WO2025155602A1 - Method of treating hemophilia a - Google Patents

Abstract

The present invention relates to methods of increasing hemoglobin in an individual suffering from severe hemophilia A by prophylactic administration of emicizumab. Pharmaceutical compositions comprising emicizumab are also provided.

Description

METHOD OF TREATING HEMOPHILIA A

CROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of and priority to U.S. Provisional Application No. 63/621,559, filed January 16, 2024, the content of which is hereby incorporated herein by reference in its entirety.

REFERENCE TO AN ELECTRONIC SEQUENCE LISTING

[0002] The contents of the electronic sequence listing (146392068640SEQLIST.xml; Size: 15,421 bytes; and Date of Creation: January 9, 2025) is herein incorporated by reference in its entirety.

FIELD OF THE INVENTION

[0003] The present invention relates to methods of increasing hemoglobin in an individual suffering from hemophilia A by prophylactic administration of emicizumab. Further provided herein are pharmaceutical compositions comprising emicizumab.

BACKGROUND OF THE INVENTION

[0004] Hemophilia A is a bleeding abnormality caused by a hereditary decrease or deficiency of blood coagulation factor VIII (FVIII) function. Hemophilia A patients are generally treated with FVIII replacement therapy, which include both purified plasma-derived FVIII proteins, and recombinant FVIII proteins. These FVIII formulation are administered to address the bleeding on-demand (e.g., episodic treatment). FVIII formulations are also additionally administered at regular intervals as necessary to prevent rebleeding. In recent years, FVIII formulations also have been administered prophylactically to prevent bleeding events (e.g., as preventive administration; see, e.g., Blood 58, 1-13 (1981) and Nature 312, 330-337 (1984), hereby both incorporated by reference in their entireties). The half-life of FVIII formulations in blood is approximately 12 to 16 hours. Therefore, for continuous prevention, FVIII formulations are administered to patients three times a week (see, e.g., Nature 312, 337-342 (1984) and Biochim. Biophys. Acta 871, 268-278 (1986), hereby both incorporated by reference in their entireties). [0005] Occasionally, anti-FVIII antibodies (i.e., inhibitors) develop in hemophilia patients, which counteract the effects of the FVIII formulations. Treatments for patients who have developed inhibitors (inhibitor patients) include bypass formulations that are not dependent on FVIII function, which involves catalyzing the activation of blood coagulation factor X (FX) by activated blood coagulation factor IX (FIXa). However, in some cases, bypass formulations cannot sufficiently stop patient bleeding (see, e.g., Kempton & Meeks (2014), Blood 124(23): 3365-3372, hereby incorporated by reference in its entirety).

[0006] Recently, antibodies that functionally substitute for FVIII and their use were disclosed (see, e.g., Blood 58, 1-13 (1981); Nature 312, 330-337 (1984); and Nature 312, 337-342 (1984)). The antibodies may be effective for acquired hemophilia in which anti-FVIII autoantibodies are present.

BRIEF SUMMARY OF THE INVENTION

[0007] The present invention provides a method of increasing the hemoglobin level in an individual diagnosed with hemophilia A comprising administering a bispecific antibody (e.g., emicizumab or a biosimilar thereof) that binds to blood coagulation factor IX and/or activated blood coagulation factor IX, and binds to blood coagulation factor X in a treatment regimen to an individual diagnosed with hemophilia A, and increasing the hemoglobin level in the individual as compared to baseline. The present invention further provides a method of treating hemophilia A comprising administering the bispecific antibody and increasing the hemoglobin level in the individual as compared to baseline.

[0008] In one aspect of the present invention, there is provided a method comprising: (a) administering a bispecific antibody that binds to blood coagulation factor IX and/or activated blood coagulation factor IX, and binds to blood coagulation factor X in a treatment regimen to an individual diagnosed with hemophilia A, wherein the bispecific antibody comprises (i) a first antibody H chain comprising a variable region comprising complementarity determining regions (CDRs) 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 3, 4 and 5, respectively; (ii) a second antibody H chain comprising a variable region comprising CDRs 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 8, 9, and 10, respectively; and (iii) identical first and second antibody L chains, each L chain comprising a variable region comprising CDRs 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 13, 14 and 15, respectively; and (b) increasing the hemoglobin level in the individual as compared to a baseline level. In some embodiments, the antibody is emicizumab. [0009] In another aspect of the present invention, there is provided a method of treating hemophilia A comprising: (a) administering a bispecific antibody that binds to blood coagulation factor IX and/or activated blood coagulation factor IX, and binds to blood coagulation factor X, to an individual diagnosed with hemophilia A, wherein the bispecific antibody comprises (i) a first antibody H chain comprising a variable region comprising complementarity determining regions (CDRs) 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 3, 4 and 5, respectively; (ii) a second antibody H chain comprising a variable region comprising CDRs 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 8, 9, and 10, respectively; and (iii) identical first and second antibody L chains, each L chain comprising a variable region comprising CDRs 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 13, 14 and 15, respectively; and (b) wherein the hemoglobin level in the individual is increased as compared to a baseline level. In some embodiments, the antibody is emicizumab. [0010] In another aspect of the present invention, there is provided a method of increasing the hemoglobin level in an individual diagnosed with hemophilia A, comprising administering a bispecific antibody that binds to blood coagulation factor IX and/or activated blood coagulation factor IX, and binds to blood coagulation factor X, to an individual diagnosed with hemophilia A, wherein the bispecific antibody comprises (i) a first antibody H chain comprising a variable region comprising complementarity determining regions (CDRs) 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 3, 4 and 5, respectively; (ii) a second antibody H chain comprising a variable region comprising CDRs 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 8, 9, and 10, respectively; and (iii) identical first and second antibody L chains, each L chain comprising a variable region comprising CDRs 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 13, 14 and 15, respectively; and wherein the hemoglobin level in the individual is increased as compared to a baseline level. In some embodiments, the antibody is emicizumab.

[0011] In another aspect of the present invention, there is provided a use of a bispecific antibody for the manufacture of a medicament for the treatment of hemophilia A in an individual, wherein the bispecific antibody binds to a) blood coagulation factor IX and/or activated blood coagulation factor IX, and b) blood coagulation factor X, wherein the bispecific antibody comprises (i) a first antibody H chain comprising a variable region comprising complementarity determining regions (CDRs) 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 3, 4 and 5, respectively; (ii) a second antibody H chain comprising a variable region comprising CDRs 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 8, 9, and 10, respectively; and (iii) identical first and second antibody L chains, each L chain comprising a variable region comprising CDRs 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 13, 14 and 15, respectively; and wherein the treatment increases the hemoglobin level in the individual as compared to a baseline level.

[0012] In another aspect of the present invention, there is provided a use of a bispecific antibody for the manufacture of a medicament for increasing the hemoglobin level in an individual diagnosed with hemophilia A, wherein the bispecific antibody binds to a) blood coagulation factor IX and/or activated blood coagulation factor IX, and b) blood coagulation factor X, wherein the bispecific antibody comprises (i) a first antibody H chain comprising a variable region comprising complementarity determining regions (CDRs) 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 3, 4 and 5, respectively; (ii) a second antibody H chain comprising a variable region comprising CDRs 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 8, 9, and 10, respectively; and (iii) identical first and second antibody L chains, each L chain comprising a variable region comprising CDRs 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 13, 14 and 15, respectively, wherein the medicament increases the hemoglobin level in the individual as compared to a baseline level.

[0013] In another aspect of the present invention, there is provided a bispecific antibody for use in the treatment of hemophilia A in an individual, wherein the bispecific antibody binds to a) blood coagulation factor IX and/or activated blood coagulation factor IX, and b) blood coagulation factor X, wherein the bispecific antibody comprises (i) a first antibody H chain comprising a variable region comprising complementarity determining regions (CDRs) 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 3, 4 and 5, respectively; (ii) a second antibody H chain comprising a variable region comprising CDRs 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 8, 9, and 10, respectively; and (iii) identical first and second antibody L chains, each L chain comprising a variable region comprising CDRs 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 13, 14 and 15, respectively; and wherein the treatment increases the hemoglobin level in the individual as compared to a baseline level.

[0014] In another aspect of the present invention, there is provided a bispecific antibody for use in increasing the hemoglobin level in an individual diagnosed with hemophilia A, wherein the bispecific antibody binds to a) blood coagulation factor IX and/or activated blood coagulation factor IX, and b) blood coagulation factor X, wherein the bispecific antibody comprises (i) a first antibody H chain comprising a variable region comprising complementarity determining regions (CDRs) 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 3, 4 and 5, respectively; (ii) a second antibody H chain comprising a variable region comprising CDRs 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 8, 9, and 10, respectively; and (iii) identical first and second antibody L chains, each L chain comprising a variable region comprising CDRs 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 13, 14 and 15, respectively, wherein the use increases the hemoglobin level in the individual as compared to a baseline level.

[0015] In some embodiments according to any of the methods, uses, or bispecific antibodies for use described above, the first antibody heavy (H) chain variable region comprises the amino acid sequence set forth in SEQ ID NO:2. In some embodiments, the second antibody H chain variable region comprises the amino acid sequence set forth in SEQ ID NO:7. In some embodiments, the identical first and second antibody light (L) chain variable regions each comprises the amino acid sequences set forth in SEQ ID NO: 12. In some embodiments, the amino acid sequence of the first antibody full-length H chain is set forth in SEQ ID NO:1. In some embodiments, the amino acid sequence of the second antibody full-length H chain is set forth in SEQ ID NO: 6. In some embodiments, the amino acid sequence of the identical first and second antibody full-length L chains each is set forth in SEQ ID NO:11. In some embodiments, the antibody is emicizumab. [0016] In some embodiments according to any of the methods described above, the bispecific antibody is administered at an initial dose of 3 mg/kg every week for one or more weeks. In some embodiments, the bispecific antibody is administered at an initial dose of 3 mg/kg every week for four weeks.

[0017] In some embodiments according to any of the methods, uses, or bispecific antibodies for use described above, the bispecific antibody is administered at a maintenance dose of 1.5 mg/kg every week for at least 12 weeks, at least 24 weeks or at least 52 weeks. In some embodiments, the bispecific antibody is administered at a maintenance dose of 3 mg/kg every two weeks for at least 12 weeks, at least 24 weeks or at least 52 weeks. In some embodiments, the bispecific antibody is administered at a maintenance dose of 6 mg/kg every 4 weeks for at least 12 weeks, at least 24 weeks or at least 52 weeks.

