acid sequence of SEQ ID NO: 44, comprising heavy chain complementarity determination region 1 (HC CDR1), heavy chain complementarity determination region 2 (HC CDR2), and heavy chain complementarity determination region 3 (HC CDR3); and / or (ii) a light chain variable region (VL) containing the amino acid sequence of SEQ ID NO: 45, comprising light chain complementarity determination region 1 (LC CDR1), light chain complementarity determination region 2 (LC CDR2), and light chain complementarity determination region 3 (LC CDR3). In some embodiments, the antibody comprises HC CDR1 of SEQ ID NO: 1, HC CDR2 of SEQ ID NO: 2, HC CDR3 of SEQ ID NO: 13, and / or LC CDR1 of SEQ ID NO: 16, LC CDR2 of SEQ ID NO: 21, and LC CDR3 of SEQ ID NO: 29. In some embodiments, the antibody comprises VH having an amino acid sequence at least 85% identical to SEQ ID NO: 44, and / or VL having an amino acid sequence at least 85% identical to SEQ ID NO: 45.

[0030] In some embodiments, the Disclosure provides an isolated antibody that binds to hemoduvelin (HJV), comprising HC CDR1, HC CDR2, and HC CDR3, which together contain 10 or fewer amino acid variations, preferably 8 or fewer, more preferably 5 or fewer, and more preferably 2 or fewer, compared to any one of the antibodies listed in Table 1; and / or provides an antibody comprising LC CDR1, LC CDR2, and LC CDR3, which together contain 10 or fewer amino acid variations, preferably 8 or fewer, compared to any one of the antibodies listed in Table 1.

[0031] In some embodiments, any antibody provided herein is a humanized antibody. In some embodiments, the humanized antibody includes humanized VH and / or humanized VL. In some embodiments, the antibody is selected from the group consisting of full-length IgG, Fab fragment, F(ab') fragment, F(ab')2 fragment, scFv, and Fv. In some embodiments, the antibody is full-length IgG. In some embodiments, the antibody includes a heavy chain constant region of isotype IgG1, IgG2, IgG3, or IgG4. In some embodiments, the antibody further includes a heavy chain constant region as described in SEQ ID NOs: 46, 48, 112, or 113. In some embodiments, the antibody further includes a light chain constant region as described in SEQ ID NOs: 47. In some embodiments, the antibody comprises: (i) a heavy chain containing at least 85% the same amino acid sequence as SEQ ID NO: 51 or 114, and / or a light chain containing at least 85% the same amino acid sequence as SEQ ID NOs: 52-56; (ii) a heavy chain containing at least 85% the same amino acid sequence as SEQ ID NO: 57 or 115, and / or a light chain containing at least 85% the same amino acid sequence as SEQ ID NO: 58; (iii) a heavy chain containing at least 85% the same amino acid sequence as SEQ ID NO: 59 or 116, and / or a light chain containing at least 85% the same amino acid sequence as SEQ ID NO: 60; and (iv) a heavy chain containing at least 85% the same amino acid sequence as SEQ ID NO: 61 or 117, and / or a light chain containing at least 85% the same amino acid sequence as SEQ ID NO: 62. (v) A light chain containing at least 85% identical amino acid sequences to SEQ ID NO: 63 or 118, and / or a light chain containing at least 85% identical amino acid sequences to SEQ ID NO: 62; (vi) A heavy chain containing at least 85% identical amino acid sequences to SEQ ID NO: 61 or 117, and / or a light chain containing at least 85% identical amino acid sequences to SEQ ID NO: 65; (vii) A heavy chain containing at least 85% identical amino acid sequences to SEQ ID NO: 66 or 119, and / or a light chain containing at least 85% identical amino acid sequences to SEQ ID NO: 67; or (viii) A heavy chain containing at least 85% identical amino acid sequences to SEQ ID NO: 68 or 120, and / or a light chain containing at least 85% identical amino acid sequences to SEQ ID NO: 69.

[0032] In some embodiments, the antibody binds to HJV with an equilibrium dissociation constant (KD) lower than 100 nM. In some embodiments, the antibody binds to HJV with an equilibrium dissociation constant (KD) lower than 1 nM.

[0033] In some embodiments, the antibody is conjugated to a molecular payload. In some embodiments, the molecular payload is a detectable drug, diagnostic agent, or therapeutic agent.

[0034] In some embodiments, the disclosure also provides nucleic acids encoding isolated antibodies described herein.

[0035] In some embodiments, the disclosure also provides a vector comprising a nucleic acid encoding an antibody described herein. In some embodiments, the vector comprises a nucleic acid sequence from among the nucleic acid sequences listed in Table 3.

[0036] In some embodiments, the disclosure also provides host cells comprising nucleic acids encoding and / or vectors comprising isolated antibodies described herein.

[0037] In some embodiments, the disclosure also provides a pharmaceutical composition comprising an anti-HJV antibody described herein and a pharmaceutically acceptable carrier.

[0038] In some embodiments, the Disclosure also provides a method for producing an anti-human hemoduvelin (HJV) antibody, comprising (i) culturing the host cells described in claim 44 under conditions that enable the expression of an antibody that binds to human hemoduvelin (HJV); and (ii) collecting the cultured host cells or culture medium for collection of the antibody that binds to human hemoduvelin (HJV). In some embodiments, the method comprises purifying the antibody that binds to human hemoduvelin (HJV).

[0039] In some embodiments, the Disclosure also provides isolated antibodies conjugated to human hemoduverin (HJV) produced by expressing in host cells (i) a heavy chain encoding nucleic acid sequence at least 90% identical to SEQ ID NO: 92; and / or (ii) a light chain encoding nucleic acid sequence at least 90% identical to SEQ ID NO: 93. In some embodiments, the Disclosure also provides isolated antibodies conjugated to human hemoduverin (HJV) produced by expressing in host cells (i) a heavy chain encoding nucleic acid sequence at least 90% identical to SEQ ID NO: 94; and / or (ii) a light chain encoding nucleic acid sequence at least 90% identical to SEQ ID NO: 93. In some embodiments, the host cells are 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, or SV40-transformed monkey kidney CV1 lineage (COS) cells.

[0040] In some embodiments, the Disclosure provides an isolated antibody for use in a method of treating subjects having myelofibrosis, wherein the antibody comprises a heavy chain variable region (VH) containing the amino acid sequence of SEQ ID NO: 38 and a light chain variable region (VL) containing the amino acid sequence of SEQ ID NO: 39.

[0041] In some embodiments, subjects with myelofibrosis have anemia resulting from hepcidin synthesis induced by pro-inflammatory cytokines, and antibody administration reduces the anemia.

[0042] In some embodiments, the pro-inflammatory cytokine includes interleukin-6 (IL-6).

[0043] The aforementioned and other embodiments, implementations, actions, functions, features, and embodiments of this teaching can be better understood from the following description in conjunction with the accompanying drawings. [Brief explanation of the drawing]

[0044] The accompanying drawings incorporated herein and constituting part of this specification illustrate certain embodiments and, together with the description provided, provide non-limiting examples of certain aspects of the compositions and methods disclosed herein. [Figure 1A] Figures 1A–1G are graphs illustrating the generation and characterization of anti-HJV antibodies. Figure 1A shows a schematic process of anti-hemoduvelin antibody generation, humanization, and affinity maturation. [Figure 1B] Figures 1A-1G are graphs showing the generation and characterization of anti-HJV antibodies. Figures 1B-1G show sensorgrams of antibodies HA, hHA-004, hHA-008, hHA-009, and hHA-011 obtained by BIAcore analysis. [Figure 1C] Figures 1A-1G are graphs showing the generation and characterization of anti-HJV antibodies. Figures 1B-1G show sensorgrams of antibodies HA, hHA-004, hHA-008, hHA-009, and hHA-011 obtained by BIAcore analysis. [Figure 1D] Figures 1A-1G are graphs showing the generation and characterization of anti-HJV antibodies. Figures 1B-1G show sensorgrams of antibodies HA, hHA-004, hHA-008, hHA-009, and hHA-011 obtained by BIAcore analysis. [Figure 1E] Figures 1A-1G are graphs showing the generation and characterization of anti-HJV antibodies. Figures 1B-1G show sensorgrams of antibodies HA, hHA-004, hHA-008, hHA-009, and hHA-011 obtained by BIAcore analysis. [Figure 1F] Figures 1A-1G are graphs showing the generation and characterization of anti-HJV antibodies. Figures 1B-1G show sensorgrams of antibodies HA, hHA-004, hHA-008, hHA-009, and hHA-011 obtained by BIAcore analysis. [Figure 1G] Figures 1A-1G are graphs showing the generation and characterization of anti-HJV antibodies. Figures 1B-1G show sensorgrams of antibodies HA, hHA-004, hHA-008, hHA-009, and hHA-011 obtained by BIAcore analysis. [Figure 2A] Figures 2A-2C are graphs illustrating the BMP reporter gene assay for anti-HJV antibodies. Figure 2A shows the general principle of the HJV BMP reporter assay. [Figure 2B] Figures 2A-2C are graphs showing the BMP reporter gene assay for anti-HJV antibodies. Figure 2B shows the effect of anti-HJV antibodies on inhibiting RGMc BMP signaling. [Figure 2C] Figures 2A-2C are graphs showing the BMP reporter gene assay for anti-HJV antibodies. Figure 2C shows the effect of anti-HJV antibodies on inhibiting RGMa BMP signaling. [Figure 3] Figure 3 is a graph showing the nonspecific binding of anti-HJV antibodies to HEK293 cells. The bars, from left to right, represent 100 μg / ml, 10 μg / ml, and 1 μg / ml for each group. [Figure 4A] Figures 4A to 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] Figures 4A to 4C are schematic diagrams showing the structure and design of hHA-008 and hHA-008-QL. Figure 4B shows the structure of hHA-008-QL. [Figure 4C] Figures 4A-4C are schematic diagrams showing the structure and design of hHA-008 and hHA-008-QL. Figure 4C shows a comparison of the antibody structures between hHA-008 and hHA-008-QL. [Figure 5] Figure 5 is a graph showing the CD4+ T cell response of peripheral blood mononuclear cells (PBMCs) loaded with hHA-008 or hHA-008-QL. [Figure 6A]Figures 6A–6C are graphs showing the PK / PD analysis of hHA-008 in rats and cynomolgus monkeys (Cynomolgus macaque; cyno). Figure 6A shows the maximum effect of hHA-008 as measured by TSAT% that occurred 4–8 days after treatment. [Figure 6B] Figures 6A–6C are graphs showing the PK / PD analysis of hHA-008 in rats and cynomolgus monkeys (Cynomolgus macaque; cyno). Figure 6B shows that hHA-008 reached its maximum effect, as measured by TSAT%, approximately 1–4 days after injection in female cynomolgus monkeys. [Figure 6C] Figures 6A–6C are graphs showing the PK / PD analysis of hHA-008 in rats and cynomolgus macaques (cyno). Figure 6C shows that hHA-008 reached its maximum effect, as measured by TSAT%, approximately 1–4 days after injection in male cynomolgus macaques, although one of the males did not respond to hHA-008 treatment. [Figure 7A] Figures 7A–7F show the PK / PD correlation in cynomolgus monkeys using a single IV dose of 6 mpk. Figure 7A shows that the maximum TSAT% increase occurred 1–4 days after injection (Tmax = 1–4 days), and one of the tested animals had a dramatic decrease in TSAT around day 34. [Figure 7B] Figures 7A-7F show the PK / PD correlation in cynomolgus monkeys using a single IV dose of 6 mpk. Figure 7B shows that plasma hepcidin-25 concentration changes over time after hHA-008 injection. [Figure 7C] Figures 7A-7F show the PK / PD correlation in cynomolgus monkeys using a single IV dose of 6 mpk. Figure 7C shows that plasma hHA-008 concentration changes over time after hHA-008 injection. [Figure 7D]Figures 7A-7F show the PK / PD correlation in cynomolgus monkeys using a single IV dose of 6 mpk. Figures 7D-7F show that hHA-008 had a robust PK / PD correlation with PK (plasma antibody concentration) against TSAT% and plasma hepcidin-25 concentration. The 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 7E] Figures 7A-7F show the PK / PD correlation in cynomolgus monkeys using a single IV dose of 6 mpk. Figures 7D-7F show that hHA-008 had a robust PK / PD correlation with PK (plasma antibody concentration) against TSAT% and plasma hepcidin-25 concentration. The 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 7F] Figures 7A-7F show the PK / PD correlation in cynomolgus monkeys using a single IV dose of 6 mpk. Figures 7D-7F show that hHA-008 had a robust PK / PD correlation with PK (plasma antibody concentration) against TSAT% and plasma hepcidin-25 concentration. The 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 8A] Figures 8A–8C show that the hHA-008 antibody modulates TSAT% in a dose-dependent manner. Figure 8A shows the TSAT% and hHA-008 concentrations after animals were treated with 0 (medium control) or 0.6 mpk of hHA-008. [Figure 8B] Figures 8A–8C show that the hHA-008 antibody modulates TSAT% in a dose-dependent manner. Figure 8B shows the TSAT% and hHA-008 concentrations after animals have been treated with 0 (medium control) or 3 mpk of hHA-008. [Figure 8C] Figures 8A–8C show that the hHA-008 antibody modulates TSAT% in a dose-dependent manner. Figure 8C shows the TSAT% and hHA-008 concentrations after animals were treated with 0 (medium control) or 60 mpk of hHA-008. [Figure 9A] Figures 9A-9C are graphs showing a comparison of PK / PD between hHA-008 and hHA-008-QL. Figure 9A shows the change in TSAT% over time in cynomolgus monkeys after treatment with hHA-008 or hHA-008-QL. [Figure 9B] Figures 9A-9C are graphs showing a comparison of PK / PD between hHA-008 and hHA-008-QL. Figure 9B shows the time course of antibody plasma concentrations in cynomolgus monkeys after treatment with hHA-008 or hHA-008-QL. [Figure 9C] Figures 9A-9C are graphs showing a comparison of PK / PD between hHA-008 and hHA-008-QL. Figure 9C shows the time course of the decrease in plasma concentrations of hHA-008 and hHA-008-QL. [Figure 10A] Figures 10A and 10B are graphs showing the FcRn binding of hHA-008 and hHA-008-QL at pH 6.0 or 7.4. Figure 10A shows the FcRn binding of hHA-008 and hHA-008-QL at pH 6.0. X-axis: Time. Y-axis: Response. [Figure 10B] Figures 10A and 10B are graphs showing the FcRn binding of hHA-008 and hHA-008-QL at pH 6.0 or 7.4. Figure 10B shows the FcRn binding of hHA-008 and hHA-008-QL at pH 7.4. X axis: Time. Y axis: Response. [Figure 11] Figure 1 shows the bone marrow proliferation cycle specific to a particular hyperhepcidin disorder. [Figure 12] Figure 2 shows the hepcidin-stimulating pathway and the physiological regulation of iron homeostasis by hepcidin. [Figure 13] Figure 13 is a graph showing that IL-6 induces hepcidin expression in cynomolgus monkeys, and that hHA-008 treatment prevents the induction of inflammation (IL-6) and the increase in hepcidin-25 in cynomolgus monkeys 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. The interaction occurs at amino acids 170, 171, 180, 182, and 183 on 3720-RG-050. [Figure 15] Figure 15 shows the interaction between 3720-RG-050 and hHA-008. The 3720-RG-050 PDB structure was generated by homology using Swiss Model software. Amino acids 170-183 of 3720-RG-050 [SSPMALGANATATR (SEQ ID NO: 121)] are shown in A, B, C, D, E: front view (A); back view (B), side view 1 (C), side view 2 (D), and top view (E) in ribbon / surface representation. F, G, H, I, J: front view (F); back view (G), side view 1 (H), side view 2 (I), and top view (J) in ribbon representation. [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. The interaction occurs at amino acids 169, 171, 180, 182; 289, 293, 294, 295, 297, and 300 on 3720-RG-050. [Figure 17] Figure 17 shows the interaction 3720-RG-050 / hHA-008-QL. The 3720-RG-050 PDB structure was generated by homology using Swiss Model software. Amino acids 169-182 [TSSPMALGANATAT (SEQ ID NO: 122)] and 289-291 (SQR) of 3720-RG-050 are shown in A, B, C, D, E: front view (A); back view (B), side view 1 (C), side view 2 (D), and top view (E) in ribbon / surface representation. F, G, H, I, J: front view (F); back view (G), side view 1 (H), side view 2 (I), and top view (J) in ribbon representation. [Figure 18]Figure 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] Figure 19 shows that the decrease in the PD response (e.g., hepcidin-25 concentration and TSAT%) was consistent with the decrease in hHA-008 serum concentration after subcutaneous administration of hHA-008 to Sprague Dolly rats (Figure 19). [Figure 20A] Figures 20A–20D show PK / PD analysis in cynomolgus monkeys after subcutaneous administration of hHA-008. Figure 20A shows that the serum concentration-time profile became indistinguishable between SC injection and IV injection four days after administration. [Figure 20B] Figures 20A–20D show PK / PD analysis in cynomolgus monkeys after subcutaneous administration of hHA-008. Figures 20B–20D show that the return of serum iron to baseline levels was consistent with the decrease in hHA-008 serum concentrations after injections of 0.3 mpk, 0.6 mpk, and 1 mpk of hHA-008, either subcutaneously or intravenously. [Figure 20C] Figures 20A–20D show PK / PD analysis in cynomolgus monkeys after subcutaneous administration of hHA-008. Figures 20B–20D show that the return of serum iron to baseline levels was consistent with the decrease in hHA-008 serum concentrations after injections of 0.3 mpk, 0.6 mpk, and 1 mpk of hHA-008, either subcutaneously or intravenously. [Figure 20D] Figures 20A–20D show PK / PD analysis in cynomolgus monkeys after subcutaneous administration of hHA-008. Figures 20B–20D show that the return of serum iron to baseline levels was consistent with the decrease in hHA-008 serum concentrations after injections of 0.3 mpk, 0.6 mpk, and 1 mpk of hHA-008, either subcutaneously or intravenously. [Modes for carrying out the invention]

[0045] This disclosure is based, at least in part, on the development of anti-hemoduvelin (HJV) antibodies, e.g., the antibodies listed in Table 1 and their variants, that exhibit high binding affinity and specificity to human hemoduvelin. The use of anti-HJV antibodies and their variants in research, diagnostic / detection, and therapeutic applications is also provided. In some embodiments, this disclosure provides anti-hemoduvelin (HJV) antibodies for targeting the hepcidin pathway, which is effective in inhibiting hepcidin function and / or reducing hepcidin expression in cells, in order to modulate iron homeostasis, particularly for the treatment of myelofibrosis and / or one or more symptoms or complications thereof. Accordingly, in relevant embodiments, this disclosure provides compositions and methods for treating myelofibrosis, including primary myelofibrosis, myelofibrosis arising from myeloproliferative neoplasms and / or one or more symptoms or complications thereof, such as myelofibrosis-associated anemia, inflammation, bone marrow failure, splenomegaly, catabolism, and / or fatigue.

[0046] In some embodiments, an anti-hemoduvelin antibody is provided that binds to RGMc as its primary mode of action (compared to RGMb and RGMMa). Thus, in some embodiments, the anti-hemoduvelin antibody preferentially binds to RGMc compared to RGMb and / or RGMMa. In some embodiments, the anti-hemoduvelin antibody binds to RGMc with an equilibrium dissociation constant (KD) lower than 100 nanomolar concentration (nM) (KD < 100 nM). However, in some embodiments, the anti-hemoduvelin antibody binds to RGMc with an affinity similar to that of RGMb and / or RGMMa.

[0047] The above and other aspects, embodiments, actions, functions, features and embodiments of this instruction can be better understood from the following description in conjunction with the accompanying drawings.

[0048] I. Definition Administration: As used herein, the term “administering” or “dosing” means providing a complex to a subject in a physiologically and / or pharmacologically useful manner (e.g., treating a condition in a subject).

[0049] Affinity-matured antibody: “Affinity-matured antibody” as used herein refers to an antibody having one or more modifications to one or more CDRs, which results in an improvement in the antibody’s affinity (i.e., KD, kd, or ka) to a target antigen compared to a parent antibody without modifications. Exemplary affinity-matured antibodies may have nanomolar or even picomolar affinity to a target antigen. Various procedures for producing affinity-matured antibodies are known in the art, including screening of combinatory 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 CDRs 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 and selective or hypermutation at constant or hypermutation sites by activity-enhancing amino acid residues is described in U.S. Patent No. 6914128B1.

[0050] Antibody: As used herein, the term “antibody” refers to a polypeptide comprising at least one immunoglobulin variable domain or at least one antigenic determinant, such as 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, an F(ab')2 fragment, an Fv fragment, or an scFv fragment. In some embodiments, the antibody is a nanobody derived from a camel 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 other embodiments, 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 (V as used herein). H (abbreviated as V), and / or light (L) chain variable region (V in this specification). L Includes (abbreviated as ). In some embodiments, the antibody includes a constant domain, e.g., an Fc region. The immunoglobulin constant domain refers to the heavy or light chain constant domain. The amino acid sequences of the human IgG heavy chain and light chain constant domains 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 include a human alpha (α), delta (Δ), epsilon (ε), gamma (γ), or mu (μ) heavy chain. In certain embodiments, the antibody described herein includes the human gamma 1 CH1, CH2, and / or CH3 domains. In some embodiments, V HThe amino acid sequence of the domain includes the amino acid sequence of the human gamma (γ) heavy chain constant region, for example, any known in the art. Non-limiting examples of human constant region sequences are described in the art; see, for example, U.S. Patent No. 5,693,780 and Kabat EA et al., (1991) above. In some embodiments, V HThe domains include 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 conjugated to one or more sugar or carbohydrate molecules. In some embodiments, one or more sugar or carbohydrate molecules are conjugated to the antibody by N-glycosylation, O-glycosylation, C-glycosylation, glyciation (GPI anchoring), and / or phosphoglycosylation. In some embodiments, one or more sugar or carbohydrate molecules are monosaccharides, disaccharides, oligosaccharides, or glycans. In some embodiments, one or more sugar or carbohydrate molecules are branched oligosaccharides or branched glycans. In some embodiments, one or more sugar or carbohydrate molecules include mannose units, glucose units, N-acetylglucosamine units, or phospholipid units. In some embodiments, the antibody is a construct comprising a polypeptide containing one or more antigen-binding fragments of the present disclosure linked to a linker polypeptide or an immunoglobulin constant domain. The linker polypeptide comprises two or more amino acid residues joined by a peptide bond and is used to link one or more antigen-binding moieties. Examples of linker polypeptides have been reported [see, for example, Holliger, P., et al. (1993) Proc. Natl. Acad. Sci. USA 90:6444-6448; Poljak, RJ, et al. (1994) Structure 2:1121-1123]. Furthermore, the antibody may be part of a larger immunoadhesion molecule formed by covalent or noncovalent bonding 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 divalent and biotinylated scFv molecules [Kipriyanov, SM, et al. (1994) Mol. Immunol. 31:1047-1058].

[0051] Approximately: As used herein, the terms “approximately” or “about” mean a value that is similar to the reference value stated, when applied to one or more of the values ​​of interest. In certain embodiments, unless otherwise stated or otherwise evident from the context, the terms “approximately” or “about” mean a range of values ​​that are within 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in any direction of the reference value stated (except where such a number would exceed 100% of the possible value).

[0052] CDR: As used herein, the term “CDR” refers to the complementarity-determining region within the antibody variable sequence. A typical antibody molecule usually contains a heavy chain variable region (VH) and a light chain variable region (VL) that are involved in antigen binding. The VH and VL regions can be further subdivided into hypervariable regions, also known as “complementarity-determining regions” (“CDR”), which are interspersed with more conserved regions known as “framework regions” (“FR”). Each VH and VL typically consists 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, IMGT definition, Chothia definition, AbM definition, and / or 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 Network), the International ImMunoGeneTics Information System(Manufacturing Network) http: / / www.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 may refer to a CDR as defined by any method known in the art. Two antibodies having the same CDR means that the two antibodies have the same amino acid sequence of the CDR, as determined by the same method, e.g., the IMGT definition.

