Methods for treating complement-mediated diseases and disorders

Abstract

To provide compounds of treating a complement-mediated disease or disorder in an individual.SOLUTION: There is provided a pharmaceutical composition comprising an anti-C1s antibody for use in the treatment of a complement-mediated disease or disorder in an individual in need thereof, comprising administering to the individual an effective amount of the anti-C1s antibody, wherein the serum level of the anti-C1s antibody after administration is at least about 20 μg / mL. The effective amount of the antibody is at least about 4 g, at least about 5 g, at least about 6 g, at least about 7 g, at least about 8 g, or at least about 9 g.SELECTED DRAWING: None

Description

[Technical field]

[0001] CROSS REFERENCE TO PRIOR-FILED APPLICATION This application claims the benefit of U.S. Provisional Application No. 62 / 471,190, filed March 14, 2017, and U.S. Provisional Application No. 62 / 553,059, filed August 31, 2017, all of which are incorporated by reference in their entireties.

[0002] Reference to an Electronically Submitted Sequence Listing The contents of the Sequence Listing submitted electronically with this application in an ASCII text file (Name: 4159.505PCO2_SeqListing.TXT; Size: 24,288 bytes; and Creation Date: March 13, 2018) are hereby incorporated by reference in their entirety. [Background technology]

[0003] The complement system is a well-known effector mechanism of the immune response that provides protection from pathogens and other harmful agents as well as recovery from injury. The complement pathway includes a number of proteins that typically exist in an inactive form in the body. The classical complement pathway is triggered by the activation of the first component of complement, called the C1 complex, which consists of the C1q, C1r, and C1s proteins. Upon binding of C1 to immune complexes or other activators, the C1s component, a diisopropyl fluorophosphate (DFP)-sensitive serine protease, cleaves complement components C4 and C2, initiating activation of the classical complement pathway. The classical complement pathway appears to play a role in many diseases and disorders. Summary of the Invention [Problem to be solved by the invention]

[0004] There is a need in the art for compounds that treat complement-mediated diseases or disorders. Additionally, there is a need for compounds that can detect or monitor such diseases or disorders. There is also a need for methods for producing and using such compounds and compositions thereof. [Means for solving the problem]

[0005] The present disclosure provides a method for treating a complement-mediated disease or disorder in an individual, and a method for inhibiting the activation of complement component C4 in an individual in need thereof.In some embodiments, the method comprises administering anti-C1s antibody to the individual at a fixed dose of 5.5g.In some embodiments, the anti-C1s antibody is administered to the individual every two weeks.

[0006] In some embodiments, the anti-C1s antibody comprises a light chain complementarity determining region (CDR) of the light chain variable region of the antibody comprising the amino acid sequence of SEQ ID NO:7 and a heavy chain CDR of the heavy chain variable region of the antibody comprising the amino acid sequence of SEQ ID NO:8.

[0007] In some embodiments, the anti-C1s antibody is humanized. In some embodiments, the humanized antibody comprises a humanized light chain framework region and / or a humanized heavy chain framework region.

[0008] In some embodiments, the anti-C1s antibody comprises: i) a light chain variable region comprising complementarity determining regions (CDRs) including CDR-L1 having the amino acid sequence of SEQ ID NO:1, CDR-L2 having the amino acid sequence of SEQ ID NO:2, CDR-L3 having the amino acid sequence of SEQ ID NO:3; and ii) CDRs including CDR-H1 having the amino acid sequence of SEQ ID NO:4, CDR-H2 having the amino acid sequence of SEQ ID NO:5, and CDR-H3 having the amino acid sequence of SEQ ID NO:6. The heavy chain variable region includes

[0009] In another embodiment, the anti-C1s antibody comprises: i) a light chain variable region comprising complementarity determining regions (CDRs) including CDR-L1 having the amino acid sequence of SEQ ID NO: 10, CDR-L2 having the amino acid sequence of SEQ ID NO: 11, and CDR-L3 having the amino acid sequence of SEQ ID NO: 3; and ii) a heavy chain variable region comprising CDRs including CDR-H1 having the amino acid sequence of SEQ ID NO: 12, CDR-H2 having the amino acid sequence of SEQ ID NO: 13, and CDR-H3 having the amino acid sequence of SEQ ID NO: 14.

[0010] In another embodiment, the anti-C1s antibody comprises: a VL region comprising the amino acid sequence set forth in SEQ ID NO:15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:18.

[0011] In another embodiment, the anti-C1s antibody comprises: a VL region comprising the amino acid sequence set forth in SEQ ID NO:15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:19.

[0012] In another embodiment, the anti-C1s antibody comprises: a VL region comprising the amino acid sequence set forth in SEQ ID NO:15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:20.

[0013] In another embodiment, the anti-C1s antibody comprises: a VL region comprising the amino acid sequence set forth in SEQ ID NO:15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:21.

[0014] In another embodiment, the anti-C1s antibody comprises: a VL region comprising the amino acid sequence set forth in SEQ ID NO:16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:18.

[0015] In another embodiment, the anti-C1s antibody comprises: a VL region comprising the amino acid sequence set forth in SEQ ID NO:16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:19.

[0016] In another embodiment, the anti-C1s antibody comprises: a VL region comprising the amino acid sequence set forth in SEQ ID NO:16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:20.

[0017] In another embodiment, the anti-C1s antibody comprises: a VL region comprising the amino acid sequence set forth in SEQ ID NO:16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:21.

[0018] In another embodiment, the anti-C1s antibody comprises: a VL region comprising the amino acid sequence set forth in SEQ ID NO:17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:18.

[0019] In another embodiment, the anti-C1s antibody comprises: a VL region comprising the amino acid sequence set forth in SEQ ID NO:17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:19.

[0020] In another embodiment, the anti-C1s antibody comprises: a VL region comprising the amino acid sequence set forth in SEQ ID NO:17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:20.

[0021] In another embodiment, the anti-C1s antibody comprises: a VL region comprising the amino acid sequence set forth in SEQ ID NO:17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:21.

[0022] In some embodiments of the present disclosure, the anti-C1s antibody comprises a heavy chain constant region of isotype IgG1, IgG2, IgG3, or IgG4. In some embodiments, the anti-C1s antibody is selected from the group consisting of a Fab fragment, a F(ab')2 fragment, a scFv, and an Fv.

[0023] In some embodiments, administration of the anti-C1s antibody is via subcutaneous, intravenous, or intramuscular administration.

[0024] In some embodiments, a method for treating a complement-mediated disease or disorder in an individual includes: a) administering a first dose of an anti-C1s antibody on day 1; b) administering a second dose of an anti-C1s antibody on day 8; and c) administering an anti-C1s antibody every other week after the dose on day 8.

[0025] The present disclosure also provides a method of inhibiting activation of complement component C4 in an individual in need thereof, comprising administering an anti-C1s antibody to the individual, where the anti-C1s antibody is administered in an amount of 5.5 g. In other embodiments, the present disclosure provides a method of inhibiting activation of complement component C4 in an individual in need thereof, comprising administering an anti-C1s antibody to the individual, where if the individual weighs less than about 75 kg, the anti-C1s antibody is administered in an amount of 6.5 g. In some embodiments, the present disclosure provides a method of inhibiting activation of complement component C4 in an individual in need thereof, comprising administering an anti-C1s antibody to the individual, where if the individual weighs about 75 kg or more, the anti-C1s antibody is administered in an amount of 7.5 g.

[0026] In some embodiments, the anti-C1s antibody is administered to the individual every two weeks.

[0027] In some embodiments, the anti-C1s antibody comprises a light chain complementarity determining region (CDR) of the light chain variable region of the antibody comprising the amino acid sequence of SEQ ID NO:7 and a heavy chain CDR of the heavy chain variable region of the antibody comprising the amino acid sequence of SEQ ID NO:8.

[0028] In some embodiments, the anti-C1s antibody is humanized. In some embodiments, the humanized antibody comprises a humanized light chain framework region and / or a humanized heavy chain framework region.

[0029] In some embodiments, the anti-C1s antibody comprises: a) a light chain variable region comprising i) complementarity determining regions (CDRs) including CDR-L1 having the amino acid sequence of SEQ ID NO:1, CDR-L2 having the amino acid sequence of SEQ ID NO:2, and CDR-L3 having the amino acid sequence of SEQ ID NO:3; and ii) a heavy chain variable region comprising CDRs including CDR-H1 having the amino acid sequence of SEQ ID NO:4, CDR-H2 having the amino acid sequence of SEQ ID NO:5, and CDR-H3 having the amino acid sequence of SEQ ID NO:6.

[0030] In some embodiments, the anti-C1s antibody comprises: i) a light chain variable region comprising complementarity determining regions (CDRs) including CDR-L1 having the amino acid sequence of SEQ ID NO: 10, CDR-L2 having the amino acid sequence of SEQ ID NO: 11, and CDR-L3 having the amino acid sequence of SEQ ID NO: 3; and ii) a heavy chain variable region comprising CDRs including CDR-H1 having the amino acid sequence of SEQ ID NO: 12, CDR-H2 having the amino acid sequence of SEQ ID NO: 13, and CDR-H3 having the amino acid sequence of SEQ ID NO: 14.

[0031] In another embodiment, the anti-C1s antibody comprises: a VL region comprising the amino acid sequence set forth in SEQ ID NO:15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:18.

[0032] In another embodiment, the anti-C1s antibody comprises: a VL region comprising the amino acid sequence set forth in SEQ ID NO:15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:19.

[0033] In another embodiment, the anti-C1s antibody comprises: a VL region comprising the amino acid sequence set forth in SEQ ID NO:15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:20.

[0034] In another embodiment, the anti-C1s antibody comprises: a VL region comprising the amino acid sequence set forth in SEQ ID NO:15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:21.

[0035] In another embodiment, the anti-C1s antibody comprises: a VL region comprising the amino acid sequence set forth in SEQ ID NO:16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:18.

[0036] In another embodiment, the anti-C1s antibody comprises: a VL region comprising the amino acid sequence set forth in SEQ ID NO:16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:19.

[0037] In another embodiment, the anti-C1s antibody comprises: a VL region comprising the amino acid sequence set forth in SEQ ID NO:16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:20.

[0038] In another embodiment, the anti-C1s antibody comprises: a VL region comprising the amino acid sequence set forth in SEQ ID NO:16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:21.

[0039] In another embodiment, the anti-C1s antibody comprises: a VL region comprising the amino acid sequence set forth in SEQ ID NO:17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:18.

[0040] In another embodiment, the anti-C1s antibody comprises: a VL region comprising the amino acid sequence set forth in SEQ ID NO:17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:19.

[0041] In another embodiment, the anti-C1s antibody comprises: a VL region comprising the amino acid sequence set forth in SEQ ID NO:17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:20.

[0042] In another embodiment, the anti-C1s antibody comprises: a VL region comprising the amino acid sequence set forth in SEQ ID NO:17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:21.

[0043] In some embodiments of the present disclosure, the anti-C1s antibody comprises a heavy chain constant region of isotype IgG1, IgG2, IgG3, or IgG4. In some embodiments of the present disclosure, the anti-C1s antibody is selected from the group consisting of a Fab fragment, a F(ab')2 fragment, a scFv, and an Fv.

[0044] In some embodiments, administration of the anti-C1s antibody is via subcutaneous, intravenous, or intramuscular administration.

[0045] In some embodiments, a method for inhibiting activation of complement component C4 in an individual in need thereof comprises: a) administering a first dose of an anti-C1s antibody on day 1; b) administering a second dose of an anti-C1s antibody on day 8; and c) administering an anti-C1s antibody every other week after the dose on day 8.

[0046] The present disclosure also provides a method of treating a complement-mediated disease or disorder in a subject in need thereof, comprising administering to the subject an effective amount of an anti-C1s antibody, wherein the serum concentration of the anti-C1s antibody following administration is at least about 20 μg / mL, at least about 25 μg / mL, at least about 30 μg / mL, at least about 35 μg / mL, at least about 40 μg / mL, at least about 45 μg / mL, at least about 50 μg / mL, at least about 55 μg / mL, at least about 60 μg / mL, at least about 65 μg / mL, at least about 70 μg / mL, at least about 75 μg / mL, at least about 80 μg / mL, at least about 85 μg / mL, at least about 90 μg / mL, at least about 95 μg / mL, or at least about 100 μg / mL.

[0047] In some embodiments, the serum concentration of anti-C1s antibody after administration is between about 20 μg / mL and about 100 μg / mL, between about 20 μg / mL and about 90 μg / mL, between about 20 μg / mL and about 80 μg / mL, between about 20 μg / mL and about 70 μg / mL, between about 20 μg / mL and about 60 μg / mL, between about 20 μg / mL and about 50 μg / mL, between about 20 μg / mL and about 40 μg / mL, or between about 20 μg / mL and about 30 μg / mL.

[0048] In some embodiments, serum concentrations of anti-C1s antibodies are measured by direct enzyme-linked immunosorbent assay (ELISA).

[0049] In some embodiments, an effective amount of an anti-C1s antibody is at least about 60 mg / kg, at least about 65 mg / kg, at least about 70 mg / kg, at least about 75 mg / kg, at least about 80 mg / kg, at least about 85 mg / kg, at least about 90 mg / kg, at least about 95 mg / kg, at least about 100 mg / kg, at least about 105 mg / kg, at least about 110 mg / kg, at least about 115 mg / kg, at least about 120 mg / kg, at least about 125 mg / kg, or at least about 130 mg / kg. In another embodiment, the effective amount of anti-C1s antibody is about 4 g to about 10 g.

[0050] In some embodiments, the effective amount is between about 60 mg / kg and about 100 mg / kg, between about 60 mg / kg and about 95 mg / kg, between about 60 mg / kg and about 90 mg / kg, between about 60 mg / kg and about 85 mg / kg, between about 60 mg / kg and about 80 mg / kg, between about 60 mg / kg and about 75 mg / kg, between about 60 mg / kg and about 70 mg / kg, or between about 60 mg / kg and about 65 mg / kg. In other embodiments, the effective amount is between about 4 g and about 10 g, between about 5 g and about 8 g, between about 5.5 g and about 7.5 g, between about 6.5 g and about 7.5 g, or between about 6.5 g and about 8.5 g. In some embodiments, an effective amount is between about 4 g and about 9 g, between about 5 g and about 8 g, between about 5.5 g and about 7.5 g, between about 6 g and about 8 g, or between about 6.5 g and about 7.5 g.

[0051] In some embodiments, the effective amount is about 60 mg / kg, about 65 mg / kg, about 70 mg / kg, about 75 mg / kg, about 80 mg / kg, about 85 mg / kg, about 90 mg / kg, about 95 mg / kg, about 100 mg / kg, about 105 mg / kg, about 110 mg / kg, about 115 mg / kg, about 120 mg / kg, about 125 mg / kg, about 130 mg / kg, about 135 mg / kg, about 140 mg / kg, about 145 mg / kg, or about 150 mg / kg. In some embodiments, the effective amount is about 4 g, about 4.5 g, about 5 g, about 5.5 g, about 6 g, about 6.5 g, about 7 g, about 7.5 g, about 8 g, about 8.5 g, about 9 g, about 9.5 g, or about 10 g.

[0052] In some embodiments, the anti-C1s antibody is administered at a dosing interval of 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or 31 days.

[0053] In some embodiments, the anti-C1s antibody is administered at dosing intervals of 1 week, 2 weeks, 3 weeks, 4 weeks, or 1 month.

[0054] In some embodiments, the anti-C1s antibody increases the number of reticulocytes in the subject's blood following administration.

[0055] The present disclosure also provides a method for increasing the number of reticulocytes in the blood of a subject in need thereof, comprising administering to the subject an effective amount of an anti-C1s antibody.

[0056] In some embodiments, the anti-C1s antibody increases the number of reticulocytes in the subject's blood by at least about 1.1-fold, at least about 1.2-fold, at least about 1.3-fold, at least about 1.4-fold, at least about 1.5-fold, at least about 1.6-fold, at least about 1.7-fold, at least about 1.8-fold, at least about 1.9-fold, at least about 2.0-fold, at least about 2.1-fold, at least about 2.2-fold, at least about 2.3-fold, at least about 2.4-fold, at least about 2.5-fold, at least about 2.6-fold, at least about 2.7-fold, at least about 2.8-fold, at least about 2.9-fold, at least about 3.0-fold, at least about 4-fold, at least about 5-fold, at least about 6-fold, at least about 7-fold, at least about 8-fold, at least about 9-fold, or at least about 10-fold after administration. In some embodiments, the anti-C1s antibody increases the number of reticulocytes in the subject's blood within about 24 hours of administration.

[0057] In some embodiments of the present disclosure, the anti-C1s antibody increases the level of hemoglobin in the subject. In some embodiments, the anti-C1s antibody increases the level of hemoglobin in the subject by at least about 1.0 g / dL, 1.1 g / dL, 1.2 g / dL, 1.3 g / dL, 1.4 g / dL, 1.5 g / dL, 1.6 g / dL, 1.7 g / dL, 1.8 g / dL, 1.9 g / dL, 2.0 g / dL, 2.1 g / dL, 2.2 g / dL, 2.3 g / dL, 2.4 g / dL, 2.5 g / dL, 2.6 g / dL, 2.7 g / dL, 2.8 g / dL, 2.9 g / dL, 3.0 g / dL, 3.1 g / dL, 3.2 g / dL, 3.3 g / dL, 3.4 g / dL, 3.5 g / dL, 3.6 g / dL, 3.7 g / dL, 3.8 g / dL, 3.9 g / dL, 4.0 g / dL, 4.1 g / dL, 4.2 g / dL, 4.3 g / dL, 4.4 g / dL, 4.5 g / dL, 4.6 g / dL, 4.7 g / dL, 4.8 g / dL, 4.9 ... g / dL, 3.4g / dL, 3.5g / dL, 3.6g / dL, 3.7g / dL, 3.8g / dL, 3.9g / dL, 4.0g / dL, 4.1g / dL, 4.2g / dL, 4.3g / dL, 4.4g / dL, 4.5g / dL, 4.6g / dL, 4.7g / d L, increase to 4.8g / dL, 4.9g / dL, 5.0g / dL, 5.1g / dL, 5.2g / dL, 5.3g / dL, 5.4g / dL, 5.5g / dL, 5.6g / dL, 5.7g / dL, 5.8g / dL, 5.9g / dL, or 6.0g / dL. In some embodiments, the hemoglobin level in the subject is increased by at least 1.6 g / dL within 7 days of administration. In some embodiments, the hemoglobin level in the subject is increased to 3.9 g / dL within 6 weeks of administration.

[0058] In some embodiments of the present disclosure, the anti-C1s antibody reduces the percentage of C3d-positive red blood cells in the blood of the subject.In some embodiments, the percentage of C3d-positive red blood cells in the blood of the subject is reduced by at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% compared to the percentage of C3d-positive red blood cells in the blood of the subject before administration.In other embodiments, the percentage of C3d-positive red blood cells in the blood of the subject is reduced to about 0%, about 1%, about 2%, about 3%, about 4%, or about 5%.

[0059] In some embodiments, the anti-C1s antibody reduces the level of bilirubin in the subject. In some embodiments, the level of bilirubin in the subject is about 2.5 mg / dL, 2. Decreased to less than 4 mg / dL, 2.3 mg / dL, 2.2 mg / dL, 2.1 mg / dL, 2.0 mg / dL, 1.9 mg / dL, 1.8 mg / dL, 1.7 mg / dL, 1.6 mg / dL, 1.5 mg / dL, 1.4 mg / dL, 1.3 mg / dL, 1.2 mg / dL, 1.1 mg / dL, 1.0 mg / dL, 0.9 mg / dL, 0.8 mg / dL, 0.7 mg / dL, 0.6 mg / dL, 0.5 mg / dL, 0.4 mg / dL, 0.3 mg / dL, 0.2 mg / dL, or 0.1 mg / dL.

[0060] In some embodiments of the present disclosure, the anti-C1s antibody comprises: a) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 18; b) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 19; c) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 20; d) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 21; e) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 18; f) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16; and the amino acid sequence set forth in SEQ ID NO: 19. VH region; g) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 20; h) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 21; i) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 18; j) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 19; k) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 20; or l) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 21.

[0061] In some embodiments of the present disclosure, the anti-C1s antibody comprises: a) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 18; b) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 19; c) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 20; d) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 21; e) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 18; f) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 19 and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 21; i) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 18; j) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 19; k) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 20; or l) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 21.

[0062] In some embodiments of the present disclosure, the anti-C1s antibody comprises: a) a light chain variable region and a heavy chain variable region comprising i) a CDR-L1 having the amino acid sequence of SEQ ID NO: 1, a CDR-L2 having the amino acid sequence of SEQ ID NO: 2, and a CDR-L3 having the amino acid sequence of SEQ ID NO: 3; and ii) a heavy chain variable region (VH) comprising a CDR-H1 having the amino acid sequence of SEQ ID NO: 4, a CDR-H2 having the amino acid sequence of SEQ ID NO: 5, and a CDR-H3 having the amino acid sequence of SEQ ID NO: 6; or b) i) a CDR-L1 having the amino acid sequence of SEQ ID NO: 10, ii) a light chain variable region comprising a CDR-L2 having the amino acid sequence of SEQ ID NO:11, a CDR-L3 having the amino acid sequence of SEQ ID NO:3; and ii) a heavy chain variable region comprising a CDR-H1 having the amino acid sequence of SEQ ID NO:12, a CDR-H2 having the amino acid sequence of SEQ ID NO:13, and a CDR-H3 having the amino acid sequence of SEQ ID NO:14.

[0063] In some embodiments of the disclosure, the anti-C1s antibody comprises: a) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 18; b) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 19; c) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 20; d) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 21; e) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 18; f) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16; and the amino acid sequence set forth in SEQ ID NO: 19 a VH region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 18; j) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 19; k) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 20; or l) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 21.

[0064] In some embodiments, the anti-C1s antibody comprises a heavy chain constant region of isotype IgG1, IgG2, IgG3, or IgG4.

[0065] In some embodiments, the anti-C1s antibody is selected from the group consisting of a Fab fragment, a F(ab')2 fragment, a scFv, and an Fv.

[0066] In some embodiments, administration is via subcutaneous, intravenous, or intramuscular administration.

[0067] Embodiment E1. A method of treating a complement-mediated disease or disorder in an individual, comprising administering to the individual an anti-C1s antibody, wherein the anti-C1s antibody is administered in an amount of 5.5 g.

[0068] E2. The method of E1, wherein the anti-C1s antibody is administered to the individual every two weeks.

[0069] E3. The method described in E1 or E2, wherein the anti-C1s antibody comprises a light chain complementarity determining region (CDR) of the antibody light chain variable region comprising the amino acid sequence of SEQ ID NO:7 and a heavy chain CDR of the antibody heavy chain variable region comprising the amino acid sequence of SEQ ID NO:8.

[0070] E4. The method of any one of E1 to E3, wherein the anti-C1s antibody is humanized.

[0071] E5. The method of E4, wherein the humanized antibody comprises a humanized light chain framework region and / or a humanized heavy chain framework region.

[0072] E6. Anti-C1s antibodies: a) a light chain variable region comprising a complementarity determining region (CDR) including i) a CDR-L1 having the amino acid sequence of SEQ ID NO:1, a CDR-L2 having the amino acid sequence of SEQ ID NO:2, and a CDR-L3 having the amino acid sequence of SEQ ID NO:3; and ii) a light chain variable region comprising a CDR having the amino acid sequence of SEQ ID NO:4. a heavy chain variable region comprising CDRs comprising CDR-H1 having the amino acid sequence of SEQ ID NO:5, CDR-H2 having the amino acid sequence of SEQ ID NO:6, and CDR-H3 having the amino acid sequence of SEQ ID NO:7; b) i) a light chain variable region comprising complementarity determining regions (CDRs) including CDR-L1 having the amino acid sequence of SEQ ID NO: 10, CDR-L2 having the amino acid sequence of SEQ ID NO: 11, and CDR-L3 having the amino acid sequence of SEQ ID NO: 3; and ii) a heavy chain variable region comprising CDRs including CDR-H1 having the amino acid sequence of SEQ ID NO: 12, CDR-H2 having the amino acid sequence of SEQ ID NO: 13, and CDR-H3 having the amino acid sequence of SEQ ID NO: 14; c) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 18; d) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 19; e) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 20; f) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 21; g) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 18; h) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 19; i) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 20; j) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 21; k) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 18; l) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 19; m) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 20; or n) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 21 The method according to any one of E1 to E5, comprising:

[0073] E7. The method of any one of E1-E6, wherein the anti-C1s antibody comprises a heavy chain constant region of isotype IgG1, IgG2, IgG3, or IgG4.

[0074] E8. The method of any one of E1-E6, wherein the anti-C1s antibody is selected from the group consisting of a Fab fragment, a F(ab')2 fragment, a scFv, and an Fv.

[0075] E9. The method of any one of E1-E8, wherein said administering is via subcutaneous, intravenous, or intramuscular administration.

[0076] E10.a) on day 1, administering a first dose of anti-C1s antibody; b) on day 8, administering a second dose of anti-C1s antibody; and c) Anti-C1s antibody administered every other week after the day 8 dose The method according to any one of E1 to E9, comprising:

[0077] E11. A method of inhibiting activation of complement component C4 in an individual in need thereof, comprising administering to the individual an anti-C1s antibody, wherein the anti-C1s antibody is administered in an amount of 5.5 g.

[0078] E12. The method of E11, wherein the anti-C1s antibody is administered to the individual every two weeks.

[0079] E13. The method described in E11 or E12, wherein the anti-C1s antibody comprises a light chain complementarity determining region (CDR) of the antibody light chain variable region comprising the amino acid sequence of SEQ ID NO:7 and a heavy chain CDR of the antibody heavy chain variable region comprising the amino acid sequence of SEQ ID NO:8.

[0080] E14. The method of any one of E11 to E13, wherein the anti-C1s antibody is humanized.

[0081] E15. The method of E14, wherein the humanized antibody comprises a humanized light chain framework region and / or a humanized heavy chain framework region.

[0082] E16. Anti-C1s antibodies: a) a light chain variable region comprising i) a complementarity determining region (CDR) comprising CDR-L1 having the amino acid sequence of SEQ ID NO:1, CDR-L2 having the amino acid sequence of SEQ ID NO:2, CDR-L3 having the amino acid sequence of SEQ ID NO:3; and ii) a heavy chain variable region comprising CDRs comprising CDR-H1 having the amino acid sequence of SEQ ID NO:4, CDR-H2 having the amino acid sequence of SEQ ID NO:5, and CDR-H3 having the amino acid sequence of SEQ ID NO:6; b) i) a light chain variable region comprising complementarity determining regions (CDRs) including CDR-L1 having the amino acid sequence of SEQ ID NO: 10, CDR-L2 having the amino acid sequence of SEQ ID NO: 11, and CDR-L3 having the amino acid sequence of SEQ ID NO: 3; and ii) a heavy chain variable region comprising CDRs including CDR-H1 having the amino acid sequence of SEQ ID NO: 12, CDR-H2 having the amino acid sequence of SEQ ID NO: 13, and CDR-H3 having the amino acid sequence of SEQ ID NO: 14; c) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 18; d) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 19; e) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 20; f) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 21; g) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 18; h) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 19; i) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 20; j) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 21; k) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 18; l) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 19; m) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 20; or n) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 21 The method according to any one of E11 to E15, comprising:

[0083] E17. The method of any one of E11-E16, wherein the anti-C1s antibody comprises a heavy chain constant region of isotype IgG1, IgG2, IgG3, or IgG4.

[0084] E18. The method of any one of E11 to E16, wherein the anti-C1s antibody is selected from the group consisting of a Fab fragment, a F(ab')2 fragment, a scFv, and an Fv.

[0085] E19. The method of any one of E11-E18, wherein said administering is via subcutaneous, intravenous, or intramuscular administration.

[0086] E20.a) on day 1, administering a first dose of anti-C1s antibody; b) on day 8, administering a second dose of anti-C1s antibody; and c) Anti-C1s antibody administered every other week after the day 8 dose The method according to any one of E11 to E19, comprising:

[0087] E21. A method of treating a complement-mediated disease or disorder in a subject in need thereof, comprising administering to the subject an effective amount of an anti-C1s antibody, wherein the serum concentration of the anti-C1s antibody following administration is at least about 20 μg / mL, at least about 25 μg / mL, at least about 30 μg / mL, at least about 35 μg / mL, at least about 40 μg / mL, at least about 45 μg / mL, at least about 50 μg / mL, at least about 55 μg / mL, at least about 60 μg / mL, at least about 65 μg / mL, at least about 70 μg / mL, at least about 75 μg / mL, at least about 80 μg / mL, at least about 85 μg / mL, at least about 90 μg / mL, at least about 95 μg / mL, or at least about 100 μg / mL.

[0088] E22. The method of E21, wherein the serum concentration of anti-C1s antibody after administration is between about 20 μg / mL and about 100 μg / mL, between about 20 μg / mL and about 90 μg / mL, between about 20 μg / mL and about 80 μg / mL, between about 20 μg / mL and about 70 μg / mL, between about 20 μg / mL and about 70 μg / mL, between about 20 μg / mL and about 60 μg / mL, between about 20 μg / mL and about 50 μg / mL, between about 20 μg / mL and about 40 μg / mL, or between about 20 μg / mL and about 30 μg / mL.

[0089] E23. The method of E21 or E22, wherein the serum concentration of anti-C1s antibody is measured by direct enzyme-linked immunosorbent assay (ELISA).

[0090] E24. An effective amount is at least about 60 mg / kg, at least about 65 mg / kg, at least about 70 mg / kg, at least about 75 mg / kg, at least about 80 mg / kg, at least about 85 mg / kg, at least about 90 mg / kg, at least about 95 mg / kg, at least about 100 mg / kg, at least about 105 mg / kg, at least about 110 mg / kg, at least about 115 mg / kg, at least about 120 mg / kg, at least about 125 mg / kg, at least about 130 mg / kg, at least about 135 mg / kg, E21-E23, wherein the amount of the bolus administered is about 1 g to about 10 g, or about 4 g to about 10 g, or about 150 mg / kg, at least about 140 mg / kg, at least about 145 mg / kg, at least about 150 mg / kg, at least about 155 mg / kg, at least about 160 mg / kg, at least about 165 mg / kg, at least about 170 mg / kg, at least about 175 mg / kg, at least about 180 mg / kg, at least about 185 mg / kg, at least about 190 mg / kg, at least about 195 mg / kg, or at least about 200 mg / kg, or about 4 g to 10 g.

[0091] E25. An effective amount is between about 60 mg / kg and about 100 mg / kg, between about 60 mg / kg and about 95 mg / kg, between about 60 mg / kg and about 90 mg / kg, between about 60 mg / kg and about 95 mg / kg, E21-E23, wherein the dose is between about 85 mg / kg, about 60 mg / kg and about 80 mg / kg, about 60 mg / kg and about 75 mg / kg, about 60 mg / kg and about 70 mg / kg, or about 60 mg / kg and about 65 mg / kg, or between about 4 g and about 10 g, between about 5 g and about 8 g, between about 5.5 g and about 7.5 g, between about 6.5 g and about 7.5 g, or between about 6.5 g and about 8.5 g.

[0092] E26. The method of E25, wherein the effective amount is about 60 mg / kg, about 65 mg / kg, about 70 mg / kg, about 75 mg / kg, about 80 mg / kg, about 85 mg / kg, about 90 mg / kg, about 95 mg / kg, about 100 mg / kg, about 105 mg / kg, about 110 mg / kg, about 115 mg / kg, about 120 mg / kg, about 125 mg / kg, about 130 mg / kg, about 135 mg / kg, about 140 mg / kg, about 145 mg / kg, or about 150 mg / kg, or 4 g, 4.5 g, 5 g, 5.5 g, 6 g, 6.5 g, 7 g, 7.5 g, 8 g, 8.5 g, 9 g, 9.5 g, or 10 g.

[0093] E27. The method of any one of E21 to E26, wherein the anti-C1s antibody is administered at an administration interval of 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or 31 days.

[0094] E28. The method of any one of E21 to E26, wherein the anti-C1s antibody is administered at administration intervals of 1 week, 2 weeks, 3 weeks, 4 weeks, or 1 month.