[0018] In some embodiments according to any of the methods, uses, or bispecific antibodies for use described above, the treatment is an episodic treatment or a prophylactic treatment. In some embodiments, the treatment comprises both an episodic treatment and a prophylactic treatment. [0019] In some embodiments according to any of the methods described above, the individual has Factor VIII inhibitors in the bloodstream. In some embodiments, the individual does not have Factor VIII inhibitors in the bloodstream. In some embodiments, the individual is at least 12 years of age.

[0020] In some embodiments according to any of the methods, uses, or bispecific antibodies for use described above, the baseline level of hemoglobin is between 80 and 100 g/L. In some embodiments, the baseline level of hemoglobin is between 100 and 120 g/L. In some embodiments, the baseline level of hemoglobin is between 120 and 140 g/L. In some embodiments, the baseline level of hemoglobin is between 140 and 220 g/L.

[0021] In some embodiments according to any of the methods, uses, or bispecific antibodies for use described above, the hemoglobin level increases by 5 g/L, 10 g/L or 15 g/L as compared to a baseline level. In some embodiments, the hemoglobin level increases by 20 g/L, 25 g/L or 30 g/L as compared to a baseline level. In some embodiments, the hemoglobin level increases by 5 -10 g/L, 10-15 g/L, 15-20 g/L, 20-25 g/L, or 25-30 g/L as compared to a baseline level.

[0022] In some embodiments according to any of the methods, uses, or bispecific antibodies for use described above, the hemoglobin level is 80-100 g/L, 100-120 g/L, or 120-140 g/L at least four weeks after administration of the initial dose. In some embodiments, the hemoglobin level is 100-120 g/L, 120-140 g/L or 140-160 g/L at least twelve weeks after administration of the initial dose. In some embodiments, the hemoglobin level is 100-120 g/L, 120-140 g/L or 140-160 g/L at least twenty -four weeks after administration of the initial dose.

[0023] In some embodiments according to any of the methods, uses, or bispecific antibodies for use described above, the hematocrit level is further measured in percentage. In some embodiments, erythrocyte mean corpuscular volume levels are further measured in 10¹²/L. In some embodiments, red blood cell count is further measured in percentage. In some embodiments, red cell distribution width levels are further measured in percentage.

BRIEF DESCRIPTION OF THE DRAWINGS

[0024] The drawings illustrate certain embodiments of the features of this disclosure. These embodiments are not intended to limit the scope of the claims in any manner.

[0025] FIGs. 1A-1D show schematic overviews of the HAVEN 1 (FIG. 1A), HAVEN 3 (FIG. IB), HAVEN 4 (FIG. 1C), and STASEY (FIG. ID) clinical trials.

[0026] FIG. 2 shows the mean change in hemoglobin levels from baseline across time up to 73- 78 weeks after first dose of emicizumab for pooled participants and baseline hemoglobin subpopulations of participants with severe hemophilia A. Subpopulations based on hemoglobin levels were grouped as follows: 80 g/L<HgB<100 g/L; 100 g/L< HgB< 120 g/L; 120 g/L< HgB< 140 g/L; and 140 g/L< HgB< 180 g/L.

DETAILED DESCRIPTION

[0027] Provided herein, in certain aspects, are methods of treating hemophilia A in an individual, wherein the treatment increases the hemoglobin level of the individual compared to baseline hemoglobin levels. The disclosure of the present application is based, at least in part, on the inventors’ unexpected discovery that the bispecific antibody described herein (i.e., emicizumab or a biosimilar thereof) shows unexpected improvement in hemoglobin levels over time, in particular for individuals with especially low baseline hemoglobin levels. Specifically, as reported in the Examples, the inventors found that the bispecific antibody significantly raised the hemoglobin levels in individuals with particularly low baseline hemoglobin levels. Emicizumab provides significant therapeutic value to patients with heavy bleeding that fails to coagulate normally. Such findings provided herein represent a significant advancement in the field.

[0028] Accordingly, in one aspect, the present invention relates to methods of treating hemophilia A, comprising administering a bispecific antibody (e.g., emicizumab or a biosimilar thereof) that binds to blood coagulation factor IX and/or activated blood coagulation factor IX, and binds to blood coagulation factor X, to an individual diagnosed with hemophilia A, wherein the hemoglobin level in the individual is increased as compared to a baseline level. [0029] In another aspect, the present invention provides a method of increasing the hemoglobin level in an individual diagnosed with hemophilia A, comprising administering a bispecific antibody (e.g., emicizumab or a biosimilar thereof) that binds to blood coagulation factor IX and/or activated blood coagulation factor IX, and binds to blood coagulation factor X, to an individual diagnosed with hemophilia A, wherein the hemoglobin level in the individual is increased as compared to a baseline level.

[0030] Also provided herein is a method comprising: administering a bispecific antibody (e.g., emicizumab or a biosimilar thereof) that binds to blood coagulation factor IX and/or activated blood coagulation factor IX, and binds to blood coagulation factor X in a treatment regimen to an individual diagnosed with hemophilia A, and increasing the hemoglobin level in the individual as compared to a baseline level.

I. Definitions

[0031] Before describing the invention in detail, it is to be understood that this invention is not limited to particular compositions or biological systems, which can, 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.

[0032] As used herein and in the appended claims, the singular forms “a,” “an,” and “the” include plural references unless indicated otherwise. For example, “a disorder” includes one or more disorders.

[0033] The phrase “comprising” as used herein is open-ended, indicating that such embodiments may include additional elements. In contrast, the phrase “consisting of’ is closed, indicating that such embodiments do not include additional elements (except for trace impurities). The phrase “consisting essentially of’ is partially closed, indicating that such embodiments may further comprise elements that do not materially change the basic characteristics of such embodiments. It is understood that aspects and embodiments of the invention described herein include “comprising,” “consisting,” and “consisting essentially of’ aspects and embodiments.

[0034] As used herein, a “baseline” level (such as baseline level for hemoglobin) in a human refers to the level before an administration of a drug (e.g., emicizumab antibody) described herein to the human.

[0035] As used herein, the term “prevention” includes providing prophylaxis with respect to occurrence or recurrence of a disorder (e.g., hemophilia A) in an individual (e.g., human subject). [0036] The term “pharmaceutical formulation” refers to a preparation which is in such form as to permit the biological activity of the active ingredient to be effective, and which contains no additional components which are unacceptably toxic to a subject to which the formulation would be administered. Such formulations are sterile. “Pharmaceutically acceptable” excipients (vehicles, additives) are those which can reasonably be administered to a subject mammal to provide an effective dose of the active ingredient employed.

[0037] As used herein, the term “treatment” refers to clinical intervention designed to alter the natural course of the individual or cell being treated during the course of clinical pathology. Desirable effects of treatment include decreasing the rate of disease progression, ameliorating or palliating the disease state, and remission or improved prognosis. For example, an individual is successfully “treated” if one or more symptoms associated with the disease (e.g., asthma) are mitigated or eliminated, including, but are not limited to, decreasing symptoms resulting from the disease, increasing the quality of life of those suffering from the disease, decreasing the dose of other medications required to treat the disease, and/or prolonging survival of individuals. [0038] An “effective amount” is at least the minimum amount required to effect a measurable improvement or prevention of a particular disorder. An effective amount herein may vary according to factors such as the disease state, age, sex, and weight of the patient, and the ability of the antibody to elicit a desired response in the individual. An effective amount is also one in which any toxic or detrimental effects of the treatment are outweighed by the therapeutically beneficial effects. For prophylactic use, beneficial or desired results include results such as eliminating or reducing the risk, lessening the severity, or delaying the onset of the disease, including biochemical, histological and/or behavioral symptoms of the disease, its complications and intermediate pathological phenotypes presenting during development of the disease. For therapeutic use, beneficial or desired results include clinical results such as decreasing one or more symptoms resulting from the disease, increasing the quality of life of those suffering from the disease, decreasing the dose of other medications required to treat the disease, enhancing effect of another medication such as via targeting, delaying the progression of the disease, and/or prolonging survival. An effective amount can be administered in one or more administrations. For purposes of this invention, an effective amount of drug, compound, or pharmaceutical composition is an amount sufficient to accomplish prophylactic or therapeutic treatment either directly or indirectly. As is understood in the clinical context, an effective amount of a drug, compound, or pharmaceutical composition may or may not be achieved in conjunction with another drug, compound, or pharmaceutical composition. Thus, an “effective amount” may be considered in the context of administering one or more therapeutic agents, and a single agent may be considered to be given in an effective amount if, in conjunction with one or more other agents, a desirable result may be or is achieved.

[0039] As used herein, “in conjunction with” or “in combination with” refers to administration of one treatment modality in addition to another treatment modality. As such, “in conjunction with” or “in combination with” refers to administration of one treatment modality before, during, or after administration of the other treatment modality to the individual.

[0040] A “disorder” is any condition that would benefit from treatment including, but not limited to, chronic and acute disorders or diseases including those pathological conditions which predispose the mammal to the disorder in question.

[0041] A “subject”, “patient” or an “individual” for purposes of treatment refers to any animal classified as a mammal, including humans, domestic and farm animals, and zoo, sports, or pet animals, such as dogs, horses, cats, cows, etc. Preferably, the mammal is human.

[0042] The term “antibody” herein is used in the broadest sense and specifically covers monoclonal antibodies (including full length monoclonal antibodies), polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments so long as they exhibit the desired biological activity.

[0043] An “isolated” antibody is one which has been identified and separated and/or recovered from a component of its natural environment. Contaminant components of its natural environment are materials which would interfere with research, diagnostic or therapeutic uses for the antibody, and may include enzymes, hormones, and other proteinaceous or nonproteinaceous solutes. In some embodiments, an antibody is purified (1) to greater than 95% by weight of antibody as determined by, for example, the Lowry method, and in some embodiments, to greater than 99% by weight; (2) to a degree sufficient to obtain at least 15 residues of N-terminal or internal amino acid sequence by use of, for example, a spinning cup sequenator, or (3) to homogeneity by SDS-PAGE under reducing or nonreducing conditions using, for example, Coomassie blue or silver stain. Isolated antibody includes the antibody in situ within recombinant cells since at least one component of the antibody’s natural environment will not be present. Ordinarily, however, isolated antibody will be prepared by at least one purification step. [0044] ‘ ‘Native antibodies” refer to naturally occurring immunoglobulin molecules with varying structures. For example, native IgG antibodies are heterotetrameric glycoproteins of about 150,000 daltons, composed of two identical light chains and two identical heavy chains that are disulfide-bonded. From N- to C-terminus, each heavy chain has a variable domain (VH), also called a variable heavy domain or a heavy chain variable region, followed by three constant heavy domains (CHI, CH2, and CH3). Similarly, from N- to C-terminus, each light chain has a variable domain (VL), also called a variable light domain or a light chain variable region, followed by a constant light (CL) domain.