[0053] Typically, there are three CDRs in each variable region of the heavy and light chains, designated as CDR1, CDR2, and CDR3 for each variable region. As used herein, the term “CDR set” refers to a group of three CDRs that occur 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 et al. [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 applicable to any variable region of an antibody, but also provides precise residue boundaries defining three CDRs. These CDRs may be called Kabat CDRs. The sub-parts of the CDRs may be denoted as L1, L2, and L3 or H1, H2, and H3, where "L" and "H" indicate the light chain and heavy chain regions, respectively. These regions may also be called Chothia CDRs and have boundaries that overlap with the Kabat CDRs. Other boundaries defining CDRs that overlap with the Kabat CDRs are described by Padlan [FASEB J. 9:133-139 (1995)] and MacCallum [J Mol Biol 262(5):732-45 (1996)) is described. Further CDR boundary definitions do not have to strictly adhere to one of the systems above, but despite overlapping with Kabat CDRs, they may be shortened or extended to take into account predictions or experimental findings that do not significantly affect antigen binding, such as specific residues, groups of residues, or even the entire CDR. The methods used herein may utilize CDRs defined according to any of these systems, but preferred embodiments use Kabat or Chothia defined CDRs.

[0054] CDR transplant antibody: The term "CDR transplant antibody" refers to an antibody that contains heavy and light chain variable region sequences from a single species, but V H and / or V LThis refers to antibodies in which one or more CDR region sequences are replaced with CDR sequences of another species, for example, antibodies having mouse heavy and light chain variable regions in which one or more mouse CDRs (e.g., CDR3) are replaced with human CDR sequences.

[0055] Chimeric antibody: The term "chimeric antibody" refers to an antibody that contains heavy chain and light chain variable region sequences from one species as well as a constant region sequence from another species, for example, an antibody having mouse heavy chain and light chain variable regions linked to a human constant region.

[0056] Complementarity: As used herein, the term “complementarity” refers to the ability of two nucleotides or two sets of nucleotides to pair precisely. In particular, complementarity is a term that characterizes the degree of hydrogen bond pairs that result in bonding 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), then the bases are considered 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., fluctuation base pairs and Hoogsteen base pairs). For example, in some embodiments, due to 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, is considered complementary by hybridizing with any of A, C, U, or T. Inosine (I) is also considered a universal base in the art and is considered complementary to any of A, C, U, or T.

[0057] Conservative amino acid substitutions: As used herein, “conservative amino acid substitutions” refer to amino acid substitutions that do not alter the relative charge or size characteristics of the protein in which the substitution is made. Variants can be prepared according to methods of modifying polypeptide sequences known to those skilled in the art, for example, references that compile such methods, e.g., 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 amino acids within 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.

[0058] Tolerance Reactivity: As used herein, and in the context of targeted agents (e.g., antibodies), the term “tolerance reactivity” refers to the property of a drug to specifically bind to one or more antigens of similar types or classes (e.g., diverse homologous, paralogous, or orthologous antigens) with similar affinity or binding activity. For example, in some embodiments, an antibody that is tolerance reactive to similar types or classes of human and non-human primate antigens (e.g., human hemoduvelin and non-human primate hemoduvelin) can bind to human and non-human primate antigens with similar affinity or binding activity. In some embodiments, the antibody is cross-reactive to similar types or classes of human and rodent antigens. In some embodiments, the antibody is cross-reactive to similar types or classes of rodent and non-human primate antigens. In some embodiments, the antibody is cross-reactive to similar types or classes of human, non-human primate, and rodent antigens.

[0059] Effective dose: As used herein, “effective dose” refers to the amount of each activator (e.g., anti-HJV antibody) required to produce a therapeutic effect on a subject, either alone or in combination with one or more other activators. In some embodiments, the therapeutic effect is a decrease in hepcidin levels or activity, an increase in transferrin saturation levels (TSAT%), a decrease in circulating transferrin levels, and / or an alleviation of the disease state (e.g., a reduction in anemia or a reduction in the progression of myelofibrosis).

[0060] Framework: As used herein, the term “framework” or “framework sequence” refers to the sequence of the remaining variable region excluding the CDRs. Since the precise definition of a CDR sequence can be determined by different systems, the meaning of a framework sequence is assumed to be correspondingly subject to 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 sub-regions (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. Without specifying a particular sub-region as FR1, FR2, FR3, or FR4, as otherwise mentioned, the framework region represents a combination of FRs within the variable region of a single, spontaneously occurring immunoglobulin chain. As used herein, FR represents one of four subregions, and FRs represents two or more of the four subregions that constitute a framework region. Human heavy and light chain acceptor sequences are known in the art. In one embodiment, acceptor sequences known in the art may be used in the antibodies disclosed herein.

[0061] Hemoduvelin (HJV): As used herein, the term “hemoduvelin (HJV)” (also known as repulsive guidance molecule C (RGMc) or type 2 hemochromatosis protein (HFE2)) refers to a membrane-bound soluble form protein that controls hepcidin production via the BMP / SMAD signaling pathway. The HFE2 gene encodes two known classes of HJV molecules: GPI-anchored and glycosylated, which are targeted to the membrane and undergo different fates. HJV exists in diverse isoforms, including two soluble isoforms and two membrane-bound isoforms. In some embodiments, the dominant membrane-bound isoform is a disulfide-bonded two-strand form consisting of an N-terminal and a C-terminal fragment. In some embodiments, the full-length single-strand isoform binds to the membrane but is released from the cell surface and accumulates in the extracellular fluid. In some embodiments, HJV may be of human (NCBI Gene ID 148738), non-human primate (e.g., NCBI Gene ID 698805), or rodent (e.g., NCBI Gene ID 69585 or NCBI Gene ID 310681) 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 as well as in cardiac and skeletal muscle.

[0062] Hepcidin: As used herein, “hepcidin” refers to an iron-regulating peptide hormone primarily produced in the liver and encoded by the HAMP gene. In some embodiments, hepcidin controls the delivery of iron to the plasma from iron-absorbing intestinal cells, from macrophages that recycle red blood cells, and from iron-storing hepatocytes. In some embodiments, hepcidin inhibits iron transport by binding to ferroportin, an iron export channel located on the basolateral surface of intestinal cells and on the plasma membrane of reticuloendothelial cells (macrophages). In some embodiments, inhibition of ferroportin prevents iron from being exported, and iron is sequestered within the cell. In some embodiments, by inhibiting ferroportin, hepcidin prevents intestinal cells from bringing iron into the hepatic portal system, thereby reducing dietary iron absorption. Hepcidin expression involves multiple embodiments, including, for example, transcription of the HAMP gene, translation of the transcribed mRNA, and post-translational processing of the hepcidin precursor into the bioactive hepcidin-25 peptide [DTHFPICIFCCGCCHRSKCGMCCKT (SEQ ID NO: 129)]. In some embodiments, hepcidin expression is modulated via the hemoduvelin-induced BMP signaling pathway. In some embodiments, hepcidin expression is modulated via the IL-6-JAK-STAT signaling pathway.

[0063] HJV-induced BMP signaling: As used herein, the term “HJV-induced BMP signaling” refers to signaling mediated by hemojuvelin (HJV), a membrane-bound coreceptor of bone morphogenetic protein (BMP) signaling, via BMP receptors. 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, for example, positively regulating 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, the induction of BMP signaling by HJV is independent of neogenin. However, in some embodiments, neogenin facilitates the 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 the iron-dependent activation of Smad signaling. In some embodiments, BMP6 is secreted from sinusoidal endothelial cells of the liver and binds to BMP receptors (BMPRs) on hepatocytes, thereby activating the SMAD signaling cascade. In such embodiments, HJV contributes as a co-receptor for such BMP6, for example, by positively regulating hepcidin levels on hepatocytes. In some embodiments, BMPs transmit 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, the constitutively active type II receptor phosphorylates the type I receptor, which in turn phosphorylates intracellular receptor-activating Smad (R-Smad), primarily Smad1, Smad5, and / or Smad8. In such embodiments, the activated R-Smad forms a complex with a common partner, Smad4, and translocates to the nucleus to regulate gene transcription, e.g., induction of hepcidin expression.

[0064] Hepcidin antagonist: As used herein, “hepcidin antagonist” refers to a drug that reduces hepcidin expression and / or hepcidin activity (directly or indirectly). In some embodiments, a hepcidin antagonist reduces hepcidin-25 levels and / or activity (directly or indirectly). In some embodiments, a hepcidin antagonist inhibits hepcidin-induced ferroportin degradation. Thus, in some embodiments, a hepcidin antagonist indirectly targets hepcidin function through the hepcidin-stimulating pathway, thereby reducing hepcidin expression. In some embodiments, a hepcidin antagonist directly targets hepcidin function, for example, by binding to a hepcidin peptide and capturing free hepcidin, or by binding to ferroportin and inhibiting the 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, as disclosed, for example, 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 respective relevant contents of which are incorporated herein by reference.

[0065] Human Antibodies: As used herein, the term “human antibody” is intended to include antibodies having variable and constant regions derived from human germline immunoglobulin sequences. Human antibodies in this disclosure may include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-directed mutagenesis in vitro or by somatic mutagenesis in vivo), for example, in CDRs, particularly CDR3. However, as used herein, the term “human antibody” is not intended to include antibodies in which germline CDR sequences of another mammalian species, such as mouse, have been transplanted into human framework sequences.

[0066] 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 V H and / or V L This refers to antibodies in which at least a portion of the sequence has been modified to be more "human-like," that is, to more closely resemble the human germline variable sequence. One type of humanized antibody is a CDR-implanted antibody, in which the human CDR sequence has been modified to be more similar to that of a non-human V H and V L The sequence is introduced and replaces the corresponding non-human CDR sequence. In one embodiment, a humanized anti-hemoduvelin antibody and antigen-binding moiety are provided. Such antibodies may be produced by obtaining a mouse anti-hemoduvelin monoclonal antibody using humanization that employs in vitro genetic engineering after classical hybridoma techniques, such as disclosed in the international application PCT2005 / 123126A2 by Kasaian et al.

[0067] Isolated antibody: As used herein, “isolated antibody” is intended to mean an antibody that substantially does not contain other antibodies with different antigenic properties (for example, an isolated antibody that specifically binds to hemoduveline substantially does not contain antibodies that specifically bind to antigens other than hemoduveline). An isolated antibody that specifically binds to hemoduveline may, however, cross-reactive to other antigens such as other repulsive guidance molecule (RGM) proteins (e.g., RGMa and / or RGMb). Furthermore, an isolated antibody may not substantially contain other cellular material and / or chemical substances.

[0068] JAK / STAT signaling: As used herein, the term “JAK-STAT signaling” refers to a cell receptor-mediated signaling pathway that recruits a Janus kinase (JAK), such as Janus kinase 1 (JAK1) or Janus kinase 2 (JAK2), to activate a signaling and transcriptional activator (STAT), such as STAT3. In some embodiments, as described in Maliken, BD, et al., The Hepcidin Circuits Act: Balancing Iron and Inflammation, Hepatology. 2011 May; 53(5): 1764-1766, JAK-STAT signaling involves the binding of the cytokine interleukin-6 (IL-6) to its homologous cell receptor, which then recruits Janus kinase 2 (JAK2) to phosphorylate STAT3. In some embodiments, STAT3 then translocates to the nucleus (after JAK2 activation / phosphorylation). In some embodiments, activated STAT3 then induces hepcidin transcription, for example, by binding to a STAT3-binding motif in the hepcidin promoter region. Thus, in some embodiments, hepcidin expression is induced via JAK-STAT signaling during inflammation induced by IL-6-mediated STAT3 activation.

[0069] Kabat numbering: The terms “Kabat numbering,” “Kabat definition,” and “Kabat labeling” are used interchangeably herein. As recognized in the art, these terms 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 in its antigen-binding moiety [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 is the range of amino acids 31-35 of CDR1, 50-65 of CDR2, and 95-102 of CDR3. Regarding the light chain variable region, the hypervariable region is located in the range of amino acids 24 to 34 of CDR1, 50 to 56 of CDR2, and 89 to 97 of CDR3.

[0070] Myelofibrosis: As used herein, the term “myelofibrosis” refers to a progressive fibrosis characterized by pathological myeloproliferation and abnormal cytokine production resulting in inflammation and / or dysfunction of the bone marrow niche of the subject. Myelofibrosis and related fibrosis often arise from a non-clonal fibroblastic response to inflammatory and fibrogenic cytokines produced by abnormal clonal myeloid cells, such as megakaryocytes. Myelofibrosis typically results in bone marrow failure, splenomegaly, catabolism, and anemia. In some embodiments, myelofibrosis occurs de novo in the subject. In such embodiments, myelofibrosis is considered “primary” myelofibrosis. However, in some embodiments, myelofibrosis arises from a pre-existing myeloproliferative neoplasm. In some embodiments, the pre-existing myeloproliferative neoplasm is polycythemia. In some embodiments, the pre-existing myeloproliferative neoplasm is essential thrombocytopenia.

[0071] Myelofibrosis-associated anemia: As used herein, the term “myelofibrosis-associated anemia” refers to a condition occurring in the context of myelofibrosis or as a complication of myelofibrosis, characterized by a deficiency in the blood’s ability to transport oxygen. In some embodiments, myelofibrosis-associated anemia is a result of a deficiency in red blood cells, a deficiency in hemoglobin, and / or a deficiency in total blood volume. In some embodiments, myelofibrosis-associated anemia is iron deficiency anemia or myeloplasia anemia. In some embodiments, myelofibrosis-associated anemia is further associated with chronic inflammatory disease. In some embodiments, myelofibrosis-associated anemia is associated with high circulating hepcidin levels in the subject. In some embodiments, myelofibrosis-associated anemia is associated with high circulating hepcidin levels induced by pro-inflammatory cytokines (e.g., IL-6) in the subject. Examples of anemia other than myelofibrosis-associated anemia include anemia associated with rheumatoid arthritis, anemia of infection, autoimmune hemolytic anemia, aplastic anemia, hypoplastic anemia, pure red cell aplasia and anemia resulting from renal failure or endocrine disorders, megaloblastic anemia, anemia resulting from heme or globin synthesis deficiencies, anemia caused by structural defects in red blood cells, such as sickle cell anemia and sideroblastic anemia, anemia associated with chronic infections, such as malaria, trypanosomiasis, HIV, hepatitis viruses or other viruses, anemia caused by bone marrow deficiency in the absence of myelofibrosis, and chemotherapy-induced anemia.

[0072] Oligonucleotides: As used herein, the term “oligonucleotide” refers to oligomeric nucleic acid compounds up to 200 nucleotides in length. Examples of oligonucleotides include, but are not limited to, RNAi oligonucleotides (e.g., siRNA, shRNA), microRNAs, gapmers, mixedmers, phosphorodiamidate morpholinos, peptide nucleic acids, aptamers, and guide nucleic acids (e.g., Cas9 guide RNA). Oligonucleotides may be single-stranded or double-stranded. In some embodiments, oligonucleotides may contain one or more modified nucleotides (e.g., 2'-O-methylglycosulfate, purine, or pyrimidine modification). In some embodiments, oligonucleotides may contain one or more modified internucleotide bonds. In some embodiments, oligonucleotides may contain one or more phosphorothioate bonds, which may have an Rp or Sp stereochemical structure.

[0073] Recombinant Antibodies: As used herein, the term “recombinant human antibodies” includes all human antibodies (as described in detail herein) prepared, expressed, created or isolated by recombinant means, such as antibodies expressed using recombinant expression vectors transfected into host cells, antibodies isolated from recombinant, combinatorial human antibody libraries [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], antibodies isolated from animals (e.g., mice) that are transgenic of human immunoglobulin genes [e.g., Taylor, LD, et al. (1992) Nucl. Acids [See 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 intended to include antibodies prepared, expressed, created or isolated by 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 if transgenic animals of human Ig sequences are used), and thus the recombinant antibody is V H and V L The amino acid sequence of the region is human germline V H and V LThe sequence is sequence-derived or related, but does not necessarily have to be naturally present in the human antibody germline repertoire in vivo. One embodiment of the present disclosure provides a complete human antibody capable of binding to human hemoduverin, which can be produced using techniques well known in the art, such as using a human Ig phage library, for example, disclosed in international application PCT2005 / 007699A2 by Jermutus et al.

[0074] Selective: As used herein, the terms “selective” or “selectively” refer 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 is distinguishable from the reference molecule in an inhibition assay or other inhibitory environment. For example, with respect to an inhibitor, the term “selectively inhibits” refers to the ability of an inhibitor to inhibit its target molecule to a degree that is distinguishable from a reference molecule that is substantially not inhibited in an inhibition assay, to the extent that it enables selective inhibition of the target molecule, as described herein. For example, the half-number inhibitory concentrations (IC50) of the target molecule and / or reference molecule can be tested by a kinase inhibition assay (e.g., kinase efficacy assay by Carna Biosciences), as described in Asshoff, 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 complete, the signal produced by the enzymatic activity to the substrate can be measured. The half-number inhibitory concentrations of the target molecule and / or 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 antagonistize target molecules. In some embodiments, the molecules described herein selectively neutralize target molecules.

[0075] Specific binding: As used herein, the term “specific binding” refers to the ability of a molecule to bind to a binding partner with a certain degree of affinity or binding strength so that the molecule can be used to distinguish the binding partner from a suitable 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 certain degree of affinity or binding strength so that the antibody can be used to distinguish the particular antigen from others to the extent that it enables selective targeting of certain cells, such as muscle cells, through binding to the antigen, as described herein, for example. In some embodiments, the antibody binds 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 to M or smaller targets D If present, the antibody binds specifically to the target. In some embodiments, the antibody binds specifically to hemoduveline.

[0076] Subject: As used herein, the term “subject” refers to mammals. In some embodiments, the subject is a non-human primate or rodent. In some embodiments, the subject is a human. In some embodiments, the subject is a patient, e.g., a human patient having or suspected of having a disease. In some embodiments, the subject is a human patient having or suspected of having myelofibrosis and / or one or more conditions resulting from myelofibrosis.

[0077] Treatment: As used herein, the terms “treating” or “treatment” refer to the application or administration of a composition comprising one or more active agents (e.g., an anti-HJV antibody) to a subject having a target disease or disorder, symptoms of a disease / disorder, or a predisposition to a disease / disorder, for the purpose of treating, curing, alleviating, reducing, altering, relieving, improving, or influencing a disorder, symptoms of a disease, or 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.

[0078] II. Anti-hemoduvelin (HJV) antibody In some embodiments, the hemoduverin antagonist binds to one or more proteins of the Repulsive Guidance Molecules (RGM) family, including RGMa, RGMb, and RGMc (HJV). In some embodiments, the hemoduverin antagonist binds more selectively to hemoduverin (RGMc) than to RGMa and RGMb. In some embodiments, the hemoduverin antagonist is an antisense oligonucleotide that reduces hemoduverin expression (see, for example, U.S. Patent No. 7,534,764; U.S. Patent Application Publication No. 2014 / 127325; and International Publication No. 2016 / 180784; these are incorporated herein by reference). In some embodiments, the hemoduverin antagonist is a small molecule compound that inhibits hemoduverin, for example, by competitive binding and / or chemical modification of hemoduverin.

[0079] In some embodiments, the hemoduverin antagonist is an antibody (e.g., HA001-HA012) specific to hemoduverin and / or one or more proteins of the RGM protein family (e.g., RGMa, RGMb). Suitable antibodies specific to hemoduverin and / or one or more RGM proteins that may be useful in certain methods provided herein are provided, for example, in U.S. Patent No. 10,118,958; and 8,507,435; U.S. Patent Application Publication 2013 / 330343; U.S. Patent Application Publication 2015 / 166672; and U.S. Patent Application Publication 2017 / 029499; and International Publication 2015 / 171691; and International Publication 2018 / 009624, which are incorporated herein by reference. In some embodiments, antibodies that bind to human hemoduverin (HJV) with high specificity and affinity are provided herein. In some embodiments, the anti-HJV antibodies described herein specifically bind to any extracellular epitope of HJV or an 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 an amino acid segment of human or non-human primate HJV.

[0080] In some embodiments, the anti-HJV antibodies described herein specifically bind to epitopes on human HJV. Human HJV is a 426-amino acid protein with a 31-amino acid predicted N-terminal signal peptide and a 45-amino acid C-terminal GPI attachment signal. An exemplary human HJV amino acid sequence is shown in SEQ ID NO: 128: MGEPGQSPSPRSSHGSPPTLSTLTLLLLLCGHAHSQCKILRCNAEYVSSTLSLRGGGSSGALRGGGGGGRGGGVGSGGLCRALRSYALCTRRTARTCRGDLAFHSAVHGIEDLMIQHNCSRQGPTAPPPPRGPALPGAGSGLPAPDPCDYEGRFSRLHGRPPGFLHCASFGDPHVRSFHHHFHTCRVQGAWPLLDNDFLFVQATSSPMALGANATATRKLTIIFKNMQECIDQKVYQAEVDNLPVAFEDGSINGGDRPGGSSLSIQTANPGNHVEIQAAYIGTTIIIRQTAGQLSFSIKVAEDVAMAFSAEQDLQLCVGGCPPSQRLSRSERNRRGAITIDTARRLCKEGLPVEDAYFHSCVFDVLISGDPNFTVAAQAALEDARAFLPDLEKLHLFPSDAGVPLSSATLLAPLLSGLFVLWLCIQ (Sequence ID 128).

[0081] In some embodiments, the anti-HJV antibodies described herein may be conjugated to fragments of human HJV. The HJV fragments may be approximately 5 to 425 amino acids, 10 to 400 amino acids, 50 to 350 amino acids, 100 to 300 amino acids, 150 to 250 amino acids, 200 to 300 amino acids, or 75 to 150 amino acids in length. The fragments may contain a consecutive number of amino acids from RGMc. An example of the amino acids in an HJV fragment is shown in SEQ ID NO: 123. This is described as QCKILRCNAEYVSSTLSLRGGGSSGALRGGGGGGRGGGVGSGGLCRALRSYALCTRRTARTCRGDLAFHSAVHGIEDLMIQHNCSRQGPTAPPPPRGPALPGAGSGLPAPDPCDYEGRFSRLHGRPPGFLHCASFGDPHVRSFHHHFHTCRVQGAWPLLDNDFLFVQATSSPMALGANATATRKLTIIFKNMQECIDQKVYQAEVDNLPVAFEDGSINGGDRPGGSSLSIQTANPGNHVEIQAAYIGTTIIIRQTAGQLSFSIKVAEDVAMAFSAEQDLQLCVGGCPPSQRLSRSERNRRGAITIDTARRLCKEGLPVEDAYFHSCVFDVLISGDPNFTVAAQAALEDARAFLPDLEKLHLFPSD (Sequence ID 123).

[0082] In some embodiments, the anti-HJV antibodies described herein bind to different epitopes within human HJV or human HJV fragments.

[0083] In some embodiments, the anti-HJV antibody interacts with the epitope within amino acids 160-190 of SEQ ID NO: 123. In some embodiments, the anti-HJV antibody interacts with the epitope having the amino acid sequence of amino acids 170-183 of SEQ ID NO: 123. In some embodiments, the anti-HJV antibody interacts with the epitope having the 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(R) 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.

[0084] In some embodiments, the anti-HJV antibody interacts with epitopes in amino acids 160-190 and / or 280-310 of SEQ ID NO: 123. In some embodiments, the anti-HJV antibody interacts with epitopes in amino acids 169-182 and / or 289-300 of SEQ ID NO: 123. In some embodiments, the anti-HJV antibody interacts with epitopes in amino acids 169-182 and 289-300 of SEQ ID NO: 123. In some embodiments, the anti-HJV antibody interacts with epitopes 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, 171-180, and 180-182 of SEQ ID NO: 123, as well as amino acids 289-293, 293-294, 294-295, 295-297, and 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.