[0095] E29. The method of any one of E21 to E28, wherein the anti-C1s antibody increases the number of reticulocytes in the subject's blood following administration.

[0096] E30. A method of increasing the number of reticulocytes in the blood of a subject in need thereof, comprising administering to the subject an effective amount of an anti-C1s antibody.

[0097] E31. The method of E29 or E30, wherein the anti-C1s antibody, upon administration, increases the number of reticulocytes in the subject's blood by at least 1.1 fold, at least 1.2 fold, at least 1.3 fold, at least 1.4 fold, at least 1.5 fold, at least 1.6 fold, at least 1.7 fold, at least 1.8 fold, at least 1.9 fold, at least 2.0 fold, at least 2.1 fold, at least 2.2 fold, at least 2.3 fold, at least 2.4 fold, at least 2.5 fold, at least 2.6 fold, at least 2.7 fold, at least 2.8 fold, at least 2.9 fold, at least 3.0 fold, at least 4 fold, at least 5 fold, at least 6 fold, at least 7 fold, at least 8 fold, at least 9 fold, or at least 10 fold.

[0098] E32. The method of any one of E29-E31, wherein the anti-C1s antibody increases the number of reticulocytes in the subject's blood within about 24 hours of administration.

[0099] E33. The method of any one of E1-E32, wherein the anti-C1s antibody increases the level of hemoglobin in the subject.

[0100] E34. The anti-C1s antibody increases the hemoglobin level in the subject to at least about 1.0 g / dL, 1.1 g / dL, 1.2 g / dL, 1.3 g / dL, 1.4 g / dL, 1.5 g / dL, 1.6 g / dL, 1.7 g / dL, 1.8 g / dL, 1.9 g / dL, 2.0 g / dL, 2.1 g / dL, 2.2 g / dL, 2.3 g / dL, 2.4 g / dL, 2.5 g / dL, 2.6 g / dL, 2.7 g / dL, 2.8 g / dL, 2.9 g / dL, 3.0 g / dL, 3.1 g / dL, 3.2 g / dL, 3.3 g / dL, 3.4 g / dL, 3.5 g / dL, 3.6 g / dL, 3.7 g / dL, 3.8 g / dL, 3.9 g / dL, 4.0 g / dL, 4.1 g / dL, 4.2 g / dL, 4.3 g / dL, 4.4 g / dL, 4.5 g / dL, 4.6 g / dL, 4.7 g / dL, 4.8 g / dL, 4.9 g / dL, 5.0 g / dL, 5.1 g / dL, 5.2 g / dL, 5.3 g / dL, 5.4 g / dL, 5.5 g / dL, 5.6 g / dL, 5.7 g / dL, 5.8 g / dL, 5.9 g / dL, 6.0 g / dL, 6.1 g / dL, 6.2 g / dL, 6.3 g / dL, 4.0 g / dL, 4.1 g / dL, 4.2 g / dL, 4.3 g / dL, 4.4 g / dL, 4.5 g / dL, 4.6 g / dL, 4.7 g / dL, 4.8 g / dL, 4.9 g / dL, 5.0 g / dL, 5. / 1 g / dL, 5.2 g / dL, 5.3 g / dL, 5.4 g / dL, 5.5 g / dL, 5.6 g / dL, 5.7 g / dL, 5.8 g / dL, 5.9 g / dL, or 6.0 g / dL.

[0101] E35. The method of E33, wherein the level of hemoglobin in the subject is increased by at least 1.6 g / dL within 7 days of administration.

[0102] E36. The method of E33, wherein the level of hemoglobin in the subject is increased to 3.9 g / dL within 6 weeks of administration.

[0103] E37. The method of any one of E1-E36, wherein the anti-C1s antibody reduces the percentage of C3d positive red blood cells in the subject, eg, in the blood.

[0104] E38. The method of E37, wherein the percentage of C3d positive red blood cells in the subject is reduced 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 about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% compared to the percentage of C3d positive red blood cells in the subject before administration.

[0105] E39. The method of any one of E1-E38, wherein the anti-C1s antibody reduces the level of bilirubin in the subject, eg, in the blood.

[0106] E40. The method of E39, wherein the level of bilirubin in the subject is reduced to less than 2.5 mg / dL, 2.4 mg / dL, 2.3 mg / dL, 2.2 mg / dL, 2.1 mg / dL, 2.0 mg / dL, 1.9 mg / dL, 1.8 mg / dL, 1.7 mg / dL, 1.6 mg / dL, 1.5 mg / dL, 1.4 mg / dL, 1.3 mg / dL, 1.2 mg / dL, 1.1 mg / dL, 1.0 mg / dL, 0.9 mg / dL, 0.8 mg / dL, 0.7 mg / dL, 0.6 mg / dL, 0.5 mg / dL, 0.4 mg / dL, 0.3 mg / dL, 0.2 mg / dL, or 0.1 mg / dL.

[0107] E41. Anti-C1s antibodies: a) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 18; b) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 19; c) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 20; d) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 21; e) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 18; f) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 19; g) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 20; h) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 21; i) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 18; j) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 19; k) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 20; or l) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 21 The method of any one of E21 to E40, wherein the antibody cross-competes with an antibody comprising:

[0108] E42. Anti-C1s antibodies: a) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 18; b) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 19; c) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 20; d) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 21; e) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 18; f) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 19; g) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 20; h) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 21; i) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 18; j) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 19; k) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 20; or l) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 21 The method of any one of E21 to E41, wherein the antibody binds to the same epitope as an antibody comprising the antibody.

[0109] E43. Anti-C1s antibodies: a) a light chain variable region and a heavy chain variable region comprising i) a CDR-L1 having the amino acid sequence of SEQ ID NO:1, a CDR-L2 having the amino acid sequence of SEQ ID NO:2, a CDR-L3 having the amino acid sequence of SEQ ID NO:3; and ii) a heavy chain variable region (VH) comprising a CDR-H1 having the amino acid sequence of SEQ ID NO:4, a CDR-H2 having the amino acid sequence of SEQ ID NO:5, and a CDR-H3 having the amino acid sequence of SEQ ID NO:6; or b) i) a light chain variable region comprising CDR-L1 having the amino acid sequence of SEQ ID NO: 10, CDR-L2 having the amino acid sequence of SEQ ID NO: 11, and CDR-L3 having the amino acid sequence of SEQ ID NO: 3; and ii) a heavy chain variable region comprising CDR-H1 having the amino acid sequence of SEQ ID NO: 12, CDR-H2 having the amino acid sequence of SEQ ID NO: 13, and CDR-H3 having the amino acid sequence of SEQ ID NO: 14. The method according to any one of E21 to E42, comprising:

[0110] E44. Anti-C1s antibodies: a) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 18; b) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 19; c) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 20; d) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 21; e) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 18; f) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 19; g) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 20; h) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 21; i) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 18; j) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 19; k) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 20; or l) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 21 The method according to any one of E21 to E43, comprising:

[0111] E45. The method of any one of E21 to E44, wherein the anti-C1s antibody comprises a heavy chain constant region of isotype IgG1, IgG2, IgG3, or IgG4.

[0112] E46. The method of any one of E21 to E45, wherein the anti-C1s antibody is selected from the group consisting of a Fab fragment, a F(ab')2 fragment, a scFv, and an Fv.

[0113] E47. The method of any one of E21-E46, wherein administration is via subcutaneous, intravenous, or intramuscular administration. [Brief description of the drawings]

[0114] [Figure 1] 1 is a table showing characteristics of cold agglutinin disease (CAD) patients who were administered anti-C1s antibodies. [Figure 2-1]2A-2C show laboratory parameters of CAD patients before and during treatment with BIVV009. FIG. 2A shows the baseline laboratory parameters of the patient before treatment with BIVV009. FIG. 2B shows the minimum and maximum laboratory parameters of the patient during treatment with BIVV009. FIG. 2C shows the maximum change in laboratory parameters during treatment with BIVV009. [Figure 2-2] Continued from Figure 2-1. [Figure 2-3] Continued from Figure 2-2. [Diagram 3] Figures 3A-3B show the pharmacokinetics and pharmacodynamics of the anti-C1s antibody, BIVV009. Figure 3A shows the concentration-response analysis of BIVV009 levels and classical pathway activity in serum samples taken from normal healthy volunteers (NHVs). Figure 3B shows the mean pharmacokinetic profile of BIVV009 in patients (n=10) with cold agglutinin disease (CAD). [Figure 4-1] Figures 4A-4C show hematological responses to BIVV009 infusion. Data are median and interquartile range of 10 patients. Figure 4A shows the levels of C3d-positive red blood cells (%) after BIVV009 administration. Figure 4B (filled squares) shows the levels of hemoglobin (g / dL) after BIVV009 administration. Open triangles in Figure 4B represent the median hemoglobin levels in the subgroup of patients with primary cold agglutinin disease using Berentsen's definition. Figure 4C shows the levels of bilirubin (mg / dL) after BIVV009 administration. Data are median and interquartile range of 10 patients. [Figure 4-2] Continued from Figure 4-1. [Diagram 5] Figure 1 shows a plot of circulating bilirubin levels versus BIVV009 concentration, where the dotted line on the x-axis represents 20 μg / mL BIVV009 in serum and the dotted line on the y-axis represents 1.2 mg / dL (upper limit of normal). [Figure 6] FIG. 1 shows a comparison of hemoglobin levels over time to BIVV009 response in patients with CAD. PRBCs are packed red blood cells. [Figure 7-1] 7A-7F show biochemical response patterns in patients with CAD upon repeated BIVV009 administration. Arrows indicate BIVV009 administration. BIVV009 dose levels are also provided above the solid bars. FIG. 7A shows reticulocyte levels (×109 / L) over time (days) after repeated BIVV009 administration. FIG. 7B shows hemoglobin levels (g / dL) over time (days) after repeated BIVV009 administration. FIG. 7C shows haptoglobin levels (mg / dL) over time (days) after repeated BIVV009 administration. FIG. 7D shows lactate dehydrogenase (LDH) levels (U / L) over time (days) after repeated BIVV009 administration. FIG. 7E shows serum classical complement pathway activity (CH50) over time (days) after repeated BIVV009 administration. FIG. 7F shows bilirubin levels (mg / dL) over time (days) after repeated BIVV009 administration. [Figure 7-2] Continued from Figure 7-1. [Figure 7-3] Continued from Figure 7-2. [Figure 8] Schematic representation of the clinical trial protocol for administration of BIVV009 to kidney transplant recipients diagnosed with late active ABMR associated with signs of classical pathway (CP) activation caused by donor-specific antibodies (DSA). Index Bx, baseline biopsy; FU Bx, follow-up biopsy; EOS, end of study. [Figure 9] FIG. 1 shows individual DSA specificities in subjects participating in clinical trials identified at the time of study inclusion decision. [Figure 10A] FIG. 1 shows the relationship between median (interquartile range) serum concentrations (log scale) of BIVV009 and % overall CP activity detected by the WIESLAB® CP assay. [Figure 10B]FIG. 1 shows the effect of BIVV009 on C3d fixation elicited by immunodominant donor-specific antibodies (DSA) in a single bead assay or with a broad panel of third party anti-HLA antibodies (patient serum as complement source) pre-coated onto mixed beads, and in parallel, the IgG mean fluorescence intensity (MFI) of immunodominant DSA and its ability to fix recombinant C1q. [Figure 11-1] 11A-11H show the effect of BIVV009 on histomorphometric and molecular biopsy results. FIG. 11A shows C4d staining in peritubular capillaries (C4d score). FIG. 11B shows the degree of microcirculation (g+ptc score). FIG. 11C shows the degree of transplant glomerulopathy (cg score). FIG. 11D shows the ABMR score. FIG. 11E shows the TCMR score. FIG. 11F shows the total rejection score. FIG. 11G shows the acute kidney injury (AKI) score. FIG. 11H shows the chronic injury (atrophy / fibrosis) score. Box plots represent median, interquartile range and range. Wilcoxon rank test was used for statistical comparison. [Figure 11-2] Continued from Figure 11-1. [Figure 11-3] Continued from Figure 11-2. [Figure 11-4] Continued from Figure 11-3. [Figure 12-1]Figures 12A-12L show the effect of BIVV009 on pathogenesis-based transcription (PBT) scores. Panels show the difference in PBT expression between index and follow-up biopsies. Figure 12A shows transcripts representative of T cell burden (TCB). Figure 12B shows transcripts representative of cytotoxic T cell infiltration (QCAT). Figure 12C shows transcripts representative of NK cell burden (NKB). Figures 12D and 12E show transcripts representative of macrophage-related transcripts (QCMAT, AMAT1). Figure 12F shows transcripts representative of gamma-interferon-related transcripts (GRIT1). Figure 12G shows transcripts associated with the presence of DSA (DSAST). Figure 12H shows transcripts associated with endothelial inflammation (ENDAT). Figure 12I shows response to DSA-related transcripts (eDSAST). Figure 12J shows transcripts associated with acute kidney injury and wound repair (IRRAT). Figures 12K and 12L show transcripts representative of healthy kidney tissue and normal function (KT1, KT2), ​​respectively. [Figure 12-2] Continued from Figure 12-1. [Figure 12-3] Continued from Figure 12-2. [Figure 12-4] Continued from Figure 12-3. [Figure 12-5] Continued from Figure 12-4. [Figure 12-6] Continued from Figure 12-5. [Figure 13] FIG. 1 shows the course of estimated glomerular filtration rate (eGFR, mL / min / 1.73 m2) and urinary protein / creatinine (P / C) ratio (mg / g) in subjects over the 50-day study period. Arrows indicate days of BIVV009 administration. [Figure 14] Flowchart of a Phase 1 clinical trial of healthy subjects treated with either BIVV009 (humanized anti-C1s monoclonal antibody) or negative control. *Subject did not receive a second infusion due to gastroenteritis in Part B (multiple infusions); the subject was not excluded from the final analysis due to minimal variability in the adjusted PK data. [Figure 15-1]15A-15B are graphical representations of the mean (+SE) serum concentrations of BIVV009 versus time following a single 60-minute intravenous infusion of BIVV009 (Part A; FIG. 15A) and the mean (+SE) serum trough concentrations of BIVV009 versus time following a weekly 60-minute intravenous infusion of BIVV009 (Part B; FIG. 15B) in healthy volunteers. [Figure 15-2] Continued from Figure 15-1. [Figure 16-1] 16A-16C are graphical representations of individual body weight versus AUClastD (μg*hours / mL / mg) (FIG. 16A), CmaxD (μg / mL / mg) (FIG. 16B), and half-life_lambda_z (hours) (FIG. 16C). AUC: area under the concentration-time curve; MAD: multiple dose escalation; Cmax: maximum serum concentration; HL: half-life. [Figure 16-2] Continued from Figure 16-1. [Figure 17-1] Figures 17A-17C are graphical representations of mean (+SE) serum classical complement pathway (CP) activity versus time following a single 60-minute intravenous infusion of BIVV009 in healthy volunteers (Figure 17A), mean (+SE) serum trough CP activity versus time following a single or weekly 60-minute intravenous infusion of BIVV009 (Figure 17B), and individual trough serum CP activity versus time following multiple weekly 60-minute intravenous infusions of BIVV009 in healthy volunteers (Figure 17C). [Figure 17-2] Continued from Figure 17-1. [Figure 17-3] Continued from Figure 17-2. [Figure 18] Figure 1. Predicted weight (kg) distribution in a Phase 3 study. Simulations were based on 631 patients with CAgD (mean (SD) = 77.0 (19.7) kg, median (min-max) = 74.8 (40.6-163.3) kg) extracted from a US electronic medical record and claims database. [Figure 19]FIG. 1 shows simulated median (90% prediction interval (PI)) BIVV009 concentrations for proposed dosing regimens. The solid line represents the median BIVV009 concentration and the shaded area represents the 90% prediction interval. The dotted line represents the BIVV009 concentration (100 μg / mL) at which target-mediated pharmacokinetics (TMDD) begins to occur. [Figure 20] 20A-20B show fluorescent microscopy images of monkey esophageal tissue incubated with serum from a patient with bullous pemphigoid in the absence or presence of anti-C1s antibodies and stained for the presence of C3d. Fluorescence indicates deposition of C3d on the cell surface. FIG. 20A shows the level of C3d deposition in the absence of anti-C1s antibodies. FIG. 20B shows the level of C3d deposition in the presence of anti-C1s antibodies. [Figure 21] 21A-21C show fluorescence microscopy images of patient skin biopsies from a bullous pemphigoid patient treated with BIVV009 antibody. Fluorescence shows C3d deposition at the dermal-epidermal junction. FIG. 21A shows the level of C3d deposition at the dermal-epidermal junction before BIVV009 treatment. FIG. 21B shows the level of C3d deposition at the dermal-epidermal junction during BIVV009 treatment. FIG. 21C shows the level of C3d deposition at the dermal-epidermal junction afterBIVV009 treatment and antibody washout. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0115] definition In order that the present disclosure may be more readily understood, certain terms are first defined. As used in this application, each of the following terms shall have the meaning set forth below, unless expressly stated otherwise herein. Additional definitions are set forth throughout this application.

[0116] The present disclosure includes embodiments in which exactly one member of the group is present in, employed in, or associated with a given product or process. The present disclosure includes embodiments in which more than one, or all of the members of the group are present in, employed in, or associated with a given product or process.

[0117] Furthermore, "and / or" when used herein shall be interpreted as a specific disclosure of each of two specified features or elements, with or without the others. Thus, when the term "and / or" is used herein in phrases such as "A and / or B", it is intended to include "A and B", "A or B", "A" (single), and "B" (single). Similarly, when the term "and / or" is used in phrases such as "A, B, and / or C", it is intended to include each of the following aspects: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (single); B (single); and C (single).

[0118] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains. See, e.g., Concise Dictionary of Biomedicine and Molecular Biology, Juo, Pei-Show, 2nd Edition, 2002, CRC Press; The Dictionary of Cell and Molec and Oxford Dictionary Of Biochemistry And Molecular Biology, 3rd ed., 1999, Academic Press; and Oxford Dictionary Of Biochemistry And Molecular Biology, Revised Edition, 2000, Oxford University Press, provide one of skill with a general dictionary of many of the terms used in this disclosure.

[0119] Whenever an embodiment is described herein using the language "comprising," other similar embodiments described with the terms "consisting of" and / or "consisting essentially of" are also provided.

[0120] Units, prefixes, and symbols are denoted in their International System of Units (SI) accepted form. Numeric ranges are inclusive of the endpoints defining the range. When a range of values ​​is recited, it is understood that each integer value between the recited upper and lower limits of the range and each fraction thereof is also specifically disclosed, including each subrange between such values. The upper and lower limits of any range may be independently included or excluded from the range, and each range including either, neither, or both of the upper and lower limits is also included in the disclosure. When values ​​are explicitly recited, it is understood that values ​​that are approximately the same quantity or amount as the recited values ​​are also within the scope of the disclosure. When combinations are disclosed, each subcombination of the elements of the combination is also specifically disclosed and is within the scope of the disclosure. Conversely, when different elements or groups of elements are individually disclosed, combinations thereof are also disclosed. Where any element of the disclosure is disclosed as having multiple alternatives, instances of that disclosure in which each alternative is excluded alone, or in any combination with the other alternatives, are also hereby disclosed; more than one element of the disclosure may have such an exclusion, and therefore all combinations of elements with such exclusions are disclosed.

[0121] Nucleotides are described by their commonly accepted single-letter codes. Unless otherwise specified, nucleic acids are written from left to right in a 5' to 3' orientation. Nucleotides are described herein by their commonly known single-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Commission. Thus, A stands for adenine, C stands for cytosine, G stands for guanine, T stands for thymine, and U stands for uracil.

[0122] Amino acids are referred to herein by either their commonly known three letter symbols or by the one-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Commission. Unless otherwise specified, amino acid sequences are written left to right in amino to carboxy orientation.

[0123] The term "about" when used in conjunction with numerical values ​​throughout this specification and claims refers to an acceptable margin of accuracy for those skilled in the art. Typically, such margin of accuracy is ±10%.

[0124] It should be noted that, as used herein and in the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, a reference to "an anti-C1s antibody" includes a plurality of such antibodies, a reference to "the complement-mediated disease" includes a reference to one or more complement-mediated diseases and their equivalents known to those skilled in the art, and so forth. It is further noted that the claims may be drafted to exclude any optional element. As such, this statement is intended to provide a general understanding of the scope of the claims and to be consistent with the principles of the present invention. It is intended to serve as a predicate for use in conjunction with a plain description or for use of a "negative" limitation.

[0125] When ranges are given, both endpoints are included. Furthermore, unless otherwise specified, or otherwise apparent from the context and the knowledge of one of ordinary skill in the art, values ​​expressed as ranges can, in different embodiments of this disclosure, envisage every specific value or subrange within the stated range, down to the tenth of the unit of the lower limit of that range, unless the context clearly dictates otherwise.

[0126] The terms "antibody" and "immunoglobulin" include antibodies or immunoglobulins of any isotype, fragments of antibodies that retain specific binding to an antigen, such as, but not limited to, Fab, Fv, scFv, and Fd fragments, chimeric antibodies, humanized antibodies, engineered antibodies, single chain antibodies (scAbs), single domain antibodies (dAbs), single domain heavy chain antibodies, single domain light chain antibodies, bispecific antibodies, multispecific antibodies, and fusion proteins that include antigen-binding (also referred to herein as antigen-binding) portions of antibodies and non-antibody proteins. Antibodies can be detectably labeled, for example, with radioisotopes, enzymes that generate detectable products, fluorescent proteins, and the like. Antibodies can be further conjugated to other moieties, such as members of specific binding pairs, such as biotin (a member of the biotin-avidin specific binding pair). Antibodies can also be bound to a solid support, such as, but not limited to, polystyrene plates or beads. Also included in the term are Fab', Fv, F(ab')2, and / or other antibody fragments that retain specific binding to an antigen, and monoclonal antibodies. A monoclonal antibody, as used herein, is an antibody produced by a group of identical cells, all produced by repeated cell replication from a single cell. That is, that clone of cells produces only a single antibody species. Monoclonal antibodies can be produced using hybridoma production techniques, although other production methods known to those skilled in the art can also be used (e.g., antibodies from antibody phage display libraries). Antibodies can be monovalent or bivalent. Antibodies can be Ig monomers, which are "Y-shaped" molecules consisting of four polypeptide chains: two heavy chains and two light chains linked by disulfide bonds.

[0127] The term "humanized immunoglobulin" as used herein refers to an immunoglobulin that contains portions of immunoglobulins of different origins, at least one portion of which contains an amino acid sequence of human origin. For example, a humanized antibody may contain a portion derived from an immunoglobulin of non-human origin, such as a mouse, with the required specificity, and a portion derived from an immunoglobulin sequence of human origin, either chemically joined by conventional techniques (e.g., synthesis) or produced as a contiguous polypeptide using genetic engineering techniques (e.g., DNA encoding the protein portion of a chimeric antibody can be expressed to produce a contiguous polypeptide chain) (e.g., chimeric immunoglobulin). Another example of a humanized immunoglobulin is an immunoglobulin that contains one or more immunoglobulin chains that contain CDRs derived from an antibody of non-human origin and framework regions derived from light and / or heavy chains of human origin (e.g., CDR-grafted antibodies with or without framework modifications). Chimeric or CDR-grafted single chain antibodies are also included in the term humanized immunoglobulin. See, e.g., Cabilly et al., U.S. Pat. No. 4,816,567; Cabilly et al., European Patent No. 0,125,023 B1; Boss et al., U.S. Pat. No. 4,816,397; Boss et al., European Patent No. 0,120,694 B1; Neuberger, MS et al., WO 86 / 01533; Neuberger, MS et al., European Patent No. 0,194,276 B1; Winter, U.S. Pat. No. 5,225,539; Winter, European Patent No. 0,239,400 B1; Padlan, EA et al., European Patent Application No. 0,519,596 A1. Also with respect to single chain antibodies, Ladner et al., U.S. Pat. See also, US Pat. No. 4,946,778; Huston, US Pat. No. 5,476,786; and Bird, RE et al., Science, 242:423-426 (1988).

[0128] For example, humanized immunoglobulins can be produced by using synthetic and / or recombinant nucleic acids to produce genes (e.g., cDNAs) encoding the desired humanized chains. For example, nucleic acid (e.g., DNA) sequences encoding humanized variable regions can be constructed by using PCR mutagenesis methods to alter DNA sequences encoding human or humanized chains, e.g., DNA templates derived from previously humanized variable regions (see, e.g., Kamman, M. et al., Nucl. Acids Res., 17:5404 (1989); Sato, K. et al., Cancer Research, 53:851-856 (1993); Daugherty, BL et al., Nucleic Acids Res., 19(9):2471-2476 (1991); and Lewis, AP and JS Crowe, Gene, 101:297-302 (1991)). Variants can also be readily produced using these or other suitable methods. For example, cloned variable regions can be mutagenized and sequences encoding variants with the desired specificity selected (e.g., from phage libraries; see, e.g., Krebber et al., U.S. Pat. No. 5,514,548; Hoogenboom et al., WO 93 / 06213, published April 1, 1993).

[0129] "Antibody fragments" include portions of an intact antibody, such as the antigen-binding or variable regions of an intact antibody. Examples of antibody fragments include Fab, Fab', F(ab')2, and Fv fragments; diabodies; linear antibodies (Zapata et al., Protein Eng. 8(10):1057-1062 (1995)); domain antibodies (dAbs; Holt et al. (2003) Trends Biotechnol. 21:484); single-chain antibody molecules; and multispecific antibodies formed from antibody fragments. Papain digestion of an antibody produces two identical antigen-binding fragments, called "Fab" fragments, each with a single antigen-binding site, and a residual "Fc" fragment, a name reflecting its ability to crystallize readily. Pepsin treatment produces an F(ab')2 fragment that has two antigen-binding sites and is still capable of cross-linking antigen.

[0130] "Fv" is the minimum antibody fragment which contains a complete antigen recognition and binding site. This region consists of a dimer of one heavy- and one light-chain variable domain in tight, non-covalent association. This configuration allows the three CDRs of each variable domain to interact to form the V H -V L An antigen-binding site is defined on the surface of the dimer. Collectively, the six CDRs confer antigen-binding specificity to the antibody. However, even a single variable domain (or half of an Fv containing only three CDRs specific for an antigen) has the ability to recognize and bind antigen, although with lower affinity than the entire binding site.

[0131] A "Fab" fragment also contains the constant domain of the light chain and the first constant domain (CH1) of the heavy chain. Fab fragments differ from Fab' fragments by the addition of a few residues at the carboxyl terminus of the heavy chain CH1 domain including one or more cysteines from the antibody hinge region. As used herein, Fab'-SH is the designation for Fab' in which the cysteine ​​residues of the constant domains bear a free thiol group. F(ab')2 antibody fragments were originally produced as pairs of Fab' fragments which have hinge cysteines between them. Other chemical couplings of antibody fragments are also known.

[0132] The "light chains" of antibodies (immunoglobulins) from any vertebrate species can be assigned to one of two clearly distinct types, called kappa and lambda, based on the amino acid sequence of their constant domains. Depending on the amino acid sequence of the constant domain of their heavy chains, the immunoglobulins can be classified into two distinct types: Immunoglobulins can be assigned to different classes. There are five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, and some of these classes can be further divided into subclasses (isotypes), e.g., IgG1, IgG2, IgG3, IgG4, IgA, and IgA2. The subclasses can be further divided into types, e.g., IgG2a and IgG2b.

[0133] "Single-chain Fv" or "sFv" or "scFv" antibody fragments are fragments of the V chain of an antibody. H and V L In some embodiments, an Fv polypeptide comprises a V domain, and these domains are present in a single polypeptide chain. H Domain and V LIt may further comprise a polypeptide linker between the domains which enables the sFv to form the desired structure for antigen binding. For a general discussion of sFvs, see Pluckthun, The Pharmacology of Monoclonal Antibodies, Vol. 113, Eds. Rosenburg and Moore, Springer-Verlag, New York, pp. 269-315 (1994).

[0134] The term "diabody" refers to a small antibody fragment with two antigen-binding sites, which fragments are bound to the same polypeptide chain (V H -V L ) in the light chain variable domain (V L ) linked to a heavy chain variable domain (V H ). By using a linker that is too short to allow pairing between the two domains on the same chain, the domains are forced to pair with complementary domains on another chain, creating two antigen-binding sites. Diabodies are described in more detail, for example, in EP 404,097; WO 93 / 11161; and Hollinger et al. (1993) Proc. Natl. Acad. Sci. USA 90:6444-6448.

[0135] The term "affinity" refers to the degree to which a binding molecule, such as an antibody, binds to an antigen in such a way as to shift the equilibrium between the antigen and the binding molecule toward the presence of a complex formed by their binding. Thus, when antigen and binding molecule are associated at relatively equal concentrations, a high affinity binding molecule will bind to available antigen in such a way as to shift the equilibrium toward a higher concentration of the resulting complex. The binding molecules, such as antibodies, or antigen-binding fragments, variants or derivatives thereof of the present disclosure can also be described or specified in terms of their binding affinity to the antigen. The affinity of a binding molecule, such as an antibody, to an antigen can be determined experimentally using any suitable method. (See, for example, Berzofsky et al., "Antibody-Antigen Interactions," In Fundamental Immunology, edited by Paul, WE, Raven Press: New York, NY (1984); Kuby, Janis Immunology, WH Freeman and Company: New York, NY (1992); and methods described therein).

[0136] The measured affinity of a particular binding molecule-antigen interaction may vary when measured under different conditions (e.g., salt concentration, pH). Thus, affinity and other antigen binding parameters (e.g., K D , K a , K d Measurements of ) are preferably made using standardized solutions of binding molecule and antigen, and standardized buffers.

[0137] "High affinity" with respect to a binding molecule, e.g., an antibody, is at least about 1×10 7 liters / mole, or at least about 1 x 10 8 liters / mole, or at least about 1 x 10 9 liters / mole, or at least about 1 x 10 10 liters / mole, or at least about 1 x 10 11 liters / mole, or at least about 1 x 10 12liters / mole, or at least about 1 x 10 13 liters / mole, or at least about 1 x 10 14 Equilibrium association constant (K aff "High affinity" binding refers to the It may be different for each isotype.

[0138] K D , equilibrium dissociation constant is a term also used to describe antibody affinity, K aff It is the reciprocal of K D is k d k a (i.e., k d / k a ) and expressed as molar concentration (M). D The K value can be determined using methods well established in the art. D Available methods for determining K include biolayer interference (BLI) assays, surface plasmon resonance, biosensor systems such as the BIACORE® system or flow cytometry, and Scatchard analysis. D When used, the term "high affinity" for an antibody refers to an antibody with a specific affinity of about 1×10 -7 Less than M or about 1×10 -8 Less than M or about 1×-10 -9 Less than M or about 1×10 -10 Less than M or about 1×10 -11 Less than M or about 1×10 -12 Less than M or about 1×10 -13 Less than M, approx. 1×10 -14 The equilibrium dissociation constant (K D )

[0139] The affinity may be at least 1-fold greater, at least 2-fold greater, at least 3-fold greater, at least 4-fold greater, at least 5-fold greater, at least 6-fold greater, at least 7-fold greater, at least 8-fold greater, at least 9-fold greater, at least 10-fold greater, at least 20-fold greater, at least 30-fold greater, at least 40-fold greater, at least 50-fold greater, at least 60-fold greater, at least 70-fold greater, at least 80-fold greater, at least 90-fold greater, at least 100-fold greater, or at least 1,000-fold greater, or even greater, than that of an antibody having an unrelated amino acid sequence. The affinity of an antibody for a target protein may be, for example, from about 100 nanomolar (nM) to about 0.1 nM, from about 100 nM to about 1 picomolar (pM), or from about 100 nM to about 1 femtomolar (fM), or even higher. The term "binding activity" as used herein refers to the reluctance of a complex of two or more substances to dissociate after dilution. The terms "immunoreactive" and "preferentially binds" are used interchangeably herein with respect to antibodies and / or antigen-binding fragments.