[0045] The term “constant domain” refers to the portion of an immunoglobulin molecule having a more conserved amino acid sequence relative to the other portion of the immunoglobulin, the variable domain, which contains the antigen binding site. The constant domain contains the CHI, CH2 and CH3 domains (collectively, CH) of the heavy chain and the CL domain of the light chain.

[0046] The “variable region” or “variable domain” of an antibody refers to the amino-terminal domains of the heavy or light chain of the antibody. The variable domain of the heavy chain may be referred to as “VH.” The variable domain of the light chain may be referred to as “VL.” These domains are generally the most variable parts of an antibody and contain the antigen-binding sites.

[0047] The term “variable” refers to the fact that certain portions of the variable domains differ extensively in sequence among antibodies and are used in the binding and specificity of each particular antibody for its particular antigen. However, the variability is not evenly distributed throughout the variable domains of antibodies. It is concentrated in three segments called hypervariable regions (HVRs) both in the light-chain and the heavy -chain variable domains. The more highly conserved portions of variable domains are called the framework regions (FR). The variable domains of native heavy and light chains each comprise four FR regions, largely adopting a beta-sheet configuration, connected by three HVRs, which form loops connecting, and in some cases forming part of, the beta-sheet structure. The HVRs in each chain are held together in close proximity by the FR regions and, with the HVRs from the other chain, contribute to the formation of the antigen -bin ding site of antibodies (see Kabat et al., Sequences of Proteins of Immunological Interest, Fifth Edition, National Institute of Health, Bethesda, Md. (1991)). The constant domains are not involved directly in the binding of an antibody to an antigen, but exhibit various effector functions, such as participation of the antibody in antibodydependent cellular toxicity.

[0048] The “light chains” of antibodies (immunoglobulins) from any mammalian species can be assigned to one of two clearly distinct types, called kappa (“K”) and lambda (“ ”), based on the amino acid sequences of their constant domains.

[0049] In the present invention, the term “H chain” refers to a heavy chain of an antibody and the term “L chain” refers to a light chain of an antibody.

[0050] In the present invention, the term “commonly shared L chain” refers to an L chain that can link with two or more different H chains, and show binding ability to each antigen. Herein, the term “different H chain(s)” preferably refers to H chains of antibodies against different antigens, but is not limited thereto, and also refers to H chains whose amino acid sequences are different from each other. Commonly shared L chain can be obtained, for example, according to the method described in WO 2006/109592.

[0051] Regarding the L chains, since diversity of L chain variable regions is lower than that of H chain variable regions, commonly shared L chains that can confer binding ability to both H chains may be obtained. The antibodies of the present invention may be different, but preferably have commonly shared L chains.

[0052] The term IgG “isotype” or “subclass” as used herein is meant any of the subclasses of immunoglobulins defined by the chemical and antigenic characteristics of their constant regions. [0053] Depending on the amino acid sequences of the constant domains of their heavy chains, antibodies (immunoglobulins) can be assigned to different classes. There are five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, and several of these may be further divided into subclasses (isotypes), e.g., IgGl, IgG2, IgG3, IgG4, IgAl, and IgA2. The heavy chain constant domains that correspond to the different classes of immunoglobulins are called a, y, E, y, and p, respectively. The subunit structures and three-dimensional configurations of different classes of immunoglobulins are well known and described generally in, for example, Abbas et al. Cellular and Mol. Immunology, 4^th ed. (W.B. Saunders, Co., 2000). An antibody may be part of a larger fusion molecule, formed by covalent or non-covalent association of the antibody with one or more other proteins or peptides.

[0054] The terms “full length antibody,” “intact antibody” and “whole antibody” are used herein interchangeably to refer to an antibody in its substantially intact form, not antibody fragments as defined below. The terms particularly refer to an antibody with heavy chains that contain an Fc region.

[0055] Furthermore, antibodies of the present invention may include not only whole antibodies but also antibody fragments and low-molecular-weight antibodies (minibodies), and modified antibodies.

[0056] “Antibody fragments” comprise a portion of an intact antibody, preferably comprising the antigen binding region thereof. In some embodiments, the antibody fragment described herein is an antigen-binding fragment. Examples of antibody fragments include Fab, Fab', F(ab')2, and Fv fragments; diabodies; linear antibodies; single-chain antibody molecules; and multispecific antibodies formed from antibody fragments.

[0057] Papain digestion of antibodies produces two identical antigen-binding fragments, called “Fab” fragments, each with a single antigen-binding site, and a residual “Fc” fragment, whose name reflects its ability to crystallize readily. Pepsin or IdeS treatment yields an F(ab')2 fragment that has two antigen-combining sites and is still capable of cross-linking antigen.

[0058] ‘ ‘Fv” is the minimum antibody fragment which contains a complete antigen-binding site. In one embodiment, a two-chain Fv species consists of a dimer of one heavy- and one light-chain variable domain in tight, non-covalent association. In a single-chain Fv (scFv) species, one heavy- and one light-chain variable domain can be covalently linked by a flexible peptide linker such that the light and heavy chains can associate in a “dimeric” structure analogous to that in a two-chain Fv species. It is in this configuration that the three HVRs of each variable domain interact to define an antigen-binding site on the surface of the VH-VL dimer. Collectively, the six HVRs confer antigen-binding specificity to the antibody. However, even a single variable domain (or half of an Fv comprising only three HVRs specific for an antigen) has the ability to recognize and bind antigen, although at a lower affinity than the entire binding site.

[0059] The Fab fragment contains the heavy- and light-chain variable domains and also contains the constant domain of the light chain and the first constant domain (CHI) of the heavy chain. Fab' fragments differ from Fab fragments by the addition of a few residues at the carboxy terminus of the heavy chain CHI domain including one or more cysteines from the antibody hinge region. Fab’-SH is the designation herein for Fab' in which the cysteine residue(s) of the constant domains bear a free thiol group. F(ab')2 antibody fragments originally were produced as pairs of Fab' fragments which have hinge cysteines between them. Other chemical couplings of antibody fragments are also known.

[0060] “Single-chain Fv” or “scFv” antibody fragments comprise the VH and VL domains of antibody, wherein these domains are present in a single polypeptide chain. Generally, the scFv polypeptide further comprises a polypeptide linker between the VH and VL domains which enables the scFv to form the desired structure for antigen binding. For a review of scFv, see, e.g. , Pluckthiin, in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds., (Springer- Verlag, New York, 1994), pp. 269-315.

[0061] The term “diabodies” refers to antibody fragments with two antigen-binding sites, which fragments comprise a heavy-chain variable domain (VH) connected to a light-chain variable domain (VL) in the same polypeptide chain (VH-VL). 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 the complementary domains of another chain and create two antigen-binding sites. Diabodies may be bivalent or bispecific. Diabodies are described more fully in, for example, EP 404,097; WO 1993/01161; Hudson et al., Nat. Med. 9:129-134 (2003); and Hollinger et al., Proc. Natl. Acad. Sci. USA 90: 6444-6448 (1993). Triabodies and tetrabodies are also described in Hudson et al., Nat. Med. 9: 129-134 (2003).

[0062] The term “monoclonal antibody” as used herein refers to an antibody obtained from a population of substantially homogeneous antibodies, e.g., the individual antibodies comprising the population are identical except for possible mutations, e.g., naturally occurring mutations, that may be present in minor amounts. Thus, the modifier “monoclonal” indicates the character of the antibody as not being a mixture of discrete antibodies. In certain embodiments, such a monoclonal antibody typically includes an antibody comprising a polypeptide sequence that binds a target, wherein the target-binding polypeptide sequence was obtained by a process that includes the selection of a single target binding polypeptide sequence from a plurality of polypeptide sequences. For example, the selection process can be the selection of a unique clone from a plurality of clones, such as a pool of hybridoma clones, phage clones, or recombinant DNA clones. It should be understood that a selected target binding sequence can be further altered, for example, to improve affinity for the target, to humanize the target binding sequence, to improve its production in cell culture, to reduce its immunogenicity in vivo, to create a multispecific antibody, etc., and that an antibody comprising the altered target binding sequence is also a monoclonal antibody of this invention. In contrast to polyclonal antibody preparations, which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody of a monoclonal antibody preparation is directed against a single determinant on an antigen. In addition to their specificity, monoclonal antibody preparations are advantageous in that they are typically uncontaminated by other immunoglobulins.

[0063] The modifier “monoclonal” indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies, and is not to be construed as requiring production of the antibody by any particular method. For example, the monoclonal antibodies to be used in accordance with the invention may be made by a variety of techniques, including, for example, the hybridoma method (e.g., Kohler and Milstein, Nature, 256:495-97 (1975); Hongo et al., Hybridoma, 14 (3): 253-260 (1995), Harlow et al., Antibodies: A Laboratory Manual, (Cold Spring Harbor Laboratory Press, 2^nd ed. 1988); Hammerling et al., in: Monoclonal Antibodies and T-Cell Hybridomas 563-681 (Elsevier, N.Y., 1981)), recombinant DNA methods (see, e.g., U.S. Pat. No. 4,816,567), phage-display technologies (see, e.g., Clackson et al., Nature, 352: 624-628 (1991); Marks et al., J. Mol. Biol. 222: 581-597 (1992); Sidhu et al., J. Mol. Biol. 338(2): 299-310 (2004); Lee et al., J. Mol. Biol. 340(5): 1073-1093 (2004); Fellouse, Proc. Natl. Acad. Sci. USA 101(34): 12467-12472 (2004); and Lee et al., J. Immunol. Methods 284(1-2): 119-132 (2004), and technologies for producing human or humanlike antibodies in animals that have parts or all of the human immunoglobulin loci or genes encoding human immunoglobulin sequences (see, e.g., WO 1998/24893; WO 1996/34096; WO 1996/33735; WO 1991/10741; Jakobovits et al., Proc. Natl. Acad. Sci. USA 90: 2551 (1993); Jakobovits et al., Nature 362: 255-258 (1993); Bruggemann et al., Year in Immunol. 7:33 (1993); U.S. Pat. Nos. 5,545,807; 5,545,806; 5,569,825; 5,625,126; 5,633,425; and 5,661,016; Marks et al., Bio/Technology 10: 779-783 (1992); Lonberg et al., Nature 368: 856-859 (1994); Morrison, Nature 368: 812-813 (1994); Fishwild et al., Nature Biotechnol. 14: 845-851 (1996); Neuberger, Nature Biotechnol. 14: 826 (1996); and Lonberg and Huszar, Intern. Rev. Immunol. 13: 65-93 (1995).