[0085] In some embodiments, the anti-HJV antibodies described herein are affinity-mature clones. In some embodiments, the anti-HJV antibodies are 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 The anti-HJV antibodies specifically bind to HJV (e.g., human or non-human primate HJV) along with (indicated by ). For example, the anti-HJV antibodies of this disclosure can bind to hemoduverin protein (e.g., human hemoduverin) with affinities of 5 pM to 500 nM, e.g., 50 pM to 100 nM, e.g., 500 pM to 50 nM. This disclosure also includes antibodies that compete with any of the antibodies described herein for binding to hemoduverin protein (e.g., human hemoduverin) and have affinities of 100 nM or lower (e.g., 80 nM or lower, 50 nM or lower, 20 nM or lower, 10 nM or lower, 500 pM or lower, 50 pM or lower, or 5 pM or lower). The affinity and binding kinetics of anti-HJV antibodies can be tested using any preferred method, including but not limited to biosensor technologies (e.g., OCTET or BIACORE). In some embodiments, the anti-HJV antibody described herein is K in the range of less than nanomolar concentrations. D It binds to HJV together with [another antibody]. In some embodiments, the anti-HJV antibodies described herein selectively bind to RGMc but not selectively bind to RGMa or RGMb.

[0086] 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 fluorescence assays). 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 P2O] 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 the bound protein as a function of the target protein concentration. The concentration of the bound protein ([bound]) is generally given by the following formula: [Bound]=[Free] / (Kd+[Free]) This relates to the concentration of free target protein ([free]).

[0087] K A It is not always necessary to make an accurate determination of K; in some cases, it is sufficient to obtain a quantitative indicator of affinity, which can be determined using methods such as ELISA or FACS analysis. A Because it is proportional to the activity, it can be used for comparison, for example, to determine whether the affinity is higher, for example, twice as high, and a qualitative indicator of affinity can be obtained, or an estimate of affinity can be obtained, for example, by functional assays, for example, activity in in vitro or in vivo assays.

[0088] Table 1 provides heavy chain (HC) and light chain (LC) sequences, heavy chain variable domain (VH) and light chain variable domain (VL), CDR sequence, and heavy chain and light chain constant region sequences for non-limiting examples of anti-HJV antibodies.

[0089] [Table 1-1] [Table 1-2] [Table 1-3] Table 1-4 Table 1-5 Table 1-6 Table 1-7 Table 1-8 Table 1-9 Table 1-10 Table 1-11 Table 1-12

[0090] 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 be spontaneously cyclized to pyroglutamic acid by post-translational modification. 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, anti-HJV antibodies having N-terminal pyroglutamic acid are impurities in the 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 some embodiments, the 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.

[0091] In some embodiments, the anti-HJV antibody of this disclosure comprises one or more 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, the anti-HJV antibody of this disclosure comprises HC CDR1, HC CDR2, and HC CDR3 provided for any one of the antibodies selected from Table 1. In some embodiments, the anti-HJV antibody of this disclosure comprises one or more 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, the anti-HJV antibody of this disclosure comprises LC CDR1, LC CDR2, and LC CDR3 provided for any one of the anti-HJV antibodies selected from Table 1.

[0092] In some embodiments, the anti-HJV antibody of this 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 chain and light chain CDR3 domains may play a particularly important role in the antibody's binding specificity / affinity to the antigen. Therefore, the anti-HJV antibody of this disclosure may comprise at least the heavy chain and / or light chain CDR3 of any one of the anti-HJV antibodies selected from Table 1.

[0093] In some embodiments, the isolated anti-HJV antibody includes a heavy chain variable region comprising heavy chain CDR1 (HC CDR1), heavy chain CDR2 (HC CDR2), and heavy chain CDR3 (HC CDR3).

[0094] In some embodiments, by Kabat definition, HC CDR1 may include the amino acid sequence X1YGMN (SEQ ID NO: 105) (where X1 may be N or Y). Alternatively or additionally, HC CDR2 may include the amino acid sequence MIYYDSSX2KHYADSVKG (SEQ ID NO: 106) (where X2 may be E or D). Alternatively or additionally, HC CDR3 may include the amino acid sequence GX3TPDX4 (SEQ ID NO: 107) (where X3 may be T or S, and X4 may be Y, V, or K).

[0095] In some embodiments, by Kabat definition, the anti-HJV antibody may include a light chain variable region comprising light chain CDR1 (LC CDR1), light chain CDR2 (LC CDR2), and light chain CDR3 (LC CDR3). In some embodiments, LC CDR1 may include the amino acid sequence 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, LC CDR2 may include X9VSX 10 RFS(Sequence ID 109)(where X9 can be E, D or A, X 10It may also contain an amino acid sequence of (which may be N, S, T, E, or H). Alternatively or additionally, LC CDR3 may be X 11 QX 12 TX 13 DPX 14 X 15 (Sequence code 110) (where X 11 It can be F or M, X 12 can be V or A, and X 13 can be H or Y, and X 14 It can be M, L, or V, X 15 It may also contain an amino acid sequence of (which can be T or S).

[0096] Also within the scope of this disclosure are any exemplary anti-HJV antibody functional variants disclosed herein. Functional variants retain 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, while exhibiting V (Voltage) compared to the reference antibody. H and / or V L It may contain one or more amino acid residue variations in one or more medium or HC CDRs and / or one or more LC CDRs.

[0097] In some embodiments, any anti-HJV antibody of this 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 positions of one or more CDRs in 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 the antibodies described herein may vary by one, two, three, four, five, or six amino acid positions, as long as immunospecific binding to hemoduvelin (e.g., human hemoduvelin) is maintained (e.g., substantially maintained at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% of the binding of the original antibody from which it originates). For example, in some embodiments, the position defining the CDR of any antibody described herein may be altered by shifting the N-terminal and / or C-terminal boundary of the CDR by one, two, three, four, five, or six amino acids compared to any one CDR position of any antibody described herein, as long as immunospecific binding to hemoduvelin (e.g., human hemoduvelin) is maintained (e.g., substantially maintaining at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% of the binding of the original antibody from which it originates). In another embodiment, the length of one or more CDRs in 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 the antibodies described herein may vary by one, two, three, four, five, or more amino acids (e.g., short or long), as long as immunospecific binding to hemoduvelin (e.g., human hemoduvelin) is maintained (e.g., substantially maintained at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% of the binding of the original antibody from which it originates).

[0098] Therefore, in some embodiments, the HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and / or LC CDR3 described herein may have one, two, three, four, five, or more amino acids shorter than one or more of the CDRs described herein (e.g., CDRs from any of the anti-HJV antibodies selected from Table 1), as long as immunospecific binding to hemoduvelin (e.g., human hemoduvelin) is maintained (e.g., substantially maintained by at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% compared to binding to 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 have one, two, three, four, five, or more amino acids longer than one or more of the CDRs described herein (e.g., CDRs from any of the anti-HJV antibodies selected from Table 1), provided that immunospecific binding to hemoduvelin (e.g., human hemoduvelin) is maintained (e.g., substantially maintained by at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% compared to the binding of the original antibody from which it is derived). In some embodiments, the amino portions of HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and / or LC CDR3 described herein may have one, two, three, four, five, or more amino acids extended compared to one or more CDRs described herein (e.g., CDRs from any of the anti-HJV antibodies selected from Table 1), provided that immunospecific binding to hemoduvelin (e.g., human hemoduvelin) is maintained (e.g., substantially maintained by at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% compared to binding to the original antibody from which it is derived).In some embodiments, the carboxyl portions of HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and / or LC CDR3 described herein may be elongated by one, two, three, four, five, or more amino acids compared to one or more CDRs described herein (e.g., CDRs from any of the anti-HJV antibodies selected from Table 1), provided that immunospecific binding to hemoduvelin (e.g., human hemoduvelin) is maintained (e.g., substantially maintained by at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% compared to binding to the original antibody from which it is derived). In some embodiments, the amino portions of HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and / or LC CDR3 described herein may be shortened by one, two, three, four, five, or more amino acids compared to one or more CDRs described herein (e.g., CDRs from any of the anti-HJV antibodies selected from Table 1), provided that immunospecific binding to hemoduvelin (e.g., human hemoduvelin) is maintained (e.g., substantially maintained by at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% compared to binding to the original antibody from which it is derived). In some embodiments, the carboxyl portions of HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and / or LC CDR3 described herein may be shortened by one, two, three, four, five, or more amino acids compared to one or more CDRs described herein (e.g., CDRs from any of the anti-HJV antibodies selected from Table 1), provided that immunospecific binding to hemoduvelin (e.g., human hemoduvelin) is maintained (e.g., substantially maintained by at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% compared to binding to the original antibody from which it is derived).Any method can be used, for example, using the binding assays and conditions described in the Art, to determine whether immunospecific binding to hemoduvelin (e.g., human hemoduvelin) is maintained.

[0099] In some examples, any anti-HJV antibody of this disclosure has one or more CDR sequences (e.g., HC CDR or LC CDR) substantially similar to any one of the anti-HJV antibodies selected from Table 1. For example, an antibody may contain one or more CDR sequences from any anti-HJV antibody selected from Table 1, containing up to five, four, three, two, or one amino acid residue variation 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), provided that immunospecific binding to hemoduverin (e.g., human hemoduverin) is maintained (e.g., substantially maintained by at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or 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 CDR provided herein may be a conserved variation. Conservation variations can be introduced into the CDR at positions where the residues would not be involved in interaction with hemoduvelin proteins (e.g., human hemoduvelin protein) if determined, for example, based on the crystal structure. Some aspects of this 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 provided herein (e.g., HC CDR1, HC CDR2, and HC CDR3), for example, any CDR-H sequence provided for any one of the anti-HJVs selected from Table 1. In some embodiments, any VL domain provided herein comprises one or more CDR-L sequences provided herein (e.g., LC CDR1, LC CDR2, and LC CDR3), for example, any LC CDR sequence provided for any one of the anti-HJV antibodies selected from Table 1.

[0100] In some embodiments, the anti-HJV antibody of this disclosure comprises any antibody comprising the heavy chain variable domain and / or light chain variable domain of any one anti-HJV antibody selected from Table 1, as well as any variants thereof. In some embodiments, the anti-HJV antibody of this disclosure comprises any antibody comprising the heavy chain variable and light chain variable pairs of any anti-HJV antibody selected from Table 1.

[0101] Aspects of this 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 includes 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 sequences and / or light-chain variable sequences of any anti-HJV antibody selected from Table 1. In some embodiments, the homologous heavy-chain variable and / or light-chain variable amino acid sequences are not different within any of the CDR sequences provided herein. For example, in some embodiments, the 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 other than any of the CDR sequences provided herein. In some embodiments, any anti-HJV antibody provided herein includes a heavy chain variable sequence and a light chain variable sequence that include a framework sequence which is at least 75%, 80%, 85%, 90%, 95%, 98%, or 99% identical to the framework sequence of any anti-HJV antibody selected from Table 1.

[0102] In some embodiments, the anti-HJV antibody of the Disclosure is a humanized antibody (e.g., a humanized variant containing one or more CDRs from Table 1). In some embodiments, the anti-HJV antibody of the Disclosure contains the same HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and LC CDR3 as shown in Table 1, and also contains a humanized heavy chain variable region and / or a humanized light chain variable region.

[0103] A humanized antibody is a human immunoglobulin (recipient antibody) in which residues from the recipient's complementarity-determining region (CDR) are replaced with residues from a non-human species (donor antibody), such as mouse, rat, or rabbit, that possess the desired specificity, affinity, and capabilities. In some embodiments, Fv framework region (FR) residues of the human immunoglobulin are replaced with corresponding non-human residues. Furthermore, the humanized antibody may also include residues that are not found in the recipient antibody or in the transferred CDR or framework sequence, but are included to further refine and optimize antibody performance. Generally, a humanized antibody includes at least one, and typically two, substantially all, variable domains, where all or substantially all of the CDR region corresponds to that of a non-human immunoglobulin, and all or substantially all of the FR region corresponds to that of the human immunoglobulin consensus sequence. The humanized antibody also optimally and typically includes at least a portion of the immunoglobulin constant region or domain (Fc) of the human immunoglobulin. The antibody may have a modified Fc region as described in International Publication No. 99 / 58572. Other forms of the humanized antibody may have one or more CDRs (one, two, three, four, five, or six) that are modified with respect to the original antibody, which are also referred to as one or more CDRs derived from one or more CDRs from the original antibody. The humanized antibody may also undergo affinity maturation.

[0104] In some embodiments, humanization is achieved by transplanting a CDR (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 antibody of this disclosure is a humanized variant comprising one or more amino acid substitutions compared to any one of the VHs listed in Table 1 (e.g., in the VH framework region) and / or one or more amino acid substitutions compared to any one of the VLs listed in Table 1 (e.g., in the VL framework region).

[0105] In some embodiments, the anti-HJV antibody of the Disclosure is a humanized antibody comprising a VH containing 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 any VH of any of the anti-HJV antibodies listed in Table 1. Alternatively or additionally, the anti-HJV antibody of the Disclosure is a humanized antibody comprising a VL containing 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 any one VL of any of the anti-HJV antibodies listed in Table 1.

[0106] In some embodiments, the anti-HJV antibody of the present disclosure comprises heavy chain variable domains HC CDR1, HC CDR2, and HC CDR3 having the amino acid sequence of SEQ ID NO: 7. Alternatively, the anti-HJV antibody of the present disclosure comprises light chain variable domains LC CDR1, LC CDR2, and LC CDR3 having the amino acid sequence of SEQ ID NO: 8.

[0107] In some embodiments, the Kabat definition system defines the anti-HJV antibody of this disclosure as comprising HC CDR1 having the amino acid sequence of SEQ ID NO: 1, HC CDR2 having the amino acid sequence of SEQ ID NO: 2, HC CDR3 having the amino acid sequence of SEQ ID NO: 3, 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.

[0108] In some embodiments, the anti-HJV antibody of the Disclosure comprises HC CDR1, HC CDR2, and HC CDR3, each containing a total of five or fewer amino acid variations (e.g., five, four, three, two, or one or fewer 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. Where used in any part of the Disclosure, "total" 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 Disclosure comprises LC CDR1, LC CDR2, and LC CDR3, each containing a total of five or fewer amino acid variations (e.g., five, four, three, two, or one or fewer 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.

[0109] In some embodiments, the anti-HJV antibody of the Disclosure comprises HC CDR1, HC CDR2, and HC CDR3, which together have at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical amino acids, 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 antibody of the Disclosure comprises LC CDR1, LC CDR2, and LC CDR3, which together have at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical amino acids, 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.

[0110] In some embodiments, the anti-HJV antibody of the present disclosure includes HC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR1 having the amino acid sequence of SEQ ID NO: 1; HC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR2 having the amino acid sequence of SEQ ID NO: 2; and / or HC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR3 having the amino acid sequence of SEQ ID NO: 3. Alternatively or additionally, the anti-HJV antibodies of this disclosure include LC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR1 having the amino acid sequence of SEQ ID NO. 4; LC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR2 having the amino acid sequence of SEQ ID NO. 5; and / or LC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR3 having the amino acid sequence of SEQ ID NO. 6.

[0111] In some embodiments, the anti-HJV antibody of the present disclosure comprises VH, which comprises the amino acid sequence of SEQ ID NO: 7. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises VL, which comprises the amino acid sequence of SEQ ID NO: 8.

[0112] In some embodiments, the anti-HJV antibody of the present disclosure comprises a VH containing 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 described in SEQ ID NO: 7. Alternatively, the anti-HJV antibody of the present disclosure comprises a VL containing 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 described in SEQ ID NO: 8.

[0113] In some embodiments, the anti-HJV antibody of the present disclosure comprises a VH having an amino acid sequence that is at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to the VH described in SEQ ID NO: 7. Alternatively, the anti-HJV antibody of the present disclosure comprises a VL having an amino acid sequence that is at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to the VL described in SEQ ID NO: 8.

[0114] In some embodiments, the anti-HJV antibody of the present disclosure comprises heavy chain variable domains HC CDR1, HC CDR2, and HC CDR3 having the amino acid sequence of SEQ ID NO: 7. Alternatively, the anti-HJV antibody of the present disclosure comprises light chain variable domains LC CDR1, LC CDR2, and LC CDR3 having the amino acid sequence of SEQ ID NO: 30.

[0115] In some embodiments, the Kabat definition system defines the anti-HJV antibody of this disclosure as comprising HC CDR1 having the amino acid sequence of SEQ ID NO: 1, HC CDR2 having the amino acid sequence of SEQ ID NO: 2, HC CDR3 having the amino acid sequence of SEQ ID NO: 3, 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.

[0116] In some embodiments, the anti-HJV antibody of the Disclosure comprises HC CDR1, HC CDR2, and HC CDR3, each containing a total of five or fewer amino acid variations (e.g., five, four, three, two, or one or fewer 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. Where used in any part of the Disclosure, "total" 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 Disclosure comprises LC CDR1, LC CDR2, and LC CDR3, each containing a total of five or fewer amino acid variations (e.g., five, four, three, two, or one or fewer 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.

[0117] In some embodiments, the anti-HJV antibody of the Disclosure comprises HC CDR1, HC CDR2, and HC CDR3, which together have at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical amino acids, 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 antibody of the Disclosure comprises LC CDR1, LC CDR2, and LC CDR3, which together have at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical amino acids, 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.

[0118] In some embodiments, the anti-HJV antibody of the present disclosure includes HC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR1 having the amino acid sequence of SEQ ID NO: 1; HC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR2 having the amino acid sequence of SEQ ID NO: 2; and / or HC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR3 having the amino acid sequence of SEQ ID NO: 3. Alternatively or additionally, the anti-HJV antibodies of this disclosure include LC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR1 having the amino acid sequence of SEQ ID NO. 4; LC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR2 having the amino acid sequence of SEQ ID NO. 49; and / or LC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR3 having the amino acid sequence of SEQ ID NO. 24.

[0119] In some embodiments, the anti-HJV antibody of the present disclosure comprises VH, which comprises the amino acid sequence of SEQ ID NO: 7. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises VL, which comprises the amino acid sequence of SEQ ID NO: 30.

[0120] In some embodiments, the anti-HJV antibody of the present disclosure comprises a VH containing 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 described in SEQ ID NO: 7. Alternatively, the anti-HJV antibody of the present disclosure comprises a VL containing 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 described in SEQ ID NO: 30.

[0121] In some embodiments, the anti-HJV antibody of the present disclosure comprises a VH having an amino acid sequence that is at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to the VH described in SEQ ID NO: 7. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises a VL having an amino acid sequence that is at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to the VL described in SEQ ID NO: 30.

[0122] In some embodiments, the anti-HJV antibody of the present disclosure comprises heavy chain variable domains HC CDR1, HC CDR2, and HC CDR3 having the amino acid sequence of SEQ ID NO: 7. Alternatively, the anti-HJV antibody of the present disclosure comprises light chain variable domains LC CDR1, LC CDR2, and LC CDR3 having the amino acid sequence of SEQ ID NO: 31.

[0123] In some embodiments, the Kabat definition system defines the anti-HJV antibody of this disclosure as comprising HC CDR1 having the amino acid sequence of SEQ ID NO: 1, HC CDR2 having the amino acid sequence of SEQ ID NO: 2, HC CDR3 having the amino acid sequence of SEQ ID NO: 3, 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.

[0124] In some embodiments, the anti-HJV antibody of the Disclosure comprises HC CDR1, HC CDR2, and HC CDR3, each containing a total of five or fewer amino acid variations (e.g., five, four, three, two, or one or fewer 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. Where used in any part of the Disclosure, "total" 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 Disclosure comprises LC CDR1, LC CDR2, and LC CDR3, each containing a total of five or fewer amino acid variations (e.g., five, four, three, two, or one or fewer 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.

[0125] In some embodiments, the anti-HJV antibody of the present disclosure comprises HC CDR1, HC CDR2, and HC CDR3, which together comprise 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, and are at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises LC CDR1, LC CDR2, and LC CDR3, which together comprise 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, and are at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical.

[0126] In some embodiments, the anti-HJV antibody of the present disclosure includes HC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR1 having the amino acid sequence of SEQ ID NO: 1; HC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR2 having the amino acid sequence of SEQ ID NO: 2; and / or HC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR3 having the amino acid sequence of SEQ ID NO: 3. Alternatively or additionally, the anti-HJV antibodies of this disclosure include LC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR1 having the amino acid sequence of SEQ ID NO. 4; LC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR2 having the amino acid sequence of SEQ ID NO. 18; and / or LC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR3 having the amino acid sequence of SEQ ID NO. 25.

[0127] In some embodiments, the anti-HJV antibody of the present disclosure comprises VH, which comprises the amino acid sequence of SEQ ID NO: 7. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises VL, which comprises the amino acid sequence of SEQ ID NO: 31.

[0128] In some embodiments, the anti-HJV antibody of the present disclosure comprises a VH containing 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 described in SEQ ID NO: 7. Alternatively, the anti-HJV antibody of the present disclosure comprises a VL containing 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 described in SEQ ID NO: 31.

[0129] In some embodiments, the anti-HJV antibody of the present disclosure comprises a VH having an amino acid sequence that is at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to the VH described in SEQ ID NO: 7. Alternatively, the anti-HJV antibody of the present disclosure comprises a VL having an amino acid sequence that is at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to the VL described in SEQ ID NO: 31.

[0130] In some embodiments, the anti-HJV antibody of the present disclosure comprises heavy chain variable domains HC CDR1, HC CDR2, and HC CDR3 having the amino acid sequence of SEQ ID NO: 7. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises light chain variable domains LC CDR1, LC CDR2, and LC CDR3 having the amino acid sequence of SEQ ID NO: 32.

[0131] In some embodiments, the Kabat definition system defines the anti-HJV antibody of this disclosure as comprising HC CDR1 having the amino acid sequence of SEQ ID NO: 1, HC CDR2 having the amino acid sequence of SEQ ID NO: 2, HC CDR3 having the amino acid sequence of SEQ ID NO: 3, 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.

[0132] In some embodiments, the anti-HJV antibody of the Disclosure comprises HC CDR1, HC CDR2, and HC CDR3, each containing a total of five or fewer amino acid variations (e.g., five, four, three, two, or one or fewer 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. Where used in any part of the Disclosure, "total" 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 Disclosure comprises LC CDR1, LC CDR2, and LC CDR3, each containing a total of five or fewer amino acid variations (e.g., five, four, three, two, or one or fewer 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.

[0133] In some embodiments, the anti-HJV antibody of the Disclosure comprises HC CDR1, HC CDR2, and HC CDR3, which together have at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical amino acids, 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 antibody of the Disclosure comprises LC CDR1, LC CDR2, and LC CDR3, which together have at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical amino acids, 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.

[0134] In some embodiments, the anti-HJV antibody of the present disclosure includes HC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR1 having the amino acid sequence of SEQ ID NO: 1; HC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR2 having the amino acid sequence of SEQ ID NO: 2; and / or HC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR3 having the amino acid sequence of SEQ ID NO: 3. Alternatively or additionally, the anti-HJV antibodies of this disclosure include LC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR1 having the amino acid sequence of SEQ ID NO. 14; LC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR2 having the amino acid sequence of SEQ ID NO. 19; and / or LC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR3 having the amino acid sequence of SEQ ID NO. 25.

[0135] In some embodiments, the anti-HJV antibody of the present disclosure comprises VH, which comprises the amino acid sequence of SEQ ID NO: 7. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises VL, which comprises the amino acid sequence of SEQ ID NO: 32.

[0136] In some embodiments, the anti-HJV antibody of the present disclosure includes a VH containing 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 described in SEQ ID NO: 7. Alternatively or additionally, the anti-HJV antibody of the present disclosure includes a VL containing 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 described in SEQ ID NO: 32.