[0140] The term "binding" refers to a direct association between two molecules, for example, by covalent, electrostatic, hydrophobic, and ionic and / or hydrogen bonding interactions, such as salt bridges and water bridges. The subject anti-C1s antibodies specifically bind to an epitope in the complement C1s protein. "Specific binding" refers to binding that is at least about 10 -7 M or greater affinity, e.g., 5×10 -7 M, 10 -8 M, 5×10 -8 "Non-specific binding" refers to binding with an affinity of about 10 M or greater. -7 Binding with an affinity of less than M, e.g., 10 -6 M, 10 -5 M, 10 -4 It refers to binding with an affinity such as M.

[0141] The term "compete" or "cross-compete" as used herein with respect to a binding molecule, e.g., an antibody, means that a first binding molecule, e.g., a first antibody or an antigen-binding portion thereof, binds to an epitope in a manner sufficiently similar to that of a second binding molecule, e.g., a second antibody or an antigen-binding portion thereof, such that the result of binding of the first binding molecule to its cognate epitope in the presence of the second binding molecule is detectably reduced compared to the binding of the first binding molecule in the absence of the second binding molecule. Alternatively, it may, but does not necessarily, occur that the binding of the second binding molecule to its epitope is also detectably reduced in the presence of the first binding molecule. That is, the first binding molecule can inhibit the binding of the second binding molecule to its epitope without the second molecule inhibiting the binding of the first binding molecule to its respective epitope. However, if each binding molecule detectably inhibits the binding of the other binding molecule to its cognate epitope, whether to the same extent, to a greater extent, or to a lesser extent, the binding molecules are said to "cross-compete" with each other for binding of their respective epitopes. Both competing and cross-competing binding molecules are included in the present disclosure.

[0142] Binding molecules, e.g., antibodies, are said to "bind to the same epitope" or "contain the same binding site" or have "essentially the same binding" characteristics if the binding molecules cross-compete so that only one antibody can bind to an epitope at a given time, i.e., one binding molecule prevents binding or modulation of the action of the other.

[0143] Competition, as used herein, means at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% greater relative inhibition, as determined, for example, by competitive ELISA assays or ForteBio assays as described in the Examples section. It may be desirable to set a higher threshold of relative inhibition as a measure of what is a suitable level of competition in a particular situation. Thus, for example, the criteria for competitive binding can be set when at least about 40% relative inhibition is detected, or at least about 45%, or at least about 50%, or at least about 55%, or at least about 60%, or at least about 65%, or at least about 70%, or at least about 75%, or at least about 80%, or at least about 85%, or at least about 90%, or at least about 95%, or even about 100% relative inhibition is detected before an antibody is considered sufficiently competitive.

[0144] The term "epitope" as used herein refers to an antigenic protein determinant (e.g., an amino acid subsequence of C1s) capable of binding to a binding molecule, e.g., an antibody. Epitopes usually consist of a chemically active surface arrangement of molecules, such as amino acids or sugar side chains, and usually have specific charge characteristics in addition to specific three-dimensional structural features. The part of an antibody or binding molecule that recognizes an epitope is called a paratope. Epitopes of protein antigens are divided into two categories based on their structure and interaction with the paratope: conformational epitopes and linear epitopes. Conformational epitopes are composed of discontinuous sections of the antigen's amino acid sequence. These epitopes interact with the paratope based on the three-dimensional surface features and shape or tertiary structure of the antigen. In contrast, linear epitopes interact with the paratope based on their primary structure. Linear epitopes are formed by a continuous sequence of amino acids from the antigen.

[0145] The term "CDR" or "complementarity determining region," as used herein, is intended to mean the non-contiguous antigen-binding sites found within the variable regions of both heavy and light chain polypeptides. CDRs are described in Kabat et al., J. Biol. Chem. 252:6609-6616 (1977); Kabat et al., US Pept. of Health and Human Services, "Sequences of proteins of immunological interest" (1991) (also referred to herein as Kabat 1991); Chothia et al., J. Mol. Biol. 196:901-917 (1987) (also referred to herein as Chothia 1987); and MacCallum et al., J. Mol. Biol. 262:732-745 (1996), in which the definitions include overlapping or subsets of amino acid residues when compared with each other. Nevertheless, application of either definition to refer to the CDRs of an antibody or grafted antibody or variants thereof is intended to be within the scope of the term as defined and used herein. The amino acid residues that comprise the CDRs, as defined in each of the above cited references, are set forth below in Table 1 for comparison. The CDRs provided in the present disclosure can be compared with those defined by Kabat et al. The definition was based on 1991.

[0146] [Table 1]

[0147] The terms "CDR-L1", "CDR-L2", and "CDR-L3" as used herein refer to the first, second, and third CDRs, respectively, in a light chain variable region. The terms "CDR-H1", "CDR-H2", and "CDR-H3" as used herein refer to the first, second, and third CDRs, respectively, in a heavy chain variable region. The terms "CDR-1", "CDR-2", and "CDR-3" as used herein refer to the first, second, and third CDRs, respectively, in the variable region of either chain.

[0148] As used herein, the term "framework", when used in reference to an antibody variable region, is intended to mean all amino acid residues outside the CDR regions in the variable region of the antibody. The framework of a variable region is generally a non-contiguous amino acid sequence of about 100-120 amino acids in length, but is intended to refer to only those amino acids outside the CDRs. The term "framework region", as used herein, is intended to mean each domain of the framework separated by the CDRs.

[0149] An "isolated" antibody is one that has been identified and separated and / or recovered from a component of its natural environment. Contaminant components of its natural environment are substances that would be expected to interfere with the diagnostic or therapeutic use of the antibody, such as enzymes, hormones, and other proteinaceous or non-proteinaceous solutes. In some embodiments, the antibody is expected to be purified (1) to greater than 90%, greater than 95%, or greater than 98%, e.g., greater than 99%, by weight of the antibody as determined by the Lowry method, (2) to a degree sufficient to obtain at least 15 residues of N-terminal or internal amino acid sequence by use of a spinning cup sequenator, or (3) to homogeneity by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under reducing or non-reducing conditions using Coomassie blue or silver stain. An isolated antibody also includes an antibody in its original location in a recombinant cell, provided that at least one component of the antibody's natural environment is no longer present. In some cases, an isolated antibody is expected to be produced by at least one purification step.

[0150] The terms "polypeptide," "peptide," and "protein" are used interchangeably herein to refer to polymeric forms of amino acids of any length, and as such include polypeptides containing genetically and non-genetically encoded amino acids, chemically or biochemically modified or derived amino acids, and polypeptides having modified peptide backbones. The term includes, but is not limited to, fusion proteins, fusion proteins with heterologous amino acid sequences, fusions with heterologous and homologous leader sequences with or without an N-terminal methionine residue; immunologically tagged proteins, and the like.

[0151] The term "identity" as used herein refers to overall monomer conservation between polymeric molecules, e.g., polypeptide molecules or polynucleotide molecules (e.g., DNA molecules and / or RNA molecules). The term "identical" without any additional modifiers, e.g., protein A is identical to protein B, means that the sequences are 100% identical (100% sequence identity). Describing two sequences as, e.g., "70% identical" is the same as stating that they have, e.g., "70% sequence identity."

[0152] The calculation of the percent identity of two polynucleotide sequences can be performed, for example, by aligning the two sequences for optimal comparison purposes (e.g., for optimal alignment, gaps can be introduced into one or both of the first and second nucleic acid sequences, and non-identical sequences can be ignored for comparison purposes). In certain embodiments, the length of the sequence aligned for comparison purposes is at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or 100% of the length of the reference sequence. The nucleotides at the corresponding nucleotide positions are then compared. If a position in the first sequence is occupied by the same nucleotide as the corresponding position in the second sequence, the molecules are identical at that position. The percent identity between two sequences is a function of the number of identical positions common to the sequences, taking into account the number of gaps that need to be introduced for optimal alignment of the two sequences and the length of each gap. The comparison of sequences and the determination of the percent identity between two sequences can be achieved using a mathematical algorithm. When comparing DNA and RNA, thymine (T) and uracil (U) can be considered equivalent.

[0153] Suitable software programs are available from various sources and are for alignment of both protein and nucleotide sequences. One suitable program for determining percent sequence identity is bl2seq, which is part of the BLAST program package available from the BLAST website of the U.S. Government's National Center for Biotechnology Information (blast.ncbi.nlm.nih.gov). Bl2seq performs comparisons between two sequences using either the BLASTN or BLASTP algorithm. BLASTN is used to compare nucleic acid sequences, while BLASTP is used to compare amino acid sequences. Other suitable programs include, for example, Needle, Stretcher, Water, or Matcher, which are part of the EMBOSS bioinformatics program package, also available from the European Bioinformatics Institute (EBI) at www.ebi.ac.uk / Tools / psa.

[0154] Sequence alignment can be performed using methods known in the art, such as MAFFT, Clustal (ClustalW, Clustal X, or Clustal Omega), MUSCLE, and the like.

[0155] Different regions in a single polynucleotide or polypeptide target sequence aligned with a polynucleotide or polypeptide reference sequence may each have their own percent sequence identity. Note that the percent sequence identity value is rounded to the nearest tenth. For example, 80.11, 80.12, 80.13, and 80.14 are: will be rounded down to 80.1, while 80.15, 80.16, 80.17, 80.18, and 80.19 will be rounded up to 80.2. Also note that length values ​​are always integers.

[0156] In certain embodiments, the identity percentage (ID)% of a first amino acid sequence (or nucleic acid sequence) to a second amino acid sequence (or nucleic acid sequence) is calculated as ID%=100×(Y / Z), where Y is the number of amino acid residues (or nucleic acid bases) scored as identical pairs in the alignment of the first and second sequences (by visual inspection or by a specific sequence alignment program), and Z is the total number of residues in the second sequence. If the length of the first sequence is longer than the second sequence, the percent identity of the first sequence to the second sequence will be higher than the percent identity of the second sequence to the first sequence.

[0157] Those skilled in the art will understand that the generation of sequence alignment for calculating percent sequence identity is not limited to a pairwise sequence-sequence comparison determined only by primary sequence data. It will also be understood that sequence alignment can be generated by integrating sequence data with data from heterogeneous sources, such as structural data (e.g., crystallographic protein structures), functional data (e.g., mutation arrangement), or phylogenetic data. A suitable program for integrating heterogeneous data to generate multiple sequence alignment is T-Coffee, which is available at www.tcoffee.org, or alternatively, for example, from EBI. It will also be understood that the final alignment used to calculate percent sequence identity can be assembled either automatically or manually.

[0158] The terms "treatment", "treating", "treating" and the like, as used herein, refer to obtaining a desired pharmacological and / or physiological effect. Such an effect may be prophylactic, in terms of completely or partially preventing a disease or its symptoms, and / or may be therapeutic, in terms of partially or completely curing a disease and / or adverse effects that may result from the disease. "Treatment", as used herein, encompasses any treatment of disease in a mammal, particularly a human, and as such includes (a) preventing the onset of a disease in a subject who may be susceptible to the disease but has not yet been diagnosed as having it; (b) inhibiting the disease, i.e., halting its progression; (c) alleviating the disease, e.g., bringing about remission of the disease; and (d) alleviating or reducing symptoms associated with the disease.

[0159] The terms "individual," "subject," "host," and "patient" are used interchangeably herein and refer to mammals, including but not limited to murines (rats, mice), non-human primates, humans, canines, felines, ungulates (e.g., horses, cows, sheep, pigs, goats), and the like. These terms also include any animal that has a complement system, such as mammals, fish, and some invertebrates. As such, these terms include mammals, fish, and invertebrate companion animals, farm animals, working animals, zoo animals, and laboratory animals that contain a complement system.

[0160] A "therapeutically effective amount," "effective amount," or "effective amount" refers to the amount of an anti-complement C1s antibody that, when administered to a mammal or other subject for treating a disease, is sufficient to effect treatment for such disease.

[0161] The term "less than" (<) means a value that is less than, but not equal to, the reference value. The term "greater than" (>) means a value that is greater than, but not equal to, the reference value. The term "less than or equal to" (≦) means a value that is less than or equal to the reference value. The term "greater than or equal to" (≧) means a value that is greater than or equal to a reference value.

[0162] Before the present disclosure is further described, it is to be understood that this disclosure is not limited to particular embodiments described, which may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present disclosure will be limited only by the appended claims.

[0163] When a range of values ​​is provided, it is understood that each intervening value between the upper and lower limits of the range, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, and all other stated or intervening values ​​within the stated range are included in the disclosure. The upper and lower limits of these smaller ranges can be independently included in the smaller ranges, which are also included in the disclosure, unless there is any specifically excluded upper or lower limit in the stated range. When a stated range includes one or both of the limits, ranges excluding either or both of the limits included in such ranges are also included in the disclosure.

[0164] Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, the preferred methods and materials are now described. All publications mentioned herein are incorporated by reference to disclose and describe the methods and / or materials, together with those cited in the publications.

[0165] It is understood that certain features of the present disclosure that are described for clarity in the context of separate embodiments can also be provided in combination in a single embodiment. Conversely, various features of the present disclosure that are briefly described in the context of a single embodiment can also be provided separately or in any suitable subcombination. All combinations of the embodiments related to the present disclosure are specifically included in the present disclosure and are disclosed herein as if each and every combination were individually and expressly disclosed. In addition, all subcombinations of the various embodiments and elements thereof are specifically included in the present disclosure and are disclosed herein as if each and every such subcombination were individually and expressly disclosed herein.

[0166] The publications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Nothing herein should be construed as an admission that the disclosure of the present invention is not entitled to antedate such publications by virtue of prior disclosure. Further, the publication dates provided may be different from the actual publication dates which may need to be independently confirmed.

[0167] Detailed Description The present disclosure provides a method for treating a complement-mediated disease or disorder in an individual, and a method for inhibiting the activation of complement component C4 in an individual in need thereof. In some embodiments, the method comprises administering to the individual an anti-C1s antibody at a fixed dose of 5.5 g. In some embodiments, the method comprises administering to the individual an anti-C1s antibody at a fixed dose of between about 4.0 g and about 10.0 g, for example, about 4 g, about 4.5 g, about 5 g, about 5.5 g, about 6 g, about 6.5 g, about 7.5 g, about 8 g, about 8.5 g, about 9 g, about 9.5 g, or about 10 g. In some embodiments, the method comprises administering to the individual an effective amount of an anti-C1s antibody, and the serum concentration of the antibody is between about 20 μg / ml and about 150 μg / ml.

[0168] Anti-C1s antibody A suitable anti-C1s antibody for the present disclosure specifically binds to a conformational epitope within amino acids 272-422 of the amino acid sequence of human C1s as follows: (SEQ ID NO:9)

[0169] Anti-C1s antibodies suitable for the present disclosure inhibit C1s-mediated cleavage of complement component C4. In some cases, anti-C1s antibodies suitable for use in the methods of the present disclosure inhibit C1s-mediated cleavage of complement component C4, but do not inhibit C1s-mediated cleavage of complement component C2. In some cases, the antibody inhibits a component of the classical complement pathway; in some cases, the component of the classical complement pathway is C1s. In some cases, the antibody does not inhibit the protease activity of C1s.

[0170] In some cases, the anti-C1s antibody suitable for the present disclosure is humanized. In some cases, the anti-C1s antibody comprises a humanized light chain framework region. In some cases, the anti-C1s antibody comprises a humanized heavy chain framework region. In some cases, the anti-C1s antibody comprises a humanized light chain framework region and a humanized heavy chain framework region. In some cases, the anti-C1s antibody suitable for the present disclosure is a humanized monoclonal antibody.

[0171] Humanization of the framework region reduces the risk that the antibody will elicit a human anti-mouse antibody (HAMA) response in humans. Methods for determining immune responses recognized in the art can be implemented to monitor HAMA responses in a particular patient or during clinical trials. Patients administered humanized antibodies can be given an assessment of immunogenicity at the outset and throughout the administration of therapy. HAMA responses are measured by detecting antibodies against the humanized therapeutic reagent in serum samples from patients using methods known to those skilled in the art, such as surface plasmon resonance technology (BIACORE) and / or solid-phase enzyme-linked immunosorbent assay (ELISA) analysis. In many cases, the subject's humanized anti-C1s antibody does not elicit a substantial HAMA response in human subjects.

[0172] Specific amino acids from the human variable region framework residues are selected for substitution based on their potential effect on CDR conformation and / or antigen binding. The non-natural juxtaposition of murine CDR regions with human variable framework regions can result in non-natural structural constraints that, unless corrected by substitution of specific amino acid residues, result in loss of binding affinity.

[0173] The selection of amino acid residues for substitution may be determined in part by computer modeling. Computer hardware and software for generating three-dimensional images of immunoglobulin molecules are known in the art. Generally, molecular models are generated starting from an analyzed structure of an immunoglobulin chain or domain thereof. The chain to be modeled is compared with the chain or domain of the analyzed three-dimensional structure for amino acid sequence similarity, and the chain or domain showing the greatest sequence similarity is selected as the starting point for building the molecular model. Chains or domains with at least 50% sequence identity are selected for modeling, e.g., those with at least 60%, at least 70%, at least 80%, at least 90% sequence identity or higher are selected for modeling. The analyzed starting structure is modified to allow for differences between the actual amino acids in the immunoglobulin chain or domain to be modeled and those of the starting structure. The modified structures are then assembled into composite immunoglobulins. Finally, the model is refined by energy minimization and further by verifying that all atoms are within appropriate distances from each other and that bond distances and angles are within chemically acceptable limits.

[0174] The CDR and framework regions were identified using the Kabat, Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, Md., 1987 and 1991). Alternative structural definitions have been proposed by Chothia et al., J. Mol. Biol. 196:901 (1987); Nature 342:878 (1989); and J. Mol. Biol. 186:651 (1989) (collectively referred to as "Chothia"). Amino acids present in murine antibodies can be selected for substitution into a humanized antibody if the framework residues as defined in Kabat, above, constitute structural loop residues as defined in Chothia, above. Residues "adjacent to a CDR region" include amino acid residues located immediately adjacent to one or more of the CDRs in the primary sequence of a humanized immunoglobulin chain, e.g., a CDR as defined in Kabat, or an amino acid residue located immediately adjacent to a CDR as defined in Chothia (see, e.g., Chothia and Lesk, 1992). (See JMB 196:901 (1987)). These amino acids are particularly likely to interact with amino acids in the CDRs and, if selected from the acceptor, are likely to distort the donor CDRs and reduce affinity. In addition, adjacent amino acids can directly interact with the antigen (Amit et al., Science 233:747 (1986)), and selecting these amino acids from the donor may be desirable to maintain all of the affinity-generating antigen contacts in the original antibody.

[0175] In some cases, an anti-C1s antibody suitable for the present disclosure comprises a variable region of a light chain (VL) comprising CDR-L1, CDR-L2, and CDR-L3 present in the VL comprising the amino acid sequence of SEQ ID NO:7. SEQ ID NO:7: QIVLTQSPAIMSASLGERVTMTCTASSSVSSSYLHWYQQKPGSSPKLWIYSTSNLASGVPARFSGSGSGTFYSLTISSMEAEDDATYYCHQYYRLPPITFGAGTKLELK

[0176] In some cases, an anti-C1s antibody suitable for the present disclosure comprises a variable region of a heavy chain (VH) comprising CDR-H1, CDR-H2, and CDR-H3 present in the VH comprising the amino acid sequence of SEQ ID NO:8. SEQ ID NO:8: EVMLVESGGALVKPGGSLKLSCAASGFTFSNYAMSWVRQI PEKRLEWVATISSGGSHTYYLDSVKGRFTISRDNARDTLYLQMSSLRSEDTALYYCARLFTGYAMDYWGQGTSVTVSS

[0177] In some cases, anti-C1s antibodies suitable for the present disclosure include: a) a light chain region comprising CDR-L1, CDR-L2, and CDR-L3 having the amino acid sequences set forth in SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3, respectively; and b) a heavy chain region comprising CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences set forth in SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6, respectively. In some of these embodiments, the anti-C1s antibodies include humanized V H and / or V L Includes framework regions. SEQ ID NO:1: SSVSSSYLHWYQ; SEQ ID NO:2: STSNLASGVP; SEQ ID NO:3: HQYYRLPPIT; SEQ ID NO:4: GFTFSNYAMSWV; SEQ ID NO:5: ISSGGSHTYY; SEQ ID NO: 6: ARLFTGGYAMDY

[0178] In some cases, anti-C1s antibodies suitable for the present disclosure include: a) a light chain region comprising CDR-L1, CDR-L2, and CDR-L3 having the amino acid sequences set forth in SEQ ID NO: 10, SEQ ID NO: 11, and SEQ ID NO: 3, respectively; and b) a heavy chain region comprising CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences set forth in SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 14, respectively. In some of these embodiments, the anti-C1s antibodies include humanized VH and / or V L Including framework regions. SEQ ID NO: 10: TASSSVSSSYLH; SEQ ID NO:11: STSNLAS; SEQ ID NO:3: HQYYRLPPIT; SEQ ID NO:12: NYAMS; SEQ ID NO:13: TISSGGSHTYYLDSVKG; SEQ ID NO: 14: LFTGHAMDY

[0179] In some cases, an anti-C1s antibody suitable for the present disclosure comprises a light chain variable region comprising an amino acid sequence that is 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO:15. SEQ ID NO:15: QIVLTQSPAILSLSPGERATMSCTASSSVSSSYLHWYQQKPGKAPKLWIYSTSNLASGVPSRFSGSGSGTFYTLTISSLQAEDFATYYCHQYYRLPPITFGQGTKLEIK

[0180] In some cases, an anti-C1s antibody suitable for the present disclosure comprises a light chain variable region comprising an amino acid sequence that is 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO:16. SEQ ID NO:16: QIVLTQSPATLSLSPGERATMSCTASSSVSSSYLHWYQQKPGKAPKLWIYSTSNLASGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCHQYYRLPPITFGQGTKLEIK

[0181] In some cases, anti-C1s antibodies suitable for the present disclosure may have an amino acid sequence that is 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO:17. The light chain variable region comprises: SEQ ID NO:17: QIVLTQSPATLSLSPGERATLSCTASSSVSSSYLHWYQQKPGKAPKLWIYSTSNLASGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCHQYYRLPPITFGQGTKLEIK

[0182] In some cases, an anti-C1s antibody suitable for the present disclosure comprises a heavy chain variable region comprising an amino acid sequence that is 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO:18. SEQ ID NO:18: EVMLVESGGGLVKPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVATISSGGSHTYYLDSVKGRFTISRDNSKDTLYLQMSSLRAEDTALYYCARLFTGYAMDYWGQGTSVTVSS

[0183] In some cases, an anti-C1s antibody suitable for the present disclosure comprises a heavy chain variable region comprising an amino acid sequence that is 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO:19. SEQ ID NO:19: EVMLVESGGGLVKPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVATISSGGSHTYYLDSVKGRFTISRDNSKDTLYLQMNSLRAEDTALYYCARLFTGYAMDYWGQGTLVTVSS

[0184] In some cases, an anti-C1s antibody suitable for the present disclosure comprises a heavy chain variable region comprising an amino acid sequence that is 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO:20. SEQ ID NO:20: EVMLVESGGGLVKPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVATISSGGSHTYYLDSVKGRFTISRDNSKDTLYLQMSSLRAEDTALYYCARLFTGYAMDYWGQGTSVTVSS

[0185] In some cases, an anti-C1s antibody suitable for the present disclosure comprises a heavy chain variable region comprising an amino acid sequence that is 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO:21. SEQ ID NO:21: EVQLVESGGGLVKPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVATISSGGSHTYYLDSVKGRFTISRDNSKNTLYLQMNSLRAEDTALYYCARLFTGYAMDYWGQGTLVTVSS

[0186] In some cases, an anti-C1s antibody suitable for the present disclosure comprises a VL region comprising the amino acid sequence set forth in SEQ ID NO:15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:18.

[0187] In some cases, an anti-C1s antibody suitable for the present disclosure comprises a VL region comprising the amino acid sequence set forth in SEQ ID NO:15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:19.

[0188] In some cases, an anti-C1s antibody suitable for the present disclosure comprises a VL region comprising the amino acid sequence set forth in SEQ ID NO:15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:20.

[0189] In some cases, an anti-C1s antibody suitable for the present disclosure comprises a VL region comprising the amino acid sequence set forth in SEQ ID NO:15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:21.

[0190] In some cases, an anti-C1s antibody suitable for the present disclosure comprises a VL region comprising the amino acid sequence set forth in SEQ ID NO:16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:18.

[0191] In some cases, an anti-C1s antibody suitable for the present disclosure comprises a VL region comprising the amino acid sequence set forth in SEQ ID NO:16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:19.

[0192] In some cases, an anti-C1s antibody suitable for the present disclosure comprises a VL region comprising the amino acid sequence set forth in SEQ ID NO:16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:20.

[0193] In some cases, an anti-C1s antibody suitable for the present disclosure comprises a VL region comprising the amino acid sequence set forth in SEQ ID NO:16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:21.

[0194] In some cases, an anti-C1s antibody suitable for the present disclosure comprises a VL region comprising the amino acid sequence set forth in SEQ ID NO:17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:18.

[0195] In some cases, an anti-C1s antibody suitable for the present disclosure comprises a VL region comprising the amino acid sequence set forth in SEQ ID NO:17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:19.

[0196] In some cases, an anti-C1s antibody suitable for the present disclosure comprises a VL region comprising the amino acid sequence set forth in SEQ ID NO:17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:20.

[0197] In some cases, an anti-C1s antibody suitable for the present disclosure comprises a VL region comprising the amino acid sequence set forth in SEQ ID NO:17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:21.

[0198] In some embodiments, an anti-C1s antibody suitable for the present disclosure is an antibody that cross-competes with a reference antibody. a) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 18; b) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 19; c) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 20; d) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 21; e) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 18; f) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 19; g) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 20; h) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 21; i) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 18; j) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 19; k) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 20; or l) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 21 Includes.

[0199] In some cases, an anti-C1s antibody suitable for the present disclosure cross-competes with an antibody comprising a variable region (VL) of a light chain region comprising CDR-L1, CDR-L2, and CDR-L3 present in the VL comprising the amino acid sequence of SEQ ID NO:7.

[0200] In some cases, an anti-C1s antibody suitable for the present disclosure cross-competes with an antibody comprising a variable region (VH) of a heavy chain region comprising CDR-H1, CDR-H2, and CDR-H3 present in the VH comprising the amino acid sequence of SEQ ID NO:8.

[0201] In some cases, anti-C1s antibodies suitable for the present disclosure cross-compete with an antibody comprising: a) a light chain region comprising CDR-L1, CDR-L2, and CDR-L3 having the amino acid sequences set forth in SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3, respectively; and b) a heavy chain region comprising CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences set forth in SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6, respectively. In some of these embodiments, the anti-C1s antibody cross-competes with an antibody comprising a humanized V H and / or V L Includes framework regions.

[0202] In some cases, anti-C1s antibodies suitable for the present disclosure cross-compete with an antibody comprising: a) a light chain region comprising CDR-L1, CDR-L2, and CDR-L3 having the amino acid sequences set forth in SEQ ID NO: 10, SEQ ID NO: 11, and SEQ ID NO: 3, respectively; and b) a heavy chain region comprising CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences set forth in SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 14, respectively. In some of these embodiments, the anti-C1s antibody cross-competes with an antibody comprising a humanized V H and / or V L Includes framework regions.

[0203] In other embodiments, an anti-C1s antibody suitable for the present disclosure is an antibody that specifically binds to the same epitope as a reference antibody. a) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 18; b) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 19; c) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 20; d) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 21; e) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 18; f) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 19; g) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 20; h) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 21; i) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 18; j) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 19; k) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 20; or l) a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 21 Includes.

[0204] In some cases, an anti-C1s antibody suitable for the present disclosure specifically binds to the same epitope as an antibody comprising a variable region (VL) of a light chain region comprising CDR-L1, CDR-L2, and CDR-L3 present in the VL comprising the amino acid sequence of SEQ ID NO:7.

[0205] In some cases, an anti-C1s antibody suitable for the present disclosure specifically binds to the same epitope as an antibody comprising a variable region (VH) of a heavy chain region comprising CDR-H1, CDR-H2, and CDR-H3 present in the VH comprising the amino acid sequence of SEQ ID NO:8.

[0206] In some cases, an anti-C1s antibody suitable for the present disclosure specifically binds to the same epitope as an antibody comprising: a) a light chain region comprising CDR-L1, CDR-L2, and CDR-L3 having the amino acid sequences set forth in SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3, respectively; and b) a heavy chain region comprising CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences set forth in SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6, respectively. In some of these embodiments, the anti-C1s antibody is a humanized V H and / or V L Includes framework regions.

[0207] In some cases, an anti-C1s antibody suitable for the present disclosure specifically binds to the same epitope as an antibody comprising: a) a light chain region comprising CDR-L1, CDR-L2, and CDR-L3 having the amino acid sequences set forth in SEQ ID NO: 10, SEQ ID NO: 11, and SEQ ID NO: 3, respectively; and b) a heavy chain region comprising CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences set forth in SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 14, respectively. In some of these embodiments, the anti-C1s antibody is a humanized V H and / or V L Includes framework regions.

[0208] In some cases, an anti-C1s antibody suitable for the present disclosure comprises a light chain variable region comprising an amino acid sequence that is at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO:15.

[0209] In some cases, an anti-C1s antibody suitable for the present disclosure may be selected from the group consisting of SEQ ID NO: 16. and a light chain variable region comprising an amino acid sequence at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a described amino acid sequence.

[0210] In some cases, an anti-C1s antibody suitable for the present disclosure comprises a light chain variable region comprising an amino acid sequence that is at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO:17.

[0211] In some cases, an anti-C1s antibody suitable for the present disclosure comprises a heavy chain variable region comprising an amino acid sequence that is at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO:18.

[0212] In some cases, an anti-C1s antibody suitable for the present disclosure comprises a heavy chain variable region comprising an amino acid sequence that is at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO:19.

[0213] In some cases, an anti-C1s antibody suitable for the present disclosure comprises a heavy chain variable region comprising an amino acid sequence that is at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO:20.

[0214] In some cases, an anti-C1s antibody suitable for the present disclosure comprises a heavy chain variable region comprising an amino acid sequence that is at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO:21.

[0215] In some cases, an anti-C1s antibody suitable for the present disclosure cross-competes with an antibody comprising a VL region comprising the amino acid sequence set forth in SEQ ID NO:15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:18.

[0216] In some cases, an anti-C1s antibody suitable for the present disclosure cross-competes with an antibody comprising a VL region comprising the amino acid sequence set forth in SEQ ID NO:15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:19.

[0217] In some cases, an anti-C1s antibody suitable for the present disclosure cross-competes with an antibody comprising a VL region comprising the amino acid sequence set forth in SEQ ID NO:15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:20.

[0218] In some cases, an anti-C1s antibody suitable for the present disclosure cross-competes with an antibody comprising a VL region comprising the amino acid sequence set forth in SEQ ID NO:15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:21.

[0219] In some cases, an anti-C1s antibody suitable for the present disclosure cross-competes with an antibody comprising a VL region comprising the amino acid sequence set forth in SEQ ID NO:16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:18.

[0220] In some cases, an anti-C1s antibody suitable for the present disclosure may be selected from the group consisting of SEQ ID NO: 16. a VL region comprising the amino acid sequence set forth in SEQ ID NO:19; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:19.

[0221] In some cases, an anti-C1s antibody suitable for the present disclosure cross-competes with an antibody comprising a VL region comprising the amino acid sequence set forth in SEQ ID NO:16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:20.

[0222] In some cases, an anti-C1s antibody suitable for the present disclosure cross-competes with an antibody comprising a VL region comprising the amino acid sequence set forth in SEQ ID NO:16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:21.

[0223] In some cases, an anti-C1s antibody suitable for the present disclosure cross-competes with an antibody comprising a VL region comprising the amino acid sequence set forth in SEQ ID NO:17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:18.

[0224] In some cases, an anti-C1s antibody suitable for the present disclosure cross-competes with an antibody comprising a VL region comprising the amino acid sequence set forth in SEQ ID NO:17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:19.

[0225] In some cases, an anti-C1s antibody suitable for the present disclosure cross-competes with an antibody comprising a VL region comprising the amino acid sequence set forth in SEQ ID NO:17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:20.

[0226] In some cases, an anti-C1s antibody suitable for the present disclosure cross-competes with an antibody comprising a VL region comprising the amino acid sequence set forth in SEQ ID NO:17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO:21.