[0064] The monoclonal antibodies herein specifically include “chimeric” antibodies in which a portion of the heavy and/or light chain is identical with or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical with or homologous to corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit the desired biological activity (see, e.g., U.S. Pat. No. 4,816,567; and Morrison et al., Proc. Natl. Acad. Sci. USA 81 :6851-6855 (1984)). Chimeric antibodies include PRIMATTZED® antibodies wherein the antigen-binding region of the antibody is derived from an antibody produced by, e.g., immunizing macaque monkeys with the antigen of interest.

[0065] ‘ ‘Humanized” forms of non-human (e.g., murine) antibodies are chimeric antibodies that contain minimal sequence derived from non-human immunoglobulin. In one embodiment, a humanized antibody is a human immunoglobulin (recipient antibody) in which residues from a HVR of the recipient are replaced by residues from a HVR of a non-human species (donor antibody) such as mouse, rat, rabbit, or nonhuman primate having the desired specificity, affinity, and/or capacity. In some instances, FR residues of the human immunoglobulin are replaced by corresponding non-human residues. Furthermore, humanized antibodies may comprise residues that are not found in the recipient antibody or in the donor antibody. These modifications may be made to further refine antibody performance. In general, a humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the hypervariable loops correspond to those of a non-human immunoglobulin, and all or substantially all of the FRs are those of a human immunoglobulin sequence. The humanized antibody optionally will also comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. For further details, see, e.g., Jones et al., Nature 321 :522-525 (1986); Riechmann et al., Nature 332:323-329 (1988); and Presta, Curr. Op. Struct. Biol. 2:593-596 (1992). See also, e.g., Vaswani and Hamilton, Ann. Allergy, Asthma & Immunol. 1 :105-115 (1998); Harris, Biochem. Soc. Transactions 23: 1035-1038 (1995); Hurle and Gross, Curr. Op. Biotech. 5:428-433 (1994); and U.S. Pat. Nos. 6,982,321 and 7,087,409.

[0066] A “human antibody” is one which possesses an amino acid sequence which corresponds to that of an antibody produced by a human and/or has been made using any of the techniques for making human antibodies as disclosed herein. This definition of a human antibody specifically excludes a humanized antibody comprising non-human antigen-binding residues. Human antibodies can be produced using various techniques known in the art, including phage-display libraries. Hoogenboom and Winter, J. Mol. Biol., 227:381 (1991); Marks et al., J. Mol. Biol., 222:581 (1991). Also available for the preparation of human monoclonal antibodies are methods described in Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p. 77 (1985); Boemer et al., J. Immunol., 147(l):86-95 (1991). See also van Dijk and van de Winkel, Curr. Opin. Pharmacol., 5: 368-74 (2001). Human antibodies can be prepared by administering the antigen to a transgenic animal that has been modified to produce such antibodies in response to antigenic challenge, but whose endogenous loci have been disabled, e.g., immunized xenomice (see, e.g., U.S. Pat. Nos. 6,075,181 and 6,150,584 regarding XENOMOUSETM technology). See also, for example, Li et al., Proc. Natl. Acad. Sci. USA, 103:3557-3562 (2006) regarding human antibodies generated via a human B-cell hybridoma technology.

[0067] A “species-dependent antibody” is one which has a stronger binding affinity for an antigen from a first mammalian species than it has for a homologue of that antigen from a second mammalian species. Normally, the species-dependent antibody “binds specifically” to a human antigen (e.g., has a binding affinity (Kd) value of no more than about 1 *10'⁷ M, preferably no more than about 1 * 1 O'⁸ M and preferably no more than about 1 * 1 O'⁹ M) but has a binding affinity for a homologue of the antigen from a second nonhuman mammalian species which is at least about 50 fold, or at least about 500 fold, or at least about 1000 fold, weaker than its binding affinity for the human antigen. The species-dependent antibody can be any of the various types of antibodies as defined above, but preferably is a humanized or human antibody.

[0068] IgG-type bispecific antibodies can be secreted from hybrid hybridomas (quadromas) produced by fusing two kinds of hybridomas that produce IgG antibodies (Milstein C et al. Nature 1983, 305: 537-540). They can also be secreted by taking the L chain and H chain genes constituting the two kinds of IgGs of interest, a total of four kinds of genes, and introducing them into cells to coexpress the genes. In this case, by introducing suitable amino acid substitutions to the CH3 regions of the H chains, IgGs having a heterogeneous combination of H chains can be preferentially secreted (Ridgway J B et al. Protein Engineering 1996, 9: 617-621; Merchant A M et al. Nature Biotechnology 1998, 16: 677-681; WO 2006/106905; Davis J H et al. Protein Eng Des Sei. 2010, 4: 195-202).

[0069] The term “hypervariable region,” “HVR,” or “HV,” when used herein refers to the regions of an antibody variable domain which are hypervariable in sequence and/or form structurally defined loops. Generally, antibodies comprise six HVRs; three in the VH (Hl, H2, H3), and three in the VL (LI, L2, L3). In native antibodies, H3 and L3 display the most diversity of the six HVRs, and H3 in particular is believed to play a unique role in conferring fine specificity to antibodies. See, e.g., Xu et al., Immunity 13:37-45 (2000); Johnson and Wu, in Methods in Molecular Biology 248:1-25 (Lo, ed., Human Press, Totowa, N.J., 2003). Indeed, naturally occurring camelid antibodies consisting of a heavy chain only are functional and stable in the absence of light chain. See, e.g., Hamers-Casterman et al., Nature 363:446-448 (1993); Sheriff et al., Nature Struct. Biol. 3:733-736 (1996).

[0070] A number of HVR delineations are in use and are encompassed herein. The Kabat Complementarity Determining Regions (CDRs) are based on sequence variability and are the most commonly used (Kabat et al., Sequences of Proteins of Immunological Interest, 5^th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991)). Chothia refers instead to the location of the structural loops (Chothia and Lesk J. Mol. Biol. 196:901-917 (1987)). The AbM HVRs represent a compromise between the Kabat HVRs and Chothia structural loops, and are used by Oxford Molecular’s AbM antibody modeling software. The

“contact” HVRs are based on an analysis of the available complex crystal structures. The residues from each of these HVRs are noted below.

Loop Kabat AbM Chothia Contact

LI L24-L34 L24-L34 L26-L32 L30-L36

L2 L50-L56 L50-L56 L50-L52 L46-L55

L3 L89-L97 L89-L97 L91-L96 L89-L96

Hl H31-H35B H26-H35B H26-H32 H30-H35B (Kabat Numbering)

Hl H31-H35 H26-H35 H26-H32 H30-H35 (Chothia Numbering)

H2 H50-H65 H50-H58 H53-H55 H47-H58

H3 H95-H102 H95-H102 H96-H101 H93-H101

[0071] HVRs may comprise “extended HVRs” as follows: 24-36 or 24-34 (LI), 46-56 or 50-56 (L2) and 89-97 or 89-96 (L3) in the VL and 26-35 (Hl), 50-65 or 49-65 (H2) and 93-102, 94- 102, or 95-102 (H3) in the VH. The variable domain residues are numbered according to Kabat etal., supra, for each of these definitions.

[0072] HVRs may comprise “extended HVRs” as follows: 24-36 or 24-34 (LI), 46-56 or 50-56 (L2) and 89-97 or 89-96 (L3) in the VL and 26-35 (Hl), 50-65 or 49-65 (H2) and 93-102, 94- 102, or 95-102 (H3) in the VH. The variable domain residues are numbered according to Kabat etal., supra, for each of these definitions. [0073] “Framework” or “FR” residues are those variable domain residues other than the HVR residues as herein defined.

[0074] The term “variable domain residue numbering as in Kabat’ ’ or “amino acid position numbering as in Kabat,” and variations thereof, refers to the numbering system used for heavy chain variable domains or light chain variable domains of the compilation of antibodies in Kabat etal., supra. Using this numbering system, the actual linear amino acid sequence may contain fewer or additional amino acids corresponding to a shortening of, or insertion into, a FR or HVR of the variable domain. For example, a heavy chain variable domain may include a single amino acid insert (residue 52a according to Kabat) after residue 52 of H2 and inserted residues (e.g., residues 82a, 82b, and 82c, etc. according to Kabat) after heavy chain FR residue 82. The Kabat numbering of residues may be determined for a given antibody by alignment at regions of homology of the sequence of the antibody with a “standard” Kabat numbered sequence.

[0075] The Kabat numbering system is generally used when referring to a residue in the variable domain (approximately residues 1-107 of the light chain and residues 1-113 of the heavy chain) (e.g. , Kabat et al., Sequences of Immunological Interest. 5 ^th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991)). The “EU numbering system” or “EU index” is generally used when referring to a residue in an immunoglobulin heavy chain constant region (e.g., the EU index reported in Kabat etal., supra,' Edelman etal., (1969), Proc Natl Acad Sci USA 63:78-85). The “EU index as in Kabat” refers to the residue numbering of the human IgGl EU antibody.

[0076] As use herein, the term “binds”, “specifically binds to” or is “specific for” refers to measurable and reproducible interactions such as binding between a target and an antibody, which is determinative of the presence of the target in the presence of a heterogeneous population of molecules including biological molecules. For example, an antibody that binds to or specifically binds to a target (which can be an epitope) is an antibody that binds this target with greater affinity, avidity, more readily, and/or with greater duration than it binds to other targets. In one embodiment, the extent of binding of an antibody to an unrelated target is less than about 10% of the binding of the antibody to the target as measured, e.g., by a radioimmunoassay (RIA). In certain embodiments, an antibody that specifically binds to a target has a dissociation constant (Kd) of < IpM, < 100 nM, < 10 nM, < 1 nM, or < 0.1 nM. In certain embodiments, an antibody specifically binds to an epitope on a protein that is conserved among the protein from different species. In another embodiment, specific binding can include, but does not require exclusive binding.

[0077] A “functional Fc region” possesses an “effector function” of a native sequence Fc region. Exemplary “effector functions” include Clq binding; CDC; Fc receptor binding; ADCC; phagocytosis; down regulation of cell surface receptors (e.g., B cell receptor; BCR), etc. Such effector functions generally require the Fc region to be combined with a binding domain (e.g., an antibody variable domain) and can be assessed using various assays as disclosed, for example, in definitions herein.