[0137] In some embodiments, the anti-HJV antibody of the present disclosure comprises a VH having at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) the same amino acid sequence as the VH described in SEQ ID NO: 7. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises a VL having at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) the same amino acid sequence as the VL described in SEQ ID NO: 32.

[0138] In some embodiments, the anti-HJV antibody of the present disclosure comprises HC CDR1, HC CDR2, and HC CDR3 heavy chain variable domains having the amino acid sequence of SEQ ID NO: 7. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises LC CDR1, LC CDR2, and LC CDR3 light chain variable domains having the amino acid sequence of SEQ ID NO: 33.

[0139] In some embodiments, the anti-HJV antibody of the present disclosure is defined by the Kabat definition system as including HC CDR1 having the amino acid sequence of SEQ ID NO: 1, HC CDR2 having the amino acid sequence of SEQ ID NO: 2, HC CDR3 having the amino acid sequence of SEQ ID NO: 3, 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.

[0140] In some embodiments, the anti-HJV antibody of the Disclosure comprises HC CDR1, HC CDR2, and HC CDR3, each containing a total of five or fewer amino acid variations (e.g., five, four, three, two, or one or fewer 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. Where used in any part of the Disclosure, "total" 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 Disclosure comprises LC CDR1, LC CDR2, and LC CDR3, each containing a total of five or fewer amino acid variations (e.g., five, four, three, two, or one or fewer 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.

[0141] In some embodiments, the anti-HJV antibody of the present disclosure comprises HC CDR1, HC CDR2, and HC CDR3, which together have at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical amino acids, 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 antibody of the present disclosure comprises LC CDR1, LC CDR2, and LC CDR3, which together have at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical amino acids, 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.

[0142] In some embodiments, the anti-HJV antibody of the present disclosure includes HC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR1 having the amino acid sequence of SEQ ID NO: 1; HC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR2 having the amino acid sequence of SEQ ID NO: 2; and / or HC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR3 having the amino acid sequence of SEQ ID NO: 3. Alternatively or additionally, the anti-HJV antibodies of this disclosure include LC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR1 having the amino acid sequence of SEQ ID NO. 15; LC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR2 having the amino acid sequence of SEQ ID NO. 20; and / or LC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR3 having the amino acid sequence of SEQ ID NO. 26.

[0143] In some embodiments, the anti-HJV antibody of the present disclosure comprises VH, which comprises the amino acid sequence of SEQ ID NO: 7. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises VL, which comprises the amino acid sequence of SEQ ID NO: 33.

[0144] In some embodiments, the anti-HJV antibody of the present disclosure includes a VH containing 20 or fewer amino acid variations compared to the VH described in SEQ ID NO: 7 (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). Alternatively or additionally, the anti-HJV antibody of the present disclosure includes a VL containing 20 or fewer amino acid variations compared to the VL described in SEQ ID NO: 33 (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).

[0145] In some embodiments, the anti-HJV antibody of the present disclosure comprises a VH having at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) the same amino acid sequence as the VH described in SEQ ID NO: 7. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises a VL having at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) the same amino acid sequence as the VL described in SEQ ID NO: 33.

[0146] In some embodiments, the anti-HJV antibody of the present disclosure comprises HC CDR1, HC CDR2, and HC CDR3 heavy chain variable domains having the amino acid sequence of SEQ ID NO: 34. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises LC CDR1, LC CDR2, and LC CDR3 light chain variable domains having the amino acid sequence of SEQ ID NO: 35.

[0147] In some embodiments, the Kabat definition system defines the anti-HJV antibody of this disclosure as comprising HC CDR1 having the amino acid sequence of SEQ ID NO: 9, HC CDR2 having the amino acid sequence of SEQ ID NO: 2, HC CDR3 having the amino acid sequence of SEQ ID NO: 3, 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.

[0148] In some embodiments, the anti-HJV antibody of the Disclosure comprises HC CDR1, HC CDR2, and HC CDR3, each containing a total of five or fewer amino acid variations (e.g., five, four, three, two, or one or fewer 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. Where used in any part of the Disclosure, "total" 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 Disclosure comprises LC CDR1, LC CDR2, and LC CDR3, each containing a total of five or fewer amino acid variations (e.g., five, four, three, two, or one or fewer 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.

[0149] In some embodiments, the anti-HJV antibody of the present disclosure comprises HC CDR1, HC CDR2, and HC CDR3, which together have at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical amino acids, 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 antibody of the present disclosure comprises LC CDR1, LC CDR2, and LC CDR3, which together have at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical amino acids, 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.

[0150] In some embodiments, the anti-HJV antibody of the present disclosure includes HC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR1 having the amino acid sequence of SEQ ID NO. 9; HC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR2 having the amino acid sequence of SEQ ID NO. 2; and / or HC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR3 having the amino acid sequence of SEQ ID NO. 3. Alternatively or additionally, the anti-HJV antibodies of this disclosure include LC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR1 having the amino acid sequence of SEQ ID NO. 16; LC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR2 having the amino acid sequence of SEQ ID NO. 21; and / or LC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR3 having the amino acid sequence of SEQ ID NO. 27.

[0151] In some embodiments, the anti-HJV antibody of the present disclosure comprises VH, which comprises the amino acid sequence of SEQ ID NO: 34. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises VL, which comprises the amino acid sequence of SEQ ID NO: 35.

[0152] In some embodiments, the anti-HJV antibody of the Disclosure comprises a VH containing 20 or fewer amino acid variations compared to the VH described in SEQ ID NO: 34 (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). Alternatively or additionally, the anti-HJV antibody of the Disclosure comprises a VL containing 20 or fewer amino acid variations compared to the VL described in SEQ ID NO: 35 (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).

[0153] In some embodiments, the anti-HJV antibody of the present disclosure comprises a VH having at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) the same amino acid sequence as the VH described in SEQ ID NO: 34. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises a VL having at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) the same amino acid sequence as the VL described in SEQ ID NO: 35.

[0154] In some embodiments, the anti-HJV antibody of the present disclosure comprises HC CDR1, HC CDR2, and HC CDR3 heavy chain variable domains having the amino acid sequence of SEQ ID NO: 36. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises LC CDR1, LC CDR2, and LC CDR3 light chain variable domains having the amino acid sequence of SEQ ID NO: 37.

[0155] In some embodiments, the anti-HJV antibody of the present disclosure is defined by the Kabat definition system as comprising HC CDR1 having the amino acid sequence of SEQ ID NO: 1, HC CDR2 having the amino acid sequence of SEQ ID NO: 10, HC CDR3 having the amino acid sequence of SEQ ID NO: 11, 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.

[0156] In some embodiments, the anti-HJV antibody of the Disclosure comprises HC CDR1, HC CDR2, and HC CDR3, each containing a total of five or fewer amino acid variations (e.g., five, four, three, two, or one or fewer 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. Where used in any part of the Disclosure, "total" 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 Disclosure comprises LC CDR1, LC CDR2, and LC CDR3, each containing a total of five or fewer amino acid variations (e.g., five, four, three, two, or one or fewer 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.

[0157] In some embodiments, the anti-HJV antibody of the present disclosure comprises HC CDR1, HC CDR2, and HC CDR3, which together comprise 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, and are at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises LC CDR1, LC CDR2, and LC CDR3, which together comprise 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, and are at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical.

[0158] In some embodiments, the anti-HJV antibody of the present disclosure includes HC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR1 having the amino acid sequence of SEQ ID NO. 1; HC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR2 having the amino acid sequence of SEQ ID NO. 10; and / or HC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR3 having the amino acid sequence of SEQ ID NO. 11. Alternatively or additionally, the anti-HJV antibodies of this disclosure include LC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR1 having the amino acid sequence of SEQ ID NO. 17; LC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR2 having the amino acid sequence of SEQ ID NO. 18; and / or LC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR3 having the amino acid sequence of SEQ ID NO. 28.

[0159] In some embodiments, the anti-HJV antibody of the present disclosure comprises VH, which comprises the amino acid sequence of SEQ ID NO: 36. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises VL, which comprises the amino acid sequence of SEQ ID NO: 37.

[0160] In some embodiments, the anti-HJV antibody of the present disclosure includes a VH containing 20 or fewer amino acid variations compared to the VH described in SEQ ID NO: 36 (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). Alternatively or additionally, the anti-HJV antibody of the present disclosure includes a VL containing 20 or fewer amino acid variations compared to the VL described in SEQ ID NO: 37 (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).

[0161] In some embodiments, the anti-HJV antibody of the present disclosure comprises a VH having at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) the same amino acid sequence as the VH described in SEQ ID NO: 36. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises a VL having at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) the same amino acid sequence as the VL described in SEQ ID NO: 37.

[0162] In some embodiments, the anti-HJV antibody of the present disclosure comprises HC CDR1, HC CDR2, and HC CDR3 heavy chain variable domains having the amino acid sequence of SEQ ID NO: 38. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises LC CDR1, LC CDR2, and LC CDR3 light chain variable domains having the amino acid sequence of SEQ ID NO: 39.

[0163] In some embodiments, the anti-HJV antibody of the present disclosure is defined by the Kabat definition system as comprising HC CDR1 having the amino acid sequence of SEQ ID NO: 1, HC CDR2 having the amino acid sequence of SEQ ID NO: 2, HC CDR3 having the amino acid sequence of SEQ ID NO: 3, 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.

[0164] In some embodiments, the anti-HJV antibody of the Disclosure comprises HC CDR1, HC CDR2, and HC CDR3, each containing a total of five or fewer amino acid variations (e.g., five, four, three, two, or one or fewer 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. Where used in any part of the Disclosure, "total" 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 Disclosure comprises LC CDR1, LC CDR2, and LC CDR3, each containing a total of five or fewer amino acid variations (e.g., five, four, three, two, or one or fewer 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.

[0165] In some embodiments, the anti-HJV antibody of the Disclosure comprises HC CDR1, HC CDR2, and HC CDR3, which together have at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical amino acids, 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 antibody of the Disclosure comprises LC CDR1, LC CDR2, and LC CDR3, which together have at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical amino acids, 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.

[0166] In some embodiments, the anti-HJV antibody of the present disclosure includes HC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR1 having the amino acid sequence of SEQ ID NO: 1; HC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR2 having the amino acid sequence of SEQ ID NO: 2; and / or HC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR3 having the amino acid sequence of SEQ ID NO: 3. Alternatively or additionally, the anti-HJV antibodies of this disclosure include LC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR1 having the amino acid sequence of SEQ ID NO. 17; LC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR2 having the amino acid sequence of SEQ ID NO. 5; and / or LC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR3 having the amino acid sequence of SEQ ID NO. 27.

[0167] In some embodiments, the anti-HJV antibody of the present disclosure comprises VH, which comprises the amino acid sequence of SEQ ID NO: 38. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises VL, which comprises the amino acid sequence of SEQ ID NO: 39.

[0168] In some embodiments, the anti-HJV antibody of the present disclosure includes a VH containing 20 or fewer amino acid variations compared to the VH described in SEQ ID NO: 38 (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). Alternatively or additionally, the anti-HJV antibody of the present disclosure includes a VL containing 20 or fewer amino acid variations compared to the VL described in SEQ ID NO: 39 (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).

[0169] In some embodiments, the anti-HJV antibody of the present disclosure comprises a VH having at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) the same amino acid sequence as the VH described in SEQ ID NO: 38. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises a VL having at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) the same amino acid sequence as the VL described in SEQ ID NO: 39.

[0170] In some embodiments, the anti-HJV antibody of this disclosure comprises HC CDR1, HC CDR2, and HC CDR3 heavy chain variable domains having the amino acid sequence of SEQ ID NO: 38. Alternatively or additionally, the anti-HJV antibody of this disclosure comprises LC CDR1, LC CDR2, and LC CDR3 light chain variable domains having the amino acid sequence of SEQ ID NO: 41.

[0171] In some embodiments, the Kabat definition system defines the anti-HJV antibody of this disclosure as comprising HC CDR1 having the amino acid sequence of SEQ ID NO: 1, HC CDR2 having the amino acid sequence of SEQ ID NO: 2, HC CDR3 having the amino acid sequence of SEQ ID NO: 3, 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.

[0172] In some embodiments, the anti-HJV antibody of the Disclosure comprises HC CDR1, HC CDR2, and HC CDR3, each containing a total of five or fewer amino acid variations (e.g., five, four, three, two, or one or fewer 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. Where used in any part of the Disclosure, "total" 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 Disclosure comprises LC CDR1, LC CDR2, and LC CDR3, each containing a total of five or fewer amino acid variations (e.g., five, four, three, two, or one or fewer 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.

[0173] In some embodiments, the anti-HJV antibody of the present disclosure comprises HC CDR1, HC CDR2, and HC CDR3, which together have at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical amino acids, 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 antibody of the present disclosure comprises LC CDR1, LC CDR2, and LC CDR3, which together have at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical amino acids, 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.

[0174] In some embodiments, the anti-HJV antibody of the present disclosure includes HC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR1 having the amino acid sequence of SEQ ID NO: 1; HC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR2 having the amino acid sequence of SEQ ID NO: 2; and / or HC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR3 having the amino acid sequence of SEQ ID NO: 3. Alternatively or additionally, the anti-HJV antibodies of this disclosure include LC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR1 having the amino acid sequence of SEQ ID NO. 50; LC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR2 having the amino acid sequence of SEQ ID NO. 22; and / or LC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR3 having the amino acid sequence of SEQ ID NO. 28.

[0175] In some embodiments, the anti-HJV antibody of the present disclosure comprises VH, which comprises the amino acid sequence of SEQ ID NO: 38. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises VL, which comprises the amino acid sequence of SEQ ID NO: 41.

[0176] In some embodiments, the anti-HJV antibody of the present disclosure includes a VH containing 20 or fewer amino acid variations compared to the VH described in SEQ ID NO: 38 (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). Alternatively or additionally, the anti-HJV antibody of the present disclosure includes a VL containing 20 or fewer amino acid variations compared to the VL described in SEQ ID NO: 41 (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).

[0177] In some embodiments, the anti-HJV antibody of the present disclosure comprises a VH having at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) the same amino acid sequence as the VH described in SEQ ID NO: 38. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises a VL having at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) the same amino acid sequence as the VL described in SEQ ID NO: 41.

[0178] In some embodiments, the anti-HJV antibody of the present disclosure comprises HC CDR1, HC CDR2, and HC CDR3 heavy chain variable domains having the amino acid sequence of SEQ ID NO: 42. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises LC CDR1, LC CDR2, and LC CDR3 light chain variable domains having the amino acid sequence of SEQ ID NO: 43.

[0179] In some embodiments, the Kabat definition system defines the anti-HJV antibody of this disclosure as comprising HC CDR1 having the amino acid sequence of SEQ ID NO: 1, HC CDR2 having the amino acid sequence of SEQ ID NO: 2, HC CDR3 having the amino acid sequence of SEQ ID NO: 12, 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.

[0180] In some embodiments, the anti-HJV antibody of the Disclosure comprises HC CDR1, HC CDR2, and HC CDR3, each containing a total of five or fewer amino acid variations (e.g., five, four, three, two, or one or fewer 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. Where used in any part of the Disclosure, "total" 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 Disclosure comprises LC CDR1, LC CDR2, and LC CDR3, each containing a total of five or fewer amino acid variations (e.g., five, four, three, two, or one or fewer 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.

[0181] In some embodiments, the anti-HJV antibody of the present disclosure comprises HC CDR1, HC CDR2, and HC CDR3, which together comprise 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, and are at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises LC CDR1, LC CDR2, and LC CDR3, which together comprise 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, and are at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical.

[0182] In some embodiments, the anti-HJV antibody of the present disclosure includes HC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR1 having the amino acid sequence of SEQ ID NO. 1; HC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR2 having the amino acid sequence of SEQ ID NO. 2; and / or HC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR3 having the amino acid sequence of SEQ ID NO. 12. Alternatively or additionally, the anti-HJV antibodies of this disclosure include LC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR1 having the amino acid sequence of SEQ ID NO. 15; LC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR2 having the amino acid sequence of SEQ ID NO. 23; and / or LC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR3 having the amino acid sequence of SEQ ID NO. 27.

[0183] In some embodiments, the anti-HJV antibody of the present disclosure comprises VH, which comprises the amino acid sequence of SEQ ID NO: 42. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises VL, which comprises the amino acid sequence of SEQ ID NO: 43.

[0184] In some embodiments, the anti-HJV antibody of the present disclosure includes a VH containing 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 described in SEQ ID NO: 42. Alternatively or additionally, the anti-HJV antibody of the present disclosure includes a VL containing 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 described in SEQ ID NO: 43.

[0185] In some embodiments, the anti-HJV antibody of the present disclosure comprises a VH having at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) the same amino acid sequence as the VH described in SEQ ID NO: 42. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises a VL having at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) the same amino acid sequence as the VL described in SEQ ID NO: 43.

[0186] In some embodiments, the anti-HJV antibody of the present disclosure comprises HC CDR1, HC CDR2, and HC CDR3 heavy chain variable domains having the amino acid sequence of SEQ ID NO: 44. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises LC CDR1, LC CDR2, and LC CDR3 light chain variable domains having the amino acid sequence of SEQ ID NO: 45.

[0187] In some embodiments, the Kabat definition system defines the anti-HJV antibody of this disclosure as comprising HC CDR1 having the amino acid sequence of SEQ ID NO: 1, HC CDR2 having the amino acid sequence of SEQ ID NO: 2, HC CDR3 having the amino acid sequence of SEQ ID NO: 13, 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.

[0188] In some embodiments, the anti-HJV antibody of the Disclosure comprises HC CDR1, HC CDR2, and HC CDR3, each containing a total of five or fewer amino acid variations (e.g., five, four, three, two, or one or fewer 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. Where used in any part of the Disclosure, "total" 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 Disclosure comprises LC CDR1, LC CDR2, and LC CDR3, each containing a total of five or fewer amino acid variations (e.g., five, four, three, two, or one or fewer 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.

[0189] In some embodiments, the anti-HJV antibody of the present disclosure comprises HC CDR1, HC CDR2, and HC CDR3, which together comprise 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, and are at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises LC CDR1, LC CDR2, and LC CDR3, which together comprise 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, and are at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical.

[0190] In some embodiments, the anti-HJV antibody of the present disclosure includes HC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR1 having the amino acid sequence of SEQ ID NO. 1; HC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR2 having the amino acid sequence of SEQ ID NO. 2; and / or HC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR3 having the amino acid sequence of SEQ ID NO. 13. Alternatively or additionally, the anti-HJV antibodies of this disclosure include LC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR1 having the amino acid sequence of SEQ ID NO. 16; LC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR2 having the amino acid sequence of SEQ ID NO. 21; and / or LC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR3 having the amino acid sequence of SEQ ID NO. 29.

[0191] In some embodiments, the anti-HJV antibody of the present disclosure comprises VH, which comprises the amino acid sequence of SEQ ID NO: 44. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises VL, which comprises the amino acid sequence of SEQ ID NO: 45.

[0192] In some embodiments, the anti-HJV antibodies of the present disclosure include a VH containing 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 containing 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.

[0193] In some embodiments, the anti-HJV antibodies of the present disclosure include a VH having an amino acid sequence that is 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, the anti-HJV antibodies of the present disclosure include a VL having an amino acid sequence that is at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to the VL set forth in SEQ ID NO: 45.

[0194] The CDRs of an antibody can 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 system annotates each amino acid of a given antibody sequence (e.g., a VH or VL sequence) with 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 of ordinary skill in the art can obtain the CDR sequences of the anti-HJV antibodies provided in Table 1 using different numbering systems.

[0195]

Table 2

[0196] In some embodiments, the anti-HJV antibody of this disclosure is a chimeric antibody that may include a heavy chain constant region and a light chain constant region from a human antibody. A chimeric antibody refers to an antibody having 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, both the light chain and heavy chain variable regions mimic the variable region of an antibody derived from one species of mammal (e.g., a non-human mammal, e.g., mouse, rabbit, and rat), and the constant region is homologous to a sequence in an antibody derived from another mammal, e.g., human. In some embodiments, amino acid modifications may be made in the variable region and / or the constant region.

[0197] In some embodiments, the anti-HJV antibodies described herein are chimeric antibodies that may include a heavy chain constant region and a light chain constant region from a human antibody. A chimeric antibody refers to an antibody having 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, both the light chain and heavy chain variable regions mimic the variable region of an antibody derived from one species of mammal (e.g., a non-human mammal, e.g., mouse, rabbit, and rat), and the constant region is homologous to a sequence in an antibody derived from another mammal, e.g., human. In some embodiments, amino acid modifications may be made in the variable region and / or the constant region.

[0198] In some embodiments, the anti-HJV antibody of this disclosure comprises a VL domain and / or VH domain of any one of the anti-HJV antibodies selected from Table 1, and 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 have been described in the Art, see, for example, Kabat EA et al., (1991) cited above. An example of a human IgG1 constant region is given below: ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(Sequence ID 103)

[0199] In some embodiments, the heavy chain of any of the anti-HJV antibodies described herein includes a mutant human IgG1 constant region. For example, the introduction of an LALA mutation in the CH2 domain of human IgG1 (a mutant derived from mAb b12 mutated to replace the lower hinge residues Leu234 Leu235 with Ala234 and Ala235) is known to reduce Fcg receptor binding [Bruhns, P., et al. (2009) and Xu, D. et al. (2000)]. Mutant human IgG1 constant regions are provided below (mutations are in bold and underlined): [ka]

[0200] In some embodiments, the heavy chain of any of the anti-HJV antibodies described herein further comprises mutations in the human IgG1 constant region, e.g., the introduction of T250Q and M248L substitutions. In some embodiments, such substitutions may affect FcRn binding and serum half-life (see International Publication Nos. 2005047307 and 2013063110). Exemplary IgG1 constant regions containing LALA and QL mutations are provided below (mutations are in bold and underlined): [ka]

[0201] In some embodiments, particularly when using Chinese hamster ovary cells (CHO cells), it can be understood that lysine at the C-terminus of the heavy chain is cleaved during antibody production. Therefore, the constant region of human IgG1 in the secreted antibody may be as follows: ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG(Sequence ID 111)

[0202] In some embodiments, mutant human IgG1 containing an LALA mutation in the secreted antibody may be as follows: [ka]

[0203] In some embodiments, mutant human IgG1 including LALA mutations and QL mutations may be as follows: [ka]

[0204] In some embodiments, any light chain of the anti-HJV antibody described herein may further include a CL, which may be any light chain constant region (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, and its sequence is provided below: RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(Sequence ID 47)

[0205] Other antibody heavy and light chain constant regions are well known in the art, and are provided, for example, in the IMGT database (www.imgt.org) or www.vbase2.org / vbstat.php, both of which are incorporated herein by reference.

[0206] In some embodiments, the anti-HJV antibodies described herein include a heavy chain containing a heavy chain constant region that is at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to any of the VHs listed in Table 1 or any variant thereof, and to SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 112, or SEQ ID NO: 113. In some embodiments, the anti-HJV antibodies described herein include a heavy chain containing a heavy chain constant region that has 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 any of the VHs listed in Table 1 or any variant thereof, and to SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 112, or SEQ ID NO: 113. In some embodiments, the anti-HJV antibody described herein comprises a heavy chain containing one of the VHs listed in Table 1 or any variant thereof and the heavy chain constant region described in SEQ ID NO: 46. In some embodiments, the anti-HJV antibody described herein comprises a heavy chain containing one of the VHs listed in Table 1 or any variant thereof and the heavy chain constant region described in SEQ ID NO: 48. In some embodiments, the anti-HJV antibody described herein comprises a heavy chain containing one of the VHs listed in Table 1 or any variant thereof and the heavy chain constant region described in SEQ ID NO: 112. In some embodiments, the anti-HJV antibody described herein comprises a heavy chain containing one of the VHs listed in Table 1 or any variant thereof and the heavy chain constant region described in SEQ ID NO: 113.