[0227] In some cases, an anti-C1s antibody suitable for the present disclosure binds to the same epitope as an antibody comprising a variable region (VL) of a light chain region comprising CDR-L1, CDR-L2, and CDR-L3 present in the VL comprising the amino acid sequence of SEQ ID NO:7.

[0228] In some cases, an anti-C1s antibody suitable for the present disclosure binds to the same epitope as an antibody comprising a variable region of a heavy chain (VH) including CDR-H1, CDR-H2, and CDR-H3 present in the VH comprising the amino acid sequence of SEQ ID NO:8.

[0229] In some cases, an anti-C1s antibody suitable for the present disclosure binds to the same epitope as an antibody comprising: a) a light chain region comprising CDR-L1, CDR-L2, and CDR-L3 having the amino acid sequences set forth in SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3, respectively; and b) a heavy chain region comprising CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences set forth in SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6, respectively. In some of these embodiments, the anti-C1s antibody is a humanized V H and / or V L Includes framework regions.

[0230] In some cases, an anti-C1s antibody suitable for the present disclosure binds to the same epitope as an antibody comprising: a) a light chain region comprising CDR-L1, CDR-L2, and CDR-L3 having the amino acid sequences set forth in SEQ ID NO: 10, SEQ ID NO: 11, and SEQ ID NO: 3, respectively; and b) a heavy chain region comprising CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences set forth in SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 14, respectively. In some of these embodiments, the anti-C1s antibody is a humanized V H and / or V L Includes framework regions.

[0231] In some cases, an anti-C1s antibody suitable for the present disclosure binds to the same epitope as an antibody comprising a light chain variable region comprising an amino acid sequence that is 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO:15.

[0232] In some cases, an anti-C1s antibody suitable for the present disclosure binds to the same epitope as an antibody comprising a light chain variable region comprising an amino acid sequence that is 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO:16.

[0233] In some cases, an anti-C1s antibody suitable for the present disclosure binds to the same epitope as an antibody comprising a light chain variable region comprising an amino acid sequence that is 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO:17.

[0234] In some cases, an anti-C1s antibody suitable for the present disclosure binds to the same epitope as an antibody comprising a heavy chain variable region comprising an amino acid sequence that is 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO:18.

[0235] In some cases, an anti-C1s antibody suitable for the present disclosure binds to the same epitope as an antibody comprising a heavy chain variable region comprising an amino acid sequence that is 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO:19.

[0236] In some cases, an anti-C1s antibody suitable for the present disclosure binds to the same epitope as an antibody comprising a heavy chain variable region comprising an amino acid sequence that is 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO:20.

[0237] In some cases, an anti-C1s antibody suitable for the present disclosure binds to the same epitope as an antibody comprising a heavy chain variable region comprising an amino acid sequence that is 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO:21.

[0238] In some cases, the anti-C1s antibody suitable for the present disclosure binds to the same epitope as an antibody comprising a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 18. In some cases, the anti-C1s antibody suitable for the present disclosure binds to the same epitope as an antibody comprising a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 19. In some cases, the anti-C1s antibody suitable for the present disclosure binds to the same epitope as an antibody comprising a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 20. In some cases, the anti-C1s antibody suitable for the present disclosure binds to the same epitope as an antibody comprising a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 21. In some cases, the anti-C1s antibody suitable for the present disclosure binds to the same epitope as an antibody comprising a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 18. In some cases, an anti-C1s antibody suitable for the present disclosure may have a VL domain comprising the amino acid sequence set forth in SEQ ID NO:16. and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 19. In some cases, an anti-C1s antibody suitable for the present disclosure binds to the same epitope as an antibody comprising a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 20. In some cases, an anti-C1s antibody suitable for the present disclosure binds to the same epitope as an antibody comprising a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 21. In some cases, an anti-C1s antibody suitable for the present disclosure binds to the same epitope as an antibody comprising a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 18. In some cases, an anti-C1s antibody suitable for the present disclosure binds to the same epitope as an antibody comprising a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 19. In some cases, an anti-C1s antibody suitable for the present disclosure binds to the same epitope as an antibody comprising a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 20. In some cases, an anti-C1s antibody suitable for the present disclosure binds to the same epitope as an antibody comprising a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17; and a VH region comprising the amino acid sequence set forth in SEQ ID NO: 21.

[0239] In some embodiments, a suitable anti-C1s antibody for the present disclosure is BIVV009. BIVV009 comprises a heavy chain region comprising the amino acid sequence set forth in SEQ ID NO:22 and a light chain region comprising the amino acid sequence set forth in SEQ ID NO:23.

[0240] SEQ ID NO: 22 Heavy chain of BIVV009: EVQLVESGGGLVKPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVATISSGGSHTYYLDSVKGRFTISRDNSKNTLYLQMNSLRAEDTALYYCARLFTGYAMDYWGQG TLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGP PCPPCPAFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKT ISKAKGQPREPQVYTLPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK

[0241] SEQ ID NO: 23 Light chain of BIVV009: QIVLTQSPATLSLSPGERATMSCTASSSVSSSYLHWYQQKPGKAPKLWIYSTSNLASGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCHQYYRLPPITFGQGTKLEI KRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0242] In some embodiments, an anti-C1s antibody suitable for the present disclosure is selected from the group consisting of an Ig monomer, a Fab fragment, a F(ab')2 fragment, an Fd fragment, a scFv, a scAb, a dAb, an Fv, a single domain heavy chain antibody, and a single domain light chain antibody.

[0243] In some cases, the anti-C1s antibody suitable for the present disclosure is an immunoglobulin The antibody may comprise a constant region (e.g., an Fc region) of a human Fc region, if present. The Fc region, if present, may be a human Fc region or an Fc region from any animal having a complement system. In some embodiments, the Fc region, if present, is a human Fc region. If a constant region is present, the antibody may comprise both a light chain constant region and a heavy chain constant region. A suitable heavy chain constant region comprises a CH1, hinge, CH2, CH3, and CH4 region. The antibodies described herein include antibodies with all types of constant regions, such as IgM, IgG, IgD, IgA, and IgE, as well as antibodies with all isotypes, such as IgG1, IgG2, IgG3, and IgG4. An example of a suitable heavy chain Fc region is an Fc of human isotype IgG1. Another example of a suitable heavy chain Fc region is an Fc of human isotype IgG2. Yet another example of a suitable heavy chain Fc region is an Fc of human isotype IgG3. The light chain constant region may be lambda or kappa. Anti-C1s antibodies suitable for use in the methods of the present disclosure may contain sequences from more than one class or isotype. Antibodies can be expressed as tetramers containing two light chains and two heavy chains, as separate heavy and light chains, as Fab, Fab', F(ab')2, and Fv, or as single chain antibodies in which the heavy and light chain variable domains are linked via a spacer.

[0244] In some cases, the heavy chain region is of isotype IgG4. In some of these embodiments, the hinge region comprises an S241P substitution. See, e.g., Angal et al. (1993) Mol. Immunol. 30:105. In some of these embodiments, the hinge region comprises an L236E (or L235E, using EU numbering; Kabat et al. (1991) Sequences of Proteins of Immunological Interest, 5th ed., USDept. Health and Human Services, Bethesda, MD, NIH Press 91-3242) substitution. See, e.g., Reddy et al. (2000) J. Immunol. 164:1925; and Klechevsky et al. (2010) Blood 116:1685. In some of these embodiments, the hinge region comprises an S241P substitution and an L236E substitution.

[0245] In other embodiments, an anti-C1s antibody suitable for the present disclosure comprises a heavy chain comprising an amino acid sequence at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO:XX. In some embodiments, an anti-C1s antibody suitable for the present disclosure comprises a light chain comprising an amino acid sequence at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO:XX. In other embodiments, an anti-C1s antibody suitable for the present disclosure comprises a heavy chain comprising an amino acid sequence at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO:XX, and a light chain comprising an amino acid sequence at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO:XX. In some embodiments, an anti-C1s antibody suitable for the present disclosure comprises a heavy chain comprising an amino acid sequence at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO:XX, and a light chain comprising an amino acid sequence at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO:XX, wherein the anti-C1s antibody is a BIV antibody. Contains the six CDRs of V009.

[0246] Anti-C1s antibodies suitable for the present disclosure may be substantially pure, e.g., at least about 80% to 85% pure, at least about 85% to 90% pure, at least about 90% to 95% pure, or 98% to 99% or more pure, e.g., free from contaminants such as dead cell debris, macromolecules other than anti-C1s antibodies, etc.

[0247] composition The anti-C1s antibody will generally be present in a composition, such as a pharmaceutical composition.

[0248] Compositions comprising anti-C1s antibodies may also include one or more of the following: a salt, e.g., NaCl, MgCl2, KCl, MgSO4, and the like; a buffer, e.g., Tris buffer, N-(2-hydroxyethyl)piperazine-N'-(2-ethanesulfonic acid) (HEPES), 2-(N-morpholino)ethanesulfonic acid (MES), 2-(N-morpholino)ethanesulfonic acid sodium salt (MES), 3-(N-morpholino)propanesulfonic acid (MOPS), N-tris[hydroxymethyl]methyl-3-aminopropanesulfonic acid (TAPS), and the like; a solubilizing agent; a detergent, e.g., a non-ionic detergent, e.g., Tween-20, and the like; a protease inhibitor; glycerol, and the like.

[0249] In carrying out the method of the present disclosure, the anti-C1s antibody can be administered to an individual using any convenient means capable of producing the desired therapeutic or diagnostic effect. Thus, the anti-C1s antibody can be incorporated into various formulations for therapeutic administration. More specifically, the anti-C1s antibody can be formulated into a pharmaceutical composition by combining with a suitable pharma-ceutically acceptable carrier, a pharma-ceutically acceptable diluent, or other pharma-ceutically acceptable excipient, or can be formulated into a solid, semi-solid, liquid, or gaseous form of preparation, such as tablets, capsules, powders, granules, ointments, liquids, suppositories, injections, inhalants, and aerosols. In some cases, the pharmaceutical composition comprises an anti-C1s antibody and a pharma-ceutically acceptable excipient.

[0250] In pharmaceutical dosage forms, the anti-C1s antibody may be administered in the form of its pharma- ceutically acceptable salt, or the anti-C1s antibody may be used alone or in appropriate association with other pharma- ceutically active compounds, as well as in combination therewith. The following methods and excipients are merely illustrative and in no way limiting.

[0251] For oral preparations, the anti-C1s antibody may be used alone or in combination with suitable excipients for making tablets, powders, granules or capsules, such as conventional excipients such as lactose, mannitol, corn starch or potato starch; binders such as crystalline cellulose, cellulose derivatives, acacia, corn starch or gelatin; disintegrants such as corn starch, potato starch or sodium carboxymethylcellulose; lubricants such as talc or magnesium stearate; and, if necessary, diluents, buffers, wetting agents, preservatives and flavorings.

[0252] Anti-C1s antibodies can be formulated into preparations for injection by dissolving, suspending or emulsifying the antibodies in aqueous or non-aqueous solvents, such as vegetable oils or other similar oils, propylene glycol, synthetic fatty acid glycerides, injectable organic esters (e.g., ethyl oleate), esters of higher fatty acids or propylene glycol; optionally with conventional additives, such as solubilizers, isotonicity agents, suspending agents, emulsifiers, stabilizers and preservatives. Parenteral vehicles include sodium chloride, sodium lauryl sulfate, sodium phosphate esters ... Intravenous vehicles include sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, lactated Ringer's solution, or fixed oils. Intravenous vehicles include fluid and nutrient replenishers, electrolyte replenishers (e.g., those based on Ringer's dextrose), and the like. In addition, the pharmaceutical compositions of the present disclosure may contain additional agents, such as dopamine or psychopharmacological drugs, depending on the intended use of the pharmaceutical composition.

[0253] Pharmaceutical compositions containing anti-C1s antibodies are prepared by mixing the subject antibody having the desired purity with optional physiologically acceptable carriers, other excipients, stabilizers, surfactants, buffers and / or isotonicity agents. Acceptable carriers, other excipients and / or stabilizers are non-toxic to recipients at the dosages and concentrations employed, and include, for example, buffers such as phosphate buffers, citrate buffers and other organic acid buffers; antioxidants such as ascorbic acid, glutathione, cysteine, methionine and citric acid; preservatives (e.g., ethanol, benzyl alcohol, phenol, m-cresol, p-chloro-m-cresol, methyl or propyl paraben, benzalkonium chloride, or combinations thereof); amino acids such as arginine, glycine, ornithine, lysine, histidine, glutamic acid, aspartic acid, isoleucine, leucine, alanine, phenanthroline, phenylalanine ... monosaccharides, disaccharides and other carbohydrates; low molecular weight (less than about 10 residues) polypeptides; proteins, such as gelatin or serum albumin; chelating agents, such as EDTA; sugars, such as trehalose, sucrose, lactose, glucose, mannose, maltose, galactose, fructose, sorbose, raffinose, glucosamine, N-methylglucosamine, galactosamine, and neuraminic acid; and / or non-ionic surfactants, such as Tween, Brij, Pluronic, Triton-X, or polyethylene glycol (PEG).

[0254] The pharmaceutical composition may be in liquid form, lyophilized form, or liquid form reconstituted from lyophilized form, and lyophilized preparations will be reconstituted with sterile solution before administration.The standard procedure for reconstituting a lyophilized composition is to add back a volume of pure water (typically equal to the volume removed during lyophilization); however, to produce pharmaceutical compositions for parenteral administration, solutions containing antibacterial agents can be used; see also Chen (1992) Drug Dev Ind Pharm 18, pp. 1311-54.

[0255] Exemplary antibody concentrations in pharmaceutical compositions suitable for use in the methods of the present disclosure may range from about 1 mg / mL to about 200 mg / mL, or from about 50 mg / mL to about 200 mg / mL, or from about 150 mg / mL to about 200 mg / mL. In some embodiments, the antibody concentration is from about 10 mg / mL to about 60 mg / mL, from about 12 mg / mL to about 58 mg / mL, from about 14 mg / mL to about 56 mg / mL, from about 16 mg / mL to about 54 mg / mL, from about 17 mg / mL to about 52 mg / mL, or from about 18 mg / mL to about 50 mg / mL. In some embodiments, the antibody concentration is 18 mg / mL. In some embodiments, the antibody concentration is 50 mg / mL.

[0256] Aqueous formulations of anti-C1s antibodies can be prepared in pH buffers, for example, at a pH ranging from about 4.0 to about 7.0, or from about 5.0 to about 6.0, or alternatively at about 5.5. Examples of buffers suitable for a pH within this range include phosphate, histidine, citrate, succinate, acetate buffers and other organic acid buffers. The concentration of the buffer can be from about 1 mM to about 100 mM, or from about 5 mM to about 50 mM, depending, for example, on the buffer and the desired tonicity of the formulation.

[0257] Tonicity agents can be included in the antibody formulation to modulate the tonicity of the formulation. Exemplary tonicity agents include sodium chloride, potassium chloride, glycerin, and any member from the group of amino acids, sugars, as well as combinations thereof.In some embodiments, the aqueous formulation is isotonic, but hypertonic or hypotonic solutions may also be suitable.The term "isotonic" refers to a solution that has the same tonicity as any other solution to which it is compared, such as saline or serum.Isotonicity agents can be used in amounts of about 5mM to about 350mM, for example, in amounts of 100mM to 350nM.

[0258] Surfactants can also be added to antibody formulations to reduce aggregation of the formulated antibody and / or minimize the formation of particulates in the formulation and / or reduce adsorption. Exemplary surfactants include polyoxyethylene sorbitan fatty acid esters (Tween), polyoxyethylene alkyl ethers (Brij), alkylphenyl polyoxyethylene ethers (Triton-X), polyoxyethylene-polyoxypropylene copolymers (Poloxamer, Pluronic), and sodium dodecyl sulfate (SDS). Examples of suitable polyoxyethylene sorbitan fatty acid esters are polysorbate 20 (sold under the trademark Tween 20™) and polysorbate 80 (sold under the trademark TWEEN 80™). Examples of suitable polyethylene-polypropylene copolymers are those sold under the names PLURONIC® F68 or POLOXAMER 188™. Examples of suitable polyoxyethylene alkyl ethers are those sold under the trademark BRIJ™. Exemplary concentrations of the surfactant may range from about 0.001% to about 1% w / v.

[0259] Lyoprotectants can also be added to protect labile active ingredients (e.g., proteins) from destabilizing conditions during the lyophilization process.For example, known lyoprotectants include sugars (such as glucose and sucrose); polyols (such as mannitol, sorbitol and glycerol); and amino acids (such as alanine, glycine and glutamic acid).Lyoprotectants are included in an amount of about 10 mM to 500 nM.

[0260] In some cases, a suitable formulation comprises an anti-C1s antibody and one or more of the agents identified above (e.g., surfactants, buffers, stabilizers, tonicity agents), and is essentially free of one or more preservatives, such as ethanol, benzyl alcohol, phenol, m-cresol, p-chloro-m-cresol, methyl or propyl paraben, benzalkonium chloride, and combinations thereof. In other embodiments, a preservative is included in the formulation, for example, at a concentration ranging from about 0.001 to about 2% (w / v).

[0261] For example, a suitable formulation may be a liquid or lyophilized formulation suitable for parenteral administration that contains: about 1 mg / mL to about 200 mg / mL of a subject antibody; about 0.001% to about 1% of at least one surfactant; about 1 mM to about 100 mM buffer; optionally about 10 mM to about 500 mM stabilizer; and about 5 mM to about 305 mM isotonicity agent; and has a pH of about 4.0 to about 7.0.

[0262] As another example, a suitable parenteral formulation is a liquid or lyophilized formulation comprising about 1 mg / mL to about 200 mg / mL of anti-C1s antibody; 0.04% Tween 20 w / v; 20 mM L-histidine; and 250 mM sucrose; having a pH of 5.5.

[0263] As another example, a subject parenteral formulation may be: 1) a lyophilized formulation comprising 15 mg / mL of an anti-C1s antibody; 0.04% Tween 20 w / v; 20 mM L-histidine; and 250 mM sucrose; with a pH of 5.5; or 2) a lyophilized formulation comprising 75 mg / mL of a subject antibody; 0.04% Tween 20 w / v; 20 mM L-histidine; and 250 mM sucrose; with a pH of 5.5; or 3) a lyophilized formulation comprising 75 mg / mL of an anti-C1s antibody; s antibody; 0.02% Tween 20 w / v; 20 mM L-histidine; and 250 mM sucrose; a lyophilized formulation having a pH of 5.5; or 4) 75 mg / mL anti-C1s antibody; 0.04% Tween 20 w / v; 20 mM L-histidine; and 250 mM trehalose; a lyophilized formulation having a pH of 5.5; or 5) 75 mg / mL anti-C1s antibody; 0.02% Tween 20 w / v; 20 mM L-histidine; and 250 mM trehalose; a lyophilized formulation having a pH of 5.5.

[0264] As another example, a suitable parenteral formulation may be: 1) a liquid formulation comprising 7.5 mg / mL anti-C1s antibody; 0.02% Tween 20 w / v; 120 mM L-histidine; and 125 mM sucrose; and having a pH of 5.5; or 2) a liquid formulation comprising 37.5 mg / mL anti-C1s antibody; 0.02% Tween 20 w / v; 10 mM L-histidine; and 125 mM sucrose; and having a pH of 5.5; or 3) a liquid formulation comprising 37.5 mg / mL anti-C1s antibody; 0.01% Tween 20 w / v; 10 mM L-histidine; and 125 mM sucrose; 10 mM sucrose; and a liquid formulation having a pH of 5.5; or 4) a liquid formulation containing 37.5 mg / mL anti-C1s antibody; 0.02% Tween 20 w / v; 10 mM L-histidine; and 125 mM trehalose; and a liquid formulation having a pH of 5.5; or 5) a liquid formulation containing 37.5 mg / mL anti-C1s antibody; 0.01% Tween 20 w / v; 10 mM L-histidine; and 125 mM trehalose; and a liquid formulation having a pH of 5.5; or 6) a liquid formulation containing 5 mg / mL anti-C1s antibody; 0.02% Tween 20 w / v; 20 mM L-histidine. and 250 mM trehalose; and a liquid formulation having a pH of 5.5; or 7) a liquid formulation containing 75 mg / mL anti-C1s antibody; 0.02% Tween 20 w / v; 20 mM L-histidine; and 250 mM mannitol; and a pH of 5.5; or 8) a liquid formulation containing 75 mg / mL anti-C1s antibody; 0.02% Tween 20 w / v; 20 mM L-histidine; and 140 mM sodium chloride; and a pH of 5.5; or 9) a liquid formulation containing 150 mg / mL anti-C1s antibody; 0.02% Tween 20 w / v; 20 mM L-histidine; and 250 mM trehalose; a liquid formulation having a pH of 5.5; or 10) 150 mg / mL anti-C1s antibody; 0.02% Tween 20 w / v; 20 mM L-histidine; and 250 mM mannitol; a liquid formulation having a pH of 5.5; or 11) 150 mg / mL anti-C1s antibody; 0.02% Tween 20 w / v; 20 mM L-histidine; and 140 mM sodium chloride; a liquid formulation having a pH of 5.5; or 12) 10 mg / mL anti-C1s antibody; 0.It is a liquid formulation containing: 0.1% Tween 20 w / v; 20 mM L-histidine; and 40 mM sodium chloride; and has a pH of 5.5.

[0265] Suitable excipient vehicles are, for example, water, saline, dextrose, glycerol, ethanol, etc., and combinations thereof. In addition, if necessary, vehicle may contain minor amounts of auxiliary substances, such as wetting or emulsifying agents or pH buffering agents. The actual method of producing such dosage forms is known or will be obvious to those skilled in the art. For example, see Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pennsylvania, 17th Edition, 1985. The composition or formulation to be administered will in any case contain an appropriate amount of the subject's antibody to achieve the desired state in the subject being treated.

[0266] Pharmaceutically acceptable excipients, such as vehicles, adjuvants, carriers or diluents, are readily available to the public. Additionally, pharma- ceutically acceptable auxiliary substances, such as pH adjusting and buffering agents, tonicity adjusting agents, stabilizers, wetting agents, and the like, are readily available to the public.

[0267] Dosage The present disclosure provides a method of treating a complement-mediated disease or disorder in an individual, comprising administering an anti-C1s antibody to the individual, wherein the anti-C1s antibody is administered in an effective amount of at least 4 g, at least 4.5 g, at least 5 g, at least 5.5 g, at least 6 g, at least 6.5 g, at least 7 g, at least 7.5 g, at least 8 g, at least 8.5 g, at least 9 g, at least 9.5 g, or at least 10 g.

[0268] In some embodiments, the anti-C1s antibody is administered in an effective amount of between about 5.5g and about 10g, between about 5.5g and about 9.5g, between about 5.5g and about 9g, between about 5.5g and about 8.5g, between about 5.5g and about 8g, between about 5.5g and about 7.5g, between about 5.5g and about 7g, between about 5.5g and about 6.5g, or between about 5.5g and about 6g. In some embodiments, the anti-C1s antibody is administered in an amount of between about 4.5g and about 8.5g, between about 4.5g and about 8g, between about 4.5g and about 7.5g, between about 4.5g and about 7g, between about 4.5g and about 6.5g, between about 4.5g and about 6g, between about 4.5g and about 5.5g, or between about 4.5g and about 5g. In some embodiments, the anti-C1s antibody is administered in an amount of between about 7.5g and about 12g, between about 7.5g and about 11.5g, between about 7.5g and about 11g, between about 7.5g and about 10.5g, between about 7.5g and about 10g, between about 7.5g and about 9.5g, between about 7.5g and about 9g, between about 7.5g and about 8.5g, or between about 7.5g and about 8g.

[0269] In one embodiment, the present disclosure provides a method of treating a complement-mediated disease or disorder in an individual, comprising administering an anti-C1s antibody to the individual, wherein the anti-C1s antibody is administered in an amount of 5.5 g. In some cases, the 5.5 g dose of anti-C1s antibody is administered to the individual every other week. In some cases, the method comprises: a) administering 5.5 g of anti-C1s antibody on day 1; b) administering 5.5 g of anti-C1s antibody on day 8; and c) administering 5.5 g of anti-C1s antibody every other week after administration on day 8. In some cases, the 5.5 g dose of anti-C1s antibody is administered to the individual every other week for a period of about 4 weeks to 1 year, e.g., about 4 weeks to about 8 weeks, about 2 months to about 6 months, or about 6 months to 1 year. In some cases, the 5.5 g dose of anti-C1s antibody is administered to the individual every other week for a period of more than 1 year. For example, in some cases, a 5.5 g dose of anti-C1s antibody is administered to an individual every two weeks for a period of 1 to 50 years, e.g., 1 to 2 years, 2 to 5 years, 5 to 10 years, 10 to 20 years, 20 to 30 years, 30 to 40 years, or 40 to 50 years.

[0270] In some embodiments, the individual in accordance with the methods of the invention weighs 75 kg or more and the anti-C1s antibody is administered in an effective amount of about 7.5 g. In other embodiments, the individual in accordance with the methods of the invention weighs 75 kg or less and the anti-C1s antibody is administered in an effective amount of about 6.5 g.

[0271] In another embodiment, the present disclosure also provides a method of treating a complement-mediated disease or disorder in an individual, comprising administering an anti-C1s antibody to the individual, wherein the anti-C1s antibody is administered in an effective amount of about 6.5 g. In some cases, the effective amount of about 6.5 g of the anti-C1s antibody is administered to the individual every other week. In some cases, the method comprises: a) administering an effective amount of about 6.5 g of the anti-C1s antibody on day 1; b) administering an effective amount of about 6.5 g of the anti-C1s antibody on day 8; and c) administering an effective amount of about 6.5 g of the anti-C1s antibody every other week after administration on day 8. In some cases, the effective amount of about 6.5 g of the anti-C1s antibody is administered to the individual every other week for a period of about 4 weeks to 1 year, e.g., about 4 weeks to about 8 weeks, about 2 months to about 6 months, or about 6 months to 1 year. In some cases, an effective amount of about 6.5 g of anti-C1s antibody is administered to an individual every other week for a period of more than one year. An effective amount of .5 g is administered to an individual every two weeks for a period of 1 to 50 years, e.g., 1 to 2 years, 2 to 5 years, 5 to 10 years, 10 to 20 years, 20 to 30 years, 30 to 40 years, or 40 to 50 years.

[0272] In another embodiment, the present disclosure also provides a method of treating a complement-mediated disease or disorder in an individual, comprising administering an anti-C1s antibody to the individual, wherein the anti-C1s antibody is administered in an effective amount of about 7.5 g. In some cases, the effective amount of about 7.5 g of the anti-C1s antibody is administered to the individual every other week. In some cases, the method includes: a) administering an effective amount of about 7.5 g of the anti-C1s antibody on day 1; b) administering an effective amount of about 7.5 g of the anti-C1s antibody on day 8; and c) administering an effective amount of about 7.5 g of the anti-C1s antibody every other week after administration on day 8. In some cases, the effective amount of about 7.5 g of the anti-C1s antibody is administered to the individual every other week for a period of about 4 weeks to 1 year, e.g., about 4 weeks to about 8 weeks, about 2 months to about 6 months, or about 6 months to 1 year. In some cases, an effective amount of about 7.5 g of anti-C1s antibody is administered to an individual every other week for a period of greater than 1 year, for example, in some cases, an effective amount of about 7.5 g of anti-C1s antibody is administered to an individual every other week for a period of 1 to 50 years, e.g., 1 to 2 years, 2 to 5 years, 5 to 10 years, 10 to 20 years, 20 to 30 years, 30 to 40 years, or 40 to 50 years.

[0273] In other embodiments, the present disclosure provides a method of treating a complement-mediated disease or disorder in an individual, comprising administering an anti-C1s antibody to the individual, wherein the anti-C1s antibody is administered in an effective amount of between about 6.5 g and about 7.5 g. In some cases, an effective amount of between about 6.5 g and about 7.5 g of the anti-C1s antibody is administered to the individual every other week. In some cases, the method comprises administering an effective amount of between about 6.5 g and about 7.5 g of the anti-C1s antibody on days 0 and 7, and then every other week thereafter. In some cases, an effective amount of between about 6.5 g and 7.5 g of the anti-C1s antibody is administered to the individual every other week for a period of about 4 weeks to 1 year, e.g., about 4 weeks to about 8 weeks, about 2 months to about 6 months, or about 6 months to 1 year. In some cases, an effective amount of between about 6.5 g and 7.5 g of the anti-C1s antibody is administered to the individual every other week for a period of more than 1 year.

[0274] The present disclosure also provides a method of treating a complement-mediated disease or disorder in a subject in need thereof, comprising administering to the subject an effective amount of an anti-C1s antibody, wherein the serum concentration of the anti-C1s antibody following administration is at least about 20 μg / mL, at least about 25 μg / mL, at least about 30 μg / mL, at least about 35 μg / mL, at least about 40 μg / mL, at least about 45 μg / mL, at least about 50 μg / mL, at least about 55 μg / mL, at least about 60 μg / mL, at least about 65 μg / mL, at least about 70 μg / mL, at least about 75 μg / mL, at least about 80 μg / mL, at least about 85 μg / mL, at least about 90 μg / mL, at least about 95 μg / mL, or at least about 100 μg / mL. In some embodiments of the present disclosure, the serum concentration of the anti-C1s antibody after administration is between about 20 μg / mL and about 100 μg / mL, between about 20 μg / mL and about 90 μg / mL, between about 20 μg / mL and about 80 μg / mL, between about 20 μg / mL and about 70 μg / mL, between about 20 μg / mL and about 70 μg / mL, between about 20 μg / mL and about 60 μg / mL, between about 20 μg / mL and about 50 μg / mL, between about 20 μg / mL and about 40 μg / mL, or between about 20 μg / mL and about 30 μg / mL. In certain embodiments, the serum concentration of the anti-C1s antibody after administration is at least about 20 μg / mL.

[0275] The serum concentration of anti-C1s antibodies in a subject can be measured using techniques known in the art. In some embodiments, anti-C1s antibodies are measured using a direct enzyme-linked immunosorbent assay (ELISA). In some embodiments, anti-C1s antibodies are measured using an indirect Measured using ELISA. In some embodiments, anti-C1s antibodies are measured using sandwich ELISA. In some embodiments, anti-C1s antibodies are measured using competitive ELISA.

[0276] The present disclosure provides a method of treating a complement-mediated disease or disorder in a subject in need thereof, comprising administering to the subject an effective amount of an anti-C1s antibody, wherein the effective amount of the anti-C1s antibody is at least about 45 mg / kg, at least about 50 mg / kg, at least about 55 mg / kg, at least about 60 mg / kg, at least about 65 mg / kg, at least about 70 mg / kg, at least about 75 mg / kg, at least about 80 mg / kg, at least about 85 mg / kg, at least about 90 mg / kg, at least about 95 mg / kg, or at least about 100 mg / kg. In a specific embodiment, the effective amount of the anti-C1s antibody is at least about 60 mg / kg.

[0277] In some embodiments, the effective amount of anti-C1s antibody is between about 60 mg / kg and about 100 mg / kg, between about 60 mg / kg and about 95 mg / kg, between about 60 mg / kg and about 90 mg / kg, between about 60 mg / kg and about 85 mg / kg, between about 60 mg / kg and about 80 mg / kg, between about 60 mg / kg and about 75 mg / kg, between about 60 mg / kg and about 70 mg / kg, or between about 60 mg / kg and about 65 mg / kg. In some embodiments, the effective amount of anti-C1s antibody is between about 45 mg / kg and about 85 mg / kg, between about 45 mg / kg and about 80 mg / kg, between about 45 mg / kg and about 75 mg / kg, between about 45 mg / kg and about 70 mg / kg, between about 45 mg / kg and about 65 mg / kg, between about 45 mg / kg and about 60 mg / kg, or between about 45 mg / kg and about 50 mg / kg. In some embodiments, an effective amount of an anti-C1s antibody is between about 85 mg / kg and about 150 mg / kg, between about 85 mg / kg and about 145 mg / kg, between about 85 mg / kg and about 140 mg / kg, between about 85 mg / kg and about 135 mg / kg, between about 85 mg / kg and about 130 mg / kg, between about 85 mg / kg and about 125 mg / kg, between about 85 mg / kg and about 125 mg / kg, between about 85 mg / kg and about 120 mg / kg, between about 85 mg / kg and about 115 mg / kg, between about 85 mg / kg and about 110 mg / kg, between about 85 mg / kg and about 105 mg / kg, between about 85 mg / kg and about 100 mg / kg, between about 85 mg / kg and about 95 mg / kg, or between about 85 mg / kg and about 90 mg / kg.