[0078] As used herein, “Percent (%) amino acid sequence identity” and “homology” with respect to a peptide, polypeptide or antibody sequence are defined as the percentage of amino acid residues in a candidate sequence that are identical with the amino acid residues in the specific peptide or polypeptide sequence, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity, and not considering any conservative substitutions as part of the sequence identity. Alignment for purposes of determining percent amino acid sequence identity can be achieved in various ways that are within the skill in the art, for instance, using publicly available computer software such as BLAST, BLAST-2, ALIGN or MEGALIGNTM (DNASTAR) software. Those skilled in the art can determine appropriate parameters for measuring alignment, including any algorithms needed to achieve maximal alignment over the full length of the sequences being compared.

II. Bi-Specific Antibodies

[0079] The present invention relates to a multispecific antigen-binding molecule (i.e., bispecific antibody) that recognizes FIX and/or FIXa, and FX, and functionally substitutes for cofactor function of FVIII. The bispecific antibody comprises a first polypeptide comprising an antigenbinding site that recognizes FIX and/or activated FIX (FIXa) and a third polypeptide comprising an antigen-binding site that recognizes FIX and/or activated FIX (FIXa), as well as a second polypeptide comprising an antigen-binding site that recognizes FX and a fourth polypeptide comprising an antigen-binding site that recognizes FX. The first polypeptide and the third polypeptide, and the second polypeptide and the fourth polypeptide each comprises an antigenbinding site of an antibody H (heavy) chain and an antigen-binding site of an antibody L (light) chain. For example, in a bispecific antibody of the present invention, the first polypeptide and the third polypeptide comprise an antigen-binding site of an H (heavy) chain and of a L (light) chain of an antibody against FIX or FIXa, respectively; and the second polypeptide and the fourth polypeptide comprise an antigen-binding site of an H chain and L chain of an antibody against FX, respectively. In some embodiments, the antigen-binding sites of the antibody L chain comprised in the first polypeptide and the third polypeptide, and the second polypeptide and the fourth polypeptide may be commonly shared L chains.

[0080] The multi-specific antigen-binding molecules of the present invention, preferably bispecific antibodies, are antibodies having specificity to two or more different antigens, or molecules comprising fragments of such antibodies. The antibodies of the present invention are not particularly limited, but are preferably monoclonal antibodies. Monoclonal antibodies used in the present invention include not only monoclonal antibodies derived from animals such as humans, mice, rats, hamsters, rabbits, sheep, camels, and monkeys, but also include artificially modified gene recombinant antibodies such as chimeric antibodies, humanized antibodies, and bispecific antibodies.

[0081] Antibodies of the present invention are preferably recombinant antibodies produced using genetic recombination techniques (See, for example, Borrebaeck C A K and Larrick J W, THERAPEUTIC MONOCLONAL ANTIBODIES, Published in the United Kingdom by MACMILLAN PUBLISHERS LTD, 1990). Recombinant antibodies can be obtained by cloning DNAs encoding antibodies from hybridomas or antibody -producing cells, such as sensitized lymphocytes, that produce antibodies, inserting them into suitable vectors, and then introducing them into hosts (host cells) to produce the antibodies.

[0082] In some embodiments, the bispecific antibody is emicizumab or a biosimilar thereof. Emicizumab (also known as RO5534262 and ACE910) is a recombinant, humanized, bispecific, immunoglobulin G4 (IgG4) monoclonal antibody that binds with moderate affinity to activated factor IX (FIXa) and factor X (FX), thereby mimicking the co-factor function of factor VIII (FVIII). In patients with hemophilia A, hemostasis can be restored irrespective of the presence of FVIII inhibitors, as emicizumab shares no sequence homology with FVIII. In some embodiments, the first polypeptide of the bispecific antibody of the present invention comprises a heavy (H) chain variable region (VH1) comprising complementarity determining regions (CDRs) 1, 2, and 3, wherein the CDR1 comprises the amino acid sequence of SEQ ID NO:3; the CDR2 comprises the amino acid sequence of SEQ ID NO:4; and the CDR3 comprises the amino acid sequence of SEQ ID NO: 5. In some embodiments, the second polypeptide of the bispecific antibody of the present invention comprises a heavy chain variable region (VH2) comprising complementarity determining regions (CDRs) 1, 2, and 3, wherein the CDR1 comprises the amino acid sequence of SEQ ID NO:8; the CDR2 comprises the amino acid sequence of SEQ ID NO:9; and the CDR3 comprises the amino acid sequence of SEQ ID NO: 10. In some embodiments, the third and/or fourth polypeptide of the bispecific antibody of the present invention comprises a light chain variable region (VL) comprising complementarity determining regions (CDRs) 1, 2, and 3, wherein the CDR1 comprises the amino acid sequence of SEQ ID NO: 13; the CDR2 comprises the amino acid sequence of SEQ ID NO: 14; and the CDR3 comprises the amino acid sequence of SEQ ID NO: 15. In some embodiments, the first polypeptide H chain variable region comprises the amino acid sequence set forth in SEQ ID NO:2. In some embodiments, the second polypeptide H chain variable region comprises the amino acid sequence set forth in SEQ ID NO:7. In some embodiments, the third and fourth polypeptide L chains comprise identical amino acid sequences. In some embodiments, the third and fourth polypeptide L chains comprise identical L chain variable regions each comprising the amino acid sequences set forth in SEQ ID NO: 12. In some embodiments, the amino acid sequence of the first polypeptide full-length H chain is set forth in SEQ ID NO:1. In some embodiments, the amino acid sequence of the second polypeptide full- length H chain is set forth in SEQ ID NO:6. In other embodiments, the amino acid sequence of each of the identical third polypeptide full-length L chain and fourth polypeptide full-length L chain is set forth in SEQ ID NO: 11. In some embodiments, the bispecific antibody comprises a first polypeptide comprising the amino acid sequence of SEQ ID NO:1, a second polypeptide comprising the amino acid sequence of SEQ ID NO:6, and a third and fourth identical polypeptides each comprising the amino acid sequence of SEQ ID NO: 11. Emicizumab is further described in WO2012/067176 and US 9,334,331, which are incorporated by reference herein in their entirety.

III. Pharmaceutical Compositions

[0083] Also provided herein are pharmaceutical compositions and formulations, e.g., for the treatment of a condition (e.g., hemophilia A) as described herein. In some embodiments, the pharmaceutical compositions and formulations further comprise a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical compositions and formulations comprise an anti-FIXa*anti-FX antibody, such as emicizumab or a biosimilar thereof, as described herein.

[0084] After preparation of the anti -FlXa* anti -FX antibody, such as emicizumab or a biosimilar thereof, the pharmaceutical formulation comprising said antibody is prepared. The therapeutically effective amount of antibody present in the formulation is determined by, e.g., taking into account the desired dose volumes and mode(s) of administration. In some embodiments, the anti -FlXa* anti -FX antibody is formulated at concentrations from about 25mg/mL to about 400 mg/mL, for example, from about 25 mg/mL to about 400 mg/mL, or from about 50 mg/mL to about 350 mg/mL, or from about 35 mg/mL to about 300 mg/mL, or from about 40 mg/mL to about 250 mg/mL, or from about 45 mg/mL to about 200 mg/mL, or from about 25 mg/mL to about 150 mg/mL, or from about 30 mg/mL to about 140 mg/mL, or from about 35 mg/mL to about 130 mg/mL, or from about 40 mg/mL to about 120 mg/mL, or from about 50 mg/mL to about 130 mg/mL, or from about 50 mg/mL to about 125 mg/mL, or from about 50 mg/mL to about 120 mg/mL, or from about 50 mg/mL to about 110 mg/mL, or from about 50 mg/mL to about 100 mg/mL, or from about 50 mg/mL to about 90 mg/mL, or from about 50 mg/mL to about 80 mg/mL, or from about 54 mg/mL to about 66 mg/mL. In some embodiments, the anti -FlXa* anti -FX antibody is formulated at concentrations of about any of 25mg/mL, 30mg/mL, 35mg/mL, 40mg/mL, 45mg/mL, 50mg/mL, 75mg/mL, lOOmg/mL, 125mg/mL, 150mg/mL, 175mg/mL, 200mg/mL, 225mg/mL, 250mg/mL, 275mg/mL, 300mg/mL, 325mg/mL, 350mg/mL, 375mg/mL, or 400mg/mL.

[0085] Pharmaceutical compositions used for therapeutic or preventive purposes, which comprise antibodies of the present invention as active ingredients, can be formulated by mixing, if necessary, with suitable pharmaceutically acceptable carriers, vehicles, and such that are inactive against the antibodies. For example, sterilized water, physiological saline, stabilizers, excipients, antioxidants (such as ascorbic acid), buffers (such as phosphate, citrate, histidine, and other organic acids), antiseptics, surfactants (such as PEG and Tween), chelating agents (such as EDTA), and binders may be used. They may also comprise other low-molecular-weight polypeptides, proteins such as serum albumin, gelatin, and immunoglobulins, amino acids such as glycine, glutamine, asparagine, glutamic acid, aspargic acid, methionine, arginine, and lysine, sugars and carbohydrates such as polysaccharides and monosaccharides, and sugar alcohols such as mannitol and sorbitol. When preparing an aqueous solution for injection, physiological saline and isotonic solutions comprising glucose and other adjuvants such as D-sorbitol, D-mannose, D-mannitol, and sodium chloride may be used, and if necessary, in combination with appropriate solubilizers such as alcohol (for example, ethanol), polyalcohols (such as propylene glycol and PEG), and nonionic surfactants (such as polysorbate 80, polysorbate 20, poloxamer 188, and HCO-50). By mixing hyaluronidase into the formulation, a larger fluid volume can be administered subcutaneously (Expert Opin Drug Deliv. 2007 July; 4(4): 427-40).

[0086] Exemplary lyophilized antibody formulations are described in US Patent No. 6,267,958. Aqueous antibody formulations include those described in US Patent No. 6,171,586 and W02006/044908, the latter formulations including a histidine-acetate buffer. E.g. , Strickley & Lambert (2021), J Pharm Sci 110:2590-2608 e2556, hereby incorporated by reference in its entirety, for further description of appropriate antibody formulations.

[0087] The composition and formulation herein may also contain more than one active ingredient as necessary for the particular indication being treated, preferably those with complementary activities that do not adversely affect each other. Such active ingredients are suitably present in combination in amounts that are effective for the purpose intended.

[0088] Active ingredients may be entrapped in microcapsules prepared, for example, by coacervation techniques or by interfacial polymerization, for example, hydroxymethylcellulose or gelatin-microcapsules and poly-(methylmethacylate) microcapsules, respectively, in colloidal drug delivery systems (for example, liposomes, albumin microspheres, microemulsions, nanoparticles and nanocapsules) or in macroemulsions. Such techniques are disclosed in Remington ’s Pharmaceutical Sciences 16^th edition, Osol, A. Ed. (1980).