[0207] In some embodiments, the anti-HJV antibodies described herein include a light chain comprising any one of the VLs listed in Table 1 or any variant thereof and a light chain constant region that is 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 include 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 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) as compared to SEQ ID NO: 47. In some embodiments, the anti-HJV antibodies described herein include a light chain comprising any one of the VLs listed in Table 1 or any variant thereof and a light chain constant region as set forth in SEQ ID NO: 47.

[0208] Examples of the IgG heavy and light chain amino acid sequences of the anti-HJV antibodies described are provided in Table 1 above.

[0209] In some embodiments, the anti-HJV antibody of this disclosure comprises a heavy chain containing 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 116, 117, 118, 119, or 120 amino acid variations) compared to the heavy chain described in any one of SEQ ID NOs. Alternatively or additionally, the anti-HJV antibodies of this disclosure include a light chain containing 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 described 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 include a heavy chain having at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) the same amino acid sequence as 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 include a light chain having at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) the same amino acid sequence as 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 include a heavy chain containing one of the amino acid sequences 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 include a light chain containing one of the amino acid sequences of SEQ ID NOs. 52, 53, 54, 55, 56, 58, 60, 62, 65, 67, or 69.

[0210] In some embodiments, the anti-HJV antibody of the present disclosure comprises a heavy chain containing 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 described in either one of SEQ ID NOs. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises a light chain containing 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 described in any one of SEQ ID NOs. In some embodiments, the anti-HJV antibody described herein comprises a heavy chain containing an amino acid sequence identical to at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) of either SEQ ID NO: 51 or 114. Alternatively or additionally, the anti-HJV antibody described herein comprises a light chain containing an amino acid sequence identical to at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) of either SEQ ID NO: 52. In some embodiments, the anti-HJV antibody described herein comprises a heavy chain containing an amino acid sequence of either SEQ ID NO: 51 or 114. Alternatively or additionally, the anti-HJV antibody described herein comprises a light chain containing an amino acid sequence of either SEQ ID NO: 52.

[0211] In some embodiments, the anti-HJV antibody of the present disclosure comprises a heavy chain containing 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 described in either one of SEQ ID NOs. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises a light chain containing 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 described in any one of SEQ ID NOs. In some embodiments, the anti-HJV antibody described herein comprises a heavy chain containing an amino acid sequence identical to at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) of either SEQ ID NO: 51 or 114. Alternatively or additionally, the anti-HJV antibody described herein comprises a light chain containing an amino acid sequence identical to at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) of any of SEQ ID NO: 53. In some embodiments, the anti-HJV antibody described herein comprises a heavy chain containing an amino acid sequence of either SEQ ID NO: 51 or 114. Alternatively or additionally, the anti-HJV antibody described herein comprises a light chain containing an amino acid sequence of any of SEQ ID NO: 53.

[0212] In some embodiments, the anti-HJV antibody of the present disclosure comprises a heavy chain containing 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 described in either one of SEQ ID NOs. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises a light chain containing 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 described in any one of SEQ ID NOs. In some embodiments, the anti-HJV antibody described herein comprises a heavy chain containing an amino acid sequence identical to at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) of either SEQ ID NO: 51 or 114. Alternatively or additionally, the anti-HJV antibody described herein comprises a light chain containing an amino acid sequence identical to at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) of any of SEQ ID NO: 54. In some embodiments, the anti-HJV antibody described herein comprises a heavy chain containing an amino acid sequence of either SEQ ID NO: 51 or 114. Alternatively or additionally, the anti-HJV antibody described herein comprises a light chain containing an amino acid sequence of any of SEQ ID NO: 54.

[0213] In some embodiments, the anti-HJV antibody of the present disclosure comprises a heavy chain containing 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 described in either one of SEQ ID NOs. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises a light chain containing 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 described in any one of SEQ ID NOs. In some embodiments, the anti-HJV antibody described herein comprises a heavy chain containing an amino acid sequence identical to at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) of either SEQ ID NO: 51 or 114. Alternatively or additionally, the anti-HJV antibody described herein comprises a light chain containing an amino acid sequence identical to at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) of either SEQ ID NO: 55. In some embodiments, the anti-HJV antibody described herein comprises a heavy chain containing an amino acid sequence of either SEQ ID NO: 51 or 114. Alternatively or additionally, the anti-HJV antibody described herein comprises a light chain containing an amino acid sequence of either SEQ ID NO: 55.

[0214] In some embodiments, the anti-HJV antibody of the present disclosure comprises a heavy chain containing 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 described in either one of SEQ ID NOs. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises a light chain containing 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 described in any one of SEQ ID NOs. In some embodiments, the anti-HJV antibody described herein comprises a heavy chain containing an amino acid sequence identical to at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) of either SEQ ID NO: 51 or 114. Alternatively or additionally, the anti-HJV antibody described herein comprises a light chain containing an amino acid sequence identical to at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) of any of SEQ ID NO: 56. In some embodiments, the anti-HJV antibody described herein comprises a heavy chain containing an amino acid sequence of either SEQ ID NO: 51 or 114. Alternatively or additionally, the anti-HJV antibody described herein comprises a light chain containing an amino acid sequence of any of SEQ ID NO: 56.

[0215] In some embodiments, the anti-HJV antibody of the present disclosure comprises a heavy chain containing 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 described in either one of SEQ ID NOs. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises a light chain containing 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 described in any one of SEQ ID NOs. In some embodiments, the anti-HJV antibody described herein comprises a heavy chain containing an amino acid sequence identical to at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) of either SEQ ID NO: 57 or 115. Alternatively or additionally, the anti-HJV antibody described herein comprises a light chain containing an amino acid sequence identical to at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) of either SEQ ID NO: 58. In some embodiments, the anti-HJV antibody described herein comprises a heavy chain containing an amino acid sequence of either SEQ ID NO: 57 or 115. Alternatively or additionally, the anti-HJV antibody described herein comprises a light chain containing an amino acid sequence of either SEQ ID NO: 58.

[0216] In some embodiments, the anti-HJV antibody of the present disclosure comprises a heavy chain containing 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 described in either one of SEQ ID NOs. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises a light chain containing 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 described in any one of SEQ ID NOs. In some embodiments, the anti-HJV antibody described herein comprises a heavy chain containing an amino acid sequence identical to at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) of either SEQ ID NO: 59 or 116. Alternatively or additionally, the anti-HJV antibody described herein comprises a light chain containing an amino acid sequence identical to at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) of any of SEQ ID NO: 60. In some embodiments, the anti-HJV antibody described herein comprises a heavy chain containing an amino acid sequence of either SEQ ID NO: 59 or 116. Alternatively or additionally, the anti-HJV antibody described herein comprises a light chain containing an amino acid sequence of any of SEQ ID NO: 60.

[0217] In some embodiments, the anti-HJV antibody of the present disclosure comprises a heavy chain containing 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 described in either one of SEQ ID NOs. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises a light chain containing 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 described in either one of SEQ ID NOs. In some embodiments, the anti-HJV antibody described herein comprises a heavy chain containing an amino acid sequence identical to at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) of either SEQ ID NO: 61 or 117. Alternatively or additionally, the anti-HJV antibody described herein comprises a light chain containing an amino acid sequence identical to at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) of either SEQ ID NO: 62. In some embodiments, the anti-HJV antibody described herein comprises a heavy chain containing an amino acid sequence of either SEQ ID NO: 61 or 117. Alternatively or additionally, the anti-HJV antibody described herein comprises a light chain containing an amino acid sequence of either SEQ ID NO: 62.

[0218] In some embodiments, the anti-HJV antibody of the present disclosure comprises a heavy chain containing 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 described in either one of SEQ ID NOs. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises a light chain containing 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 described in any one of SEQ ID NOs. In some embodiments, the anti-HJV antibody described herein comprises a heavy chain containing an amino acid sequence identical to at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) of either SEQ ID NO: 63 or 118. Alternatively or additionally, the anti-HJV antibody described herein comprises a light chain containing an amino acid sequence identical to at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) of either SEQ ID NO: 62. In some embodiments, the anti-HJV antibody described herein comprises a heavy chain containing an amino acid sequence of either SEQ ID NO: 63 or 118. Alternatively or additionally, the anti-HJV antibody described herein comprises a light chain containing an amino acid sequence of either SEQ ID NO: 62.

[0219] In some embodiments, the anti-HJV antibody of the present disclosure comprises a heavy chain containing 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 described in either one of SEQ ID NOs. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises a light chain containing 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 described in any one of SEQ ID NOs. In some embodiments, the anti-HJV antibody described herein comprises a heavy chain containing an amino acid sequence identical to at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) of either SEQ ID NO: 61 or 117. Alternatively or additionally, the anti-HJV antibody described herein comprises a light chain containing an amino acid sequence identical to at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) of either SEQ ID NO: 65. In some embodiments, the anti-HJV antibody described herein comprises a heavy chain containing an amino acid sequence of either SEQ ID NO: 61 or 117. Alternatively or additionally, the anti-HJV antibody described herein comprises a light chain containing an amino acid sequence of either SEQ ID NO: 65.

[0220] In some embodiments, the anti-HJV antibody of the present disclosure comprises a heavy chain containing 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 described in either one of SEQ ID NOs. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises a light chain containing 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 described in either one of SEQ ID NOs. In some embodiments, the anti-HJV antibody described herein comprises a heavy chain containing an amino acid sequence identical to at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) of either SEQ ID NO: 66 or 119. Alternatively or additionally, the anti-HJV antibody described herein comprises a light chain containing an amino acid sequence identical to at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) of any of SEQ ID NO: 67. In some embodiments, the anti-HJV antibody described herein comprises a heavy chain containing an amino acid sequence of either SEQ ID NO: 66 or 119. Alternatively or additionally, the anti-HJV antibody described herein comprises a light chain containing an amino acid sequence of any of SEQ ID NO: 67.

[0221] In some embodiments, the anti-HJV antibody of the present disclosure comprises a heavy chain containing 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 described in either SEQ ID NO: 68 or 120. Alternatively or additionally, the anti-HJV antibody of the present disclosure comprises a light chain containing 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 described in any one of SEQ ID NO: 69. In some embodiments, the anti-HJV antibody described herein comprises a heavy chain containing an amino acid sequence identical to at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) of either SEQ ID NO: 68 or 120. Alternatively or additionally, the anti-HJV antibody described herein comprises a light chain containing an amino acid sequence identical to at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) of any of SEQ ID NO: 69. In some embodiments, the anti-HJV antibody described herein comprises a heavy chain containing an amino acid sequence of either SEQ ID NO: 68 or 120. Alternatively or additionally, the anti-HJV antibody described herein comprises a light chain containing an amino acid sequence of any of SEQ ID NO: 69.

[0222] The anti-HJV antibodies described herein may be in any antibody form, including but not limited to intact (i.e., full-length) antibodies, their antigen-binding fragments (e.g., Fab, F(ab'), F(ab')2, Fv), single-chain antibodies, bispecific antibodies, or nanobodies. 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).

[0223] In some embodiments, conservative mutations may be introduced into the antibody sequence (e.g., CDR or framework sequence) at a position where the residue is unlikely to be involved in interaction with the target antigen (e.g., hemoduvelin), for example, when determined based on the crystal structure. In some embodiments, one, two or more mutations (e.g., amino acid substitutions) are introduced in the Fc region of the anti-HJV antibody described herein [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] to alter one or more functional properties of the antibody, such as serum half-life, complement fixation, Fc receptor binding and / or antigen-dependent cytotoxicity.

[0224] In some embodiments, for example, as described in U.S. Patent No. 5,677,425, one, two, or more mutations (e.g., amino acid substitutions) are introduced into the hinge region of the Fc region (CH1 domain), resulting in a change (e.g., an increase or decrease) in the number of cysteine ​​residues in the hinge region. The number of cysteine ​​residues in the hinge region of the CH1 domain may be changed, for example, to facilitate the 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.

[0225] In some embodiments, one, two, or more mutations (e.g., amino acid substitutions) are introduced into the Fc region of muscle-targeting antibodies described herein [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] to increase or decrease the affinity of the antibody to the Fc receptor on the surface of effector cells (e.g., activated Fc receptor) [e.g., using numbering by Kabat's numbering system (e.g., the EU index in Kabat)]. Mutations in the Fc region of an antibody that decrease or increase the affinity of the antibody to the Fc receptor and techniques for introducing such mutations into the Fc receptor or fragments thereof are known to those skilled in the art. Examples of mutations in the Fc receptor of an antibody that may be made to alter the affinity of the antibody to 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 International Publication Nos. 02 / 060919; 98 / 23289; and 97 / 34631, which are incorporated herein by reference.

[0226] 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 (preferably an Fc or hinge-Fc domain fragment) to alter (e.g., decrease or increase) the half-life of the antibody in vivo. For example, see International Publication No. 02 / 060919; International Publication No. 98 / 23289; and International Publication No. 97 / 34631; and U.S. Patents No. 5,869,046, No. 6,121,022, No. 6,277,375 and No. 6,165,745.

[0227] 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 (preferably 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 its FcRn binding fragment (preferably Fc or hinge-Fc domain fragment) to increase the half-life of the antibody in vivo. In some embodiments, the antibody may 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 numbering by the EU index in Kabat [Kabat EA et al., (1991) cited above]. In some embodiments, the constant region of IgG1 of the antibodies described herein includes a substitution of methionine (M) to tyrosine (Y) at position 252, serine (S) to threonine (T) at position 254, and a substitution of threonine (T) to glutamic acid (E) at position 256, as numbered by the EU index in Kabat. See U.S. Patent No. 7,658,921, incorporated herein by reference. This type of mutant IgG, referred to as the “YTE mutant,” has been shown to exhibit a four-fold increase in half-life compared to the wild-type version of the same antibody [Dall'Acqua WF et al., (2006) J Biol Chem 281: 23514-24]. In some embodiments, the antibody comprises an IgG constant domain containing 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.

[0228] In some embodiments, one, two, or more amino acid substitutions are introduced into the IgG constant domain Fc region to alter the effector function of an anti-HJV antibody. The effector ligand whose affinity to it is altered may be, for example, the Fc receptor or the C1 component of complement. This approach is described in more detail in U.S. Patents 5,624,821 and 5,648,260. In some embodiments, deletion or inactivation of the constant domain (through point mutation or other means) can reduce the Fc receptor binding of the circulating antibody, thereby increasing tumor localization. See, for example, U.S. Patents 5,585,097 and 8,591,886 for descriptions of mutations that delete or inactivate the constant domain, thereby increasing tumor localization. In some embodiments, one or more amino acid substitutions may be introduced into the Fc region of the antibodies described herein to remove potential glycosylation sites on the Fc region that may reduce Fc receptor binding [see, for example, Shields RL et al., (2001) J Biol Chem 276: 6591-604].

[0229] In some embodiments, one or more amino acids in the constant region of the anti-HJV antibody described herein may be replaced with different amino acid residues so that the antibody has modified Clq binding and / or reduced or absent complement-dependent cytotoxicity (CDC). This approach is described in more detail in U.S. Patent 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 antibody described herein are modified to alter the antibody's ability to immobilize complement. This approach is described in more detail in International Publication No. 94 / 29351. In some embodiments, the Fc region of the antibody described herein is modified to increase the antibody's ability to mediate antibody-dependent cytotoxicity (ADCC) and / or increase its affinity for the Fcγ receptor. This approach is described in more detail in International Publication No. 00 / 42072.

[0230] In some embodiments, the heavy and / or light chain variable domain sequences of the antibodies provided herein may be used, for example, to generate CDR-implanted, chimeric, humanized, or compound human antibodies or antigen-binding fragments as described elsewhere herein. As will be understood by those skilled in the art, any variant, CDR-implanted, chimeric, humanized, or compound antibody derived from any antibody provided herein may be useful in the compositions and methods described herein, maintaining the ability to specifically bind to hemoduverin, and as a result, the variant, CDR-implanted, chimeric, humanized, or compound antibody has at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or higher binding to hemoduverin compared to the original antibody from which it is derived.

[0231] In some embodiments, the antibodies provided herein include mutations that confer desired properties to the antibody. For example, to avoid potential complications resulting from Fab-arm exchange, which are known to occur with native IgG4 mAbs, the antibodies provided herein may include 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], where serine 228 (EU numbering; residue 241, Kabat numbering) is converted to proline, resulting in an IgG1-like hinge sequence. Thus, any of the antibodies may include a stabilizing "Adair" mutation.

[0232] 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, one or more sugar or carbohydrate molecules are conjugated to the antibody by N-glycosylation, O-glycosylation, C-glycosylation, glycieation (GPI anchoring), and / or phosphoglycosylation. In some embodiments, one or more sugar or carbohydrate molecules are monosaccharides, disaccharides, oligosaccharides, or glycans. In some embodiments, one or more sugar or carbohydrate molecules are branched oligosaccharides or branched glycans. In some embodiments, one or more sugar or carbohydrate molecules include mannose units, glucose units, N-acetylglucosamine units, N-acetylgalactosamine units, galactose units, fucose units, or phospholipid units. In some embodiments, there are about 1 to 10, about 1 to 5, about 5 to 10, about 1 to 4, about 1 to 3, or about 2 sugar molecules. In some embodiments, the glycosylated antibody is completely or partially glycosylated. In some embodiments, the antibody is glycosylated by chemical reaction or enzymatic means. In some embodiments, the antibody is glycosylated in vitro or inside a cell, and the cell may lack an enzyme in the N- or O-glycosylation pathway, such as glycosyltransferase. In some embodiments, the antibody is functionalized with a sugar or carbohydrate molecule, as described in International Patent Application Publication No. 2014065661, published on May 1, 2014, titled "Modified Antibody, Antibody-Conjugate and Method for Preparation thereof".

[0233] 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 chain and / or light chain sequence. In some embodiments, the anti-HJV antibody described herein includes any one of the VH and VL sequences described herein, any one of the IgG heavy chain and light chain sequences, or any one of the F(ab') heavy chain and light chain sequences, and further includes a signal peptide (e.g., an N-terminal signal peptide). In some embodiments, the signal peptide includes the amino acid sequence MEFGLSWLFLVAILKGVQC (SEQ ID NO: 104).

[0234] III. Preparation of anti-HJV antibodies The antibodies capable of binding to hemoduveline described herein can be prepared by any method known in the art. See, for example, Harlow and Lane, (1998) Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, New York.

[0235] In some embodiments, antibodies specific to a target antigen (e.g., HJV) may be produced by conventional hybridoma techniques. A full-length target antigen or a fragment thereof, which may be linked to a carrier protein, e.g., KLH, may be used to immunize a host animal to produce antibodies that bind to that antigen. The routes and schedules for immunization of the host animal are generally carried out according to established prior art for antibody stimulation and production, as further described herein. General techniques for mouse, humanization, and human antibody production are known in the art and are described herein. Any mammalian subject, including humans, or antibody-producing cells therefrom, are assumed to be engineered to serve as a basis for the production of mammalian hybridoma cell lines, including human hybridoma cell lines. Typically, the host animal is inoculated intraperitoneally, intramuscularly, orally, subcutaneously, in the sole of the foot, and / or intradermally, in amounts of immunogen, including those described herein.

[0236] If desired, the antibody of interest (monoclonal or polyclonal) (e.g., produced by a hybridoma) may be sequenced, and the polynucleotide sequence may then be cloned into a vector for expression or reproduction. The sequence encoding the antibody of interest may be maintained in the vector in host cells, 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 antibody’s affinity (affinity maturation) or other characteristics. For example, the constant region may be engineered to more closely resemble the human constant region so that the antibody avoids an immune response when 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 against the target antigen. It is evident to those skilled in the art that one or more polynucleotide changes can be made to an antibody while still maintaining its binding specificity to the target antigen.

[0237] In other embodiments, fully human antibodies may be obtained by using commercially available mice engineered to express specific human immunoglobulin proteins. Transgenic animals designed to produce a more desirable (e.g., fully human antibodies) or more robust immune response may also be used for humanization or the production of human antibodies. Examples of such techniques include XenomouseRTM from Amgen, Inc. (Fremont, CA) and HuMAb-MouseRTM and TC Mouse™ from Medarex, Inc. (Princeton, NJ) or H2L2 mice from Harbour Antibodies BV (Holland). Alternatively, antibodies may be produced by recombination using phage display or yeast techniques. See, for example, 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 the immunoglobulin variable (V) domain gene repertoire from non-immunized donors.

[0238] Antigen-binding fragments of intact antibodies (full-length antibodies) can be prepared by routine methods. For example, F(ab')2 fragments can be produced by pepsin digestion of the antibody molecule, and Fab fragments can be generated by reducing the disulfide crosslinks of the F(ab')2 fragment. Genetically engineered antibodies, such as humanized antibodies, chimeric antibodies, single-chain antibodies, and bispecific antibodies, can be produced, for example, by conventional recombination techniques. In one example, DNA encoding a monoclonal antibody specific to a target antigen can be readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that have the ability to specifically bind to the genes encoding the heavy and light chains of the monoclonal antibody). Hybridoma cells serve as a preferred source of such DNA. Once isolated, the DNA may be placed in 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 otherwise do not produce immunoglobulin proteins, to obtain the synthesis of monoclonal antibodies in recombinant host cells. See, for example, PCT International Publication 87 / 04462. The DNA can then be modified, for example, by substituting coding sequences of human heavy and light chain constant domains at the location of 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 this manner, genetically modified antibodies with binding specificity to target antigens, such as “chimeric” or “hybrid” antibodies, can be prepared.

[0239] Single-chain antibodies can be prepared via recombinant techniques by linking nucleotide sequences encoding a heavy-chain variable region and a light-chain variable region. Preferably, a flexible linker is incorporated between the two variable regions.

[0240] Alternatively, phage or yeast scFv libraries can be produced by applying techniques described for the production of single-chain antibodies (U.S. Patents No. 4,946,778 and 4,704,692), and HJV-specific scFv clones can be identified from the library using routine procedures. Positive clones can then be subjected to further screening to identify clones with high HJV binding affinity.

[0241] Antibodies obtained according to methods known in the art and described herein can be characterized using methods known in the art. For example, one method is to identify the epitopes to which the antigen binds, or "epitope mapping." There are many methods known in the art for mapping and characterizing the location of epitopes on proteins, including, for example, elucidation of the crystalline structure of antibody-antigen complexes, competitive assays, gene fragment expression assays, and synthetic peptide-based assays, as described in 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 sequences to which the antibody binds. Epitopes can be linear epitopes, i.e., those contained within a single stretch of amino acids, or conformational epitopes formed by three-dimensional interactions of amino acids, which do not necessarily have to be contained within a single stretch (linear primary structure sequence). Peptides of variable length (e.g., at least 4-6 amino acids) may be isolated or synthesized (e.g., by recombination) and used for antibody-binding assays. In another example, the epitope to which the antibody binds may be determined in a systematic screening by using overlap peptides derived from the target antigen sequence and determining antibody binding. According to a gene fragment expression assay, an open reading frame encoding the target antigen is fragmented randomly or by a specific genetic composition, and the reactivity of the expressed fragments of the antigen with the antibody to be tested is determined. The gene fragments may be produced, for example, by PCR, then transcribed in vitro and translated into proteins in the presence of radioactive amino acids. Antibody binding to the radiolabeled antigen fragments is then determined by immunoprecipitation and gel electrophoresis.A particular epitope can also be identified by using a large library of random peptide sequences presented on the surface of phage particles (a phage library). Alternatively, a defined library of overlapping peptide fragments can be tested for binding to a test antibody in a simple binding assay. In additional examples, antigen-binding domain mutagenesis, domain swapping experiments, and alanine scanning mutagenesis may be performed to identify the sufficient and / or required residues for epitope binding. Alternatively, a competitive assay may be performed using other antibodies known to bind to the same antigen to determine whether an antibody binds to the same epitope as another antibody. Competitive assays are well known to those skilled in the art.