[0278] In some embodiments, the effective amount for the methods of the present invention is about 45 mg / kg, about 50 mg / kg, about 55 mg / kg, about 60 mg / kg, about 65 mg / kg, about 70 mg / kg, about 75 mg / kg, about 80 mg / kg, about 85 mg / kg, about 90 mg / kg, about 95 mg / kg, about 100 mg / kg, about 105 mg / kg, about 110 mg / kg, about 115 mg / kg, about 120 mg / kg, about 125 mg / kg, about 130 mg / kg, about 135 mg / kg, about 140 mg / kg, about 145 mg / kg, or about 150 mg / kg.

[0279] The present disclosure provides a method of treating a complement-mediated disease or disorder in a subject in need thereof, comprising administering to the subject an effective amount of an anti-C1s antibody, wherein the anti-C1s antibody is administered at a dosing interval of 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or 31 days.

[0280] In some embodiments, the anti-C1s antibody is administered at a dosing interval of 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 2 months, 3 months, or 4 months. In some embodiments, after administration of the anti-C1s antibody, the anti-C1s antibody increases the number of reticulocytes in the subject's blood.

[0281] In some embodiments, the anti-C1s antibody is administered in one or more loading doses followed by spaced doses, which can be administered about 7 days apart, about 14 days apart, about 21 days apart, about 28 days apart, about 2 months apart, about 3 months apart, or about 4 months apart. In some embodiments, the loading dose for the present disclosure is about 45mg / kg, about 50mg / kg, about 55mg / kg, about 60mg / kg, about 65mg / kg, about 70mg / kg, about 75mg / kg, about 80mg / kg, about 85mg / kg, about 90mg / kg, about 95mg / kg, about 100mg / kg, about 105mg / kg, about 110mg / kg, about 115mg / kg, about 120mg / kg, about 125mg / kg, about 130mg / kg, about 135mg / kg, about 140mg / kg, about 145mg / kg, or about 150mg / kg. In some embodiments, the loading dose is a different dosage from the dose administered at the interval. In some embodiments, the loading dose is the same dosage as the dose administered at the interval. In one embodiment, the anti-C1s antibody is administered at a loading dose of 60 mg / kg twice weekly, followed by doses of 60 mg / kg every other week.

[0282] The present disclosure provides a method for increasing the number of reticulocytes in the blood of a subject in need thereof, comprising administering to the subject an effective amount of an anti-C1s antibody. In some embodiments, the anti-C1s antibody increases the number of reticulocytes in the blood of the subject by at least about 1.1-fold, at least about 1.2-fold, at least about 1.3-fold, at least about 1.4-fold, at least about 1.5-fold, at least about 1.6-fold, at least about 1.7-fold, at least about 1.8-fold, at least about 1.9-fold, at least about 2.0-fold, at least about 2.1-fold, at least about 2.2-fold, at least about 2.3-fold, at least about 2.4-fold, at least about 2.5-fold, at least about 2.6-fold, at least about 2.7-fold, at least about 2.8-fold, at least about 2.9-fold, at least about 3.0-fold, at least about 4-fold, at least about 5-fold, at least about 6-fold, at least about 7-fold, at least about 8-fold, at least about 9-fold, or at least about 10-fold after administration.

[0283] In some embodiments, the anti-C1s antibody reduces the number of reticulocytes in the subject's blood within about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 9 hours, about 10 hours, about 11 hours, about 12 hours, about 13 hours, about 14 hours, about 15 hours, about 16 hours, about 17 hours, about 18 hours, about 19 hours, about 20 hours, about 21 hours, about 22 hours, or within about 24 hours of administration. The increase is within about 23 hours, within about 24 hours, within about 1 day, within about 2 days, within about 3 days, within about 4 days, within about 5 days, within about 6 days, within about 7 days, within about 8 days, within about 9 days, within about 10 days, within about 11 days, within about 12 days, within about 13 days, within about 14 days, within about 2 weeks, within about 3 weeks, within about 4 weeks, within about 5 weeks, within about 6 weeks, within about 7 weeks, within about 8 weeks, within about 9 weeks, within about 10 weeks, within about 11 weeks, or within about 12 weeks.

[0284] The present disclosure provides a method for increasing the number of reticulocytes in the blood of a subject in need thereof, comprising administering to the subject an effective amount of an anti-C1s antibody, wherein the effective amount of the anti-C1s antibody is at least about 45 mg / kg, at least about 50 mg / kg, at least about 55 mg / kg, at least about 60 mg / kg, at least about 65 mg / kg, at least about 70 mg / kg, at least about 75 mg / kg, at least about 80 mg / kg, at least about 85 mg / kg, at least about 90 mg / kg, at least about 95 mg / kg, or at least about 100 mg / kg.

[0285] In some embodiments, an effective amount of an anti-C1s antibody is from about 60 mg / kg to about 100 mg / kg, from about 60 mg / kg to about 95 mg / kg, from about 60 mg / kg to about 90 mg / kg, from about 60 mg / kg to about 85 mg / kg, from about 60 mg / kg to about 80 mg / kg, from about Between about 60 mg / kg to about 75 mg / kg, about 60 mg / kg to about 70 mg / kg, or about 60 mg / kg to about 65 mg / kg. In some embodiments, the effective amount of anti-C1s antibody is between about 45 mg / kg to about 85 mg / kg, about 45 mg / kg to about 80 mg / kg, about 45 mg / kg to about 75 mg / kg, about 45 mg / kg to about 70 mg / kg, about 45 mg / kg to about 65 mg / kg, about 45 mg / kg to about 60 mg / kg, or about 45 mg / kg to about 50 mg / kg. In some embodiments, the effective amount of an anti-C1s antibody is between about 85 mg / kg to about 150 mg / kg, about 85 mg / kg to about 145 mg / kg, about 85 mg / kg to about 140 mg / kg, about 85 mg / kg to about 135 mg / kg, about 85 mg / kg to about 130 mg / kg, about 85 mg / kg to about 125 mg / kg, about 85 mg / kg to about 125 mg / kg, about 85 mg / kg to about 120 mg / kg, about 85 mg / kg to about 115 mg / kg, about 85 mg / kg to about 110 mg / kg, about 85 mg / kg to about 105 mg / kg, about 85 mg / kg to about 100 mg / kg, about 85 mg / kg to about 95 mg / kg, or about 85 mg / kg to about 90 mg / kg.

[0286] In some embodiments, the effective amount for the methods of the present invention is about 45 mg / kg, about 50 mg / kg, about 55 mg / kg, about 60 mg / kg, about 65 mg / kg, about 70 mg / kg, about 75 mg / kg, about 80 mg / kg, about 85 mg / kg, about 90 mg / kg, about 95 mg / kg, about 100 mg / kg, about 105 mg / kg, about 110 mg / kg, about 115 mg / kg, about 120 mg / kg, about 125 mg / kg, about 130 mg / kg, about 135 mg / kg, about 140 mg / kg, about 145 mg / kg, or about 150 mg / kg.

[0287] The present disclosure also provides a method for increasing the number of reticulocytes in the blood of a subject in need thereof, comprising administering an anti-C1s antibody to the individual, wherein the anti-C1s antibody is administered in an amount of at least about 4 g, at least about 4.5 g, at least about 5 g, at least about 5.5 g, at least about 6 g, at least about 6.5 g, at least about 7 g, at least about 7.5 g, at least about 8 g, at least about 8.5 g, at least about 9 g, at least about 9.5 g, or at least about 10 g.

[0288] In some embodiments, the anti-C1s antibody is administered in an amount between about 5.5g to about 10g, about 5.5g to about 9.5g, about 5.5g to about 9g, about 5.5g to about 8.5g, about 5.5g to about 8g, about 5.5g to about 7.5g, about 5.5g to about 7g, about 5.5g to about 6.5g, or about 5.5g to about 6g. In some embodiments, the anti-C1s antibody is administered in an amount between about 4.5g to about 8.5g, about 4.5g to about 8g, about 4.5g to about 7.5g, about 4.5g to about 7g, about 4.5g to about 6.5g, about 4.5g to about 6g, about 4.5g to about 5.5g, or about 4.5g to about 5g. In some embodiments, the anti-C1s antibody is administered in an amount between about 7.5g to about 12g, about 7.5g to about 11.5g, about 7.5g to about 11g, about 7.5g to about 10.5g, about 7.5g to about 10g, about 7.5g to about 9.5g, about 7.5g to about 9g, about 7.5g to about 8.5g, or about 7.5g to about 8g.

[0289] The present disclosure provides a method of increasing hemoglobin levels in a subject in need thereof, comprising administering to the subject an effective amount of an anti-C1s antibody. In some embodiments, the anti-C1s antibody increases hemoglobin levels in a subject after administration to at least 1.0 g / dL, 1.1 g / dL, 1.2 g / dL, 1.3 g / dL, 1.4 g / dL, 1.5 g / dL, 1.6 g / dL, 1.7 g / dL, 1.8 g / dL, 1.9 g / dL, 2.0 g / dL, 2.1 g / dL, 2.2 g / dL, 2.3 g / dL, 2.4 g / dL, 2.5 g / dL, 2.6 g / dL, 2.7 g / dL, 2.8 g / dL, 2.9 g / dL, 3.0 g / dL, 3.1 g / dL, 3.2 g / dL, 3.3 g / dL, 3.4 g / dL, 3.6 g / dL, 3.8 g / dL, 3.9 g / dL, 4.0 g / dL, 4.1 g / dL, 4.2 g / dL, 4.3 g / dL, 4.4 g / dL, 4.5 g / dL, 4.6 g / dL, 4.7 g / dL, 4.8 g / dL, 4.9 g / dL, 5.0 g / dL, 5.1 g / dL, 5.2 g / dL, 5.3 g / dL, 5.4 g / dL, 5.5 g / dL, 5.6 g / dL, 5.7 g / dL, 5.8 g / dL, 5.9 g / dL, 6.0 g / dL, 6.1 g / dL, 6.2 g / dL, 6.3 g / dL, 6.4 g / dL, 6.5 g / dL, / dL, 3.5g / dL, 3.6g / dL, 3.7g / dL, 3.8g / dL, 3.9g / dL, 4.0g / dL, 4.1g / dL, 4.2g / dL, 4.3g / dL, 4.4g / dL, or 4.5g / dL.

[0290] In some embodiments, the anti-C1s antibody, following administration, increases total hemoglobin levels in the subject to at least 10.0 g / dL, at least 10.1 g / dL, at least 10.2 g / dL, at least 10.3 g / dL, at least 10.4 g / dL, at least 10.5 g / dL, at least 10.6 g / dL, at least 10.7 g / dL, at least 10.8 g / dL, at least 10.9 g / dL, at least 11.0 g / dL, at least 11.1 g / dL, at least 11.2 g / dL, at least 11.3 g / dL, at least 11.4 g / dL, at least 11.6 g / dL, at least 11.8 ...4 g / dL, at least 11.6 g / dL, at least 11.8 g / dL, at least 11.8 g / dL, at least 11.9 g / dL, at least 11.8 g / dL, at least 11.9 g / dL, at least 11.8 g / dL, at least 11.9 g / dL, at least 11 g / dL, at least 11.5g / dL, at least 11.6g / dL, at least 11.7g / dL, at least 11.8g / dL, at least 11.9g / dL, at least 12.0g / dL, at least 12.1g / dL, at least 12.2g / dL, at least 12.3g / dL, at least 12.4g / dL, at least 12.5g / dL, at least 12.6g / dL, at least 12.7g / dL, at least 12.8g / dL, at least 12.9g / dL, at least 13.0g / dL, at least 13.1g / dL, at least at least 13.2g / dL, at least 13.3g / dL, at least 13.4g / dL, at least 13.5g / dL, at least 13.6g / dL, at least 13.7g / dL, at least 13.8g / dL, at least 13.9g / dL, at least 14.0g / dL, at least 14.1g / dL, at least 14.2g / dL, at least 14.3g / dL, at least 14.4g / dL, at least 14.5g / dL, at least 14.6g / dL, at least 14.7g / dL, at least 14.8g / dL, at least 14.9g / dL , at least 15.0g / dL, at least 15.1g / dL, at least 15.2g / dL, at least 15.3g / dL, at least 15.4g / dL, at least 15.5g / dL, at least 15.6g / dL, at least 15.7g / dL, at least 15.8g / dL, at least 15.9g / dL, at least 16.0g / dL, at least 16.1g / dL, at least 16.2g / dL, at least 16.3g / dL, at least 16.4g / dL, at least 16.5g / dL, at least 16.6g / dL, at least 16.7 g / dL, at least 16.8 g / dL, at least 16.9 g / dL, at least 17.0 g / dL, at least 17.1 g / dL, at least 17.2 g / dL, at least 17.3 g / dL, at least 17.4 g / dL, at least 17.5 g / dL, at least 17.6 g / dL, at least 17.7 g / dL, at least 17.8 g / dL, at least 17.9 g / dL, or at least 18.0 g / dL.

[0291] In some embodiments, the anti-C1s antibody increases hemoglobin levels in a subject within about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 9 hours, about 10 hours, about 11 hours, about 12 hours, about 13 hours, about 14 hours, about 15 hours, about 16 hours, about 17 hours, about 18 hours, about 19 hours, about 20 hours, about 21 hours, about 22 hours, or about 24 hours after administration. In some embodiments, the increase is within about 1 hour, within about 23 hours, within about 24 hours, within about 1 day, within about 2 days, within about 3 days, within about 4 days, within about 5 days, within about 6 days, within about 7 days, within about 8 days, within about 9 days, within about 10 days, within about 11 days, within about 12 days, within about 13 days, within about 14 days, within about 2 weeks, within about 3 weeks, within about 4 weeks, within about 5 weeks, within about 6 weeks, within about 7 weeks, within about 8 weeks, within about 9 weeks, within about 10 weeks, within about 11 weeks, or within about 12 weeks.

[0292] In certain embodiments, the present disclosure provides a method for increasing the level of hemoglobin, e.g., in the blood, in a subject in need thereof, comprising administering to the subject an effective amount of an anti-C1s antibody, wherein the level of hemoglobin in the subject, e.g., in the blood, is increased by at least 1.6 g / dL within 7 days of administration. Hemoglobin levels in the sham group increased to 3.9 g / dL within 6 weeks of administration.

[0293] The present disclosure provides a method for increasing hemoglobin levels in a subject in need thereof, comprising administering to the subject an effective amount of an anti-C1s antibody, wherein the effective amount of the anti-C1s antibody is at least about 45 mg / kg, at least about 50 mg / kg, at least about 55 mg / kg, at least about 60 mg / kg, at least about 65 mg / kg, at least about 70 mg / kg, at least about 75 mg / kg, at least about 80 mg / kg, at least about 85 mg / kg, at least about 90 mg / kg, at least about 95 mg / kg, or at least about 100 mg / kg.

[0294] In some embodiments, the effective amount of anti-C1s antibody is between about 60 mg / kg and about 100 mg / kg, between about 60 mg / kg and about 95 mg / kg, between about 60 mg / kg and about 90 mg / kg, between about 60 mg / kg and about 85 mg / kg, between about 60 mg / kg and about 80 mg / kg, between about 60 mg / kg and about 75 mg / kg, between about 60 mg / kg and about 70 mg / kg, or between about 60 mg / kg and about 65 mg / kg. In some embodiments, the effective amount of anti-C1s antibody is between about 45 mg / kg and about 85 mg / kg, between about 45 mg / kg and about 80 mg / kg, between about 45 mg / kg and about 75 mg / kg, between about 45 mg / kg and about 70 mg / kg, between about 45 mg / kg and about 65 mg / kg, between about 45 mg / kg and about 60 mg / kg, or between about 45 mg / kg and about 50 mg / kg. In some embodiments, an effective amount of an anti-C1s antibody is between about 85 mg / kg and about 150 mg / kg, between about 85 mg / kg and about 145 mg / kg, between about 85 mg / kg and about 140 mg / kg, between about 85 mg / kg and about 135 mg / kg, between about 85 mg / kg and about 130 mg / kg, between about 85 mg / kg and about 125 mg / kg, between about 85 mg / kg and about 125 mg / kg, between about 85 mg / kg and about 120 mg / kg, between about 85 mg / kg and about 115 mg / kg, between about 85 mg / kg and about 110 mg / kg, between about 85 mg / kg and about 105 mg / kg, between about 85 mg / kg and about 100 mg / kg, between about 85 mg / kg and about 95 mg / kg, or between about 85 mg / kg and about 90 mg / kg.

[0295] In some embodiments, the effective amount is about 45 mg / kg, about 50 mg / kg, about 55 mg / kg, about 60 mg / kg, about 65 mg / kg, about 70 mg / kg, about 75 mg / kg, about 80 mg / kg, about 85 mg / kg, about 90 mg / kg, about 95 mg / kg, about 100 mg / kg, about 105 mg / kg, about 110 mg / kg, about 115 mg / kg, about 120 mg / kg, about 125 mg / kg, about 130 mg / kg, about 135 mg / kg, about 140 mg / kg, about 145 mg / kg, or about 150 mg / kg.

[0296] The present disclosure also provides a method of increasing hemoglobin levels in a subject in need thereof, comprising administering an anti-C1s antibody to the individual, wherein the anti-C1s antibody is administered in an amount of at least 4 g, at least 4.5 g, at least 5 g, at least 5.5 g, at least 6 g, at least 6.5 g, at least 7 g, at least 7.5 g, at least 8 g, at least 8.5 g, at least 9 g, at least 9.5 g, or at least 10 g.

[0297] In some embodiments, the anti-C1s antibody is administered in an amount of between about 5.5g and about 10g, between about 5.5g and about 9.5g, between about 5.5g and about 9g, between about 5.5g and about 8.5g, between about 5.5g and about 8g, between about 5.5g and about 7.5g, between about 5.5g and about 7g, between about 5.5g and about 6.5g, or between about 5.5g and about 6g. In some embodiments, the anti-C1s antibody is administered in an amount of between about 4.5g and about 8.5g, between about 4.5g and about 8g, between about 4.5g and about 7.5g, between about 4.5g and about 7g, between about 4.5g and about 6.5g, between about 4.5g and about 6g, between about 4.5g and about 5.5g, or between about 4.5g and about 6g. In some embodiments, the anti-C1s antibody is administered in an amount between about 7.5 g and about 12, between about 7.5 g and about 11.5 g, between about 7.5 g and about 11 g, between about 7.5 g and about 10.5 g, between about 7.5 g and about 10 g, between about 7.5 g and about 9.5 g, between about 7.5 g and about 9 g, between about 7.5 g and about 8.5 g, or between about 7.5 g and about 8 g.

[0298] The present disclosure provides a method of reducing the percentage of C3d-positive red blood cells in the blood of a subject in need thereof, comprising administering to the subject an effective amount of an anti-C1s antibody. In some embodiments, the anti-C1s antibody reduces the percentage of C3d-positive red blood cells in the subject's blood 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%, or 100%, compared to the percentage of C3d-positive red blood cells in the subject's blood before administration.

[0299] In some embodiments, the anti-C1s antibody increases the percentage of C3d positive red blood cells in the subject's blood within about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 9 hours, about 10 hours, about 11 hours, about 12 hours, about 13 hours, about 14 hours, about 15 hours, about 16 hours, about 17 hours, about 18 hours, about 19 hours, about 20 hours, about 21 hours, about The decrease is within 22 hours, within about 23 hours, within about 24 hours, within about 1 day, within about 2 days, within about 3 days, within about 4 days, within about 5 days, within about 6 days, within about 7 days, within about 8 days, within about 9 days, within about 10 days, within about 11 days, within about 12 days, within about 13 days, within about 14 days, within about 2 weeks, within about 3 weeks, within about 4 weeks, within about 5 weeks, within about 6 weeks, within about 7 weeks, within about 8 weeks, within about 9 weeks, within about 10 weeks, within about 11 weeks, or within about 12 weeks.

[0300] The present disclosure provides a method for reducing the percentage of C3d-positive red blood cells in a subject in need thereof, comprising administering to the subject an effective amount of an anti-C1s antibody, wherein the effective amount of the anti-C1s antibody is at least about 45 mg / kg, at least about 50 mg / kg, at least about 55 mg / kg, at least about 60 mg / kg, at least about 65 mg / kg, at least about 70 mg / kg, at least about 75 mg / kg, at least about 80 mg / kg, at least about 85 mg / kg, at least about 90 mg / kg, at least about 95 mg / kg, or at least about 100 mg / kg.

[0301] In some embodiments, the effective amount of anti-C1s antibody is between about 60 mg / kg and about 100 mg / kg, between about 60 mg / kg and about 95 mg / kg, between about 60 mg / kg and about 90 mg / kg, between about 60 mg / kg and about 85 mg / kg, between about 60 mg / kg and about 80 mg / kg, between about 60 mg / kg and about 75 mg / kg, between about 60 mg / kg and about 70 mg / kg, or between about 60 mg / kg and about 65 mg / kg. In some embodiments, the effective amount of anti-C1s antibody is between about 45 mg / kg and about 85 mg / kg, between about 45 mg / kg and about 80 mg / kg, between about 45 mg / kg and about 75 mg / kg, between about 45 mg / kg and about 70 mg / kg, between about 45 mg / kg and about 65 mg / kg, between about 45 mg / kg and about 60 mg / kg, or between about 45 mg / kg and about 50 mg / kg. In some embodiments, an effective amount of an anti-C1s antibody is between about 85 mg / kg and about 150 mg / kg, between about 85 mg / kg and about 145 mg / kg, between about 85 mg / kg and about 140 mg / kg, between about 85 mg / kg and about 135 mg / kg, between about 85 mg / kg and about 130 mg / kg, between about 85 mg / kg and about 125 ... 0 mg / kg, between about 85 mg / kg and about 115 mg / kg, between about 85 mg / kg and about 110 mg / kg, between about 85 mg / kg and about 105 mg / kg, between about 85 mg / kg and about 100 mg / kg, between about 85 mg / kg and about 95 mg / kg, or between about 85 mg / kg and about 90 mg / kg.

[0302] In some embodiments, the effective amount is about 45 mg / kg, about 50 mg / kg, about 55 mg / kg, about 60 mg / kg, about 65 mg / kg, about 70 mg / kg, about 75 mg / kg, about 80 mg / kg, about 85 mg / kg, about 90 mg / kg, about 95 mg / kg, about 100 mg / kg, about 105 mg / kg, about 110 mg / kg, about 115 mg / kg, about 120 mg / kg, about 125 mg / kg, about 130 mg / kg, about 135 mg / kg, about 140 mg / kg, about 145 mg / kg, or about 150 mg / kg.

[0303] The present disclosure also provides a method for reducing the percentage of C3d-positive red blood cells in the blood of a subject in need thereof, comprising administering an anti-C1s antibody to the individual, wherein the anti-C1s antibody is administered in an amount of at least about 4 g, at least about 4.5 g, at least about 5 g, at least about 5.5 g, at least about 6 g, at least about 6.5 g, at least about 7 g, at least about 7.5 g, at least about 8 g, at least about 8.5 g, at least about 9 g, at least about 9.5 g, or at least about 10 g.

[0304] In some embodiments, the anti-C1s antibody is administered in an amount of between about 5.5g and about 10g, between about 5.5g and about 9.5g, between about 5.5g and about 9g, between about 5.5g and about 8.5g, between about 5.5g and about 8g, between about 5.5g and about 7.5g, between about 5.5g and about 7g, between about 5.5g and about 6.5g, or between about 5.5g and about 6g. In some embodiments, the anti-C1s antibody is administered in an amount of between about 4.5g and about 8.5g, between about 4.5g and about 8g, between about 4.5g and about 7.5g, between about 4.5g and about 7g, between about 4.5g and about 6.5g, between about 4.5g and about 6g, between about 4.5g and about 5.5g, or between about 4.5g and about 5g. In some embodiments, the anti-C1s antibody is administered in an amount of between about 7.5g and about 12g, between about 7.5g and about 11.5g, between about 7.5g and about 11g, between about 7.5g and about 10.5g, between about 7.5g and about 10g, between about 7.5g and about 9.5g, between about 7.5g and about 9g, between about 7.5g and about 8.5g, or between about 7.5g and about 8g.

[0305] The present disclosure provides a method of reducing the level of bilirubin in, for example, blood, in a subject in need thereof, comprising administering to the subject an effective amount of an anti-C1s antibody. In some embodiments, the anti-C1s antibody reduces the bilirubin level in the subject to less than 2.5 mg / dL, less than 2.4 mg / dL, less than 2.3 mg / dL, less than 2.2 mg / dL, less than 2.1 mg / dL, less than 2.0 mg / dL, less than 1.9 mg / dL, less than 1.8 mg / dL, less than 1.7 mg / dL, less than 1.6 mg / dL, less than 1.5 mg / dL, less than 1.4 mg / dL, less than 1.8 mg / dL, less than 1.7 mg / dL, less than 1.8 ... Reduce to less than 1.3 mg / dL, less than 1.2 mg / dL, less than 1.1 mg / dL, less than 1.0 mg / dL, less than 0.9 mg / dL, less than 0.8 mg / dL, less than 0.7 mg / dL, less than 0.6 mg / dL, less than 0.5 mg / dL, less than 0.4 mg / dL, less than 0.3 mg / dL, less than 0.2 mg / dL, or less than 0.1 mg / dL.

[0306] In some embodiments, the anti-C1s antibody reduces bilirubin levels in a subject, e.g., in the blood, within about 2 hours, within about 3 hours, within about 4 hours, within about 5 hours, within about 6 hours, within about 7 hours, within about 8 hours, within about 9 hours, within about 10 hours, within about 11 hours, within about 12 hours, within about 13 hours, within about 14 hours, within about 15 hours, within about 16 hours, within about 17 hours, within about 18 hours, within about 19 hours, within about 20 hours, within about 21 hours, within about 22 hours, within about 23 hours, within about 24 hours, within about 1 day, within about 2 days, within about 3 days, or within about 4 days of administration. The amount of erythrocyte sedimentation is reduced within about 4 days, within about 5 days, within about 6 days, within about 7 days, within about 8 days, within about 9 days, within about 10 days, within about 11 days, within about 12 days, within about 13 days, within about 14 days, within about 2 weeks, within about 3 weeks, within about 4 weeks, within about 5 weeks, within about 6 weeks, within about 7 weeks, within about 8 weeks, within about 9 weeks, within about 10 weeks, within about 11 weeks, or within about 12 weeks.

[0307] The present disclosure provides a method for reducing bilirubin levels, for example in the blood, in a subject in need thereof, comprising administering to the subject an effective amount of an anti-C1s antibody, wherein the effective amount of the anti-C1s antibody is at least about 45 mg / kg, at least about 50 mg / kg, at least about 55 mg / kg, at least about 60 mg / kg, at least about 65 mg / kg, at least about 70 mg / kg, at least about 75 mg / kg, at least about 80 mg / kg, at least about 85 mg / kg, at least about 90 mg / kg, at least about 95 mg / kg, or at least about 100 mg / kg.

[0308] In some embodiments, the effective amount of anti-C1s antibody is between about 60 mg / kg and about 100 mg / kg, between about 60 mg / kg and about 95 mg / kg, between about 60 mg / kg and about 90 mg / kg, between about 60 mg / kg and about 85 mg / kg, between about 60 mg / kg and about 80 mg / kg, between about 60 mg / kg and about 75 mg / kg, between about 60 mg / kg and about 70 mg / kg, or between about 60 mg / kg and about 65 mg / kg. In some embodiments, the effective amount of anti-C1s antibody is between about 45 mg / kg and about 85 mg / kg, between about 45 mg / kg and about 80 mg / kg, between about 45 mg / kg and about 75 mg / kg, between about 45 mg / kg and about 70 mg / kg, between about 45 mg / kg and about 65 mg / kg, between about 45 mg / kg and about 60 mg / kg, or between about 45 mg / kg and about 50 mg / kg. In some embodiments, an effective amount of an anti-C1s antibody is between about 85 mg / kg and about 150 mg / kg, between about 85 mg / kg and about 145 mg / kg, between about 85 mg / kg and about 140 mg / kg, between about 85 mg / kg and about 135 mg / kg, between about 85 mg / kg and about 130 mg / kg, between about 85 mg / kg and about 125 mg / kg, between about 85 mg / kg and about 125 mg / kg, between about 85 mg / kg and about 120 mg / kg, between about 85 mg / kg and about 115 mg / kg, between about 85 mg / kg and about 110 mg / kg, between about 85 mg / kg and about 105 mg / kg, between about 85 mg / kg and about 100 mg / kg, between about 85 mg / kg and about 95 mg / kg, or between about 85 mg / kg and about 90 mg / kg.

[0309] In some embodiments, the effective amount for the methods of the present invention is about 45 mg / kg, about 50 mg / kg, about 55 mg / kg, about 60 mg / kg, about 65 mg / kg, about 70 mg / kg, about 75 mg / kg, about 80 mg / kg, about 85 mg / kg, about 90 mg / kg, about 95 mg / kg, about 100 mg / kg, about 105 mg / kg, about 110 mg / kg, about 115 mg / kg, about 120 mg / kg, about 125 mg / kg, about 130 mg / kg, about 135 mg / kg, about 140 mg / kg, about 145 mg / kg, or about 150 mg / kg.

[0310] The present disclosure also provides a method for reducing bilirubin levels, for example in the blood, in a subject in need thereof, comprising administering an anti-C1s antibody to the individual, wherein the anti-C1s antibody is administered in an effective amount of at least about 4 g, at least about 4.5 g, at least about 5 g, at least about 5.5 g, at least about 6 g, at least about 6.5 g, at least about 7 g, at least about 7.5 g, at least about 8 g, at least about 8.5 g, at least about 9 g, at least about 9.5 g, or at least about 10 g.

[0311] In some embodiments, the anti-C1s antibody is between about 5.5g and about 10g, between about 5.5g and about 9.5g, between about 5.5g and about 9g, between about 5.5g and about 8.5g, between about 5.5g and about 5.5g. In some embodiments, the anti-C1s antibody is administered in an effective amount of between about 4.5g and about 8.5g, between about 4.5g and about 8g, between about 4.5g and about 7.5g, between about 5.5g and about 7g, between about 5.5g and about 6.5g, or between about 5.5g and about 6g. In some embodiments, the anti-C1s antibody is administered in an amount of between about 4.5g and about 8.5g, between about 4.5g and about 8g, between about 4.5g and about 7.5g, between about 4.5g and about 7g, between about 4.5g and about 6.5g, between about 4.5g and about 6g, between about 4.5g and about 5.5g, or between about 4.5g and about 5g. In some embodiments, the anti-C1s antibody is administered in an amount of between about 7.5g and about 12g, between about 7.5g and about 11.5g, between about 7.5g and about 11g, between about 7.5g and about 10.5g, between about 7.5g and about 10g, between about 7.5g and about 9.5g, between about 7.5g and about 9g, between about 7.5g and about 8.5g, or between about 7.5g and about 8g.

[0312] The present disclosure provides a method of inhibiting cleavage of complement component C4 in an individual, comprising administering an anti-C1s antibody to the individual, wherein the anti-C1s antibody is administered in an effective amount of about 5.5 g. In some cases, the effective amount of about 5.5 g of the anti-C1s antibody is administered to the individual once every two weeks. In some cases, the method comprises: a) administering an effective amount of about 5.5 g of the anti-C1s antibody on day 1; b) administering an effective amount of about 5.5 g of the anti-C1s antibody on day 8; and c) administering an effective amount of about 5.5 g of the anti-C1s antibody every other week after administration on day 8. In some cases, the effective amount of about 5.5 g of the anti-C1s antibody is administered to the individual every other week for a period of about 4 weeks to 1 year, e.g., about 4 weeks to about 8 weeks, about 2 months to about 6 months, or about 6 months to 1 year. In some cases, an effective amount of about 5.5 g of anti-C1s antibody is administered to an individual every other week for a period of greater than 1 year, for example, in some cases, an effective amount of about 5.5 g of anti-C1s antibody is administered to an individual every other week for a period of 1 to 50 years, e.g., 1 to 2 years, 2 to 5 years, 5 to 10 years, 10 to 20 years, 20 to 30 years, 30 to 40 years, or 40 to 50 years.