[0089] Sustained-release preparations may be prepared. Suitable examples of sustained-release preparations include semipermeable matrices of solid hydrophobic polymers containing the antibody, which matrices are in the form of shaped articles, e.g., films, or microcapsules. The formulations to be used for in vivo administration are generally sterile. Sterility may be readily accomplished, e.g., by filtration through sterile filtration membranes. Methods for preparing the pharmaceutical agents as sustained-release pharmaceutical agents are also well known, and such methods may be applied to the antibodies of the present invention (Langer et al., J. Biomed.

Mater. Res. 15: 267-277 (1981); Langer, Chemtech. 12: 98-105 (1982); U.S. Pat. No. 3,773,919; European Patent Application Publication No. EP 58,481; Sidman et al., Biopolymers 22: 547- 556 (1983); EP 133,988).

IV. Method of Treating Hemophilia A

[0090] Provided herein are methods of treating hemophilia A, wherein an individual (e.g., a human) diagnosed with hemophilia A is administered a bispecific antibody (e.g., emicizumab or biosimilar thereof) that binds to blood coagulation factor IX and/or activated blood coagulation factor IX, and binds to blood coagulation factor X, to an individual diagnosed with hemophilia A, wherein the bispecific antibody comprises: (i) a first antibody H chain comprising a variable region comprising complementarity determining regions (CDRs) 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 3, 4 and 5, respectively; (ii) a second antibody H chain comprising a variable region comprising CDRs 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 8, 9, and 10, respectively; and (iii) identical first and second antibody L chains, each L chain comprising a variable region comprising CDRs 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 13, 14 and 15, respectively; and wherein the hemoglobin level in the individual is increased as compared to a baseline level. In some embodiments, the first polypeptide H chain variable region comprises the amino acid sequence set forth in SEQ ID NO:2. In some embodiments, the second polypeptide H chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 7. In some embodiments, the third and fourth polypeptide L chains comprise identical sequences. In some embodiments, the third and fourth polypeptide L chains comprise identical L chain variable regions each comprising the amino acid sequences set forth in SEQ ID NO: 12. In some embodiments, the amino acid sequence of the first antibody full-length H chain is set forth in SEQ ID NO:1. In some embodiments, the amino acid sequence of the second antibody full- length H chain is set forth in SEQ ID NO:6. In other embodiments, the amino acid sequence of each of the identical third polypeptide full-length L chain and fourth polypeptide full-length L chain is set forth in SEQ ID NO:11. In some embodiments, the bispecific antibody comprises a first polypeptide comprising the amino acid sequence of SEQ ID NO:1, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 6, and third and fourth polypeptides each comprising the amino acid sequence of SEQ ID NO: 11.

[0091] In some embodiments, the hemophilia A is severe. In some embodiments, the hemophilia A is moderate. In some embodiments, the hemophilia A is mild.

[0092] Also provided herein are methods of increasing the hemoglobin level in an individual (e.g., a human) diagnosed with hemophilia A, comprising administering a bispecific antibody (e.g., emicizumab or biosimilar thereof) that binds to blood coagulation factor IX and/or activated blood coagulation factor IX, and binds to blood coagulation factor X, to an individual diagnosed with hemophilia A, wherein the bispecific antibody comprises: (i) a first antibody H chain comprising a variable region comprising complementarity determining regions (CDRs) 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 3, 4 and 5, respectively; (ii) a second antibody H chain comprising a variable region comprising CDRs 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 8, 9, and 10, respectively; and (iii) identical first and second antibody L chains, each L chain comprising a variable region comprising CDRs 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 13, 14 and 15, respectively; and wherein the hemoglobin level in the individual is increased as compared to a baseline level. In some embodiments, the first polypeptide H chain variable region comprises the amino acid sequence set forth in SEQ ID NO:2. In some embodiments, the second polypeptide H chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 7. In some embodiments, the third and fourth polypeptide L chains comprise identical sequences. In some embodiments, the identical first and second antibody L chain variable regions each comprises the amino acid sequences set forth in SEQ ID NO: 12. In some embodiments, the amino acid sequence of the first antibody full-length H chain is set forth in SEQ ID NO: 1. In some embodiments, the amino acid sequence of the second antibody full-length H chain is set forth in SEQ ID NO: 6. the amino acid sequence of the identical first and second antibody full-length L chain each is set forth in SEQ ID NO: 11. In some embodiments, the bispecific antibody comprises a first polypeptide comprising the amino acid sequence of SEQ ID NO: 1, a second polypeptide comprising the amino acid sequence of SEQ ID NO:6, and a third and fourth polypeptide each comprising the amino acid sequence of SEQ ID NO: 11.

[0093] In some embodiments, the hemophilia A is severe. In some embodiments, the hemophilia A is moderate. In some embodiments, the hemophilia A is mild. In some embodiments, the human subject or patient is an adolescent or an adult.

[0094] In some embodiments, the treatment is an episodic treatment. In other embodiments, the treatment is a prophylactic treatment. In other embodiments, the treatment consists of both an episodic and a prophylactic treatment. In other embodiments, the treatment comprises both an episodic and a prophylactic treatment. In other embodiments, the treatment further comprises treatment with one or more therapeutics to treat hemophilia.

Patient Population

[0095] In some embodiments, the individual is a human. In some embodiments, the individual is a human subject or patient. In some embodiments, the individual is an adolescent or an adult. In some embodiments, the individual has been, is, or will be diagnosed with hemophilia A. In some embodiments, the hemophilia A is severe. In some embodiments, the hemophilia A is moderate. In some embodiments, the hemophilia A is mild.

[0096] In some embodiments, the individual is 10 years, 11 years, 12 years, 13 years, 14 years, 15 years, 16 years, 17 years, 18 years, 19 years, 20 years, 25 years, 30 years, 35 years, 40 years, 45 years, 50 years, 55 years, 60 years, 65 years, 70 years, 75 years, 80 years, or more years of age. In other embodiments, the individual is between the ages of 12-15 years, 16-18 years, 19-21 years, 22-23 years, 24-26 years, 27-29 years, or older than 29 years of age. i. Hemoglobin Levels

[0097] Hemoglobin (Hb or hgb) is a protein in red blood cells that is responsible for delivery of oxygen to the tissues. Normal hemoglobin levels vary by age, and in general, are about 110-160 g/L for children; about 120-150 g/L for non-pregnant adult females; and about 135-180 g/L for adult males. Hemoglobin levels in patients with hemophilia A can be lower than normal.

[0098] In some embodiments, the individual displays a baseline hemoglobin level of less than about 250 g/L. For examples, in some embodiments, the individual displays a baseline hemoglobin level of less than about any of 240 g/L, 230 g/L, 220 g/L, 210 g/L, 200 g/L, 190 g/L, 180 g/L, 170 g/L, 160 g/L, 150 g/L, 140 g/L, 130 g/L, 120 g/L, 110 g/L, 100 g/L, 90 g/L, 80 g/L, 75 g/L, 70g/L, 65 g/L, 60 g/L, 55 g/L, 50 g/L, or less. In some embodiments, the individual displays a baseline hemoglobin level between about 50 g/L and about 250 g/L, for example about 50 g/L to about 70 g/L, about 60 g/L to about 80 g/L, about 70 g/L to about 90 g/L, about 80 g/L to about 100 g/L, about 90 g/L to about 110 g/L, about 100 g/L to about 120 g/L, about 110 g/L to about 130 g/L, about 120 g/L to about 140 g/L, about 130 g/L to about 150 g/L, about 140 g/L to about 160 g/L, about 150 g/L to about 170 g/L, about 160 g/L to about 180 g/L, about 170 g/L to about 190 g/L, about 180 g/L to about 200 g/L, about 190 g/L to about 210 g/L, about 200 g/L to about 220 g/L, about 210 g/L to about 230 g/L, about 220 g/L to about 240 g/L, about 230 g/L to about 250 g/L, about 50 g/L to about 85 g/L, about 75 g/L to about 105 g/L, about 85 g/L to about 125 g/L, about 115 g/L to about 175 g/L, about 140 g/L to about 220 g/L, about 150 g/L to about 250 g/L, about 200 g/L to about 235 g/L, or about 215 g/L to about 250 g/L.

[0099] In some embodiments, the hemoglobin level in an individual administered the bispecific antibody described herein increases by at least about any of 5 g/L, 10 g/L, 15 g/L, 20 g/L, 25 g/L, 30 g/L, 35 g/L, 40 g/L, 45 g/L, 50 g/L, or more as compared to a baseline hemoglobin level. In some embodiments, the hemoglobin level in an individual administered the bispecific antibody described herein increases by about 5 g/L to about 10 g/L, about 10 g/L to about 15 g/L, about 15 g/L to about 20 g/L, about 20 g/L to about 25 g/L, about 25 g/L to about 30 g/L, about 30 g/L to about 35 g/L, about 35 g/L to about 40 g/L, about 40 g/L to about 45 g/L, about 45 g/L to about 50 g/L or more as compared to a baseline hemoglobin level.

[0100] In some embodiments, the hemoglobin level is at least about 80 g/L after administration of the initial dose of the bispecific antibody described herein. In another embodiment, the hemoglobin level is at least about 80 g/L at least two weeks, at least four weeks, at least six weeks, at least eight weeks, or at least ten weeks after administration of the initial dose of the bispecific antibody described herein. For example, the hemoglobin level is about 80 g/L to about 100 g/L, about 90 g/L to about 110 g/L, about 100 g/L to about 120 g/L, about 110 g/L to about 130 g/L, about 120 g/L to about 140 g/L, about 130 g/L to about 150 g/L, about 140 g/L to about

160 g/L, about 150 g/L to about 170 g/L, about 160 g/L to about 180 g/L, about 170 g/L to about

190 g/L, about 180 g/L to about 200 g/L, about 190 g/L to about 210 g/L, about 200 g/L to about

220 g/L, about 210 g/L to about 230 g/L, about 220 g/L to about 240 g/L, about 230 g/L to about

250 g/L, about 75 g/L to about 105 g/L, about 85 g/L to about 125 g/L, about 115 g/L to about 175 g/L, about 140 g/L to about 220 g/L, about 150 g/L to about 250 g/L, about 200 g/L to about 235 g/L, or about 215 g/L to about 250 g/L at least two weeks, at least four weeks, at least six weeks, at least eight weeks, at least ten weeks, or at least twelve weeks after administration of the initial dose of the bispecific antibody described herein.