[0242] In some cases, anti-HJV antibodies are prepared by recombinant techniques as illustrated below. The nucleic acids encoding the heavy and light chains of the anti-HJV antibodies described herein can be cloned into a single expression vector, with each nucleotide sequence operably ligated to a suitable promoter. In one example, each of the nucleotide sequences encoding the heavy and light chains is operably ligated to a separate promoter. Alternatively, the nucleotide sequences encoding the heavy and light chains can be operably ligated to a single promoter, resulting in both the heavy and light chains being expressed from the same promoter. If necessary, an internal ribosome entry site (IRES) can be inserted between the heavy chain and light chain sequences.

[0243] In some cases, the nucleotide sequences encoding the two chains of an antibody are cloned into two vectors, and the two vectors can be introduced into the same or different cells. If the two chains are expressed in different cells, each of them can be isolated from the host cell expressing it, and the isolated heavy and light chains can be mixed and incubated under suitable conditions to enable antibody formation.

[0244] Generally, nucleic acid sequences encoding one or all of an antibody chain can be cloned into a suitable expression vector operably linked to a suitable promoter using methods known in the art. For example, the nucleotide sequence and vector can be contacted with restriction enzymes under suitable conditions, thereby creating complementary ends on each molecule that can pair with each other and ligate together with the ligase. Alternatively, synthetic nucleic acid linkers can be ligated to the ends of the gene. These synthetic linkers contain nucleic acid sequences corresponding to specific restriction sites in the vector. The choice of expression vector / promoter depends on the type of host cell for use in antibody production.

[0245] Various promoters are available for the expression of the antibodies described herein, including, but not limited to, the cytomegalovirus (CMV) initial promoter, viral LTRs such as Rous sarcoma virus LTR, HIV-LTR, HTLV-1 LTR, Simian virus 40 (SV40) initial promoter, Escherichia coli lac UV promoter, and herpes simplex virus tk promoter.

[0246] Adjustable promoters may also be used. Such adjustable promoters include those that use a lac repressor from E. coli as a transcription modulator to regulate transcription from mammalian cell promoters containing a lac operator [Brown, M. et al., Cell, 49:603-612 (1987)], and those that use a 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 estradiol, RU486, diphenol murislerone, or FK506 dimer, VP16, or p65 using rapamycin. Inducible systems are available from several companies, including Invitrogen, Clontech, and Ariad.

[0247] A moduloable promoter containing a repressor may be used with the operon. In one embodiment, a lac repressor from E. coli can function as a transcription modulator to regulate transcription from a mammalian cell promoter possessing a lac operator [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)]. A tetracycline repressor (tetR) is combined with a transcription activator (VP 16) to create a tetR-mammalian cell transcription activator fusion protein, tTa(tetR-VP 16), tetO, which has a minimal promoter derived from a 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. Rather than a tetR-mammalian cell transcription factor fusion derivative, the tetracycline repressor (tetR) alone can function as a potent transmodulator for regulating gene expression in mammalian cells when the tetracycline operator is appropriately positioned 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, in order to achieve its tunable effect, it does not require the use of a tetracycline repressor-mammalian cell transactivator or repressor fusion protein, which can be toxic to cells in some cases [Gossen 5 et al., Natl. Acad. Sci. USA, 89:5547-5551 (1992); Shockett et al., Proc. Natl. Acad. Sci. USA, 92:6522-6526 (1995)].

[0248] Additionally, the vector may contain some or all of the following, for example: a selection marker gene, e.g., the neomycin gene for selection of stable or transient transfectants in mammalian cells; an enhancer / promoter sequence from the earliest gene of human CMV for high levels of transcription; transcription termination and RNA processing signals from SV40 for mRNA stability; the SV40 polyomatous replication origin 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 the transgene 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, the human collagen I polyadenylation signal, the human collagen II polyadenylation signal, and the SV40 polyadenylation signal.

[0249] 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 host cells suitable for antibody production. 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 lineage (COS) cells. In some embodiments, the host cells expressing the anti-HJV antibody are CHO cells. The host cells may be cultured under conditions suitable for the expression of the antibody or any polypeptide chain thereof. Such antibodies or their polypeptide chains can be recovered by cultured cells (e.g., from cells or culture supernatant) via conventional methods, such as affinity purification. If necessary, the polypeptide chain of the antibody can be incubated under suitable conditions for a suitable period of time that allows for antibody production. In some embodiments, the host cell contains nucleic acid encoding the heavy chain of the anti-HJV antibody. In some embodiments, the host cell contains nucleic acid encoding the light chain of the anti-HJV antibody. In some embodiments, the host cell contains nucleic acid encoding both the heavy chain and the light chain.

[0250] In some embodiments, the antibody preparation methods described herein involve a recombinant expression vector encoding both the heavy and light chains of an anti-HJV antibody, as also described herein. The recombinant expression vector can be introduced into suitable host cells (e.g., dhfr-CHO cells) by conventional methods, such as calcium phosphate-mediated transfection. Positive transformant host cells are selected and can be cultured under suitable conditions that allow 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 cells can be incubated under suitable conditions that allow antibody formation.

[0251] 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 recombinant expression vectors can be introduced into suitable host cells (e.g., dhfr-CHO cells) by conventional methods, such as calcium phosphate-mediated transfection.

[0252] Alternatively, each expression vector can be introduced into a suitable host cell. A positive transformant can be selected and cultured under suitable conditions that allow expression of the antibody polypeptide chain. If 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 chain can be recovered from the host cell or culture medium and then incubated under suitable conditions that allow antibody formation. If two expression vectors are introduced into different host cells, each of them can be recovered from the corresponding host cell or corresponding culture medium. The two polypeptide chains can then be incubated under suitable conditions for antibody formation.

[0253] Standard molecular biology techniques are used to prepare recombinant expression vectors, transfect host cells, select transformants, culture the host cells, and recover antibodies from the culture medium. For example, some antibodies can be isolated by affinity chromatography using a protein A or protein G linked matrix.

[0254] Any of the nucleic acids encoding the heavy chain, light chain, or both of the anti-HJV antibody described herein (for example, provided in Table 3), vectors containing the same (for example, expression vectors), or host cells containing the vector are within the scope of this disclosure.

[0255] [Table 3-1] [Table 3-2] [Table 3-3] [Table 3-4] [Table 3-5] [Table 3-6] [Table 3-7] [Table 3-8] [Table 3-9] [Table 3-10] [Table 3-11]

[0256] In some embodiments, the anti-HJV described herein is produced by (i) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 71 and / or (ii) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 73.

[0257] In some embodiments, the anti-HJV described herein is produced by (i) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 71, and / or (ii) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 75.

[0258] In some embodiments, the anti-HJV described herein is produced by (i) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 71 and / or (ii) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 77.

[0259] In some embodiments, the anti-HJV described herein is produced by (i) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 71 and / or (ii) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 79.

[0260] In some embodiments, the anti-HJV described herein is produced by (i) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 71, and / or (ii) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 81.

[0261] In some embodiments, the anti-HJV described herein is produced by (i) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 84, and / or (ii) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 85.

[0262] In some embodiments, the anti-HJV described herein is produced by (i) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 88, and / or (ii) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 89.

[0263] In some embodiments, the anti-HJV described herein is produced by (i) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 92, and / or (ii) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 93.

[0264] In some embodiments, the anti-HJV described herein is produced by (i) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 94, and / or (ii) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 93.

[0265] In some embodiments, the anti-HJV described herein is produced by (i) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 92, and / or (ii) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 96.

[0266] In some embodiments, the anti-HJV described herein is produced by expressing (i) nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of sequence number 99, and / or (ii) nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of sequence number 100.

[0267] In some embodiments, the anti-HJV described herein is produced by (i) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 40 and / or (ii) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 64.

[0268] In some embodiments, the anti-HJV antibodies described herein may be used to deliver a molecular payload to target cells or target tissues (e.g., cells or tissues expressing HJV). Thus, the anti-HJV antibodies described herein may be conjugated to a molecular payload. The conjugates described herein may be used in a variety of applications, such as diagnostic or therapeutic applications.

[0269] In some embodiments, the complexes described herein are 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 the gene, protein, and / or nucleic acid. The molecular payload may be a small molecule, protein, nucleic acid, oligonucleotide, or any molecular entity having the ability to modulate the activity or function of the gene, protein, and / or nucleic acid in a cell. In some embodiments, the molecular payload is an oligonucleotide that targets disease-associated repeats in muscle cells.

[0270] IV. Pharmaceutical Compositions In addition to the antibodies described herein, encoding nucleic acids or sets of nucleic acids, vectors containing them, or host cells containing vectors may be mixed with pharmaceutically acceptable carriers (excipients) to form a pharmaceutical composition for use in the treatment of a target disease. “Acceptable” means that the carrier must be compatible with (and preferably have the ability to stabilize) the active ingredient of the composition and must not be 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.

[0271] The anti-HJV antibody-containing pharmaceutical compositions disclosed herein may further comprise suitable buffers. A buffer is a weak acid or base used to maintain the pH of the solution near a selected value after the addition of another acid or base. In some examples, the buffers disclosed herein may be buffers capable of maintaining physiological pH regardless of changes in carbon dioxide concentration (generated by cellular respiration). Exemplary buffers 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 buffers may comprise disodium hydrogen phosphate and sodium chloride, or potassium dihydrogen phosphate and potassium chloride.

[0272] In some embodiments, the buffer in the pharmaceutical composition 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 lower than 7, for example, about 7, 6.8, 6.5, 6.3, 6, 5.8, 5.5, 5.3, or 5.

[0273] The pharmaceutical compositions described herein comprise one or more preferred 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.)]. Since salts consist of a corresponding number of cations (positively charged ions) and anions (negative ions), the products are electrically neutral (have no net charge).

[0274] In some embodiments, the pharmaceutical composition may contain pharmaceutically acceptable carriers, excipients, or stabilizers in the form of a lyophilized formulation or 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.

[0275] Pharmaceutical compositions used for in vivo administration must be sterile. This can be easily achieved, for example, by filtration through a sterile filtration membrane. Therapeutic antibody compositions are generally placed in containers with sterile access ports, such as intravenous or subcutaneous solution bags or vials with stoppers that can be penetrated by a subcutaneous injection needle.

[0276] V. How to use Aspects of this disclosure relate to compositions and methods for treating myelofibrosis and / or one or more conditions resulting from myelofibrosis in a subject.

[0277] Figure 12 depicts the hepcidin-stimulated pathway and the physiological regulation of iron homeostasis by hepcidin. As commonly shown, hepcidin functions by binding to the iron exporter ferroportin in iron-releasing target cells (e.g., hepatocytes, duodenal cells, tissue macrophages, and other cell types). Hepcidin binding blocks iron efflux and induces ubiquitination, internalization, and lysosomal degradation of ferroportin. This results in a decrease in intracellular iron retention and ultimately a decrease in systemic iron levels.

[0278] The HAMP gene encodes a hepcidin precursor protein, which is expressed primarily in hepatocytes of the liver at lower levels than in other cells of extrahepatic tissues. The precursor protein is subsequently cleaved to produce physiologically active hepcidin. In some embodiments, the hepcidin antagonists of this disclosure are HAMP antagonists that antagonistize hepcidin function by binding to HAMP or its transcripts or translation products, or by inhibiting HAMP transcription or translation regulators to reduce HAMP expression. Examples of HAMP transcription regulators include, but are not limited to, SMAD1 / 5 / 8 (e.g., the BMP-SMAD signaling pathway) and STAT3 (e.g., the JAK-STAT signaling pathway). Thus, in some embodiments, the HAMP antagonist is a BMP-SMAD signaling pathway inhibitor or a JAK-STAT signaling pathway inhibitor.

[0279] An increase in serum or tissue iron triggers the transcriptional induction of hepcidin via the BMP-SMAD signaling pathway. The mechanism involves the secretion of bone morphogenetic protein 6 (BMP6) from hepatic sinusoidal endothelial cells, which binds to BMP receptors (BMPRs) on hepatocytes, thereby activating the SMAD signaling cascade. Examples of BMP receptors include type I (e.g., ALK2, ALK3, ALK6) and type II (e.g., ActRIIA, BMPRII) BMP receptors. The central regulator of the BMP-SMAD signaling pathway for hepcidin stimulation is the BMP co-receptor, hemoduvelin (HJV). Thus, in some embodiments, this disclosure relates to hemoduvelin antagonists (e.g., anti-HJV antibodies and their compositions) for targeting hepcidin and modulating iron homeostasis, for example, for the treatment of myelofibrosis and / or one or more conditions resulting from myelofibrosis. In some embodiments, the disclosure provides a method for modulating iron homeostasis for the treatment of myelofibrosis, involving direct inhibition of hepcidin binding to the iron exporter ferroportin. In some embodiments, the hepcidin antagonist is an anti-HJV antibody.

[0280] Myelofibrosis (MF) is a myeloproliferative disorder characterized by the abnormal proliferation of hematopoietic stem cells that result in fibrosis of the bone marrow. The production of healthy blood cells (megakaryocytes and erythrocytes, which are responsible for platelet production) is impaired. MF can be classified into primary MF (PMF) and secondary MF (SMF). PMF and SMF have similar clinical profiles, including common main symptoms such as anemia, fatigue, and splenomegaly. Primary myelofibrosis (PMF) is characterized as MF that occurs on its own. In some embodiments, PMF may be associated with elevated levels of pro-inflammatory cytokines (e.g., IL-6) in the subject [see, e.g., Cokic et al. Proinflammatory Cytokine IL-6 and JAK-STAT Signaling Pathway in Myeloproliferative Neoplasms, Mediators Inflamm. 2015; 2015: 453020]. Secondary myelofibrosis (SMF) occurs as a result of scar tissue in the bone marrow as a complication of another disease, such as an autoimmune disease. In some embodiments, the subject matter described herein has or is suspected to have PMF. In some embodiments, the subject matter described herein has or is suspected to have SMF.

[0281] In some embodiments, subjects who have or are suspected of having myelofibrosis (e.g., PMF and / or SMF) have one or more genes (e.g., JAK2 gene, thrombopoietin receptor (MPL) gene, calreticulin (CALR) gene, lymphocyte-specific adapter protein Lnk (LNK) gene, ASXL transcription regulator 1 (ASXL1) gene, serine-arginine-rich splicing factor 2 (SRSF2) gene, protein phosphatase, Mg2 + / Mn2 + Dependence gene 1D (PPM1D), isocitrate dehydrogenase (NADP) (+) )1 / Isocitrate dehydrogenase (NADP (+)This includes mutations in one or more of the following genes: )2(IDH1 / 2) gene, Tet methylcytosine dioxygenase 2 (TET2) gene, Zeste 2 Polycomb repression complex 2 subunit enhancer (EZH2) gene, U2 nuclear small RNA cofactor 1 (U2AF1) gene, nuclear factor erythrocyte 2 (NFE2) gene, SH2B adapter protein 3 (SH2B3) gene, splicing factor 3b subunit 1 (SF3B1) gene, Cbl proto-oncogene (CBL) gene, or combinations thereof.

[0282] In some embodiments, the subject has one or more mutations in the JAK2 gene. JAK2 plays a crucial role in signaling from receptors involved in the proliferation of myeloid cells mediated by EPO, TPO, and / or G-CSF [see, e.g., Alshemmari et al., Molecular Pathogenesis and Clinical Significance of Driver Mutations in Primary Myelofibrosis: A Review, Med Princ Pract, 2016;25(6):501-509]. In some embodiments, the subject contains a human JAK2 gene with a triggering mutation in exon 12 or exon 14. In some embodiments, the triggering mutation in the JAK2 gene is in exon 14, resulting in a V617F substitution. In some embodiments, the V617F mutation leads to hyperactivation of JAK2 and its associated signaling pathways. In some embodiments, hyperactivation of JAK2 leads to myelofibrosis (e.g., PMF and / or SMF).

[0283] In some embodiments, the subjects have one or more mutations in the thrombopoietin receptor (MPL) gene. MPL is a cognate receptor of thrombopoietin (TPO), and mutations that result in increased function of the MPL gene lead to impaired megakaryocyte production. In some embodiments, the subjects include the W515L / K mutation in MPL. In some embodiments, subjects with one or more mutations in the MPL gene are more likely to develop anemia compared to subjects with MF as a whole (e.g., more than 10%, more than 20%, more than 30%, more than 40%, more than 50%, more than 60%, more than 70%, more than 80%, more than 90%, more than twice, more than three times, more than four times, more than five times, more than six times, more than seven times, more than eight times, more than nine times, or more than ten times) (Guglielmelli P et al., Anemia characterises patients with myelofibrosis harboring Mpl mutation. Br J Haematol 2007; 137: 244-247).

[0284] In some embodiments, subjects have one or more mutations in the calreticulin (CALR) gene. The CALR gene encodes calreticulin protein, a multifactorial protein that regulates calcium homeostasis, cell signaling, gene expression, cell adhesion, autoimmunity, and apoptosis. Approximately 140 CALR mutations have been identified in 19 variants and are associated with MF. In some embodiments, subjects have or are suspected of having MF with an exon 9 mutation in the CALR gene.

[0285] Further mutations in other genes associated with MF have been identified. Non-exclusive examples of genes associated with MF include, for example, JAK2, MPL, CLAR, LNK, ASXL1, SRSF2, PPM1D, IDH1 / 2, TET2, EZH2, U2AF1, NFE2, SH2B3, SF3B1, or CBL. In some embodiments, subjects have or are suspected of having MF, which includes one or more mutations in one or more of the genes described herein.

[0286] In some embodiments, the subject has one or more mutations in genes involved in epigenetic regulation or splicing. In some embodiments, the one or more mutations in genes involved in epigenetic regulation or splicing are ASXL1, DNMT3A, TET2, SRSF2, U2AF1, EZH2, or SF3B1. In some embodiments, the subject has mutations in IDH1 / 2, which are associated with a risk of progression to MBN-BP.

[0287] In some embodiments, the disclosure relates to compositions (e.g., anti-HJV antibodies and their compositions) and methods for treating myelofibrosis in subjects. In some embodiments, subjects treated according to the disclosure may be identified based on appropriate diagnostic or prognostic methods. For example, the Dynamic International Prognostic Scoring System (DIPSS) and age-adjusted DIPSS provide models of patient outcomes based on several patient-specific variables, including age, hemoglobin levels, leukocyte count, peripheral hemoblasts, and systemic symptoms [see, for example, Passamonti, F., et al. Blood. 2010 Mar 4;115(9):1703-8, incorporated herein by reference]. The DIPSS model enables the calculation of a DIPSS score and the assignment of patients to risk categories for prognostic purposes. A DIPSS score of 0 indicates a “low-risk” patient, a DIPSS score of 1–2 indicates an “intermediate-1 risk” patient, a DIPSS score of 3–4 indicates an “intermediate-2 risk” patient, and a DIPSS score of 5–6 indicates a “high-risk” patient. Therefore, in some embodiments, subjects requiring treatment according to this application may have a DIPSS score of at least 1. In some embodiments, subjects have a DIPSS score of 1–4 (e.g., 1, 2, 3, or 4). In some embodiments, the DIPSS score is 5 or 6 (e.g., 5 or 6).

[0288] In some embodiments, the subjects treated in accordance with this disclosure may be evaluated by appropriate diagnostic or prognostic methods. For example, the Myeloproliferative Neoplasm-Symptom Assessment Form Total Symptom Score (MPN-SAF TSS) provides a 10-item tool designed to assess most representative and clinically relevant symptoms among patients with MPN. The tool records a patient's assessment of the incidence and severity of these disease-related symptoms. It can be used to track symptoms over time and guide subsequent management decisions [see, for example, Emanuel RM, et al. Myeloproliferative neoplasm (MPN) symptom assessment form total symptom score: prospective international assessment of an abbreviated symptom burden scoring system among patients with MPNs, J Clin Oncol. 2012;30(33):4098-4103, incorporated herein by reference]. The MPN-SAF TSS includes symptoms such as fatigue, early satiety, lethargy, lack of concentration, abdominal discomfort, night sweats, bone pain, itching, unintentional weight loss, and fever. Each symptom is rated on a severity scale from 0 (none / best possible) to 10 (worst possible / worst possible). The MPN-SAF TSS has a possible range of 0 to 100, with 100 representing the highest level of symptom severity. In some embodiments, the Myelofibrosis Symptom Assessment Form (MFSAF) is derived from the MPN-SAF TSS. The MFSAF is a means of measuring symptoms reported by more than 10% of MF patients and includes quality of life (QoL) measurements.The MFSAF includes a comprehensive assessment of fatigue, an assessment of splenomegaly and associated organic symptoms, and an assessment of other symptoms such as night sweats, itching (pruritus), bone pain, fever, unintentional weight loss, and overall quality of life [see, for example, Mesa et al., The Myelofibrosis Symptom Assessment Form (MFSAF): An Evidence-based Brief Inventory to Measure Quality of Life and Symptomatic Response to Treatment in Myelofibrosis, Leuk Res. 2009 Sep; 33(9): 1199-1203, incorporated herein by reference]. The MFSAF may be used to track symptoms over time and to guide decisions regarding subsequent management. In some embodiments, the patient has an MPN-SAF TSS of 0-100 (e.g., any score between 0 and 100), 10-100 (e.g., any score between 10 and 100), 20-100 (e.g., any score between 20 and 100), 30-100 (e.g., any score between 30 and 100), 40-100 (e.g., any score between 40 and 100), 50-100 (e.g., any score between 50 and 100), 60-100 (e.g., any score between 60 and 100), 70-100 (e.g., any score between 70 and 100), 80-100 (e.g., any score between 80 and 100), or 90-100 (e.g., any score between 00 and 100).

[0289] In some embodiments, subjects with MF (e.g., PMF or SMF) develop anemia. Anemia in MF is a result of a multifactorial process.

[0290] In some embodiments, anemia in MF may be treatment-related. In some embodiments, MF patients have been previously treated with a JAK inhibitor (e.g., ruxolitinib or fedratinib). In some embodiments, patients receiving a JAK inhibitor (e.g., ruxolitinib or fedratinib) are more likely to develop MF-associated anemia. Inhibition of the JAK-STAT signaling pathway results in inhibition of erythropoietin-mediated JAK2 signaling, which is essential for erythropoiesis. In some embodiments, initial anemia has been identified as a major adverse event associated with JAK inhibitor (e.g., ruxolitinib) treatment. [For example, the entire contents of each of the following are incorporated herein by reference: Verstovsek S, Kantarjian H, Mesa RA, et al. Safety and efficacy of INCB018424, a JAK1 and JAK2 inhibitor, in myelofibrosis. N Engl J Med. 2010;363(12):1117-1127; Verstovsek S, Mesa RA, Gotlib J, et al. A doubleblind, placebo-controlled trial of ruxolitinib for myelofibrosis. N Engl J Med. 2012;366(9):799-807; Parganas E, Wang D, Stravopodis D, et al. Jak2 is essential for signaling through a variety of cytokine receptors. Cell.] 1998;93(3):385-395;Neubauer H, Cumano A, Müller M, Wu H, Huffstadt U, Pfeffer K. Jak2 deficiency defines an essential developmental checkpoint in definitive hematopoiesis. Cell. 1998;93(3):397-409].

[0291] In some embodiments, subjects have or are at risk of having systemic or microvascular symptoms associated with myeloproliferative neoplasms (MPN). In some embodiments, subjects have or are at risk of having thromboembolic or hemorrhagic complications. In some embodiments, subjects have or are at risk of having acute myeloid leukemia (AML) in the acute transition phase of MPN. In some embodiments, subjects exhibit ribosomal disease in megakaryocytes. In some embodiments, subjects exhibit decreased GATA1 expression, particularly in megakaryocytes. In some embodiments, subjects exhibit a defect in megakaryocyte function or maturation. In some embodiments, subjects do not have nutritional iron deficiency. In some embodiments, subjects exhibit thrombocytopenia, anemia, and / or neutropenia.