[0313] The present disclosure provides a method of inhibiting cleavage of complement component C4 in an individual, comprising administering an anti-C1s antibody to the individual, wherein the anti-C1s antibody is administered in an effective amount of about 6.5 g. In some cases, a dose of about 6.5 g of the effective amount of the anti-C1s antibody is administered to the individual once every two weeks. In some cases, the method comprises: a) administering an effective amount of about 6.5 g of the anti-C1s antibody on day 1; b) administering an effective amount of about 6.5 g of the anti-C1s antibody on day 8; and c) administering an effective amount of about 6.5 g of the anti-C1s antibody every other week after administration on day 8. In some cases, the effective amount of about 6.5 g of the anti-C1s antibody is administered to the individual every other week for a period of about 4 weeks to 1 year, e.g., about 4 weeks to about 8 weeks, about 2 months to about 6 months, or about 6 months to 1 year. In some cases, an effective amount of about 6.5 g of anti-C1s antibody is administered to an individual every other week for a period of greater than 1 year, e.g., in some cases, an effective amount of about 6.5 g of anti-C1s antibody is administered to an individual every other week for a period of 1 to 50 years, e.g., 1 to 2 years, 2 to 5 years, 5 to 10 years, 10 to 20 years, 20 to 30 years, 30 to 40 years, or 40 to 50 years.

[0314] The present disclosure provides a method of inhibiting cleavage of complement component C4 in an individual, comprising administering an anti-C1s antibody to the individual, wherein the anti-C1s antibody is administered in an effective amount of about 7.5 g. In some cases, the effective amount of about 7.5 g of the anti-C1s antibody is administered to the individual once every two weeks. In some cases, the method comprises: a) administering an effective amount of about 7.5 g of the anti-C1s antibody on day 1; b) administering an effective amount of about 7.5 g of the anti-C1s antibody on day 8; and c) administering an effective amount of about 7.5 g of the anti-C1s antibody every other week after administration on day 8. In some cases, the effective amount of about 7.5 g of the anti-C1s antibody is administered to the individual every other week for a period of about 4 weeks to 1 year, e.g., about 4 weeks to about 8 weeks, about 2 months to about 6 months, or about 6 months to 1 year. In some cases, An effective amount of about 7.5 g of anti-C1s antibody is administered to an individual every other week for a period of greater than 1 year, e.g., in some cases, an effective amount of about 7.5 g of anti-C1s antibody is administered to an individual every other week for a period of 1 to 50 years, e.g., 1 to 2 years, 2 to 5 years, 5 to 10 years, 10 to 20 years, 20 to 30 years, 30 to 40 years, or 40 to 50 years.

[0315] Route of administration The anti-C1s antibodies are administered to an individual using any available method and route suitable for drug delivery, including in vivo and ex vivo methods, as well as systemic and local administration routes.

[0316] Conventional and pharma- ceutically acceptable routes of administration include intranasal, intramuscular, intratracheal, intrathecal, intracranial, subcutaneous, intradermal, topical, intravenous, intraperitoneal, intraarterial (e.g., via the carotid artery), spinal or brain delivery, rectal, nasal, oral, and other enteral and parenteral routes of administration. Routes of administration may be combined as necessary or adjusted depending on the antibody and / or the desired effect. The anti-C1s antibody composition may be administered in a single dose or in multiple doses. In some cases, the anti-C1s antibody is administered orally. In some cases, the anti-C1s antibody is administered subcutaneously. In some cases, the anti-C1s antibody is administered intramuscularly. In some cases, the anti-C1s antibody is administered intravenously.

[0317] The anti-C1s antibody can be administered to a host using any available conventional method and route suitable for delivery of conventional drugs, such as systemic or local routes. In general, routes of administration contemplated by the present disclosure include, but are not necessarily limited to, enteral, parenteral, or by inhalation.

[0318] Parenteral routes of administration other than inhalation administration include, but are not necessarily limited to, topical, transdermal, subcutaneous, intramuscular, intraorbital, intracapsular, intraspinal, intrasternal, intrathecal, and intravenous routes, i.e., any route of administration other than via the digestive tract. Parenteral administration can be performed to achieve systemic or localized delivery of the subject antibody. When systemic delivery is desired, administration typically involves invasive or systemically absorbed topical or mucosal administration of pharmaceutical preparations.

[0319] "Treatment" refers to at least the alleviation of symptoms associated with a pathological condition afflicting a host, where alleviation is used broadly to refer to at least a reduction in the magnitude of a parameter, e.g., a symptom associated with the pathological condition being treated, e.g., a complement-mediated disease or disorder. As such, treatment also includes situations in which the pathological condition, or at least the symptoms associated therewith, are completely inhibited, e.g., prevented from occurring, or arrested, e.g., terminated, such that the host is no longer afflicted with the pathological condition, or at least the symptoms characteristic of the pathological condition.

[0320] In some cases, anti-C1s antibody is administered by injection and / or delivery, for example, to a site in the cerebral artery or directly to brain tissue.Anti-C1s antibody can also be administered directly to target site, or can be administered to target site by, for example, gene gun delivery.

[0321] A variety of hosts (wherein the term "host" is used interchangeably herein with the terms "subject," "individual," and "patient") can be treated according to the present invention. Typically, such hosts are "mammals" or "mammals," where these terms generally refer to organisms within the class Mammalia, such as carnivores (e.g., cats), herbivores (e.g., cows, The term "host" is used to describe animals such as mammals, fish, and horses, omnivores (e.g., dogs, goats, and pigs), rodents (e.g., mice, guinea pigs, and rats), and primates (e.g., humans, chimpanzees, and monkeys), etc. In some embodiments, the host is an individual having a complement system, such as a mammal, fish, or invertebrate. In some cases, the host is a mammal, fish, or invertebrate companion animal, farm animal, working animal, zoo animal, or laboratory animal that includes a complement system. In some cases, the individual is a human.

[0322] Complement-Mediated Diseases and Disorders In some cases, complement-mediated diseases or disorders are characterized by the presence of elevated (higher than normal) amounts of C1s or elevated levels of complement C1s activity in cells, tissues, or body fluids. For example, in some cases, complement-mediated diseases or disorders are characterized by the presence of elevated amounts and / or activity of C1s in brain tissue and / or cerebrospinal fluid. A "higher than normal" amount of C1s in a cell, tissue, or body fluid indicates that the amount of C1s in the cell, tissue, or body fluid is higher than a normal control level, e.g., higher than a normal control level of an individual or a population of individuals of the same age group. A "higher than normal" level of C1s activity in a cell, tissue, or body fluid indicates that the proteolytic cleavage carried out by C1s in the cell, tissue, or body fluid is higher than a normal control level, e.g., higher than a normal control level of an individual or a population of individuals of the same age group. In some cases, an individual with a complement-mediated disease or disorder exhibits one or more additional symptoms of such disease or disorder.

[0323] In other cases, the complement-mediated disease or disorder is characterized by the presence of a lower than normal amount of C1s or a lower level of complement C1s activity in cells, tissues, or body fluids. For example, in some cases, the complement-mediated disease or disorder is characterized by the presence of a lower amount and / or activity of C1s in brain tissue and / or cerebrospinal fluid. A "lower than normal" amount of C1s in a cell, tissue, or body fluid indicates that the amount of C1s in the cell, tissue, or body fluid is lower than a normal control level, e.g., lower than a normal control level for an individual or a population of individuals of the same age group. A "lower than normal" level of C1s activity in a cell, tissue, or body fluid indicates that the proteolytic cleavage carried out by C1s in the cell, tissue, or body fluid is lower than a normal control level, e.g., lower than a normal control level for an individual or a population of individuals of the same age group. In some cases, an individual with a complement-mediated disease or disorder exhibits one or more additional symptoms of such disease or disorder.

[0324] A complement-mediated disease or disorder is a disease or disorder in which the amount or activity of complement C1s causes the disease or disorder in an individual. In some embodiments, the complement-mediated disease or disorder is selected from the group consisting of an autoimmune disease, a cancer, a blood disease, an infectious disease, an inflammatory disease, an ischemia-reperfusion injury, a neurodegenerative disease, a neurodegenerative disorder, an eye disease, a kidney disease, a transplant rejection, a vascular disease, and a vasculitic disease. In some cases, the complement-mediated disease or disorder is an autoimmune disease. In some cases, the complement-mediated disease or disorder is a cancer. In some cases, the complement-mediated disease or disorder is an infectious disease. In some cases, the complement-mediated disease or disorder is an inflammatory disease. In some cases, the complement-mediated disease or disorder is a blood disease. In some cases, the complement-mediated disease or disorder is an ischemia-reperfusion injury. In some cases, the complement-mediated disease or disorder is an eye disease. In some cases, the complement-mediated disease or disorder is a kidney disease. In some cases, the complement-mediated disease or disorder is transplant rejection. In some cases, the complement-mediated disease or disorder is antibody-mediated transplant rejection. In some cases, the complement-mediated disease or disorder is a vascular disease. In some cases, the complement-mediated disease or disorder is a vascular disease. In some cases, the complement mediated disease or disorder is a vasculitic disorder. In some cases, the complement mediated disease or disorder is a neurodegenerative disease or disorder. In some cases, the complement mediated disease or disorder is a neurodegenerative disease. In some cases, the complement mediated disease or disorder is a neurodegenerative disorder. In some cases, the complement mediated disease or disorder is a tauopathy.

[0325] Examples of complement-mediated diseases or disorders include, but are not limited to, age-related macular degeneration, Alzheimer's disease, amyotrophic lateral sclerosis, anaphylaxis, argyrophilic grain dementia, arthritis (e.g., rheumatoid arthritis), asthma, atherosclerosis, atypical hemolytic uremic syndrome, autoimmune diseases, Barraquer-Simons syndrome, Behcet's disease, British amyloid angiopathy, bullous pemphigoid, Burger's disease, C1q nephropathy, chronic inflammatory demyelinating polyneuropathy, cancer, fulminant antiphospholipid syndrome, cerebral amyloid angiopathy, cold agglutinin disease (primary cold agglutinin disease and secondary cerebrovascular disease (including myeloid cold agglutinin disease), corticobasal degeneration, Creutzfeldt-Jakob disease, Crohn's disease, cryoglobulinemic vasculitis, dementia pugilistica, dementia with Lewy bodies (DLB), diffuse neurofibrillary tangles with calcifications, discoid lupus erythematosus, Down's syndrome, focal segmental glomerulosclerosis, formal thought disorder, frontotemporal dementia (FTD), frontotemporal dementia with parkinsonism linked to chromosome 17, frontotemporal lobar degeneration, Gerstmann-Sträussler-Scheinker disease, Guillain-Barré syndrome, Hallervorden-Spatz disease, hemolytic uraemic syndrome, hereditary Angioedema, hypophosphastasis, idiopathic pneumonia syndrome, immune complex disease, inclusion body myositis, infectious diseases (e.g., diseases caused by bacteria (e.g., Neisseria meningitidis or Streptococcus spp.), diseases caused by viruses (e.g., human immunodeficiency virus (HIV)), or diseases caused by other infectious agents), inflammatory diseases, ischemia / reperfusion injury, mild cognitive impairment, immune thrombocytopenic purpura (ITP), molybdenum cofactor deficiency (MoCD) type A, membranoproliferative glomerulonephritis (MPGN) type I, membranoproliferative glomerulonephritis (MPGN) type I, membranoproliferative glomerulonephritis (MPGN) type II, membranoproliferative glomerulonephritis (MPGN) type III, membranoproliferative glomerulonephritis (MPGN) type V ... MPGN type II (dense deposit disease), membranous nephritis, multi-infarct dementia, lupus (e.g., systemic lupus erythematosus (SLE)), glomerulonephritis, Kawasaki disease, mucous membrane pemphigoid, cicatricial pemphigoid, multifocal motor neuropathy, multiple sclerosis, multiple system atrophy, myasthenia gravis, myocardial infarction, myotonic dystrophy, neuromyelitis optica, Niemann-Pick disease type C, non-Guam motor neuron disease with neurofibrillary tangles, Parkinson's disease, Parkinson's disease with dementia, paroxysmal nocturnal hemoglobinuria, pemphigus vulgaris, Pick's disease, postencephalitic parkinsonism, polymyositis,Prion protein cerebral amyloid angiopathy, progressive subcortical gliosis, progressive supranuclear palsy, psoriasis, sepsis, Shiga-toxin E coli (STEC)-HuS, spinal muscular atrophy, stroke, subacute sclerosing panencephalitis, neurofibrillary tangle-predominant dementia, transplant rejection, vasculitis (e.g., ANCA-associated vasculitis), Wegener's granulomatosis, sickle cell disease, cryoglobulinemia, mixed cryoglobulinemia, idiopathic mixed cryoglobulinemia, type II mixed cryoglobulinemia, type III mixed cryoglobulinemia, nephritis, drug-induced thrombocytopenia, lupus nephritis, bullous pemphigoid, epidermolysis bullosa acquisita, delayed hemolytic transfusion reaction, hypocomplementemic urticarial vasculitis syndrome, pseudophakic bullous keratopathy, and platelet refractoriness.

[0326] In some cases, the method of the present invention includes the treatment of primary CAgD in a subject in need thereof, comprising administering an effective amount of between about 6.5g and about 7.5g of an anti-C1s antibody, e.g., BIVV009, for example, about 6.5g for subjects weighing less than 75kg, and 7.5g for subjects weighing more than 75kg. In some embodiments, the method of the present invention is not limited to use in relation to severity of anemia, transfusion history, or previous treatment experience. In some embodiments, there is no requirement for a REMS prior to administration; patients are vaccinated according to local guidelines prior to treatment initiation to reduce the risk of serious infection. In some embodiments, doses are administered as an intravenous infusion over 1 hour on day 0, day 7, and then every 14 days ± 2 days starting on day 21. Intravenous infusion can be performed in a clinical setting or in a home environment. As a result of treatment, the anti-C1s antibody can improve anemia and associated clinical symptoms, eliminate transfusions, prevent hemolysis, rapid onset of action, improve fatigue and quality of life, and / or any combination thereof. In other embodiments, the treatment shows no drug-related serious or severe adverse events; no discontinuation due to adverse events, no serious infections; no requirement for REMS, most frequently reported adverse events were similar to placebo, or any combination thereof. In other embodiments, the anti-C1s antibody can prevent chronic hemolysis as a result of treatment, resulting in improvement of anemia, elimination of transfusions, improved quality of life, and ultimately reduced risk of life-threatening thromboembolic events, morbidity and mortality, and reduced utilization of medical facilities.

[0327] In some cases, the complement mediated disease or disorder is bullous pemphigoid. In some cases, the complement mediated disease or disorder is antibody mediated rejection of organ transplants. In some cases, the complement mediated disease or disorder is cold agglutinin disease. In some cases, the complement mediated disease or disorder is warm autoimmune hemolytic anemia. In some cases, the complement mediated disease or disorder is antibody mediated transplant rejection. In some cases, the complement mediated disease or disorder is immune thrombocytopenic purpura. In some cases, the complement mediated disease or disorder is neuromyelitis optica.

[0328] In some cases, the complement mediated disease or disorder is multifocal motor neuropathy (MMN). In some cases, the complement mediated disease or disorder is myasthenia gravis. In some cases, the complement mediated disease or disorder is chronic inflammatory demyelinating polyneuropathy. In some cases, the complement mediated disease or disorder is lupus nephritis. In some cases, the complement mediated disease or disorder is mucous membrane pemphigoid. In some cases, the complement mediated disease or disorder is cicatricial pemphigoid. In some cases, the complement mediated disease or disorder is ocular pemphigoid. In some cases, the complement mediated disease or disorder is antineutrophil cytoplasmic autoantibody (ANCA) associated vasculitis.

[0329] In other embodiments, the complement-mediated disease or disorder is an autoantibody-mediated peripheral neuropathy, such as, but not limited to, Guillain-Barre syndrome, myasthenia gravis, acute inflammatory demyelinating polyneuropathy (AIDP), chronic inflammatory demyelinating polyneuropathy (CIDP), acute motor axonal neuropathy (AMAN), acute motor sensory axonal neuropathy (AMSAN), pharyngocervical brachial Miller Fisher syndrome, or any combination thereof. In some embodiments, the complement-mediated disease or disorder is Guillain-Barre syndrome, which manifests as rapidly developing muscle weakness that begins in the feet and hands and spreads to the arms and upper body. Guillain-Barre syndrome can lead to death during the acute phase, as respiratory failure can occur and other autonomic nervous system functions (e.g., heart rate) can also be affected. Approximately 7.5% of all cases are fatal. Incidence: 1-2 / 100,000.

[0330] In another embodiment, the complement-mediated disease or disorder is myasthenia gravis, which manifests as weakness, fatigue, and progressive worsening during periods of physical activity, typically beginning with eye weakness; progressing to a more severe form characterized by weakness of the limbs and weakness in performing basic vital functions (chewing, swallowing, breathing). In myasthenic crisis, respiratory paralysis occurs and assisted ventilation is required to sustain life.

[0331] In another embodiment, the complement-mediated disease or disorder is multifocal motor neuropathy (MMN), which is an inflammatory autoimmune disease of the lower nervous system. MMN is a pure It is a motor neuropathy with a mean age of onset of 40 years. MMN is characterized by slowly progressive, asymmetric distal limb weakness; conduction block (CB) often affecting the ulnar, median, radial or tibial nerves; and / or muscle atrophy. Other clinical features include muscle spasms, fasciculations, and increased weakness in cold conditions. GM1-specific IgM antibodies are present in the serum of nearly half of all patients, and their titer correlates with their in vitro complement activation capacity and disease severity. Intravenous immunoglobulin (IVIg) is effective in MMN. Nevertheless, patients still suffer from slowly progressive axonal degeneration and muscle weakness that cannot be completely prevented with chronic IVIg therapy.

[0332] In another embodiment, the complement-mediated disease or disorder useful for treatment is neuromyelitis optica (NMO).NMO is caused by anti-aquaporin-4 IgG autoantibody (NMO-IgG), which activates complement, kills astrocytes, and causes the death of oligodendrocytes that myelinate optic nerve and spinal cord.Attacks are followed by loss of vision and paralysis.

[0333] In another embodiment, the complement-mediated disease or disorder useful for treatment is systemic lupus erythematosus (SLE). Systemic lupus erythematosus (SLE) is an autoimmune disease that affects 0.04% of the population in developed countries. SLE is thought to result from a defect in the body's waste removal system, in which complement plays a major role. In humans, congenital deficiency of complement proteins in the C1 complex as well as C2 and C4 is associated with an increased risk of developing SLE. However, a significant number of patients with SLE develop hypocomplementemia, which involves depletion of C1q and other components of the classical pathway: for example, complement deposition on RBCs and / or C1q deposition in affected tissues.

[0334] In another embodiment, the complement-mediated disease or disorder useful for treatment is lupus nephritis (LN). LN is a renal manifestation of SLE, which occurs in 25-50% of patients and is a major cause of morbidity and mortality. C1q antibodies are closely associated with renal involvement and are frequently a precursor to and present during acute exacerbations. Active LN is rarely observed in the absence of C1q Ab. Multiple studies have shown a negative correlation between C1q Ab titers and serum C1q, and a positive correlation with C1q deposition in the glomeruli of patients with LN.

[0335] In some embodiments, the complement-mediated disease or disorder useful for treatment is membranoproliferative glomerulonephritis (Type I) (mixed cryoglobulinemia). Mixed cryoglobulinemia is a systemic vasculitis mediated by immune complexes (IC). It is most commonly seen in the setting of chronic infection (HCV accounts for 80% of MC cases). Clinically, cryoglobulinemia itself presents with symptoms such as weakness and joint pain, as well as variable skin and visceral organ involvement. Some patients successfully suppress inflammation with steroids, but often require additional plasmapheresis to remove circulating cryoglobulins and immunosuppressive treatment to inhibit new cryoglobulin formation.

[0336] In some cases, the disclosed method includes administering to a subject with bullous pemphigoid an effective amount of between about 6.5 g and about 7.5 g of an anti-C1s antibody, e.g., BIVV009. In some cases, the disclosed method includes administering to a subject with antibody-mediated rejection of an organ transplant an effective amount of between about 6.5 g and about 7.5 g of an anti-C1s antibody, e.g., BIVV009. In some cases, the disclosed method includes administering to a subject with cold agglutinin disease an effective amount of between about 6.5 g and about 7.5 g of an anti-C1s antibody, e.g., BIVV009. In some cases, the disclosed method includes administering to a subject with warm autoimmune hemolytic anemia an effective amount of between about 6.5 g and about 7.5 g of an anti-C1s antibody, e.g., BIVV009. In some cases, the disclosed method includes administering to a subject with immune thrombocytopenic purpura an effective amount of between about 6.5 g and about 7.5 g of an anti-C1s antibody, e.g., BIVV009. For example, administering an effective amount of between about 6.5 g and about 7.5 g of BIVV009 to a subject with neuromyelitis optica. In some cases, the methods of the present disclosure include administering to a subject with neuromyelitis optica an effective amount of between about 6.5 g and about 7.5 g of an anti-C1s antibody, for example, BIVV009.

[0337] In some cases, the disclosed method includes administering to a subject with multifocal motor neuropathy (MMN) an effective amount of between about 6.5 g and about 7.5 g of an anti-C1s antibody, such as BIVV009. In some cases, the disclosed method includes administering to a subject with myasthenia gravis an effective amount of between about 6.5 g and about 7.5 g of an anti-C1s antibody, such as BIVV009. In some cases, the disclosed method includes administering to a subject with chronic inflammatory demyelinating polyneuropathy an effective amount of between about 6.5 g and about 7.5 g of an anti-C1s antibody, such as BIVV009. In some cases, the disclosed method includes administering to a subject with lupus nephritis an effective amount of between about 6.5 g and about 7.5 g of an anti-C1s antibody, such as BIVV009. In some cases, the disclosed method includes administering to a subject with mucous membrane pemphigoid an effective amount of between about 6.5 g and about 7.5 g of an anti-C1s antibody, such as BIVV009. In some cases, the disclosed method includes administering to a subject with cicatricial pemphigoid an effective amount of between about 6.5 g and about 7.5 g of an anti-C1s antibody, such as BIVV009. In some cases, the disclosed method includes administering to a subject with ocular pemphigoid an effective amount of between about 6.5 g and about 7.5 g of an anti-C1s antibody, such as BIVV009. In some cases, the disclosed method includes administering to a subject with antineutrophil cytoplasmic autoantibody (ANCA)-associated vasculitis an effective amount of between about 6.5 g and about 7.5 g of an anti-C1s antibody, such as BIVV009.

[0338] In some embodiments, administering an anti-C1s antibody of the present disclosure results in: (a) a reduction in complement activation; (b) improved cognitive function; (c) a reduction in neuronal loss; (d) a reduction in phospho-tau levels in neurons; (e) a reduction in glial cell activation; (f) a reduction in lymphocyte infiltration; (g) a reduction in macrophage infiltration; (h) a reduction in antibody deposition; (i) a reduction in glial cell loss; (j) a reduction in oligodendrocyte loss; (k) a reduction in dendritic cell infiltration; (l) a reduction in neutrophil infiltration; (m) a reduction in red blood cell lysis; (n) a reduction in erythrocyte phagocytosis; (o) a reduction in platelet phagocytosis; (p) a reduction in platelet (q) improved survival of transplanted grafts; (r) reduced macrophage-mediated phagocytosis; (s) improved vision; (t) improved motor control; (u) reduced thrombus formation; (x) reduced antibody-mediated complement activation; (y) reduced autoantibody-mediated complement activation; (z) improved anemia; (aa) reduced demyelination; (ab) reduced eosinophilia; (ac) reduced C3 deposition on red blood cells (e.g., reduced C3b, iC3b, etc. deposition on RBCs); and (ad) reduced C3 deposition on platelets (e.g., reduced C3b, iC3b, etc. deposition on platelets); and (ae ) reduction of anaphylatoxin toxin production;(af) reduction of autoantibody-mediated blister formation;(ag) reduction of autoantibody-induced pruritus;(ah) reduction of autoantibody-induced lupus erythematosus;(ai) reduction of autoantibody-mediated skin erosions;(aj) reduction of red blood cell destruction in transfusion reactions;(ak) reduction of alloantibody-induced red blood cell lysis;(al) reduction of hemolysis in transfusion reactions;(am) reduction of alloantibody-mediated platelet lysis;(an) reduction of platelet lysis in transfusion reactions;(ao) reduction of mast cell activation;(ap) reduction of mast cell histamine release reduction;(aq) reduction in vascular permeability;(ar) reduction in edema;(as) reduction in complement deposition on the endothelium of transplanted grafts;(at) reduction in anaphylatoxin production in the endothelium of transplanted grafts;(au) reduction in separation of the dermal-epithelial junction;(av) reduction in anaphylatoxin production at the dermal-epithelial junction;(aw) reduction in alloantibody mediated complement activation in the endothelium of transplanted grafts;(ax) reduction in antibody mediated neuromuscular junction loss;(ay) reduction in complement activation at the neuromuscular junction;(az) reduction in anaphylatoxin production at the neuromuscular junction;(ba) reduced complement deposition at the neuromuscular junction; (bb) reduced paralysis; (bc) reduced anesthesia; (bd) increased bladder control; (be) increased bowel control; (bf) reduced autoantibody associated mortality; (bg) reduced autoantibody associated morbidity; and (bh) reduced conduction block.

[0339] In some cases, the anti-C1s antibody is effective to achieve and maintain a serum concentration of anti-C1s antibody of at least 100 μg / ml when administered to an individual having a complement-mediated disease or disorder at a dose of 5.5 g, and when administered as a monotherapy or in one or more doses in a combination therapy. In some cases, the anti-C1s antibody is effective to achieve and maintain a serum concentration of anti-C1s antibody of at least 100 μg / ml when administered to an individual having a complement-mediated disease or disorder at a dose of 6.5 g, and when administered as a monotherapy or in one or more doses in a combination therapy. In some cases, the anti-C1s antibody is effective to achieve and maintain a serum concentration of anti-C1s antibody of at least 100 μg / ml when administered to an individual having a complement-mediated disease or disorder at a dose of 7.5 g, and when administered as a monotherapy or in one or more doses in a combination therapy.

[0340] In some cases, the anti-C1s antibody is sufficient to inhibit the classical complement pathway (CP) by at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% when administered to an individual with a complement-mediated disease or disorder at a dose of 5.5 g, and when administered as a monotherapy or in one or more doses in a combination therapy. In some cases, the anti-C1s antibody is effective to inhibit the classical complement pathway (CP) by at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% when administered to an individual with a complement-mediated disease or disorder at a dose of 5.5 g, and when administered as a monotherapy or in one or more doses in a combination therapy. In some cases, the anti-C1s antibody is effective to inhibit the classical complement pathway (CP) by at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% when administered to an individual with a complement-mediated disease or disorder at a dose of 6.5 g, and when administered as a monotherapy or in one or more doses in a combination therapy. In some cases, the anti-C1s antibody is effective to inhibit CP by 90% when administered to an individual with a complement-mediated disease or disorder at a dose of 6.5 g, and when administered as a monotherapy or in one or more doses in a combination therapy. In some cases, the anti-C1s antibody is effective to inhibit the classical complement pathway (CP) by at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% when administered to an individual with a complement-mediated disease or disorder at a dose of 7.5 g, and when administered as a monotherapy or in one or more doses in a combination therapy. In some cases, the anti-C1s antibody is effective to inhibit CP by 90% when administered to an individual with a complement-mediated disease or disorder at a dose of 7.5 g, and when administered as a monotherapy or in one or more doses in a combination therapy.

[0341] In some cases, anti-C1s antibodies, when administered at a dose of 5.5 g to individuals with a complement-mediated disease or disorder, and when administered as monotherapy or in one or more doses in combination therapy, have been shown to reduce or eliminate the following outcomes: (a) complement activation; (b) cognitive decline; (c) neuronal loss; (d) phospho-tau levels in neurons; (e) glial cell activation; (f) lymphocyte infiltration; (g) macrophage infiltration; (h) antibody deposition; (i) glial cell loss; and (j) oligodendrocyte activation. loss;(k) dendritic cell infiltration;(l) neutrophil infiltration;(m) erythrocyte lysis;(n) erythrophagocytosis;(o) thrombocytopenia;(p) thrombolysis;(q) graft rejection;(r) macrophage-mediated phagocytosis;(s) vision loss;(t) antibody-mediated complement activation;(u) autoantibody-mediated complement activation;(v) demyelination;(w) eosinophilia;(x) blister formation;(y) pruritus;(z) skin rash;(ab) skin erosion;(ac) petechiae;(ad) bleeding time;(ae) one or more of the following: anti-inflammatory, ... It is effective to achieve a reduction of at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or greater than 90%, compared to the level or extent of the outcome in the individual before treatment with the C1s antibody.

[0342] In some cases, anti-C1s antibodies, when administered at a dose of 6.5 g to individuals with a complement-mediated disease or disorder, and when administered as monotherapy or in one or more doses in combination therapy, have been shown to reduce or eliminate the following outcomes: (a) complement activation; (b) cognitive decline; (c) neuronal loss; (d) phospho-tau levels in neurons; (e) glial cell activation; (f) lymphocyte infiltration; (g) macrophage infiltration; (h) antibody deposition; (i) glial cell loss; (j) oligodendrocyte loss; (k) dendritic cell infiltration; (l) neutrophil infiltration; (m) erythrocyte lysis; (n) erythrocyte phagocytosis; (o) thrombocytosis; (p) thrombolysis; (q) graft rejection; and (r) macrophage-mediated phagocytosis. ; (s) vision loss; (t) antibody-mediated complement activation; (u) autoantibody-mediated complement activation; (v) demyelination; (w) eosinophilia; (x) blistering; (y) pruritus; (z) skin rash; (ab) skin erosions; (ac) petechiae; (ad) bleeding time; (ae) conduction block, compared to the level or degree of the outcome in the individual before treatment with the anti-C1s antibody, and is effective to achieve a reduction of at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or greater than 90%.

[0343] In some cases, anti-C1s antibodies, when administered at a dose of 7.5 g to individuals with a complement-mediated disease or disorder, and when administered as monotherapy or in one or more doses in combination therapy, have been shown to reduce or eliminate the following outcomes: (a) complement activation; (b) cognitive decline; (c) neuronal loss; (d) phospho-tau levels in neurons; (e) glial cell activation; (f) lymphocyte infiltration; (g) macrophage infiltration; (h) antibody deposition; (i) glial cell loss; (j) oligodendrocyte loss; (k) dendritic cell infiltration; (l) neutrophil infiltration; (m) erythrocyte lysis; (n) erythrocyte phagocytosis; (o) thrombocytosis; (p) thrombolysis; (q) graft rejection; and (r) macrophage-mediated phagocytosis. ; (s) vision loss; (t) antibody-mediated complement activation; (u) autoantibody-mediated complement activation; (v) demyelination; (w) eosinophilia; (x) blistering; (y) pruritus; (z) skin rash; (ab) skin erosions; (ac) petechiae; (ad) bleeding time; (ae) conduction block, compared to the level or degree of the outcome in the individual before treatment with the anti-C1s antibody, and is effective to achieve a reduction of at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or greater than 90%.

[0344] In some cases, the anti-C1s antibody, when administered at a dose of 5.5 g to an individual having a complement-mediated disease or disorder, and when administered as a monotherapy or in one or more doses in a combination therapy, exhibits an improvement in one or more of the following outcomes: a) cognitive function; b) transplant graft survival; c) vision; d) motor control; e) thrombosis (reduced clot formation); f) coagulation (reduced clotting); g) renal function; h) hematocrit (red blood cell count); i) pruritus; j) blister formation; k) skin rash; l) petechiae; m) platelet count; n) bleeding time; o) conduction block; and p) inflammation (reduced inflammation), compared to the level or degree of the outcome in the individual prior to treatment with the anti-C1s antibody. %, at least about 90%, or greater than 90% improvement.