[0101] In some embodiments, the hemoglobin level is at least about 100 g/L after administration of the initial dose of the bispecific antibody described herein. In another embodiment, the hemoglobin level is at least about 100 g/L at least two weeks, at least four weeks, at least six weeks, at least eight weeks, at least ten weeks, at least twelve weeks, at least sixteen weeks, at least twenty weeks, at least twenty -four weeks, at least twenty-eight weeks, at least thirty weeks, at least forty weeks, at least fifty weeks, at least sixty weeks, or at least eighty weeks, after administration of the initial dose of the bispecific antibody described herein. For example, the hemoglobin level is about 100 g/L to about 120 g/L, about 110 g/L to about 130 g/L, about 120 g/L to about 140 g/L, about 130 g/L to about 150 g/L, about 140 g/L to about 160 g/L, about 150 g/L to about 170 g/L, about 160 g/L to about 180 g/L, about 170 g/L to about 190 g/L, about 180 g/L to about 200 g/L, about 190 g/L to about 210 g/L, about 200 g/L to about 220 g/L, about 210 g/L to about 230 g/L, about 220 g/L to about 240 g/L, about 230 g/L to about 250 g/L, about 115 g/L to about 175 g/L, about 140 g/L to about 220 g/L, about 150 g/L to about 250 g/L, about 200 g/L to about 235 g/L, or about 215 g/L to about 250 g/L at least two weeks, at least four weeks, at least six weeks, at least eight weeks, at least ten weeks, at least twelve weeks, at least sixteen weeks, at least twenty weeks, at least twenty -five weeks, at least thirty weeks, at least forty weeks, at least fifty weeks, at least sixty weeks, at least seventy weeks, or at least eighty weeks, after administration of the initial dose of the bispecific antibody described herein.

[0102] In some embodiments, the hemoglobin level is at least about 120 g/L after administration of the initial dose of the bispecific antibody described herein. In another embodiment, the hemoglobin level is at least about 120 g/L at least two weeks, at least four weeks, at least six weeks, at least eight weeks, at least ten weeks, at least twelve weeks, at least sixteen weeks, at least twenty weeks, at least twenty -four weeks, at least twenty-eight weeks, at least thirty weeks, at least forty weeks, at least fifty weeks, at least sixty weeks, or at least eighty weeks, after administration of the initial dose of the bispecific antibody described herein. For example, the hemoglobin level is about 100 g/L to about 120 g/L, about 110 g/L to about 130 g/L, about 120 g/L to about 140 g/L, about 130 g/L to about 150 g/L, about 140 g/L to about 160 g/L, about 150 g/L to about 170 g/L, about 160 g/L to about 180 g/L, about 170 g/L to about 190 g/L, about 180 g/L to about 200 g/L, about 190 g/L to about 210 g/L, about 200 g/L to about 220 g/L, about 210 g/L to about 230 g/L, about 220 g/L to about 240 g/L, about 230 g/L to about 250 g/L, about 115 g/L to about 175 g/L, about 140 g/L to about 220 g/L, about 150 g/L to about 250 g/L, about 200 g/L to about 235 g/L, or about 215 g/L to about 250 g/L at least two weeks, at least four weeks, at least six weeks, at least eight weeks, at least ten weeks, at least twelve weeks, at least sixteen weeks, at least twenty weeks, at least twenty -five weeks, at least thirty weeks, at least forty weeks, at least fifty weeks, at least sixty weeks, at least seventy weeks, or at least eighty weeks, after administration of the initial dose of the bispecific antibody described herein. ii. FVIII Inhibitors

[0103] The development of neutralizing antibodies (inhibitors) to Factor VIII (FVIII) or Factor IX (FIX) is the most significant complication of hemophilia treatment, occurring in up to 33% of patients with severe hemophilia A and in 13% of those with non-severe hemophilia A (see, e.g., Kempton & Meeks (2014), Blood 124(23): 3365-3372). The incidence and prevalence of inhibitors is higher in patients with severe hemophilia A. [0104] In some embodiments, the individual has Factor VIII inhibitors in the bloodstream. In some embodiments, the individual does not have Factor VIII inhibitors in the bloodstream. In some embodiments, the presence of Factor VIII inhibitors in the bloodstream does not or only minimally impacts the efficacy of the bispecific antibody (e.g., emicizumab or biosimilars thereof) described herein.

[0105] In some embodiments, individuals that have Factor VIII inhibitors in the bloodstream may have previously received or are currently receiving bypassing treatments. In some embodiments, bypassing agents may have previously been administered or are currently administered on an episodic bleeding basis or as a prophylaxis against future bleeding episodes. Bypassing agents treat bleeding by producing thrombin via pathways that do not require FVIII or FIX. Bypassing agents that are currently available include recombinant Factor Vila (rFVIIa, Novoseven RT; NovoNordisk) and activated prothrombin complex concentrates (aPCC, FEIBA VH; Baxter; see, e.g., Kempton & Meeks (2014), Blood 124(23): 3365-3372). iii. Additional Measures

[0106] Hematocrit is defined as the fractional volume of whole blood occupied by red blood cells (RBCs), expressed as a percentage. Normal hematocrit levels vary based on age, sex, and race. For example, in adult males, normal levels range from about 41%-50%. In adult females, normal levels range from about 36%-44%. Hematocrit levels that fall below the normal range can be indicative of various disorders, such as anemia. In some embodiments, the hematocrit levels at baseline are less than about 50%, for example less than about any of 45%, 40%, 35%, 30%, 29%, 28%, 27%, 26%, 25%, 24%, 23%, 22%, 21%, 20%, or less. In some embodiments, hematocrit level is further measured in percentage. In some embodiments, the hematocrit level in an individual administered the bispecific antibody described herein increases by about any of 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, or more compared to levels at baseline.

[0107] Mean corpuscular volume (MCV) measures the average size and volume of a red blood cell. It has utility in helping determine the etiology of anemia. The calculation of the MCV value is by multiplying the percent hematocrit by ten divided by the erythrocyte count. In some embodiments, erythrocyte mean corpuscular volume levels are further measured in 10¹²/L. In some embodiments, the erythrocyte MCV in an individual administered the bispecific antibody described herein increases by about any of 1 fL, 2 fL, 3 fL, 4 fL, 5 £L, 6 fL, 7 fL, 8 £L, 9 fL, 10 fL, 11 fL, 12 fL, 13 fL, 14 fL, 15 fL, or more compared to levels at baseline.

[0108] Red blood cells carry hemoglobin, which binds to oxygen and is responsible for oxygenating tissues throughout the body. For adult males, the normal range of red blood cells is generally about 4.35 to about 5.65 million red blood cells per microliter (mL) of blood. For adult females, the normal range of red blood cells is generally about 3.92 to about 5.13 million red blood cells per mL of blood. In some embodiments, red blood cell count (RBC) is further measured in percentage. In some embodiments, the red blood cell count in an individual administered the bispecific antibody described herein increases by about any of 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, or more compared to levels at baseline.

[0109] Red cell distribution width (ROW) tests the differences in the volume and size of red blood cells. A high ROW is indicative of an increase in variation in the size of red blood cells compared to normal. A high ROW may be indicative of anemia or a related condition. In some embodiments, red cell distribution width levels are further measured in percentage. In some embodiments, the red cell distribution width level in an individual administered the bispecific antibody described herein increases by about any of 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, or more compared to levels at baseline. iv. Other Treatments

[0110] In some embodiments, the treatment comprises further therapeutics to treat hemophilia A. Such therapeutics include, but are not limited to, FVIII replacement therapies, such as plasma-derived Factor VIII concentrates, or recombinant FVIII concentrates. Other therapeutics can include, for example, desmopressin acetate and epsilon amino caproic acid.

[OHl] Such therapeutics can be administered prior to, during or after treatment according to the methods described herein.

Patient Dosins

[0112] In some embodiments, the bispecific antibody (e.g., emicizumab or biosimilars thereof) described herein is administered as a prophylactic treatment. In some embodiments, the bispecific antibody is administered subcutaneously. In some embodiments, the bispecific antibody is administered intravenously. [0113] In some embodiments, the bispecific antibody is administered at an initial dose every week for at least about one or more weeks. In some embodiments, the initial dose is at or about 1 mg/kg, 1.5 mg/kg, 2 mg/kg, 2.5 mg/kg, 3 mg/kg, 3.5 mg/kg, 4 mg/kg, 5 mg/kg, 6 mg/kg, 7 mg/kg, 8 mg/kg, 9 mg/kg, 10 mg/kg, or more. In some embodiments, the bispecific antibody is administered at an initial dose of about 1.5 mg/kg every week for at least about one or more weeks. In other embodiments, the bispecific antibody is administered at an initial dose of about 3 mg/kg every week for at least about one or more weeks. In some embodiments, the bispecific antibody is administered at an initial dose of about 6 mg/kg every week for at least about one or more weeks. For example, the bispecific antibody is administered at an initial dose of about 1.5 mg/kg, 3 mg/kg, or 6 mg/kg every week for at least about one week, about two weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, or longer. In particular embodiments, the bispecific antibody is administered at an initial dose of about 3 mg/kg every week for about four weeks.

[0114] In some embodiments, the bispecific antibody is further administered as part of a maintenance regimen. Maintenance doses are administered to maintain a therapeutic drug concentration, i.e., a therapeutic concentration of the bispecific antibody described herein. In some embodiments, the maintenance dose is at or about 1 mg/kg, 1.5 mg/kg, 2 mg/kg, 2.5 mg/kg, 3 mg/kg, 3.5 mg/kg, 4 mg/kg, 5 mg/kg, 6 mg/kg, 7 mg/kg, 8 mg/kg, 9 mg/kg, 10 mg/kg, or more. Maintenance doses may be administered at regular intervals, for example at once a week, once every 2 weeks, once every 3 weeks, once every 4 weeks, once every 5 weeks, once every 6 weeks, once every 7 weeks, once every 8 weeks, or longer. In some embodiments, the maintenance dose is administered at regular intervals for at least about 12 weeks, including, for example, at least about any of 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 35 weeks, 40 weeks, 45 weeks, 50 weeks, 55 weeks, 60 weeks, 65 weeks, 70 weeks, 75 weeks, 80 weeks, 85 weeks, 90 weeks, 95 weeks, 100 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 13 months, 14 months, 15 months, 16 months, 18 months, 20 months, 22 months, 24 months, 26 months, 28 months, 1 year, 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, or longer. [0115] In some embodiments, the bispecific antibody is administered at a maintenance dose of 1.5 mg/kg every week for at least about 12 weeks, at least about 24 weeks, or at least about 52 weeks. In some embodiments, the bispecific antibody is administered at a maintenance dose of 3 mg/kg every two weeks for at least about 12 weeks, at least about 24 weeks, or at least about 52 weeks. In some embodiments, the bispecific antibody is administered at a maintenance dose of 6 mg/kg every 4 weeks for at least 12 weeks, at least 24 weeks or at least 52 weeks.