[0292] In some embodiments, myelofibrosis-associated anemia is caused by ineffective erythrocyte production due to bone marrow suppression and deficiencies in iron metabolism, increased destruction of erythrocytes due to splenomegaly, increased plasma volume, an abnormal pro-inflammatory environment in the bone marrow, or a combination thereof.

[0293] In some embodiments, myelofibrosis-associated anemia is associated with abnormal iron metabolism. In some embodiments, the abnormal iron metabolism in MF patients is functional iron deficiency (FID). FID represents a state of iron-limited erythropoiesis characterized by an imbalance between the iron required for effective erythropoiesis and readily available serum iron. In FID, iron is sequestrated and unavailable for erythropoiesis, even when the body has sufficient or increased systemic iron stores. In some embodiments, FID is caused by an increase in hepcidin compared to iron storage levels. In some embodiments, the increase in hepcidin expression is caused by the upregulation of pro-inflammatory cytokines. For example, IL-6 has been reported to be higher in anemic MF patients [Birgegard et al., Inflammatory Functional Iron Deficiency Common in Myelofibrosis, Contributes to Anemia and Impairs Quality of Life. From the Nordic MPN Study Group, Eur J Haematol. 2019 Mar;102(3):235-240].

[0294] In some embodiments, myelofibrosis-associated anemia is characterized by high serum pro-inflammatory cytokine levels.

[0295] In myelofibrosis (e.g., PMF), pro-inflammatory cytokines that induce hepcidin synthesis, such as IL-6 and oncostatin-M, are typically elevated and associated with iron sequestration, iron loading of macrophages, as well as myeloproliferation and macrophage activation. These elevated hepcidin levels can lead to anemia.

[0296] In some embodiments, the pro-inflammatory cytokine is IL-6. The normal range for IL-6 (e.g., in disease-free subjects) is equal to or lower than 1.8 pg / ml. In some embodiments, subjects have higher-than-normal serum IL-6 levels, e.g., serum IL-6 levels higher than 1.8 pg / ml.

[0297] In some embodiments, the disclosure provides a method for treating subjects having myelofibrosis. In some embodiments, the subjects have anemia resulting from hepcidin synthesis induced by a pro-inflammatory cytokine. In some embodiments, the pro-inflammatory cytokine that increases hepcidin synthesis is IL-6. In some embodiments, administration of an anti-HJV antibody to the subject reduces hepcidin synthesis induced by a pro-inflammatory cytokine (e.g., IL-6). In some embodiments, administration of an anti-HJV antibody relieves FID associated with increased hepcidin synthesis induced by a pro-inflammatory cytokine (e.g., IL-6).

[0298] Determining whether the amount of antibody (e.g., anti-HJV antibody) has achieved a therapeutic effect is obvious to those skilled in the art based on the teachings provided herein. The effective dose will vary, as recognized by those skilled in the art, depending on the specific condition being treated, the severity of the condition, individual patient parameters including age, health status, size, sex, and weight, the duration of treatment, the nature of any concomitant therapies, the specific route of administration, and factors within the knowledge and expertise of the healthcare professional. These factors are well known to those skilled in the art and can be addressed simply by routine experimentation. The specific administration regimens used in the methods described herein, i.e., dose, timing, and frequency, will depend on the specific subject and their medical history, as described herein.

[0299] Empirical considerations, such as time to maximum effect, half-life, and / or time above a certain concentration, will usually contribute to the determination of the dosage. For example, antibodies compatible with the human immune system, such as humanized antibodies or fully human antibodies, may be used to prolong the antibody half-life and prevent the antibody from being attacked by the host immune system. Other reasons for dose adjustment include differences in sex, age, individual response, polymorphism at the antibody target, and / or pharmacokinetic or pharmacodynamic responses driven by receptors involved in antibody clearance. The frequency of administration may, but not necessarily, be determined and adjusted over the course of therapy, but generally based on the treatment and / or suppression and / or remission and / or delay of the target disease / disorder. Alternatively, sustained-release formulations of antibodies may be appropriate. Various formulations and devices for achieving sustained release are known in the art.

[0300] In some embodiments, the antibody dosage described herein may be empirically determined in an individual given one or more doses of the antibody. The individual is given an increasing dose of the antagonist. Disease / impairment indicators may be followed to evaluate the efficacy of the antagonist.

[0301] The frequency of administration may vary according to the claimed method. In some embodiments, the composition may be administered once. In some embodiments, the composition may be administered on multiple occasions. In some embodiments, the frequency of administration may be weekly, every two weeks, every three weeks, every four weeks, every five weeks, every six weeks, every seven weeks, every eight weeks, every nine weeks, or every ten weeks; or once a month, every two months, or every three months, or more. In some embodiments, the composition may be administered daily, twice a week, once a week, twice a month, once a month, or at any time interval that provides a suitable (e.g., maximum) efficacy while minimizing the risk to the subject's safety. In general, efficacy and the risks of treatment and safety may be monitored throughout the course of treatment.

[0302] In some embodiments, administration of anti-HJV antibodies results in a decrease in circulating hepcidin-25 concentration and / or an increase in serum TSAT% (see, for example, Figures 7D-7E), and in some embodiments, these effects persist over a period of time (e.g., one month or longer). Thus, in some embodiments, the timing and frequency of anti-HJV antibody administration may be determined by monitoring one or more biomarkers, e.g., criteria for assessing iron utilization or determining potential iron overload. For example, in some embodiments, anti-HJV antibodies are administered intermittently or according to the level of a specific biomarker, e.g., circulating hepcidin-25 level, circulating transferrin level, or transferrin saturation percentage (TSAT%). In some embodiments, biomarker levels may be used to determine whether a subject is a candidate for treatment. However, in some embodiments, biomarkers may be used to determine whether treatment should be continued, restarted, or stopped, e.g., whether treatment with anti-HJV antibodies should be stopped.

[0303] For example, in some embodiments, if the subject's TSAT% is 70% or higher, 75% or higher, 80% or higher, 85% or higher, 90% or higher, or 95% or higher, the subject may not be considered a candidate for treatment. In some cases, if the subject's TSAT% is 70% or higher, 75% or higher, 80% or higher, 85% or higher, 90% or higher, or 95% or higher, ongoing treatment with anti-HJV antibodies may be stopped or temporarily stopped, for example, to prevent iron overload. In other embodiments, administration of anti-HJV antibodies may be carried out if the subject's TSAT% is 95% or lower, 90% or lower, 80% or lower, 70% or lower, 65% or lower, 60% or lower, 55% or lower, 50% or lower, 45% or lower, 40% or lower, 35% or lower, or 30% or lower. Therefore, in some embodiments, the target TSAT% may be monitored, for example, continuously or periodically, while the patient may, for example, receive or be treated for anemia and be evaluated to prevent iron overload or to determine if further treatment is appropriate. However, naturally, other suitable markers (including, for example, ferritin levels, serum iron levels, creatinine levels, etc.) may be monitored to determine the dosage and frequency of administration according to the methods provided herein.

[0304] In some embodiments, a subject may be administered a composition provided herein (e.g., an anti-HJV antibody) at intervals of one or more for a certain period of time. In some cases, the period during which the subject is administered the composition may be divided by the 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 be determined based on the judgment of the treating physician. For example, in some embodiments, over the course of a year, a subject may be administered the composition weekly, bi-weekly, or monthly for two months, followed by a 10-month hiatus. In some embodiments, over the course of a year, a subject may be administered the composition weekly, bi-weekly, or monthly for three months, followed by a 9-month hiatus. In some embodiments, over the course of a year, a subject may be administered the composition weekly, bi-weekly, or monthly for four months, followed by an 8-month hiatus. In some embodiments, over the course of a year, a subject may be administered the composition weekly, bi-weekly, or monthly for five months, followed by a 7-month hiatus. In some embodiments, over the course of a year, the subject may be administered the composition weekly, bi-weekly, or monthly for 6 months, followed by a 6-month suspension of administration. In some embodiments, over the course of a year, the subject may be administered the composition weekly, bi-weekly, or monthly for 7 months, followed by a 5-month suspension of administration. In some embodiments, over the course of a year, the subject may be administered the composition weekly, bi-weekly, or monthly for 8 months, followed by a 4-month suspension of administration. In some embodiments, over the course of a year, the subject may be administered the composition weekly, bi-weekly, or monthly for 9 months, followed by a 3-month suspension of administration. In some embodiments, over the course of a year, the subject may be administered the composition weekly, bi-weekly, or monthly for 10 months, followed by a 2-month suspension of administration. In some embodiments, over the course of a year, the subject may be administered the composition weekly, bi-weekly, or monthly for 2 months, followed by 2 months of no administration; or administered for 3 months, followed by 3 months of no administration; or administered for 4 months, followed by 4 months of no administration.

[0305] Generally, for the administration of any of the antibodies described herein, the dose may be approximately 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, 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.

[0306] In some embodiments, the dosage of 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 1 mg / kg, mg / kg, up to 2 mg / kg, up to 3 mg / kg, up to 4 mg / kg, up to 5 g / 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, up to 80 mg / kg, up to 90 mg / kg, up to 100 mg / kg or more.

[0307] However, in some embodiments, the dose of anti-HJV antibody may be within 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, 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.

[0308] 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 greater), for example, an amount sufficient to inhibit HJV-induced BMP signaling. In other embodiments, the antibody is 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 greater).

[0309] In some embodiments, the antibody can be administered parenterally. For example, the composition for parenteral administration can be administered by subcutaneous, intradermal, intravenous, intraperitoneal, intratumoral, intramuscular, intra-articular, intra-arterial, or infusion techniques. Further, it can be administered to the subject via an injectable depot route of administration, such as using an injectable depot or biodegradable substance and method for 1, 3, or 6 months.

[0310] In some embodiments, the antibody (e.g., anti-HJV antibody) is administered intravenously. In some embodiments, the antibody (e.g., anti-HJV antibody) is administered subcutaneously. In some embodiments, subcutaneous administration of the anti-HJV antibody results in similar bioavailability compared to intravenous administration of the same antibody at the same dose.

[0311] In some embodiments, subcutaneous administration of the anti-HJV antibody results in equivalent pharmacodynamic effects (e.g., decrease in circulating hepcidin-25 level, increase in TSAT%, and / or increase in serum iron level) at a lower maximum concentration (C max ) of the anti-HJV antibody compared to intravenous administration of the same antibody. C max is the maximum (or peak) serum concentration of the drug (e.g., anti-HJV antibody) after the drug has been administered and before the administration of a second dose. In some embodiments, a low C maxAchieving this minimizes undesirable increases in the serum iron response and / or the potential for off-target effects of the antibody (e.g., binding to RGMa). In some embodiments, blunting C is achieved by subcutaneous administration of anti-HJV antibody. max This avoids an undesirable and sharp increase in the serum iron response. In some embodiments, subcutaneous administration of anti-HJV antibody slows down C max This reduces the off-target effects of the antibody. In some embodiments, this is achieved by subcutaneous administration. max This is achieved by intravenous administration of anti-HJV antibodies. max Lower by 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%.

[0312] For intravenous injection, water-soluble antibodies can be administered by drip infusion, in which a pharmaceutical formulation containing the antibody and physiologically acceptable excipients 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, dimethylacetamide, dimethyformamide, ethyl lactate, ethyl carbonate, isopropyl myristate, ethanol, and polyols (such as glycerol, propylene glycol, and liquid polyethylene glycol). In some cases, antibody preparations in preferred soluble salt forms, such as sterile formulations, can be administered dissolved in pharmaceutical excipients, such as water for injection, 0.9% saline, or 5% glucose solution.

[0313] In one embodiment, the antibody is administered via site-specific or targeted local delivery technology. Examples of site-specific or targeted local delivery technologies include various implantable depot sources or local delivery catheters for the antibody, such as infusion catheters, indwelling catheters, or needle catheters, synthetic grafts, outer membrane wraps, shunts and stents, or other implantable devices, site-specific carriers, direct injection, or direct application. See, for example, PCT International Publication No. 00 / 53211 and U.S. Patent No. 5,981,568.

[0314] In some embodiments, more than one antibody, or a combination of an antibody and another suitable therapeutic agent, may be administered to the subject requiring treatment. Antibodies may also be used in combination with other agents that help enhance and / or complement the efficacy of the agent. The efficacy of the treatment against the target disease / disorder may be evaluated by methods well known in the art.

[0315] The anti-HJV antibodies and associated treatment methods described herein may be used in combination with other types of therapies for the target diseases or disorders disclosed herein. In this context, the antibody compositions and therapeutic agents may be administered either concurrently or sequentially. Examples include chemotherapy, immunotherapy (e.g., therapy with other hepcidin antagonists), surgery, radiation, gene therapy, or anti-infective therapy. Such therapies may be administered concurrently or sequentially (in any order) with the treatments described herein.

[0316] For example, combination therapy may include anti-HJV antibodies and pharmaceutical compositions described herein, formulated and / or administered in combination with at least one additional therapeutic agent. Such combination therapy may advantageously utilize lower doses of the therapeutic agents administered, thereby preventing potential toxicities or complications associated with various monotherapies. Furthermore, the additional therapeutic agents disclosed herein may act in addition to or separately from the hepcidin / BMP pathway, thereby enhancing and / or cooperating with the effects of the anti-HJV antibody.

[0317] In some embodiments, the disclosure relates to compositions (e.g., anti-HJV antibodies and compositions thereof) and methods for treating myelofibrosis-associated anemia. Myelofibrosis-associated anemia may be characterized as mild to moderate or severe anemia according to appropriate diagnostic threshold parameters. For example, in some embodiments, myelofibrosis-associated anemia is characterized based on hemoglobin (Hgb) levels, and the severity of anemia increases with decreasing Hgb levels. In some embodiments, mild to moderate anemia is associated with Hgb levels of at least 8 g / dL and below the lower limit of normal (e.g., between about 8 g / dL and about 14 g / dL, between about 8 g / dL and about 12 g / dL, between about 8 g / dL and about 10 g / dL, between about 10 g / dL and about 14 g / dL, or between about 10 g / dL and about 12 g / dL). In some embodiments, severe anemia is associated with Hgb levels of approximately 8 g / dL or less (e.g., between approximately 2 g / dL and approximately 8 g / dL, between approximately 4 g / dL and approximately 8 g / dL, or between approximately 6 g / dL and approximately 8 g / dL). In some embodiments, severe anemia is associated with red blood cell transfusion dependence. In some embodiments, severe anemia is associated with red blood cell transfusion independence resulting from a therapeutic intervention (e.g., therapeutic recovery from a transfusion-dependent state), and the subject depends on ongoing therapeutic procedures to maintain transfusion independence.

[0318] In some embodiments, the Disclosure provides compositions and methods for treating subjects known to or suspected to have a blood disorder characterized by low systemic iron levels. In some embodiments, the subjects have myelofibrosis and / or one or more conditions resulting from myelofibrosis as otherwise described herein. In some embodiments, the subjects are red blood cell transfusion dependent. In some embodiments, the subjects are red blood cell transfusion independent.

[0319] Therefore, in some embodiments, subjects requiring treatment in accordance with this disclosure have previously received therapeutic intervention for a blood disorder. In some embodiments, subjects have previously undergone surgical procedures to treat one or more blood disorders. In some embodiments, subjects have previously undergone splenectomy. In some embodiments, subjects have previously received therapeutic agents to treat one or more blood disorders.

[0320] In some embodiments, the subject is an erythropoietin stimulant that has been previously provided. In some embodiments, the erythropoietin stimulant is selected from the group consisting of danazol, prednisone, thalidomide, lenalidomide, and pomalidomide.

[0321] In some embodiments, the subject is a JAK-STAT pathway inhibitor, which has been previously given. 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 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, momerotinib, pacritinib, INCB039110, AG490, and PpYLKTK.

[0322] In some embodiments, the subject has previously received a growth factor ligand trap. In some embodiments, the growth factor ligand trap is a transformed growth factor beta (TGF-β) ligand trap. In some embodiments, the TGF-β ligand trap is sotatercept or raspatercept. In some embodiments, the subject has previously received an antifibrotic agent. In some embodiments, the antifibrotic agent is PRM-151.

[0323] In some embodiments, subjects requiring treatment according to the Disclosure continue to receive therapeutic treatment for blood disorders. The Disclosure therefore provides compositions and methods for treating myelofibrosis and / or one or more conditions resulting from myelofibrosis by administering a hepcidin antagonist (e.g., an anti-HJV antibody and its compositions) to subjects requiring it in combination with one or more therapeutic treatments for blood disorders.

[0324] In some embodiments, subjects are administered a hemoduveline antagonist (e.g., an anti-HJV antibody and its composition) in combination with an erythropoietin stimulant. In some embodiments, the erythropoietin stimulant is selected from the group consisting of danazol, prednisone, thalidomide, lenalidomide, and pomalidomide.

[0325] In some embodiments, subjects are administered a hemoduvelin antagonist (e.g., an anti-HJV antibody and its composition) 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 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, momerotinib, pacritinib, INCB039110, AG490, and PpYLKTK. In some embodiments, subjects are administered a hemoduvelin antagonist (e.g., an anti-HJV antibody and its composition) in combination with ruxolitinib.

[0326] In some embodiments, hemoduvelin antagonists (e.g., anti-HJV antibodies and their compositions) reduce the degree to which a 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 monotherapy may be characterized by a deficiency in the ability of the blood to transport oxygen, a deficiency in red blood cells, a deficiency in hemoglobin, and / or a deficiency in total blood volume compared to the subject's pre-treatment state. Thus, in some embodiments, hemoduvelin antagonists (e.g., anti-HJV antibodies and their compositions) reduce the degree to which a subject exhibits an anemic response to a JAK-STAT pathway inhibitor selected from the group consisting of ruxolitinib, fedratinib, momerotinib, pacritinib, INCB039110, AG490, and PpYLKTK. In some embodiments, a hemoduvelin antagonist (e.g., an anti-HJV antibody and its composition) reduces the degree to which a subject exhibits an anemic response to ruxolitinib administration.

[0327] In some embodiments, subjects are administered a hemoduvelin antagonist (e.g., an anti-HJV antibody and its composition) 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, subjects are administered a hemoduvelin antagonist (e.g., an anti-HJV antibody and its composition) in combination with an antifibrotic agent. In some embodiments, the antifibrotic agent is PRM-151.

[0328] The success of the treatment of a subject according to this 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 its composition) treatment is evaluated based on the hepcidin level (e.g., circulating hepcidin-25 level) in the subject. For example, in some embodiments, the baseline hepcidin level (e.g., circulating hepcidin-25 level) in the subject is determined (e.g., before treatment with anti-HJV antibody or in the absence of anti-HJV antibody treatment at any other time of determination) and compared to the pre-treatment hepcidin level (e.g., circulating hepcidin-25 level) in the subject. In some embodiments, the subject is successfully treated, and the anti-HJV antibody reduces the hepcidin level (e.g., circulating hepcidin-25 level) in the subject by about 1 ng / mL to about 300 ng / mL. In some embodiments, an anti-HJV antibody reduces hepcidin levels (e.g., circulating hepcidin-25 levels) in a subject by approximately 1 ng / mL to approximately 200 ng / mL, approximately 1 ng / mL to approximately 100 ng / mL, approximately 1 ng / mL to approximately 50 ng / mL, approximately 1 ng / mL to approximately 10 ng / mL, approximately 10 ng / mL to approximately 100 ng / mL, or approximately 10 ng / mL to approximately 50 ng / mL. In some embodiments, the disclosure provides a method for reducing hepcidin (e.g., circulating hepcidin-25 levels) in a subject having myelofibrosis. In some embodiments, the administration reduces 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 after administration.In some embodiments, the administration reduces hepcidin (e.g., circulating hepcidin-25 level) 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 99%, or 100% of hepcidin (e.g., circulating hepcidin-25 level) compared to the hepcidin (e.g., circulating hepcidin-25 level) in the subject before administration.

[0329] In some embodiments, anti-HJV antibody treatment is evaluated based on serum ferritin levels in the subject. For example, in some embodiments, baseline serum ferritin levels in the subject are determined (e.g., before treatment with anti-HJV antibody or in the absence of anti-HJV antibody treatment at other time points) and compared to post-treatment serum ferritin levels in the subject. In some embodiments, the subject is successfully treated, and the anti-HJV antibody reduces serum ferritin levels in the subject by approximately 1 ng / mL to approximately 200 ng / mL. In some embodiments, the anti-HJV antibody reduces serum ferritin levels in the subject by approximately 1 ng / mL to approximately 100 ng / mL, approximately 1 ng / mL to approximately 50 ng / mL, approximately 1 ng / mL to approximately 25 ng / mL, approximately 1 ng / mL to approximately 10 ng / mL, approximately 10 ng / mL to approximately 100 ng / mL, or approximately 10 ng / mL to approximately 50 ng / mL.

[0330] In some embodiments, anti-HJV antibody treatment is evaluated based on the serum hemoglobin level of the subject. For example, in some embodiments, the baseline serum hemoglobin level of the subject is determined (e.g., before treatment with anti-HJV antibody or in the absence of anti-HJV antibody treatment at any other time) and compared to the serum hemoglobin level of the subject after treatment. In some embodiments, the subject is successfully treated, and the anti-HJV antibody reduces the serum hemoglobin level of the subject by approximately 0.01 g / dL to approximately 5 g / dL. In some embodiments, anti-HJV antibodies reduce serum ferritin levels in subjects by approximately 0.01 g / dL to 1 g / dL, 0.1 g / dL to 5 g / dL, 1 g / dL to 5 g / dL, 0.01 g / dL to 0.1 g / dL, 0.5 g / dL to 2.5 g / dL, or 0.1 g / dL to 1 g / dL.

[0331] In some embodiments, other hepcidin antagonists may be hepcidin inhibitors. In some embodiments, the hepcidin inhibitor is a molecule that specifically binds to hepcidin (e.g., an antibody, antikalin, or aptamer). Examples of molecules that specifically bind to hepcidin include, but are not limited to, PRS-080, LY2787106, NOX-H94 (Lexaptepid Pegol), 12B9m, LS-B4534, lipocalin mutant proteins, and hNGAL mutant proteins (see also U.S. Patent Nos. 8,629250; 9315577; 9051382; 9657098; 9610356; 8530619, and U.S. Patent Publication Nos. 2015 / 0291675, 2018 / 0057812, and 2017 / 0247448, incorporated herein by reference).

[0332] In some embodiments, other hepcidin antagonists may be molecules that specifically bind to ferroportin (e.g., antibodies, antikalin, or aptamers). In some embodiments, the molecule that specifically binds to ferroportin is LY2928057. Molecules that bind to ferroportin and inhibit hepcidin binding without affecting ferroportin activity are described (see also U.S. Patent No. 8,183,346, incorporated herein by reference). Hepcidin inhibitors are chemical modifier compounds that modify hepcidin or ferroportin to inhibit the hepcidin-ferroportin binding interaction. For example, in some embodiments, the hepcidin inhibitor is fursultiamine [see, e.g., Fung and Nemeth. Haematologica. 2013 Nov; 98(11):1667-76].

[0333] In further embodiments, other hepcidin antagonists of the present disclosure may be HAMP antagonists that bind to HAMP or its transcription or translation product. Examples of such HAMP antagonists include, but are not limited to, antisense oligonucleotides, small molecule inhibitor compounds, and antibodies, antikalins, or aptamers that are specific to HAMP transcription or translation product. Antisense oligonucleotides may be single-stranded or double-stranded. As a non-limiting set of examples, antisense oligonucleotides may be, but are not limited to, small molecule interfering RNA (siRNA), microRNA (miRNA), short hairpin RNA (shRNA), dicer substrate interfering RNA (dsiRNA), short siRNA, or single-stranded siRNA. In some embodiments, double-stranded antisense oligonucleotides are RNAi oligonucleotides. Specific examples of HAMP antisense oligonucleotides include, but are not limited to, siHepcidin and XEN701 (see also U.S. Patent Application Publication 2016 / 0186172 and U.S. Patent Application Publication 2012 / 0115930, incorporated herein by reference).