[0345] In some cases, anti-C1s antibodies, when administered at a dose of 6.5 g to individuals with a complement-mediated disease or disorder, and when administered as monotherapy or in one or more doses in combination therapy, have been shown to improve the following outcomes: a) cognitive function; b) transplant graft survival; c) vision; d) motor control; e) thrombosis (reduced blood clot formation); f) coagulation (reduced clotting); g) renal function; h) hematocrit (red blood cell count); i) pruritus; j) blister formation; k) skin rash; l) petechiae; m) blood It is effective to achieve at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or more than 90% improvement in one or more of the following outcomes in the individual prior to treatment with the anti-C1s antibody: platelet count; n) bleeding time; o) conduction block; and p) inflammation (reduction in inflammation).

[0346] In some cases, anti-C1s antibodies, when administered at a dose of 7.5 g to individuals with a complement-mediated disease or disorder, and when administered as monotherapy or in one or more doses in combination therapy, have been shown to improve the following outcomes: a) cognitive function; b) transplant graft survival; c) vision; d) motor control; e) thrombosis (reduced blood clot formation); f) coagulation (reduced clotting); g) renal function; h) hematocrit (red blood cell count); i) pruritus; j) blister formation; k) skin rash; l) petechiae; m) blood It is effective to achieve at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or more than 90% improvement in one or more of the following outcomes in the individual prior to treatment with the anti-C1s antibody: platelet count; n) bleeding time; o) conduction block; and p) inflammation (reduction in inflammation).

[0347] In some cases, the anti-C1s antibody, when administered at a dose of 5.5 g to an individual having a complement-mediated disease or disorder, and when administered as a monotherapy or in one or more doses in combination therapy, is effective in reducing complement activation in the individual by at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or greater than 90%, compared to complement activation in the individual before treatment with the anti-C1s antibody.

[0348] In some cases, the anti-C1s antibody, when administered at a dose of 6.5 g to an individual having a complement-mediated disease or disorder, and when administered as a monotherapy or in one or more doses in combination therapy, is effective in reducing complement activation in the individual by at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or greater than 90%, compared to complement activation in the individual before treatment with the anti-C1s antibody.

[0349] In some cases, the anti-C1s antibody, when administered at a dose of 7.5 g to an individual having a complement-mediated disease or disorder, and when administered as a monotherapy or in one or more doses in combination therapy, is effective in reducing complement activation in the individual by at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or greater than 90%, compared to complement activation in the individual before treatment with the anti-C1s antibody.

[0350] In some cases, the anti-C1s antibody, when administered at a dose of 5.5 g to an individual having a complement-mediated disease or disorder, and when administered as a monotherapy or in one or more doses in combination therapy, is effective in inhibiting cleavage of complement component C4 in the individual by at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or more than 90%, compared to the level of C4 cleavage in the individual before treatment with the anti-C1s antibody.

[0351] In some cases, the anti-C1s antibody, when administered at a dose of 6.5 g to an individual having a complement-mediated disease or disorder, and when administered as a monotherapy or in one or more doses in combination therapy, is effective in inhibiting cleavage of complement component C4 in the individual by at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or more than 90%, compared to the level of C4 cleavage in the individual before treatment with the anti-C1s antibody.

[0352] In some cases, the anti-C1s antibody, when administered at a dose of 7.5 g to an individual having a complement-mediated disease or disorder, and when administered as a monotherapy or in one or more doses in combination therapy, is effective in inhibiting cleavage of complement component C4 in the individual by at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or more than 90%, compared to the level of C4 cleavage in the individual before treatment with the anti-C1s antibody.

[0353] In some cases, the anti-C1s antibody, when administered in an effective amount to an individual having a complement-mediated disease or disorder, and when administered in one or more doses as a monotherapy or in a combination therapy, is effective to achieve and maintain a serum concentration of anti-C1s antibody of at least about 20 μg / mL, at least about 25 μg / mL, at least about 30 μg / mL, at least about 35 μg / mL, at least about 40 μg / mL, at least about 45 μg / mL, at least about 50 μg / mL, at least about 55 μg / mL, at least about 60 μg / mL, at least about 65 μg / mL, at least about 70 μg / mL, at least about 75 μg / mL, at least about 80 μg / mL, at least about 85 μg / mL, at least about 90 μg / mL, at least about 95 μg / mL, or at least about 100 μg / mL.

[0354] In some cases, the anti-C1s antibody, when administered in an effective amount to an individual with a complement-mediated disease or disorder, and when administered in one or more doses as a monotherapy or in a combination therapy, achieves and maintains a serum concentration of the anti-C1s antibody that inhibits the classical complement pathway (CP) by at least 50%, at least 60%, at least 70%, at least 80%, or at least 90%. In some cases, the anti-C1s antibody, when administered in a dose of 5.5 g to an individual with a complement-mediated disease or disorder, and when administered in one or more doses as a monotherapy or in a combination therapy, is effective to inhibit the CP by 90%. In some cases, the anti-C1s antibody, when administered in a dose of 6.5 g to an individual with a complement-mediated disease or disorder, and when administered in one or more doses as a monotherapy or in a combination therapy, is effective to inhibit the CP by 90%. In some cases, the anti-C1s antibody, when administered in a dose of 7.5 g to an individual with a complement-mediated disease or disorder, further When administered in one or more doses as monotherapy or in combination therapy, it is effective in inhibiting CP by 90%.

[0355] In some cases, the anti-C1s antibody, when administered in an effective amount to an individual having a complement-mediated disease or disorder, and when administered as monotherapy or in one or more doses in combination therapy, achieves and maintains serum concentrations of anti-C1s antibody to improve or prevent the following outcomes: (a) complement activation; (b) cognitive decline; (c) neuronal loss; (d) phospho-tau levels in neurons; (e) glial cell activation; (f) lymphocyte infiltration; (g) macrophage infiltration; (h) antibody deposition; (i) glial cell loss; (j) oligodendrocyte loss; (k) dendritic cell infiltration; (l) neutrophil infiltration; (m) erythrocyte lysis; (n) erythrophagocytosis; (o) thrombocytopenia; (p) thrombolysis; (q) graft rejection; (r) macrophage infiltration; (p) thrombocytopenia ... Achieve a reduction of at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or greater than 90%, compared to the level or extent of the outcome in the individual prior to treatment with the anti-C1s antibody.

[0356] In some cases, the anti-C1s antibodies, when administered in an effective amount to an individual having a complement-mediated disease or disorder, and when administered as monotherapy or in one or more doses in combination therapy, achieve and maintain serum concentrations of anti-C1s antibodies to improve the following outcomes: a) cognitive function; b) transplant graft survival; c) vision; d) motor control; e) thrombosis (reduced blood clot formation); f) coagulation (reduced clotting); g) renal function; h) hematocrit (red blood cell count); i) pruritus; j) blister formation; k) skin. Achieve at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or greater than 90% improvement in one or more of the following outcomes in the individual prior to treatment with the anti-C1s antibody: rash; l) petechiae; m) platelet count; n) bleeding time; o) conduction block; and p) inflammation (reduction in inflammation).

[0357] In some cases, the anti-C1s antibody, when administered in an effective amount to an individual having a complement-mediated disease or disorder, and when administered in one or more doses as a monotherapy or in a combination therapy, achieves and maintains a serum concentration of the anti-C1s antibody that reduces complement activation in the individual by at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or greater than 90%, compared to complement activation in the individual before treatment with the anti-C1s antibody.

[0358] In some cases, the anti-C1s antibody, when administered in an effective amount to an individual having a complement-mediated disease or disorder, and when administered as a monotherapy or in one or more doses in a combination therapy, achieves and maintains a serum concentration of the anti-C1s antibody that inhibits cleavage of complement component C4 in the individual by at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or more than 90%, compared to the level of cleavage of C4 in the individual before treatment with the anti-C1s antibody.

[0359] In some cases, the effective amount of anti-C1s antibody is at least about 45 mg / kg, at least about 50 mg / kg, at least about 55 mg / kg, at least about 60 mg / kg, at least about 65 mg / kg, at least about 70 mg / kg, at least about 75 mg / kg, at least about 80 mg / kg, at least about 85 mg / kg, at least about 90 mg / kg, at least about 95 mg / kg, or at least about 100 mg / kg.

[0360] In some cases, the effective amount of anti-C1s antibody is between about 60 mg / kg and about 100 mg / kg, between about 60 mg / kg and about 95 mg / kg, between about 60 mg / kg and about 90 mg / kg, between about 60 mg / kg and about 85 mg / kg, between about 60 mg / kg and about 80 mg / kg, between about 60 mg / kg and about 75 mg / kg, between about 60 mg / kg and about 70 mg / kg, or between about 60 mg / kg and about 65 mg / kg. In some embodiments, the effective amount of anti-C1s antibody is between about 45 mg / kg and about 85 mg / kg, between about 45 mg / kg and about 80 mg / kg, between about 45 mg / kg and about 75 mg / kg, between about 45 mg / kg and about 70 mg / kg, between about 45 mg / kg and about 65 mg / kg, between about 45 mg / kg and about 60 mg / kg, or between about 45 mg / kg and about 50 mg / kg. In some cases, the effective amount of anti-C1s antibody is between about 85 mg / kg and about 150 mg / kg, between about 85 mg / kg and about 145 mg / kg, between about 85 mg / kg and about 140 mg / kg, between about 85 mg / kg and about 135 mg / kg, between about 85 mg / kg and about 130 mg / kg, between about 85 mg / kg and about 125 mg / kg, between about 85 mg / kg and about 125 mg / kg, between about 85 mg / kg and about 120 mg / kg, between about 85 mg / kg and about 115 mg / kg, between about 85 mg / kg and about 110 mg / kg, between about 85 mg / kg and about 105 mg / kg, between about 85 mg / kg and about 100 mg / kg, between about 85 mg / kg and about 95 mg / kg, or between about 85 mg / kg and about 90 mg / kg.

[0361] In some cases, the effective amount is about 45 mg / kg, about 50 mg / kg, about 55 mg / kg, about 60 mg / kg, about 65 mg / kg, about 70 mg / kg, about 75 mg / kg, about 80 mg / kg, about 85 mg / kg, about 90 mg / kg, about 95 mg / kg, about 100 mg / kg, about 105 mg / kg, about 110 mg / kg, about 115 mg / kg, about 120 mg / kg, about 125 mg / kg, about 130 mg / kg, about 135 mg / kg, about 140 mg / kg, about 145 mg / kg, or about 150 mg / kg.

[0362] In some cases, the anti-C1s antibody is administered in an effective amount of at least 4 g, at least 4.5 g, at least 5 g, at least 5.5 g, at least 6 g, at least 6.5 g, at least 7 g, at least 7.5 g, at least 8 g, at least 8.5 g, at least 9 g, at least 9.5 g, or at least 10 g.

[0363] In some cases, the anti-C1s antibody is administered in an effective amount of between about 5.5g and about 10g, between about 5.5g and about 9.5g, between about 5.5g and about 9g, between about 5.5g and about 8.5g, between about 5.5g and about 8g, between about 5.5g and about 7.5g, between about 5.5g and about 7g, between about 5.5g and about 6.5g, or between about 5.5g and about 6g. In some embodiments, the anti-C1s antibody is administered in an amount of between about 4.5g and about 8.5g, between about 4.5g and about 8g, between about 4.5g and about 7.5g, between about 4.5g and about 7g, between about 4.5g and about 6.5g, between about 4.5g and about 6g, between about 4.5g and about 5.5g, or between about 4.5g and about 5g. In some embodiments, the anti-C1s antibody is administered in an amount of between about 7.5g and about 12g, between about 7.5g and about 11.5g, between about 7.5g and about 11g, between about 7.5g and about 10.5g, between about 7.5g and about 10g, between about 7.5g and about 9.5g, between about 7.5g and about 9g, between about 7.5g and about 8.5g, or between about 7.5g and about 8g.

[0364] Having now described the present disclosure in detail, it is believed the present invention will be more clearly understood by reference to the following examples, which are included herein for illustrative purposes only and are not intended to limit the disclosure. EXAMPLES

[0365] Anti-C1s antibodies rapidly arrest hemolysis and correct severe anemia in a patient with transfusion-dependent primary cold agglutinin disease This example provides BIVV009 (also known as TNT009), a humanized anti-C1s antibody that provides clinical benefit for patients with cold agglutinin disease. This example provides clinical evidence that BIVV009 can rapidly stop hemolysis and restore normal hemoglobin levels in patients with cold agglutinin disease.

[0366] method: Study design: The study protocol and its amendments were approved by the national competent authorities and the ethical committee of the Medical University of Vienna, and the study is registered with ClinicalTrials.gov (NCT02502903) and EUDRACT (EUDRA-CT2014-003881-26). This is a first-in-human study with a pooled protocol design, studying single and multiple ascending doses (MAD) of BIVV009 in a randomized placebo-controlled setting in healthy volunteers (Phase 1a), as well as in a prospective open-label study design (Phase 1b) in four different diseases with a common underlying pathophysiology, namely antibody-mediated complement activation. This example focuses on the observed efficacy data of BIVV009 in a group of patients affected by cold agglutinin disease (treated from January to December 2016), augmented by confirmation of these findings upon re-exposure to the drug under a specified patient program.

[0367] Patients: Inclusion criteria included age 18 years or older, history of or willingness to be vaccinated against encapsulated pathogens (Neisseria meningitidis, Haemophilus influenzae, and Streptococcus pneumoniae); ability to understand and give informed consent; ability to cooperate, and confirmed diagnosis of cold agglutinin disease (cold agglutinin titer >1:32) within 3 months of prior enrollment. Exclusion criteria were: active infection or history of infection within the previous month; autoimmune disorders other than cold agglutinin disease; other known complement-mediated disorders; known malignancies (except for locally confined basal cell carcinoma of the skin with previous surgical removal, lymphoproliferative disorders not causally related to the complement-mediated disease under study, etc.); clinically significant hepatobiliary disorders; history of transfusion hypersensitivity; allergic or anaphylactic reactions to other therapeutic proteins; substance abuse; psychiatric illness; females of childbearing age not using contraception; concomitant therapy with other experimental drugs or participation in another clinical trial with any investigational drug within 30 days prior to the start of treatment, and weight >98 kg.

[0368] Treatment: BIVV009 is a humanized anti-C1s IgG4 monoclonal antibody with orphan drug designation in the European Union and the United States. Patients were screened and could begin drug infusion as early as 14 days after vaccination against Neisseria meningitidis, Haemophilus influenzae, and Streptococcus pneumoniae. As required by the Austrian Agency for Health and Food Safety (AGES), an initial 10 mg / kg intravenous (IV) "test" dose of BIVV009 was infused in case of unexpected adverse effects from the first infusion. One to four days later, patients received a full 60 mg / kg dose, followed by three additional 60 mg / kg infusions at weekly intervals. Patients were under constant medical observation and monitored by pulse oximetry and regular blood pressure monitoring during the 1-hour infusion. Patients were monitored with regular blood pressure readings. Patients followed the protocol for a total of 49 to 53 days.

[0369] Laboratory analysis: All laboratory parameters were measured in a fully automated manner in the central laboratory of the Medical University of Vienna. A technician additionally performed a microscopic differential blood count. All samples were collected by fresh venipuncture into pre-warmed 37°C evacuated blood collection tubes and transported in pre-warmed steel blocks to the central laboratory. In some cases, strong agglutination and ex vivo hemolysis occurred after blood collection, which prevented accurate measurement of secondary outcome parameters. Direct antiglobulin tests (DAT) were analyzed using LISS / Coombs Gelcards (Bio-Rad GmbH, Vienna, Austria). Serum BIVV009 levels were determined using a direct binding ELISA in which free BIVV009 was captured using C1s-coated plates and detected using a goat anti-human HRP-conjugated secondary antibody and developed with the colorimetric substrate 3,3',5,5'-tetramethylbenzidine (TMB). The pharmacodynamic activity of BIVV009 in serum samples was evaluated using the WIESLAB® (WL CP), an ELISA that detects deposition of membrane attack complex mediated by the classical pathway of the complement system ex vivo (Euro-Diagnostica, Malmo, Sweden). Flow cytometry to detect C3d on patients' erythrocytes was performed as described in Shi et al., Blood, 123(26):4015-22 (2014). Figures 2A-2C show a summary of the laboratory parameters measured before treatment and the maximum changes during treatment.

[0370] Statistical Analysis: For this pilot study in CAD patients, no sample size calculations were performed because no previous data were available to estimate effect size and its variability. The primary outcome variable of interest in CAD patients is the hemoglobin level, as it determines symptoms, circulatory instability, and is the main initiation factor for transfusions. Changes in hemoglobin are expressed as 95% confidence intervals. Even with sampling tubes prewarmed to 37°C, strong ex vivo red blood cell aggregation sometimes resulted in non-measurable values ​​for reticulocyte count, lactate dehydrogenase, and DAT. However, there was no data loss. Clinical response was defined as an increase in hemoglobin of ≥2 g / dL. No inferential statistical tests were planned, but Friedman's ANOVA was used for time courses and Wilcoxon test for individual effect differences between baseline and maximum. Other markers such as DAT, bilirubin, reticulocyte count, haptoglobin, lactate dehydrogenase, total complement activity (CH50) and C4 are non-independent secondary outcome parameters. Data are summarized descriptively using median and range. In cases where values ​​were below the detection limit of the assay, the value of the detection limit minus 1 was assigned to the graph (e.g., instead of haptoglobin < 12 mg / dL, haptoglobin = 11; similarly, 1:2048 was assigned to cold agglutinin titer > 1:1024). To increase the level of confidence regarding causality and exclude regression to the mean, we studied the reversal of the effect (i.e. recurrence of anemia and hemolysis) when the drug was washed out, and its repetition when rechallenged. The concept was adapted from the Clinical Trials in Small Populations guideline (CHMP / EWP / 83561 / 2005) and represents a series of non-randomized n-of-1 trials with multiple crossover periods from mid-treatment to post-treatment. The duration of the post-treatment period was maintained mainly due to the recurrence of hemolysis and severe anemia after treatment interruption, while avoiding unnecessary transfusions between periods.

[0371] result: Study population: Patient characteristics are shown in Figure 1. Thirteen patients were screened; three women were excluded due to iron deficiency anemia and inactive cold agglutinin disease, negative cold agglutinin titers, or Hb levels >11 g / dL. Ten patients were included, three from the UK and one from the UK. The final included patients were one from Canada / Spain (8 Caucasian, 1 Spanish, 1 Indian). Median disease duration was 5 years (range: 1-12 years). Three patients were studied with a moderate dose of steroids (10-25 mg / day), which was reduced to less than 10 mg prednisolone on the first study day and tapered and discontinued within the first week of BIVV009.

[0372] Pharmacokinetics and pharmacodynamics: A single dose-escalation portion of the study performed in healthy volunteers demonstrated that BIVV009 undergoes non-linear elimination at concentrations below approximately 100 μg / mL; this behavior is frequently observed with other monoclonal antibodies, suggesting that a target-mediated elimination process is involved. Using an ex vivo readout of serum classical pathway activity (WL CP ELISA, see Methods), we were able to assess the relationship between serum BIVV009 concentrations and serum classical pathway activity. Based on modeling in NHV subjects, a steep concentration-effect relationship was observed for knockdown of classical pathway activity, reaching a maximum effect (>90% inhibition of classical pathway activity) at a BIVV009 concentration of approximately 20 μg / mL (Figure 3A). NHV (2073 μg / mL and 234612 μg / mL, respectively) showed a significant increase in BIVV009 activity compared with NHV (2073 μg / mL and 234612 μg / mL, respectively). * hours / mL) and CAD patients (1885 μg / mL and 209996 μg * Mean C after four weekly doses of 60 mg / kg (hours / mL) maxAs demonstrated by the AUC and RR results, the pharmacokinetics of BIVV009 are similar regardless of disease status. As can be seen from the mean BIVV009 concentration-time profiles in CAD patients (Figure 3B), BIVV009 concentrations remained well above 20 μg / mL levels in patients receiving 60 mg / kg once weekly for four doses, and only began to approach this pharmacodynamic threshold 672 hours (28 days) after the last dose, indicating that this dosing regimen is sufficient to maintain long-acting complement inhibition above the threshold for clinical effect.

[0373] Consistent with the results observed using the WL CP ELISA, CH50, a measure of serum classical pathway activity, was significantly reduced from pretreatment levels within 24 hours of BIVV009 administration (p=0.0209). Plasma levels of complement component C4, the first substrate cleaved by C1s, gradually increased over the course of the study, resulting in a median 3.8-fold increase (p=0.0077). Flow cytometry analysis revealed that C3d-positive red blood cell counts significantly decreased from 40% (IQR: 27-49%) to a nadir of 21% (IQR: 14-27%) 5 weeks after the first dose (p<0.0172; Figure 4A). These measures of BIVV009 pharmacodynamic activity in vivo at C4 levels, and more importantly at the red blood cell surface, are consistent with its mechanism of action and suggest that BIVV009 inhibits the classical complement pathway in patients with CAD.

[0374] The present examples further show that in CAD patients, BIVV009 increases hemoglobin levels and rapidly inhibits hemolysis. BIVV009 infusion resulted in a median hemoglobin increase of 1.6 g / dL within the first week of treatment (p=0.0069; n=10), leading to a median best response of 3.9 g / dL after 6 weeks (IQR: 1.3-4.5; 95% CI: 2.1-4.5; p=0.0050; n=10) (Figure 4B). Furthermore, hemoglobin normalized completely (≧12 g / dL) during the limited study duration in four patients, and five patients experienced an increase of more than 4 g / dL. Figure 6 shows hemoglobin data over time (pre-BIVV009 administration) for one patient, demonstrating the clinical efficacy of BIVV009 in chronically anemic CAD patients. PRBC means when transfusion support was provided. These longitudinal hemoglobin data include hemoglobin values ​​over 4 years prior to enrollment in the study and demonstrate that hemoglobin levels in this patient had never reached the lower limit of normal (12 g / dL, dotted line) until BIVV009 was provided. The hematological data in Figure 7 demonstrate the clinical benefit provided by BIVV009 in the same patient over the course of treatment (represented by the solid horizontal bar) and after 12 h of treatment (represented by the solid horizontal bar). Figure 7A shows that BIVV009 administration results in an immediate increase in reticulocytes, suggesting that BIVV009 prevents the destruction of reticulocytes. Figure 7B shows that BIVV009 increased hemoglobin levels to 3.8 g / dL. Figure 7C shows that BIVV009 increases haptoglobin levels to within the normal range compared to pre-treatment when haptoglobin is below the limit of detection. Figure 7D shows that LDH levels, a marker of intravascular hemolysis, were reduced with BIVV009 treatment. Figure 7E shows that BIVV009 reduced CH50 levels, a measure of serum classical pathway activity. Finally, Figure 7F shows that BIVV009 reduced bilirubin levels, suggesting that the drug stopped extravascular hemolysis. The modulation of all these markers was reversed after BIVV009 washout (period indicated by no bars) and recurred upon retreatment (solid horizontal bars). Together, these data demonstrate that BIVV009 prevents cold agglutinin-mediated complement destruction of red blood cells and reticulocytes in CAD patients.

[0375] Reticulocyte counts increased to a median of 41% within the first 24 hours (p=0.0381, n=10) and then gradually declined into the normal range as expected with increasing hemoglobin levels. All five patients who were transfusion-dependent prior to enrollment did not receive blood transfusions during their BIVV009 treatment course. Figure 6 shows hemoglobin levels over time in patients with CAD who received packed red blood cell (PRBC) transfusions prior to beginning treatment with BIVV009. Figures 7A-7F show the biochemical response patterns of reticulocytes, hemoglobin, haptoglobin, lactate dehydrogenase (LDH), total complement activity (CH50), and bilirubin levels in patients with CAD who received repeated BIVV009 doses.

[0376] Haptoglobin was below detection levels (<11 mg / dL) in all patients prior to treatment but normalized within 1–2 weeks in four patients, confirming complete inhibition of hemolysis. Pretreatment bilirubin levels were found to be elevated in seven CAD patients, suggesting that increased red blood cells are reversed by the mononuclear phagocyte system. BIVV009 administration resulted in a 61% reduction in median bilirubin levels (p=0.0068, n=10; Fig. 4C) within 24 hours of the first infusion, normalizing in six patients. Similarly, upon BIVV009 washout, bilirubin levels increased significantly, demonstrating recurrence of hemolysis. The rapidity of reduction and normalization of circulating bilirubin upon BIVV009 treatment, in addition to its reproducibility after washout, provided a disease-associated biomarker to test the relationship between BIVV009 and extravascular hemolysis. At BIVV009 concentrations higher than 20 μg / mL, the threshold above which the drug completely inhibits serum classical pathway activity (Figure 3A), bilirubin levels were almost exclusively within the normal range. Conversely, at bilirubin levels below 20 μg / mL, bilirubin showed a trend toward exceeding the normal range (Figure 5).

[0377] Response analysis: Seven of the 10 CAD patients derived clinical benefit, defined as a hemoglobin increase of >2 g / dL, including patients who failed to respond or relapsed after rituximab (C1002), rituximab + bendamustine (C1001; C1010), or eculizumab (C1010). Three patients did not respond adequately to treatment with BIVV009 (hemoglobin increase of 0.5-1.3 g / dL). One patient (C1011) had repeatedly positive Coombs tests for both C3d and IgG (>1+), suggesting that the patient had both cold and warm autoimmune hemolytic anemia (i.e., mixed autoimmune hemolytic anemia). Two other patients had active lymphoma and showed lymphocytic bone marrow infiltration of 70% and 15% (C1003 and C1013, respectively), and one further patient showed 60% bone marrow infiltration and only a partial response (C1009). LDH levels also normalized, whereas LDH increased 2- to 3-fold in 2 of 3 nonresponders.

[0378] Repeated responses after BIVV009 washout and rechallenge: Approximately 3-4 weeks after the last dose of BIVV009, when BIVV009 levels fell below the pharmacodynamic threshold of 20 μg / mL, complement deposition on red blood cells, anemia, and hemolysis recurred in all responders (Figure 3B, Figure 4). Therefore, responders were given the opportunity to participate in a designated patient program to prove causation with consecutive treatments in a series of n-of-1 studies in six patients. Patients showing partial responses were preferred to continue without treatment (as patients traveled to and from the UK), and patients' hemoglobin levels decreased from 8.7 g / dL, while BIVV009 was at 6-6.5 g / dL during follow-up. In the remaining six patients, re-exposure to BIVV009 repeated the onset of immediate effects and rapid and complete inhibition of hemolysis.

[0379] Because pharmacokinetic analysis suggesting drug accumulation demonstrated that the weekly dose of 60 mg / kg resulted in increased trough levels of BIVV009 (Figure 2), alternative doses and dosing regimens were explored in a designated patient program. Two patients received BIVV009 at a dose of 45 mg / kg weekly for four infusions, followed by 45 mg / kg every other week. This was excluded because laboratory parameters indicated breakthrough hemolysis after seven infusions. The breakthrough was accompanied by recovery of serum CH50 activity and undetectable circulating BIVV009, confirming that the breakthrough was the result of insufficient trough concentrations. Patients were then further maintained with BIVV009 at a loading dose of 60 mg / kg weekly for two infusions, followed by 60 mg / kg every other week, again resulting in breakthrough hemolysis in two patients after eight infusions. The dose was increased to 65 mg / kg every other week or to a fixed dose of 5.5 g to prevent further breakthrough events. All five transfusion-dependent patients achieved normal hemoglobin levels (>12 g / dL) after at least one BIVV009 treatment and remained transfusion-free during treatment. Two patients discontinued the designated patient program for reasons other than drug safety or efficacy and became transfusion-dependent again, requiring transfusion support approximately every other week.

[0380] Safety: All infusions were well tolerated without premedication and without any significant drug-related adverse effects. Adverse events were few during the study; all were mild or moderate and were considered unrelated or unlikely to be related to the study drug. Patient C1001 had nausea and vomiting on two occasions, once with diarrhea. Patient C1002 complained of night sweats and a new vertebral fracture in addition to a pre-existing fracture due to long-term steroid therapy. This ultimately led to planned hospitalization for treatment of bone pain several weeks after the patient's participation in the study ended. Patient C1004 complained of pruritus and had a rash that was transient despite continued BIVV009 exposure.

[0381] Conclusion: These data indicate that C1s blockade with the anti-C1s monoclonal antibody BIVV009 rapidly corrects severe transfusion-dependent anemia in patients with primary cold agglutinin disease. EXAMPLES

[0382] Anti-C1s monoclonal antibody in late antibody-mediated renal allograft rejection-Results from a first-in-human phase 1 trial This example provides BIVV009 (also known as TNT009), a humanized anti-C1s antibody that provides clinical benefit for kidney transplant patients with antibody-mediated rejection (ABMR). This example demonstrates that BIVV009 inhibits allogeneic antibody rejection in kidney allografts. To provide clinical evidence that it effectively blocks classical complement pathway (CP) activation triggered by the body.

[0383] This single-arm Phase 1b study was designed to investigate the safety / tolerability profile and complement inhibitory activity of BIVV009 in limited-course kidney transplant recipients on long-term immunosuppression. Here, we describe the CP blocking capacity of BIVV009 and histomorphological and molecular evaluation of systematic follow-up biopsies from patients with active ABMR, as well as evidence of CP activation (in vivo C4d staining and / or detection of donor-specific antibodies (DSA) that fix complement in serum) with BIVV009 treatment.

[0384] material and method: Study Design and Objectives. This prospective Phase 1 study enrolled a single cohort of 10 kidney transplant recipients diagnosed with late acute or chronic active ABMR. The study was part of a Phase 1 basket study designed to evaluate the safety, tolerability and potential efficacy of the humanized monoclonal anti-C1s antibody BIVV009 (formerly TNT009; Bioverativ Therapeutics, Inc., South San Francisco, CA) in healthy volunteers and patients with a variety of diseases believed to be mediated by CP (cold agglutinin disease, warm autoimmune hemolytic anemia, bullous pemphigoid and ABMR). The study was registered with ClinicalTrials.gov (NCT02502903) and EUDRACT (EUDRACT number: 2014-003881-26). We hypothesized that BIVV009 is expected to be safe and well tolerated and can effectively block CP activity in patients with ABMR. The study was performed at the Department of Clinical Pharmacology (Medical University of Vienna). No classical sample size estimation was performed. The study was approved by the Ethics Committee of the Medical University of Vienna and was carried out in accordance with the guidelines of the Good Clinical Practice of Pharmaceuticals and in accordance with the principles of the Declaration of Helsinki and the Declaration of Istanbul. Figure 8 illustrates the design of the study.

[0385] STUDY PATIENTS:Ten adult kidney transplant recipients diagnosed with late stage ABMR were enrolled in this study. Subjects were recruited at the Nephrology Outpatient Clinic of the Medical University of Vienna between December 2015 and September 2016, and the study was concluded in November 2016. All participants provided written informed consent prior to enrollment. The main inclusion criteria were the ability to understand and perform informed consent, age ≥18 years, functional allograft with estimated glomerular filtration rate (eGFR) ≥20 mL / min / 1.73 m2 at or after 180 days post-transplant, detection of one or more anti-HLA class I and / or II DSA in serum, late ABMR (acute or chronic) showing histomorphological features of an active rejection process (g score >0, ptc score >0) proven by biopsy, molecular biopsy signature of ABMR determined by Molecular Microscopy Diagnostic System 18 (MMDx; molecular ABMR score ≥0.20), and signs of CP activation (complement fixing DSA and / or C4d deposits in the index biopsy). Female subjects had to be postmenopausal, surgically infertile, or willing to use highly effective birth control methods throughout the study and for 30 days after the last study visit. The main exclusion criteria were acute allograft failure and / or any treatment for rejection within 4 weeks prior to study inclusion, a diagnosis of TCMR, or contraindication to antibiotic prophylaxis with oral ciprofloxacin. Other exclusion criteria were: active acute or chronic viral, bacterial, fungal, or mycobacterial infection or history of such within the previous month, autoimmune disorder or known malignancy, clinically significant hepatobiliary disorders, history of transfusion hypersensitivity, allergic or anaphylactic reactions to other therapeutic proteins, substance abuse, psychiatric illness or other reasons that would make the subject less than adequately compliant with the study procedures, women who were pregnant, breastfeeding, or potentially unreliable regarding contraceptive practices, weight >98 kg, and procedures. Participation in another clinical trial within 30 days prior to initiation.

[0386] Study medication: Patients were admitted to the research unit (Department of Clinical Pharmacology, Medical University of Vienna) for drug administration. For safety reasons, patients received an initial test dose of 10 mg / kg of BIVV009 one day before the first full dose. Treatment consisted of four weekly doses of 60 mg / kg. BIVV009 was administered via the intravenous route over 60 minutes. Prior to administration, all subjects were vaccinated against encapsulated bacteria (Neisseria meningitidis, Haemophilus influenzae, Streptococcus pneumoniae) followed by prophylaxis with ciprofloxacin (250 mg orally twice daily) for the entire study period.

[0387] Outcome measures: Study endpoints were assessed up to day 50 (last study visit). Primary endpoints were to assess the safety and tolerability of BIVV009, the incidence and severity of adverse events (AEs) defined and classified according to the International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH) guidelines for good clinical practice for pharmaceuticals. The protocol did not include interim analyses. However, the study was governed by a continuous safety review process. Safety reviews were performed in consultation with an independent Data Safety Management Committee, and the option of study discontinuation was taken in case of unexpected clinical and laboratory findings raising safety concerns. Secondary endpoints included the pharmacokinetics of BIVV009, the ability of BIVV009 to block CP in serum (overall CP activity and DSA-induced CP activation) and transplanted kidneys (capillary C4d deposition), the effect of BIVV009 on the mean fluorescent activity of DSA and the fixation capacity of C1q, as well as eGFR (Mayo's equation) and spot urine protein / creatinine ratio. Patients underwent follow-up protocol biopsies on day 32 to evaluate the effect of BIVV009 on antibody-induced inflammation / injury and gene expression patterns.

[0388] Antibody and complement detection: Antibody and complement assays were performed on samples taken on 12 consecutive occasions: day 0 (1 hour before administration of the first test dose of BIVV009), days 1, 8, 15 and 22 (1 hour before each full dose of BIVV009), and days 29, 36, 43 and 50 (end of study visit). Serum was aliquoted and stored at -80°C without repeated freezing and thawing until analysis. To avoid variability in results between studies, serum from all time points was assayed retrospectively after the end of the study.

[0389] For Luminex-based detection of IgG-type DSA, we used the LABScreen HLA class I and II single antigen flow bead (SAFB) assay ( Sera were heat inactivated (56°C for 30 min) to prevent complement-dependent interference. The threshold for DSA positivity was set at an MFI value of ≥ 1,000. Alloreactivity patterns were analyzed using HLA Fusion 3.0 software (One Lambda) and donor specificity was assessed in terms of serology and / or low- or high-resolution donor / recipient HLA typing (HLA-A, -B, -Cw, -DR, -DQ and / or DP) results retrieved from local HLA laboratories or Eurotransplant databases. For each sample, the individual DSAs and the DSA with the highest IgG MFI (MFI_max) were recorded.

[0390] The ability of detected DSA to bind recombinant C1q was assessed in SAFB using the C1qScreen assay according to the manufacturer's instructions (One Lambda), with an MFI threshold of ≥500 for C1q positivity.

[0391] The activation of the main component C3 caused by DSA was investigated using an earlier protocol. Complement profiling was assessed by measuring the deposition of the C3 complement breakdown product C3d onto SAFB (patient serum as a complement source) according to the protocol. Briefly, patient serum was incubated with SAFB for 30 min at room temperature and then incubated with a biotin-conjugated monoclonal antibody against human C3d (4 μg / mL; Quidel, San Diego, CA, USA) for an additional 30 min. Phycoerythrin-conjugated streptavidin (1 μg / mL; eBioscience, San Diego, CA, USA) was then added for 30 min. The threshold for C3d positivity was set at MFI ≥ 100.

[0392] For the evaluation of overall CP complement activity, two different assay principles were applied. The WIESLAB® assay of the classical pathway of the complement system to evaluate the activation of the membrane attack complex caused by CP was performed according to the manufacturer's instructions (Euro-Diagnostica, Malmo, Sweden). In parallel, the ability of sera from patients administered BIVV009 to deposit complement in an earlier described solid-phase assay that specifically detects C3 activation caused by third-party HLA antibodies was evaluated. Briefly, patient sera were incubated with a mixture of LABSCREEN mixed beads (One Lambda) coated with HLA haplotypes spiked with high levels of complement-activating HLA antibodies (preincubation with a pool of heat-inactivated sera obtained from three extensively sensitized patients, each of whom had a virtual panel reactivity of >99% in the SAFB assay). After washing, the beads were stained with biotin-conjugated anti-C3d antibodies and PE-conjugated streptavidin as described above. Assay results were recorded as normalized C3d MFI by subtracting the as-measured MFI obtained with heat-inactivated non-binding negative control serum from the MFI obtained with patient serum MFI (normalized MFI). For each test serum, we calculated the average normalized C3d MFI recorded for 4 of 12 HLA class I and 5 HLA class II bead populations in the LABSCREEN mixed bead panel [one HLA class II bead population (ID25) was excluded due to consistently negative C3d staining, MFI<100].

[0393] Biopsies. Renal allograft biopsies were performed using a 16-gauge needle. Two cores were obtained for light microscopy, electron microscopy, and gene expression analysis. For immunohistochemical C4d staining, we applied a polyclonal anti-C4d antibody (BI-RC4D, Biomedica, Vienna, Austria). C4d was scored 0 (negative), 1 (minimal), 2 (focal), and 3 (diffuse), respectively. Minimal staining (C4d1) along the peritubular capillaries (PTCs) was considered positive. For gene expression analysis, 3 mm proportioned biopsy cores were placed in RNAlater, stored at -20°C, and transported at room temperature to the Alberta Transplant Applied Genomics Centre (ATAGC, University of Alberta, Edmonton, AB, Canada). RNA extraction and gene expression analysis were performed using PrimeView GeneChip arrays (Affymetrix) as previously described in detail. A series of tests were performed using a CT scan of the CT scanner (Santa Clara, CA, USA). Classifiers for rejection (ABMR, TCMR, all rejection) or acute kidney injury (AKI scores) were generated based on a reference set of 1208 biopsy samples21. In addition, scores of various etiology-based transcription (PBT) representing major biological events occurring in experimental cell culture studies, mouse transplantation studies and human kidney transplantation and shown to be involved in various annotated pathological processes (e.g., cytotoxic T cell infiltration, gamma-interferon action, natural killer cell burden, epithelial damage) were evaluated as previously described in detail. According to the 2013 update of the Banff classification (Haas et al., Am J Transplant, 14(2):272-283 (2014)), ABMR was classified according to histomorphological, immunohistochemical (C4d), ultrastructural (PTC basement membrane multilayering), serological (DSA detection) criteria, respectively, and a thoroughly validated molecular classifier for ABMR (molecular ABMR score ≥ 100). 0.2)18 is defined based on

[0394] Statistical analysis. Continuous data are presented as median, interquartile range (IQR) and range. Discrete data are presented as number and percentage. For paired sample comparisons, Wilcoxon signed rank test was used. A two-sided p value <0.05 was considered statistically significant. Analyses were performed using GraphPad Prism 6.0 (GraphPad Software Inc., San Diego, CA, USA) and IBM SPSS Statistics 24 (IBM Corporation, Armonk, NY, USA).

[0395] result: This phase 1 pilot study enrolled 10 kidney transplant recipients diagnosed with late anti-HLA DSA-positive active ABMR associated with signs of antibody-induced CP activation (complement fixation DSA and / or C4d staining in PTC). ABMR was diagnosed after a median of 4.3 years post-transplant and the first study visit was performed a median of 38 (IQR: 28-45) days after index biopsy. As shown in Figure 8, all enrolled patients received BIVV009 once at a test dose of 10 mg / kg followed by four weekly doses of 60 mg / kg and underwent a follow-up biopsy 32 days after the first infusion. Baseline characteristics and data are provided in Table 2 below.

[0396] [Table 2]

[0397] Two of the study patients were living donor transplant recipients, one of whom was ABO incompatible. Three recipients were subjected to a peri-transplant immunoadsorption protocol for preformed DSA. At study inclusion, nine patients were receiving tacrolimus mycophenolate and steroids. The median eGFR was 46 mL / min / 1.73 m2 and the urinary protein / creatinine ratio was 399 mg / g. Table 3 provides the immunological, histomorphological and molecular results obtained at baseline.

[0398] [Table 3]

[0399] Immunodominant DSA was primarily directed against HLA class II antigens (7 patients; anti-HLA DQ reactivity: n=5). Six of the patients had a history of immunodominant DSA. 1 showed significant C1q binding and 7 showed C3d binding. Individual DSA specificities identified at the time of study inclusion decision are detailed in Figure 9. Nine study patients showed chronic / active ABMR and 1 showed acute / active ABMR. Eight ABMR cases were C4d positive. No patient had T cell-mediated rejection and one index biopsy showed borderline changes. The combined glomerulitis and peritubular capillaritis score (g+ptc score) was within a median range of 4 and the transplant glomerulopathy (cg) score was 2. Median molecular ABMR and overall rejection scores were 0.78 and 0.75, respectively (Table 3).

[0400] Effect of BIVV009 on CP activity in serum: BIVV009 treatment resulted in a complete and sustained blockade of serum CP activity detected by the WIESLAB® assay (formation of membrane attack complex triggered by CP) (Figure 10A). Four weeks after the last infusion, the median CP activity was still below 50%. Thus, as shown in Figure 10A, CP inhibition was closely related to serum concentrations of BIVV009. A comparable effect was observed in the bead assay evaluating C3 cleavage triggered by HLA antibodies. As illustrated in Figure 10B, BIVV009 did not affect the MFI of immunodominant DSA or its ability to fix C1q. However, C3 activation triggered by DSA was virtually completely inhibited.

[0401] Impact of BIVV009 on C4d deposition in PTC: Follow-up biopsies performed 32 days after the first infusion showed a significant decrease in the median C4d score: 2 (IQR: 2-3) in the index versus 0 (0-1) in the follow-up biopsies (p=0.016) (Figure 11A). Of the five recipients, three showed a diffuse staining pattern (C4d3) and became completely C4d negative, and two showed only minimal staining (C4d1) in their follow-up biopsies. However, in a single case of focal staining (C4d2) in an ABO-incompatible allograft, no change in C4d staining was observed.

[0402] Impact of BIVV009 on histomorphology and molecular biopsy results: As shown in Figure 11B, the degree of microcirculatory inflammation did not change [g+ptc score: 4 (IQR: 4-5) in index vs. 5 (3-5) in follow-up biopsy; p>0.99]. As shown in Figure 11C, transplant glomerulopathy [cg score: 2 (1-3) vs. 2 (1-3); p=0.38] remained unchanged. Molecular classifiers for rejection: ABMR score: 0.78 (0.49-0.96) vs. 0.82 (0.58-0.96); p=0.67 (Figure 11D); total rejection score: 0.75 (0.62-0.85) vs. 0.71 (0.59-0.87); p>0.99 (Figure 11E); TCMR score: 0.01 (0.0-0.03) vs. 0.02 (0.0-0.01); p=0.44 (Figure 11F)], acute kidney injury [AKI score: 0.19 (0.02-0.67) vs. 0.24 (0.11-0.5 7); p=0.57 (Figure 11G)], or chronic injury (atrophy / fibrosis score: 0.41 (0.31-0.69) vs. 0.37 (0.16-0.51); p=0.43 (Figure 11H). To evaluate the impact of CP blockade on various transcriptional subsets with annotations related to distinct molecular pathogenesis, we compared changes in selected PBT scores, as exemplified in Figures 12A-12L. When indices were compared to follow-up biopsies, we found no significant differences (Figures 12A-12L).

[0403] Renal function and safety outcomes: As illustrated in Figure 13, there was no change in median eGFR [46 (IQR: 27-61) vs. 42 (27-65); p=0.85] and protein / creatinine ratio [399 (IQR: 181-672) vs. 310 (141-1,222); p=0.88] from baseline to day 50. Table 4 provides adverse events recorded during the study period.

[0404] [Table 4]

[0405] Treatment was well tolerated. All subjects had one or more AEs, but no serious adverse events (SAEs) occurred. Six patients had mild AEs and four had moderate AEs. The most frequent events recorded during the study were headache (n=3), peripheral edema (n=3), and fatigue (n=2). No AEs were considered treatment related. There were no cases of bacterial or fungal infection, although one recipient developed asymptomatic CMV viremia (up to 3000 copies / mL) 6 weeks after study initiation, which was reversible with a course of oral valganciclovir.

[0406] These results indicate that BIVV009 is able to block CP in both serum and tissues. EXAMPLES

[0407] Safety, Tolerability, and Pharmacokinetics and Pharmacodynamics of Multiple Doses of BIVV009 in Patients with Chronic Immune Thrombocytopenia (ITP) This example provides BIVV009 (also known as TNT009), a humanized anti-C1s antibody that provides clinical benefit for patients with chronic immune thrombocytopenia (ITP). The objective of this Phase 1 study is to investigate the safety, preliminary clinical benefit, and activity of BIVV009 in patients with chronic immune thrombocytopenia. This study will be an interventional study with single group assignment. Ten patients will be enrolled in this study.

[0408] [Table 5]

[0409] Outcome measures Primary outcome measures: 1. Incidence of treatment-emergent adverse events. An AE is any untoward medical occurrence in a participant participating in a clinical study, but not necessarily causally related to the medicinal / biological agent under investigation. A serious adverse event (SAE) is any AE that results in death, causes permanent or significant disability / incapacity, requires patient hospitalization or extension of current hospitalization, is a life-threatening experience, is a congenital malformation / birth abnormality, and may endanger the participant and / or require medical or surgical intervention to prevent one of the outcomes listed above. [Time frame: time from first dose to last study visit, assessed up to approximately 13 weeks].

[0410] 2. Number of participants who discontinued early. The number of participants who discontinued early will be evaluated. [Time frame: up to day 91].

[0411] 3. Number of patients with abnormal clinical laboratory values. The abnormal laboratory values included hematology, clinical chemistry panel, coagulation safety panel, urine tests, and antibodies to platelet antigens. [Time slot: Until day 91].

[0412] Secondary outcome measures: 4. Percentage of participants (CR) in full responses. Complete responses in accordance with evidence and practice guidelines for immune thrombocytopenia: 100 platelet count, measured in two cases with at least 7 days apart. * 10 9 greater than or equal to (≥) / liter, and no bleeding. [Time frame: baseline to end of treatment (9 weeks)].

[0413] 5. Percentage of participants who showed a response (R). Response. Platelet count ≥ 30 * 10 9 / liters, and two cases measured at least 7 days apart, greater than 2-fold increase from baseline, and no bleeding. [Time frame: from baseline to the end of treatment (9 weeks)].

[0414] 6. Percentage of participants who did not respond (NR): Non-responder (NR): platelet count ≥ 30 × 10 9 / liter (<30 * 10 9 / liter), or less than a two-fold increase from baseline, or bleeding. Platelet counts were measured on two occasions more than 1 day apart. [Time frame: baseline to end of treatment (9 weeks)].

[0415] 7. Percentage of participants without a complete response. Loss of complete response: Platelet count <100 measured on two occasions >1 day apart * 10 9 / liter, and / or bleeding. [Time frame: baseline to end of treatment (9 weeks)].

[0416] 8. Percentage of participants who do not respond. Loss of response: Platelet count < 30 * 10 9 / L or less than two-fold increase in platelet count from baseline, or bleeding. Platelet counts must be measured in two cases, longer than one day. [Time frame: from baseline to the end of treatment (9 weeks)].

[0417] 9. Plasma concentrations of BIVV009. Plasma concentrations of BIVV009 will be evaluated. [Time frame: up to day 91].

[0418] 10. Maximum observed plasma concentration (Cmax) of BIVV009. The maximum concentration of BIVV009 in plasma will be evaluated. [Time frame: up to day 91].

[0419] 11. Time to reach the observed maximum plasma concentration of BIVV009 (Tmax). The time (Tmax) at which the observed maximum plasma concentration of BIVV009 is reached will be evaluated. [Time slot: Until day 91]

[0420] 12. Area under the concentration-time curve (AUC) from time 0 over the dosing interval of BIVV009 (AUC[0-tau]). The area under the concentration-time curve (AUC) (AUC[0-tau]) from time 0 over the dosing interval of BIVV009 will be evaluated. [Time slot: Until day 91]

[0421] 13. Number of participants with anti-drug antibodies (ADA) to BIVV009. A blood sample will be collected to determine the number of participants with anti-drug antibodies (ADA) against BIVV009. [Time slot: Until day 91].

[0422] 14. Classical pathway levels of the complement system as measured by the WIESLAB® assay. BIVV009 inhibition of the classical pathway of the complement system is measured by the WIESLAB® assay. [Time frame: up to day 91].

[0423] 15. Total complement (CH50) level. Complement CH50 is a blood test that helps determine whether abnormalities and deficiencies of proteins in the complement system are involved in any increase in autoimmune activity. It will be evaluated using a complement assay. [Time slot: Until day 91].

[0424] 16. Total Complement Factor C4 Levels. Total C4 levels in plasma will be assessed using a complement assay. [Time Frame: up to Day 91].

[0425] 17. Components of the C1 complex: C1q and C1s levels. Complement assays will be used to assess levels of C1q and C1s in plasma. [Time frame: up to day 91].

[0426] 18. Number of participants with autoantibodies (GPIIb / IIIa and GPIb / IX) to platelet antigens. Autoantibodies (GPIIb / IIIa and GPIb / IX) against platelet antigens will be evaluated. [Time slot: Until day 91].

[0427] Eligibility Inclusion Criteria: · ITP treatments: Platelet count <30 in participants who are not responding to at least two of the following ITP treatments: corticosteroids, rituximab, thrombopoietin agonists, azathioprine, danazole, cyclosporine A, or mycophenolate mofetils * 10 9 Chronic ITP that is refractory to standard therapy, as defined by a mean blood volume of 100 / liter. ·Normal prothrombin time (PT / INR) and activated partial thromboplastin time (aPTT). · No history of coagulation disorders. · Hemoglobin levels above 10 g / deciliter (g / dL) (>10 g / deciliter) (subsequently transfusions are allowed) and normal leukocytes (WBC) and neutrophil counts (an increase in WBC / absolute neutrophil count [ANC] due to steroid treatment is allowed). Eastern Cooperative Oncology Group (ECOG) activity grade less than or equal to 2 (≦2). ·Have been vaccinated or willing to receive the vaccine against encapsulated pathogens (Menococcus, Meningitis B, Haemolytic influenzae, and Streptococcus pneumoniae). If the final vaccination is less than five years before registration, reimmunization with meningococcal conjugates is required. - Adequate intravenous (IV) access.

[0428] Exclusion criteria: · A clinically significant medical history or ongoing chronic disease that is expected to risk participant safety or impair the quality of the data obtained by participation in this study. Any kind of clinically relevant infection within the month prior to enrollment. History of venous or arterial thrombosis within the year prior to enrollment. Use of aspirin, nonsteroidal anti-inflammatory drugs (NSAIDs), or blood thinners within 1 week of enrollment. · History of lupus or other autoimmune disorders associated with antinuclear antibodies (ANA) at the time of screening. Secondary immune thrombocytopenia of any cause including lymphoma, chronic lymphocytic leukemia, and drug-induced thrombocytopenia. ·Positive hepatitis panel (e.g., hepatitis B surface antigen and / or hepatitis C virus antibody) before or at the time of screening. · Positive human immunodeficiency virus (HIV) antibodies before or at the time of screening. EXAMPLES

[0429] A randomized, first-in-human, healthy volunteer trial of BIVV009, a humanized antibody for specific inhibition of the classical complement pathway This example provides a first-in-human, double-blind, randomized, placebo-controlled, dose-escalation study of BIVV009 in healthy adults.

[0430] Healthy female and male subjects aged 18 years or older were eligible for enrollment. Subjects had to have either been vaccinated or willing to be vaccinated against encapsulated pathogens (meningococcus, haemophilus influenzae, and streptococcus pneumoniae) (at least 14 days prior to study drug administration). Subjects weighing more than 98 kg were excluded (for all subjects in all dose cohorts except the 100 mg / kg dose cohort in Part A, where the upper weight limit was 58 kg).

[0431] Study Design: In Part A, subjects were randomized to receive a single dose of BIVV009 (0.3, 1, 3, 10, 30, 60, or 100 mg / kg) or placebo in a 3:1 ratio (0.3 and Part B was administered intravenously over a period of approximately 60 minutes (1 mg / kg: n=4 / group; remaining: n=8 / group). The lowest dose of BIVV009 given was based on 1 / 300th of the no observed adverse effect level (NOAEL) in non-human primates (NHPs) that was predicted not to inhibit the classical pathway. In part B, 16 subjects (8 subjects / dose group) were given four repeat doses of BIVV009 (30 or 60 mg / kg) or placebo in a 3:1 ratio once a week, with an additional observation period of 2 weeks. Infusion of BIVV009 or placebo followed a stepwise dose escalation procedure. Part B was initiated after confirming the tolerability and safety of the maximum dose step in part A. In Part A, safety (adverse events, vital signs), pharmacokinetic (PK) profile and pharmacodynamic (PD) response were monitored 1 hour before initiation of infusion, and 0.5, 1, 4, 8 and 24 hours after initiation of infusion, and 2, 3, 4, 7 and 14 days after dosing. In Part B, safety, PK and PD were monitored at the following time points: 1 hour before initiation of the first infusion, and 0.5, 1, 4 and 8 hours after initiation of the first infusion, and daily for the next 4 days; 1 hour before initiation of the second and third infusions, and 4 hours after initiation of the second and third infusions; 1 hour before initiation of the last / fourth infusion, and 0.5, 1, 4 and 8 hours after initiation of the last / fourth infusion, and daily for the next 4 days; and 1 and 2 weeks after the last / fourth infusion.

[0432] Pharmacokinetics (PK): Maximum concentration (C max ), half-life (t 1 / 2 ), the time to reach the maximum concentration (t max ), the area under the concentration-time curve (AUC) up to the last time point showing a concentration above the lower limit of quantification (AUC ∞ ), and the area under the concentration-time curve to the last time point showing a concentration above the lower limit of quantification (AUC last ) was determined from serum concentrations. Serum concentrations of BIVV009 were determined using a validated immunoassay by a GLP-certified laboratory (Vela Laboratories, Vienna, Austria).

[0433] Pharmacodynamics (PD): Activities of the classical complement pathway were measured semiquantitatively in serum as previously published by the use of a commercially available enzyme immunoassay (Classical Complement System Pathway WIESLAB; Euro Diagnostica AB, Malmo, Sweden) (Roos, A. and Wieslander, J., Methods Mol. Biol. 1100:11-23 (2014)).

[0434] Pharmacokinetics / Pharmacodynamics (PK / PD): First, the relationship between BIVV009 concentration and CP activity was investigated to assess possible delayed responses (ie, hysteresis). Based on preliminary analysis, the concentration-activity relationship of BIVV009 and CP activity was investigated using various PK / PD models. PK / PD modeling was performed using Phoenix NLME (V7).

[0435] Safety: Safety measures were assessed by adverse events, vital signs, physical examination, electrocardiogram, and laboratory tests. The severity of adverse events was graded using the National Cancer Institute's Common Terminology Criteria for Adverse Events (CTCAE, v4.03). Laboratory tests were determined in accredited routine laboratories and consisted of hematology, blood chemistry and coagulation tests, urinalysis, and immunoassays for systemic lupus erythematosus (SLE)-associated autoantibodies.

[0436] Immunogenicity: BIVV009 antibodies (anti-drug antibodies [ADA]) were analyzed using a validated immunoassay by a GLP-certified laboratory (Vela Laboratories, Vienna, Austria) in a two-step approach (screening assay followed by confirmatory assay and determination of absolute A...

Claims

1. 1. A pharmaceutical composition for use in treating a complement-mediated disease or disorder in a subject in need thereof, the pharmaceutical composition comprising an anti-C1s antibody comprising light chain complementarity-determining regions (CDRs) of an antibody light chain variable region comprising the amino acid sequence of SEQ ID NO: 16 and heavy chain CDRs of an antibody heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 21, wherein an effective amount is about 4 grams to about 10 g, about 5 grams to about 10 g, about 6 grams to about 10 g, about 7 grams to about 10 g, about 8 grams to about 10 g, or about 9 grams to about 10 g of the anti-C1s antibody.

2. 2. The pharmaceutical composition for use of claim 1, wherein the effective amount of the antibody is between about 4 g and about 10 g, between about 5 g and about 9 g, between about 5 g and about 8 g, between about 6 g and about 8 g, between about 5.5 g and about 8.5 g, between about 6 g and about 8 g, between about 6.5 g and about 8 g, between about 6.5 g and about 7.5 g, between about 6 g and about 7.5 g, between about 7 g and about 8 g, or between about 7 g and about 7.5 g.

3. 3. The pharmaceutical composition for use according to claim 1 or 2, wherein the effective amount of the antibody is between about 6.5 g and about 7.5 g.

4. The pharmaceutical composition for use according to any one of claims 1 to 3, wherein the anti-C1s antibody increases the number of reticulocytes in the blood of the subject.

5. The pharmaceutical composition for use according to any one of claims 1 to 4, wherein the anti-C1s antibody increases the level of hemoglobin in the subject.

6. The pharmaceutical composition for use according to any one of claims 1 to 5, wherein the anti-C1s antibody reduces the percentage of C3d-positive red blood cells in the subject, for example in the blood.

7. The pharmaceutical composition for use according to any one of claims 1 to 6, wherein the anti-C1s antibody reduces the level of bilirubin in the subject, for example in the blood.

8. The pharmaceutical composition for use according to any one of claims 1 to 7, wherein the anti-C1s antibody comprises a light chain complementarity determining region 1 (CDR1) comprising the amino acid sequence of SEQ ID NO: 10, a light chain complementarity determining region 2 (CDR2) comprising the amino acid sequence of SEQ ID NO: 11, and a light chain complementarity determining region 3 (CDR3) comprising the amino acid sequence of SEQ ID NO: 3; and a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 12, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 13, and a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO:

14.

9. The pharmaceutical composition for use according to any one of claims 1 to 8, wherein the anti-C1s antibody comprises: a light chain variable region comprising the amino acid sequence of SEQ ID NO: 16; and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:

21.

10. The pharmaceutical composition for use according to any one of claims 1 to 9, wherein the anti-C1s antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO:23 and a heavy chain comprising the amino acid sequence of SEQ ID NO:

22.

11. The pharmaceutical composition for use according to any one of claims 1 to 10, wherein the effective amount is between 6.5 grams and 7.5 grams.

12. 12. The pharmaceutical composition for use according to claim 11, wherein the effective amount is 6.5 grams.

13. 13. The pharmaceutical composition for use according to claim 12, wherein the subject weighs less than 75 kilograms.

14. 12. The pharmaceutical composition for use according to claim 11, wherein the effective amount is 7.5 grams.

15. 15. The pharmaceutical composition for use according to claim 14, wherein the subject weighs 75 kilograms or more.

16. The pharmaceutical composition for use according to any one of claims 1 to 15, wherein the pharmaceutical composition further comprises sterile saline.

17. The pharmaceutical composition for use according to any one of claims 1 to 16, wherein the effective amount is administered every other week.

18. 18. The pharmaceutical composition for use according to any one of claims 1 to 17, wherein the effective amount is administered on days 0, 7, and then every 14 days from day 21 onwards.

19. The pharmaceutical composition for use according to any one of claims 1 to 18, wherein the effective amount is administered via subcutaneous administration, intravenous administration, or intramuscular administration.

20. The pharmaceutical composition for use according to any one of claims 1 to 19, wherein the effective amount is administered via intravenous infusion over a period of 1 hour.

21. Complement-mediated diseases or disorders include age-related macular degeneration, Alzheimer's disease, amyotrophic lateral sclerosis, anaphylaxis, argyrophilic grain dementia, arthritis (e.g., rheumatoid arthritis), asthma, atherosclerosis, atypical hemolytic uremic syndrome, autoimmune diseases, Barraquel-Simons syndrome, Behcet's disease, British amyloid angiopathy, bullous pemphigoid, Burger's disease, C1q nephropathy, chronic inflammatory demyelinating polyneuropathy, cancer, fulminant antiphospholipid syndrome, cerebral amyloid angiopathy, cold agglutinin disease (including primary cold agglutinin disease and secondary cold agglutinin disease), corticobasal degeneration, Creutzfeldt-Jakob disease, Crohn's disease, cryoglobulinemic vasculitis, dementia pugilistica, dementia with Lewy bodies (DLB), and diseases associated with calcification. Diffuse neurofibrillary tangle disease, discoid lupus erythematosus, Down's syndrome, focal segmental glomerulosclerosis, formal thought disorder, frontotemporal dementia (FTD), frontotemporal dementia with parkinsonism linked to chromosome 17, frontotemporal lobar degeneration, Gerstmann-Straussler-Scheinker disease, Guillain-Barré syndrome, Hallervorden-Spatz disease, hemolytic uremic syndrome, hereditary angioedema, hypophosphastasis, idiopathic pneumonia syndrome, immune complex disease, inclusion body myositis, infectious diseases (e.g., diseases caused by bacteria (e.g., meningococcus or streptococcus), diseases caused by viruses (e.g., human immunodeficiency virus (HIV)), or diseases caused by other infectious agents), inflammatory diseases, ischemia / reperfusion injury, mild cognitive impairment, immune thrombocytopenic purpura (ITP) P), molybdenum cofactor deficiency (MoCD) type A, membranoproliferative glomerulonephritis (MPGN) type I, membranoproliferative glomerulonephritis (MPGN) type II (dense deposit disease), membranous nephritis, multi-infarct dementia, lupus (e.g., systemic lupus erythematosus (SLE)), glomerulonephritis, Kawasaki disease, mucous membrane pemphigoid, cicatricial pemphigoid, multifocal motor neuropathy, multiple sclerosis, multiple system atrophy, myasthenia gravis, myocardial infarction, myotonic dysplasia trophy, neuromyelitis optica, Niemann-Pick disease type C, non-Guam motor neuron disease with neurofibrillary tangles, Parkinson's disease, Parkinson's disease with dementia, paroxysmal nocturnal hemoglobinuria, pemphigus vulgaris, Pick's disease, postencephalitic parkinsonism, polymyositis, prion protein cerebral amyloid angiopathy, progressive subcortical gliosis, progressive supranuclear palsy, psoriasis, sepsis, Shiga toxin-producing Escherichia coli (Shiga-toxin E)21. The pharmaceutical composition for use according to any one of claims 1 to 20, wherein the disease is selected from: E. coli (STEC)-HuS, spinal muscular atrophy, stroke, subacute sclerosing panencephalitis, neurofibrillary tangle-predominant dementia, transplant rejection, vasculitis (e.g., ANCA-associated vasculitis), Wegener's granulomatosis, sickle cell disease, cryoglobulinemia, mixed cryoglobulinemia, idiopathic mixed cryoglobulinemia, type II mixed cryoglobulinemia, type III mixed cryoglobulinemia, nephritis, drug-induced thrombocytopenia, lupus nephritis, bullous pemphigoid, epidermolysis bullosa acquisita, delayed hemolytic transfusion reaction, hypocomplementemic urticarial vasculitis syndrome, pseudophakic bullous keratopathy, and platelet refractoriness.

22. 22. The pharmaceutical composition for use according to any one of claims 1 to 21, wherein the complement-mediated disease or disorder is selected from multifocal motor neuropathy (MMN), chronic inflammatory demyelinating polyneuropathy (CIDP), myasthenia gravis (MG), neuromyelitis optica (NMO), systemic lupus erythematosus (SLE), lupus nephritis (LN), membranoproliferative glomerulonephritis (MPGN).

23. 23. The pharmaceutical composition for use according to any one of claims 1 to 22, wherein the complement-mediated disease or disorder is selected from cold agglutinin disease, antibody-mediated renal transplant rejection, immune thrombocytopenia, bullous pemphigoid, multifocal motor neuropathy, chronic inflammatory demyelinating polyneuropathy, myasthenia gravis, neuromyelitis optica, systemic lupus erythematosus, lupus nephritis, and membranoproliferative glomerulonephritis.

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