[0334] In some embodiments, HAMP antagonists bind to or downmodulate molecules involved in the BMP-SMAD and / or JAK-STAT signaling pathways. In some embodiments, HAMP antagonists are antisense oligonucleotides, small molecule compounds, antibodies, antikalins, or aptamers. Examples of molecules targeted by HAMP antagonists of this disclosure include, but are not limited to, HFE, BMP6, BMP receptor, transferrin, transferrin receptor, SMAD1 / 5 / 8 and SMAD4, IL-6, IL-6 receptor, JAK1 / 2, and STAT3. In some embodiments, hepcidin antagonists of this disclosure are antagonists (e.g., inhibitors) of any one or more of these molecules.

[0335] In some embodiments, other hepcidin antagonists of the present disclosure may be BMP6 antagonists that reduce hepcidin function by binding to BMP6 and / or BMP receptors and inhibiting the activation of the BMP-SMAD signaling pathway. In some embodiments, the BMP6 antagonist is an ALK2 inhibitor, such as LDN-212854, LDN-193189, LDN-214117, BLU-782, and others disclosed in Hudson, L. et al., Novel Quinazolinone Inhibitors of ALK2 Flip between Alternate Binding Modes: Structure-Activity Relationship, Structural Characterization, Kinase Profiling, and Cellular Proof of Concept. Med. Chem. 2018, 61, 16, 7261-7272 and Carvalho D, et al., ALK2 inhibitors display beneficial effects in preclinical models of ACVR1 mutant diffuse intrinsic pontine glioma. Communications Biology volume 2, Article number: 156 (2019) (the relevant content of each of these is incorporated herein by reference). In some embodiments, the BMP6 antagonist is a soluble HJV polypeptide, including fragments and / or fusion molecules (e.g., HJV-Fc) described, for example, in U.S. Patent Application Publication No. 7,534,764; U.S. Patent Application Publication No. 2012 / 064076; and International Publication Nos. 2008 / 124768 and International Publication Nos. 2012 / 150973, which are incorporated herein by reference.Additional examples of BMP6 antagonists include, but are not limited to, TP-0184, FKBP12, twisted gastrulation protein, dolsomorphin, noggin, chordin, ventroptin, follistatin, follistatin-related gene (FLRG), heparin, sulfated glycosaminoglycan, and sclerostin domain-containing protein 1 (SOSTDC1). Additional examples of BMP6 antagonists that may be useful in certain ways provided herein are, for example, U.S. Patents No. 8,318,167; No. 9,556,251; No. 9,862,764; No. 10,202,356; No. 9,682,983; No. 8,507,501; No. 9,738,636; and No. 8,795,665; U.S. Patent Application Publication 2010 / 0093760; U.S. Patent Application Publication 2014 / 0199314; U.S. Patent Application Publication 2014 / 0086919; and U.S. Patent Application Publication 2016 / 026311, which are incorporated herein by reference. Provided in No. 7; U.S. Patent Application Publication No. 2016 / 0115167; U.S. Patent Application Publication No. 2017 / 0197968; U.S. Patent Application Publication No. 2017 / 0190705; U.S. Patent Application Publication No. 2017 / 0305883; U.S. Patent Application Publication No. 2018 / 0021340; and U.S. Patent Application Publication No. 2018 / 0118835; and PCT International Publication No. 2017 / 216724; International Publication No. 2018 / 136634; International Publication No. 2018 / 053234; International Publication No. 2018 / 185341; International Publication No. 2016 / 146651; and International Publication No. 2018 / 200855.

[0336] Efficient iron signaling via the BMP-SMAD signaling pathway involves cofactors, such as the diiron (Tf) sensor transferrin receptor 2 (TfR) to stimulate hepcidin expression (Figure 12). In some embodiments, the hepcidin antagonist of this disclosure is a transferrin antagonist that antagonizes hepcidin function by binding to transferrin and / or transferrin receptor 2, thereby inhibiting activation of the BMP-SMAD signaling pathway. In some embodiments, the transferrin antagonist is an antisense oligonucleotide that targets Tf and / or TfR, such as siTfR2 (see, for example, U.S. Patent No. 9,228,188, incorporated herein by reference).

[0337] As detailed above, STAT3 is a transcriptional regulator of HAMP expression. Therefore, in some embodiments, the HAMP antagonist is a JAK-STAT signaling pathway inhibitor. In some embodiments, the JAK-STAT signaling 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. Examples of JAK1 / 2 and STAT3 inhibitors include, but are not limited to, ruxolitinib, fedratinib, momerotinib, pacritinib, INCB039110, AG490, and PpYLKTK.

[0338] The JAK-STAT3 signaling pathway is activated by the inflammatory cytokine IL-6. Binding of IL-6 to the IL-6 receptor (IL-6R) triggers receptor dimerization on hepatocytes, leading to activation of the JAK-STAT3 signaling pathway (Figure 12). Therefore, in some embodiments, JAK-STAT signaling pathway inhibitors are IL-6 antagonists that bind to IL-6 and / or the IL-6 receptor, thereby antagonizing hepcidin function by inhibiting activation of the JAK-STAT3 signaling pathway. In some embodiments, the IL-6 antagonist is selected from the group consisting of infliximab, curcumin, 3,3'-diindolylmethane, tocilizumab, and siltuximab. Additional examples of IL-6 and IL-6R inhibitors that may be useful in certain methods provided herein are provided, for example, in U.S. Patent Application Publication 2017 / 0029499, incorporated herein by reference.

[0339] Any anti-HJV antibody disclosed herein may also be used to detect the presence of HJV (e.g., soluble HJV) in vitro or in vivo. Results obtained from such detection methods may be used for diagnostic purposes (e.g., diagnosis of HJV-associated diseases) or for scientific research purposes (e.g., identification of novel HJV-secreting cell types, study of the bioactivity and / or regulation of secreted HJV). For assay applications such as diagnostic applications, the anti-HJV antibodies described herein may be conjugated with a detectable label (e.g., an imaging agent, e.g., a contrast agent) for detecting the presence of HJV (e.g., soluble HJV) in vivo or in vitro. Where used herein, “conjugated” or “attached” means that two entities are associated with sufficient affinity to realize the therapeutic / diagnostic benefit of the association between the two entities. The association between the two entities may be direct or mediated by a linker, e.g., a polymer linker.

[0340] What is conjugated or attached can include, in addition to covalent or non-covalent bonds, other forms of association, such as capture, for example, capture of one entity on or within another entity, or capture of either or both entities on or within a third entity, such as a micelle.

[0341] In other embodiments, the anti-HJV antibodies described herein can be attached to a detectable label, which is a compound having the ability to emit a detectable signal either directly or indirectly, such that the aptamer can be detected, measured, and / or qualitatively evaluated in vitro or in vivo. Examples of such "detectable labels" are intended to include, but are not limited to, fluorescent labels, chemiluminescent labels, colorimetric labels, enzyme markers, radioisotopes, and affinity tags such as biotin. Such labels can be conjugated to the aptamer directly or indirectly by conventional methods.

[0342] In some embodiments, the detectable label is an agent suitable for detecting HJV-expressing cells in vitro and can be a radioactive molecule, radiopharmaceutical, or iron oxide particles. Suitable radioactive molecules for in vivo imaging are 122 I, 123 I, 124 I, 125 I, 131 I, 18 F, 75 Br, 76 Br, 77 Br, 211 At, 225 Ac, 177 Lu, 153 Sm, 186 Re, 188 Re, 67 Cu, 213 Bi, 212 Bi, 212 Pb, and 67 Ga, including but not limited to these. Exemplary radiopharmaceuticals suitable for in vivo imaging are 111 In oxyquinoline, 131 I sodium iodide,99 mTc mebrophenin, and 99 mTc red blood cells, 123 I. Sodium iodide, 99 mTC exametagym, 99 mTc large aggregated albumin, 99 mTc medronate, 99 mTC melthiatide, 99 mTc oxide, 99 mTC pentetate, 99 mTc Pertecnetate, 99 mTc sestamibi, 99 mTc sulfur colloid, 99 Contains mTc tetrophosmine, thallium-201, or xenon-133.

[0343] The reporting agent may also be a dye, such as a fluorophore, which is useful in detecting diseases mediated by HJV-expressing cells in tissue samples.

[0344] To perform an in vitro diagnostic assay, an anti-HJV antibody can be brought into contact with a sample suspected of containing HJV, such as HJV-expressing cells or soluble HJV in the disease microenvironment. The antibody and sample may be incubated for a suitable period of time under suitable conditions to allow the antibody to bind to the HJV antigen. Such an interaction can then be detected by routine methods, such as ELISA, histological staining, or FACS.

[0345] To perform an in vivo diagnostic assay, a suitable amount of anti-HJV antibody conjugated with a label (e.g., an imaging agent or contrast agent) may be administered to the subject requiring testing. The presence of the labeled antibody may be detected by routine methods based on the signal released from the label.

[0346] For scientific research assays, anti-HJV antibodies may be used to study the biological activity of HJV, to detect the presence of HJV in cells, and / or to modulate the effects of HJV. For example, a suitable amount of anti-HJV can be contacted with a sample suspected of producing HJV (e.g., a novel cell type not previously identified as an HJV-producing cell). The cells are permeabilized before contact with the anti-HJV antibody. The antibody and sample may be incubated for a suitable period of time under suitable conditions to allow the antibody to bind to the HJV antigen. Such interactions can then be detected by routine methods, e.g., ELISA, histological staining, or FACS.

[0347] VI. Kits for therapeutic and diagnostic applications This disclosure also provides kits for therapeutic or diagnostic applications disclosed herein. Such kits may include one or more containers containing an anti-HJV antibody, for example, one of those described herein.

[0348] In some embodiments, the kit may include instructions for use by any of the methods described herein. The included instructions may include instructions for administering anti-HJV antibodies to treat, delay the onset of, or mitigate the target diseases described herein. The kit may further include instructions for selecting individuals suitable for treatment based on the identification of whether the individuals have the target disease. In yet other embodiments, the instructions may include instructions for administering antibodies to individuals at risk of the target disease.

[0349] Instructions for the use of anti-HJV antibodies generally include information regarding the dosage, dosage schedule, and route of administration for the intended treatment. Containers may be unit doses, bulk packages (e.g., multi-dose packages), or partial unit doses. Instructions supplied in the kit of the present invention are typically written instructions on labels or package inserts (e.g., paper sheets included in the kit), but machine-readable instructions (e.g., instructions on magnetic or optical storage disks) are also acceptable.

[0350] Labels or package inserts indicate that the composition is used to treat a disease or disorder, delay the onset of the disease, and / or alleviate it. Instructions may be provided for carrying out any of the methods described herein.

[0351] The kit of the present invention is in suitable packaging. Suitable packaging includes, but is not limited to, vials, bottles, jars, and flexible packaging (e.g., sealed Mylar or plastic bags).

[0352] Packages for use in combination with specific devices, such as injection devices, such as minipumps, are also envisioned. The kit may have a sterile access port (for example, the container may be an intravenous solution bag or vial with a stopper that can be pierced by a subcutaneous needle). The container may also have a sterile access port (for example, the container may be an intravenous solution bag or vial with a stopper that can be pierced by a subcutaneous needle). At least one activator in the composition is an anti-HJV antibody as described herein.

[0353] The kit may provide additional components, such as buffers and decision information. Typically, the kit includes a container and labels or package inserts on or accompanying the container. In some embodiments, the present invention provides a product comprising the contents of the kit described above.

[0354] Kits for use in the detection of hemoduveline in a sample are also provided herein. Such kits may include any of the anti-HJV antibodies described herein. In some cases, the anti-HJV antibody may be conjugated with a detectable label described herein. Where used herein, “conjugated” or “attached” means that two entities are associated with sufficient affinity to realize the therapeutic / diagnostic benefit of the association between the two entities. The association between the two entities may be direct or mediated by a linker, such as a polymer linker. Conjugation or attachment may include other forms of association besides covalent or non-covalent bonding, such as capture, e.g., capture of one entity on or within another entity, or capture of either or both entities on or within a third entity, e.g., a micelle.

[0355] Alternatively or additionally, the kit may include a secondary antibody having the ability to bind to the anti-HJV antibody. The kit may further include instructions for using the anti-HJV antibody to detect hemoduvelin. [Examples]

[0356] [Example 1] Generation and characterization of anti-HJV antibodies Hybridoma clones capable of binding to human hemoduverin were identified from rats immunized with human hemoduverin. Anti-hemoduverin monoclonal antibodies (mAbs containing rat IgG1 / κ) were humanized by CDR transplantation (hHA antibodies with a constant region of hIgG1 possessing L234A and L235A mutations). Affinity-matured anti-HJV (e.g., HA-001 to HA-011) were subjected to in vitro yeast display assays. A general method for generating humanized affinity-matured anti-HJV antibodies is shown in Figure 1A.

[0357] The binding affinity of anti-HJV antibodies to soluble human RGMa, rat RGMa, and human RGMac was measured by BIAcore analysis. Table 4 shows the affinity of anti-hemoduvelin antibodies to human RGMa. Table 5 shows the affinity of anti-hemoduvelin antibodies to rat RGMa. Table 6 shows the affinity of anti-hemoduvelin antibodies to human RGMac.

[0358] [Table 4]

[0359] [Table 5]

[0360] [Table 6]

[0361] 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 of antibodies HA, hHA-004, hHA-008, hHA-009, and hHA-011 by BIAcore analysis are shown in Figures 1B-1G. Further testing of hHA-008 for its binding affinity to human RGMa, cynomolgus monkey RGMa, rat RGMa, human RGMc, cynomolgus monkey RGMc, rat RGMc, and human RGMb is shown in Table 7. hHA-008 showed high-affinity binding to human RGMa and RGMc, along with strong cross-reactivity to cynomolgus monkeys and rodent species.

[0362] [Table 7]

[0363] The RGMc(HJV) BMP reporter gene assay was used to screen anti-HJV antibodies and characterize their efficacy 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, which is responsible for the induction of iron hormone hepcidin gene expression. The principle of the HJV BMP reporter assay is shown in Figure 2A.

[0364] Using the BRE-Luc reporter gene vector first described by Korchynskyi and Dijke (J. Biol. Chem. 2002; 277:4883c), porcine kidney epithelial cell line LLC-PK1 was transiently transfected with or without co-transfection with the RGMa expression vector described 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 via the Smad1 / 5 / 8 signaling pathway, consistent with the role of RGMa as a BMP co-receptor. All RGM members (RGMa, RGMb, and RGMc) act as BMP co-receptors 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.

[0365] 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)]. 9 × 10⁶ cells were prepared to transfect the cells. 6100 HEK293 cells were plated in a 10 cm dish and incubated at 37°C and 5% CO2 for 6 hours. The cells were then transfected at 37°C and 5% CO2 for 16 hours using 20 μl PolyJet (SignaGen, catalog # SL100688) with a 2:1 PolyJet (μL):DNA (μg) ratio, specifically 5 μg pGL[luc2P / BRE / Hygro]DNA (Abbvie) and 5 μg pcDNA-hRGMc (Abbvie). The medium was replaced with fresh HEK293 growth medium after 6 hours. The cells were then trypsinized using TrpLE (ThermoFisher, catalog # 12605010), counted, and placed in a 96-well white assay plate (Thermo Scientific Nunclon delta F96, catalog # 136102) at a rate of 1 × 10⁶ cells. 5 Cells were plated in a cell / well. Cells were treated with anti-RGMc mAb, anti-BMP2 / 4 mAb (R&D Systems, catalog # MAB3552) (as a positive control), or isotype control mAb at 37°C and 5% CO2 for 16 hours. Luciferase activity was detected using the One-Glo Luciferase kit (Promega, catalog # E6120) and 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 antibody tested for inhibition of RGMc signaling in the BMP reporter assay is shown in Table 8.

[0366] [Table 8]

[0367] Furthermore, anti-HJV antibodies were tested for their ability to inhibit RGMa signaling in a BMP reporter assay. hHA-004 and hHA-011 showed potent inhibition of membrane-bound human RGMa in the RGMa BMP reporter gene assay, while hHA-008 and hHA-009 showed no inhibition 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 lower degree compared to hHA-004 and hHA-011. Figure 2C. Table 9 shows the IC50 (nM) of each antibody tested for inhibition of RGMa signaling in a BMP reporter assay.

[0368] [Table 9]

[0369] The data above demonstrates that the efficacy of anti-HJV antibodies in neutralizing membrane RGMc in BMP reporter assays correlates with their binding affinity to soluble RGMc. Table 10 summarizes the binding affinity of anti-HJV antibodies to soluble human RGMc, as well as their ability to inhibit membrane-bound RGMc and RGMa signaling.

[0370] [Table 10]

[0371] Antibodies containing hHA, hHA-004, hHA-008, hHA-008-QL, hHA-009, and hHA-011 were tested for nonspecific cell binding to HEK293 cells by FACS analysis. The data showed that hHA, as well as its sub-sub [Example 2]

[0372] Generation of hHA-008-QL IgG serum half-life (t 1 / 2 hHA-008-QL was developed to prolong the IgG half-life. Previous studies have shown that the neonatal Fc receptor (FcRn) protects IgG from catabolism, thereby increasing the serum half-life of IgG. Therefore, the Fc portion of hHA-008-QL was modified to have T250Q and M428L mutations (QL mutations) to enhance 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.

[0373] First, hHA-008-QL was tested for its RGMc binding ability, and the data showed that hHA-008-QL has comparable binding affinity to RGMc to hHA-008 (Table 11).

[0374] [Table 11] [Example 3]

[0375] Immunogenicity studies of hHA-008 and hHA-008-QL In a CD4+ T-cell response assay, hHA-008 and hHA-008-QL were tested for their immunogenicity using peripheral blood mononuclear cells (PBMCs) from 50 donors (representing all major HLA0DR and HLA-DQ haplotypes) loaded with either hHA-008 or hHA-008-QL. The results showed that only 4% of the 50 donors (2 / 50) exhibited a T-...

Claims

1. A pharmaceutical composition for use in the treatment of anemia in subjects with myelofibrosis, comprising an antibody that binds to human hemoduvelin (HJV), wherein the antibody comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 117 and a light chain containing the amino acid sequence of SEQ ID NO:

62.

2. A pharmaceutical composition for use in the treatment of anemia in subjects with myelofibrosis, comprising an antibody that binds to human hemoduvelin (HJV), wherein the antibody comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 61 and a light chain containing the amino acid sequence of SEQ ID NO:

62.

3. The pharmaceutical composition according to claim 1 or 2, wherein the pharmaceutical composition is formulated for subcutaneous injection.

4. The pharmaceutical composition according to any one of claims 1 to 3, wherein the anemia results from hepcidin expression induced by at least one pro-inflammatory cytokine.

5. A pharmaceutical composition according to any one of claims 1 to 4, wherein anemia in a subject is reduced by administration of the pharmaceutical composition.

6. The pharmaceutical composition according to claim 4, wherein at least one pro-inflammatory cytokine is interleukin-6 (IL-6).

7. The pharmaceutical composition according to any one of claims 1 to 6, wherein the target has a myelofibrosis-inducing mutation in JAK2, MPL, ASXL1, TET2, NFE2, SH2B3, SF3B1, or CALR.

8. A pharmaceutical composition according to any one of claims 1 to 7, wherein the target is primary myelofibrosis, myelofibrosis after essential thrombocythemia (ET), or myelofibrosis after polycythemia.

9. A pharmaceutical composition according to any one of claims 1 to 8, wherein the subject has a Dynamic International Prognostic Scoring System (DIPSS) score higher than 1.

10. A pharmaceutical composition according to any one of claims 1 to 8, wherein the subject has a DIPSS score of 3 to 4.

11. A pharmaceutical composition according to any one of claims 1 to 8, wherein the subject has a DIPSS score of at least 5.

12. The pharmaceutical composition according to any one of claims 1 to 11, wherein the subject exhibits a serum hemoglobin level lower than 10 g / dL.

13. A pharmaceutical composition according to any one of claims 1 to 12, wherein the subject exhibits a serum hemoglobin level lower than 8 g / dL.

14. A pharmaceutical composition according to any one of claims 1 to 13, wherein the target is red blood cell transfusion dependent.

15. The pharmaceutical composition according to any one of claims 1 to 14, wherein the subject has previously received a selective JAK-STAT inhibitor selected from the group consisting of ruxolitinib, fedratinib, pacritinib, INCB039110, AG490, and PpYLKTK.

16. The pharmaceutical composition according to any one of claims 1 to 15, wherein administration of the pharmaceutical composition reduces hepcidin-25 in a subject within 4 hours, 6 hours, 8 hours, 12 hours, 18 hours, 24 hours, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, or 2 weeks after administration.

17. The pharmaceutical composition according to any one of claims 1 to 16, wherein administration of the pharmaceutical composition reduces the hepcidin-25 in the subject by at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% compared to the hepcidin-25 in the subject before administration.

18. A pharmaceutical composition according to any one of claims 1 to 17, wherein the pharmaceutical composition is administered to a subject once a month.

19. (i) an antibody that binds to human hemoduverin (HJV), comprising a heavy chain containing the amino acid sequence of SEQ ID NO: 61 and a light chain containing the amino acid sequence of SEQ ID NO: 62; and / or (ii) An antibody that binds to human hemoduverin (HJV), comprising a heavy chain containing the amino acid sequence of SEQ ID NO: 117 and a light chain containing the amino acid sequence of SEQ ID NO:

62. A pharmaceutical composition for use in the treatment of anemia in subjects with myelofibrosis, including the following.

20. The pharmaceutical composition according to claim 19, wherein the pharmaceutical composition is formulated for subcutaneous injection.

21. The pharmaceutical composition according to claim 19 or 20, wherein the subject has anemia resulting from hepcidin expression induced by at least one pro-inflammatory cytokine.

22. A pharmaceutical composition according to any one of claims 19 to 21, wherein anemia in a subject is reduced by administration of the pharmaceutical composition.

23. The pharmaceutical composition according to claim 21, wherein the pro-inflammatory cytokine is interleukin-6 (IL-6).

24. The pharmaceutical composition according to any one of claims 19 to 23, wherein the target has a myelofibrosis-inducing mutation in any of JAK2, MPL, ASXL1, TET2, NFE2, SH2B3, SF3B1, or CALR.

25. The pharmaceutical composition according to any one of claims 19 to 24, wherein the subject has primary myelofibrosis, myelofibrosis after essential thrombocythemia (ET), or myelofibrosis after polycythemia.

26. The pharmaceutical composition according to any one of claims 19 to 25, wherein the subject has a Dynamic International Prognostic Scoring System (DIPSS) score higher than 1.

27. A pharmaceutical composition according to any one of claims 19 to 26, wherein the subject exhibits a serum hemoglobin level lower than 10 g / dL.

28. The pharmaceutical composition according to any one of claims 19 to 27, wherein the subject exhibits a serum hemoglobin level lower than 8 g / dL.

29. A pharmaceutical composition according to any one of claims 19 to 28, wherein the target is red blood cell transfusion dependent.

30. The pharmaceutical composition according to any one of claims 19 to 29, wherein the subject has previously received a selective JAK-STAT inhibitor selected from the group consisting of ruxolitinib, fedratinib, pacritinib, INCB039110, AG490, and PpYLKTK.

31. The pharmaceutical composition according to any one of claims 19 to 30, wherein administration of the pharmaceutical composition reduces hepcidin-25 within 4 hours, 6 hours, 8 hours, 12 hours, 18 hours, 24 hours, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, or 2 weeks after administration.

32. The pharmaceutical composition according to any one of claims 19 to 31, wherein administration of the pharmaceutical composition reduces hepcidin-25 in the subject by at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% compared to hepcidin-25 in the subject before administration.

33. A pharmaceutical composition according to any one of claims 19 to 32, wherein the pharmaceutical composition is administered to a subject once a month.