[0116] Provided with the description of methods of treatment are alternate formats thereof. For example, in some embodiments, provided is a bispecific antibody described herein, such as emicizumab or a biosimilar thereof, for use in a method of increasing the hemoglobin level in an individual diagnosed with hemophilia A. In some embodiments, provided herein is a bispecific antibody described herein, such as emicizumab or a biosimilar thereof, for use in the manufacture of a medicament for increasing the hemoglobin level in an individual diagnosed with hemophilia A.

V. Kits and Articles of Manufacture

[0117] The present application further provides articles of manufacture comprising the compositions (such as pharmaceutical compositions) described herein in suitable packaging. Suitable packaging for compositions (such as pharmaceutical compositions) described herein are known in the art, and include, for example, vials (such as sealed vials), vessels, ampules, bottles, jars, flexible packaging (e.g., sealed Mylar or plastic bags), and the like. These articles of manufacture may further be sterilized and/or sealed.

[0118] The present disclosure also provides kits for use in the methods described above. In some embodiments, the kits, comprise: i) a pharmaceutical composition comprising a bispecific antibody described herein, such as emicizumab or a biosimilar thereof; and ii) instructions for administration of the pharmaceutical composition by subcutaneous injection to a human subject suffering from a condition as described herein (e.g., hemophilia A). In some embodiments, the pharmaceutical composition is an aqueous solution further comprising L-arginine hydrochloride, L-histidine, and L-histidine hydrochloride monohydrate. In some embodiments, the pharmaceutical composition further comprises sucrose. In some embodiments, the pharmaceutical composition is present in a pre-filled syringe. In some embodiments, the pharmaceutical composition is a lyophilized powder present in a vial, and the kit further comprises instructions for reconstituting the lyophilized powder in water for injection. [0119] Other additives may be included such as stabilizers, buffers (e.g., a block buffer or lysis buffer), and the like. The relative amounts of the various reagents may be varied widely to provide for concentrations in solution of the reagents which substantially optimize the antibody formulation. Particularly, the reagents may be provided as dry powders, usually lyophilized, including excipients which on dissolution will provide a reagent solution having the appropriate concentration.

EXEMPLARY EMBODIMENTS

[0120] Embodiment 1. A method comprising:

(a) administering a bispecific antibody that binds to blood coagulation factor IX and/or activated blood coagulation factor IX, and binds to blood coagulation factor X in a treatment regimen to an individual diagnosed with hemophilia A, wherein the bispecific antibody comprises

(i) a first antibody H chain comprising a variable region comprising complementarity determining regions (CDRs) 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 3, 4 and 5, respectively;

(ii) a second antibody H chain comprising a variable region comprising CDRs 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 8, 9, and 10, respectively; and

(iii) identical first and second antibody L chains, each L chain comprising a variable region comprising CDRs 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 13, 14 and 15, respectively; and

(b) increasing the hemoglobin level in the individual as compared to a baseline level.

[0121] Embodiment 2, A method of treating hemophilia A comprising: administering a bispecific antibody that binds to blood coagulation factor IX and/or activated blood coagulation factor IX, and binds to blood coagulation factor X, to an individual diagnosed with hemophilia A, wherein the bispecific antibody comprises

(i) a first antibody H chain comprising a variable region comprising complementarity determining regions (CDRs) 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 3, 4 and 5, respectively; (ii) a second antibody H chain comprising a variable region comprising CDRs 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 8, 9, and 10, respectively; and

(iii) identical first and second antibody L chains, each L chain comprising a variable region comprising CDRs 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 13, 14 and 15, respectively; and wherein the hemoglobin level in the individual is increased as compared to a baseline level.

[0122] Embodiment 3, A method of increasing the hemoglobin level in an individual diagnosed with hemophilia A, comprising: administering a bispecific antibody that binds to blood coagulation factor IX and/or activated blood coagulation factor IX, and binds to blood coagulation factor X, to an individual diagnosed with hemophilia A, wherein the bispecific antibody comprises

(i) a first antibody H chain comprising a variable region comprising complementarity determining regions (CDRs) 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 3, 4 and 5, respectively;

(ii) a second antibody H chain comprising a variable region comprising CDRs 1,

2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 8, 9, and 10, respectively; and

(iii) identical first and second antibody L chains, each L chain comprising a variable region comprising CDRs 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 13, 14 and 15, respectively; and wherein the hemoglobin level in the individual is increased as compared to a baseline level.

[0123] Embodiment 4, The method of any one of embodiments 1-3, wherein the first antibody H chain variable region comprises the amino acid sequence set forth in SEQ ID NO:2.

[0124] Embodiment 5, The method of any one of embodiments 1-4, wherein the second antibody H chain variable region comprises the amino acid sequence set forth in SEQ ID NO:7. [0125] Embodiment 6, The method of any one of embodiments 1-5, wherein the identical first and second antibody L chain variable regions each comprises the amino acid sequences set forth in SEQ ID NO: 12.

[0126] Embodiment 7, The method of any one of embodiments 1-6, wherein the amino acid sequence of the first antibody full-length H chain is set forth in SEQ ID NO: 1.

[0127] Embodiment 8, The method of any one of embodiments 1-7, wherein the amino acid sequence of the second antibody full-length H chain is set forth in SEQ ID NO:6.

[0128] Embodiment 9, The method of any one of embodiments 1-8, wherein the amino acid sequence of the identical first and second antibody full-length L chains each is set forth in SEQ ID NOT E

[0129] Embodiment 10. The method of any one of embodiments 1-9, wherein the bispecific antibody is administered at an initial dose of 3 mg/kg every week for one or more weeks.

[0130] Embodiment 11. The method of embodiment 10, wherein the bispecific antibody is administered at an initial dose of 3 mg/kg every week for four weeks.

[0131] Embodiment 12, The method of any one of embodiments 1-11, wherein the bispecific antibody is administered at a maintenance dose of 1.5 mg/kg every week for at least 12 weeks, at least 24 weeks or at least 52 weeks.

[0132] Embodiment 13, The method of any one of embodiments 1-11, wherein the bispecific antibody is administered at a maintenance dose of 3 mg/kg every two weeks for at least 12 weeks, at least 24 weeks or at least 52 weeks.

[0133] Embodiment 14, The method of any one of embodiments 1-11, wherein the bispecific antibody is administered at a maintenance dose of 6 mg/kg every 4 weeks for at least 12 weeks, at least 24 weeks or at least 52 weeks.

[0134] Embodiment 15, The method of any one of embodiments 1-14, wherein the treatment is an episodic treatment or a prophylactic treatment. [0135] Embodiment 16, The method of any one of embodiments 1-14, wherein the treatment comprises both an episodic treatment and a prophylactic treatment.

[0136] Embodiment 17, The method of any one of embodiments 1-16, wherein the individual has Factor VIII inhibitors in the bloodstream.

[0137] Embodiment 18, The method of any one of embodiments 1-16, wherein the individual does not have Factor VIII inhibitors in the bloodstream.

[0138] Embodiment 19, The method of any one of embodiments 1-18, wherein the individual is at least 12 years of age.

[0139] Embodiment 20, The method of any one of embodiments 1-19, wherein the baseline level of hemoglobin is between 80 and 100 g/L.

[0140] Embodiment 21, The method of any one of embodiments 1-19, wherein the baseline level of hemoglobin is between 100 and 120 g/L.

[0141] Embodiment 22, The method of any one of embodiments 1-19, wherein the baseline level of hemoglobin is between 120 and 140 g/L.

[0142] Embodiment 23, The method of any one of embodiments 1-19, wherein the baseline level of hemoglobin is between 140 and 220 g/L.

[0143] Embodiment 24, The method of any one of embodiments 1-23, wherein the hemoglobin level increases by 5 g/L, 10 g/L or 15 g/L as compared to a baseline level.

[0144] Embodiment 25, The method of any one of embodiments 1-23, wherein the hemoglobin level increases by 20 g/L, 25 g/L or 30 g/L as compared to a baseline level.

[0145] Embodiment 26, The method of any one of embodiments 1-23, wherein the hemoglobin level increases by 5 -10 g/L, 10-15 g/L, 15-20 g/L, 20-25 g/L, or 25-30 g/L as compared to a baseline level. [0146] Embodiment 27, The method of any one of embodiments 1-26, wherein the hemoglobin level is 80-100 g/L, 100-120 g/L, or 120-140 g/L at least four weeks after administration of the initial dose.

[0147] Embodiment 28, The method of any one of embodiments 1-26, wherein the hemoglobin level is 100-120 g/L, 120-140 g/L or 140-160 g/L at least twelve weeks after administration of the initial dose.

[0148] Embodiment 29, The method of any one of embodiments 1-26, wherein the hemoglobin level is 100-120 g/L, 120-140 g/L or 140-160 g/L at least twenty-four weeks after administration of the initial dose.

[0149] Embodiment 30, The method of any one of embodiments 1-29, wherein the hematocrit level is further measured in percentage.

[0150] Embodiment 31 , The method of any one of embodiments 1-30, wherein erythrocyte mean corpuscular volume levels are further measured in 10¹²/L.

[0151] Embodiment 32, The method of any one of embodiments 1-31, wherein red blood cell count is further measured in percentage.

[0152] Embodiment 33, The method of any one of embodiments 1-32, wherein red cell distribution width levels are further measured in percentage.

[0153] Embodiment 34, The method of any one of embodiments 1-33, wherein the antibody is emicizumab.

[0154] Embodiment 35, Use of a bispecific antibody for the manufacture of a medicament for the treatment of hemophilia A in an individual, wherein the bispecific antibody binds to a) blood coagulation factor IX and/or activated blood coagulation factor IX, and b) blood coagulation factor X, wherein the bispecific antibody comprises

(i) a first antibody H chain comprising a variable region comprising complementarity determining regions (CDRs) 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 3, 4 and 5, respectively; (ii) a second antibody H chain comprising a variable region comprising CDRs 1,

2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 8, 9, and 10, respectively; and

(iii) identical first and second antibody L chains, each L chain comprising a variable region comprising CDRs 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 13, 14 and 15, respectively; and wherein the treatment increases the hemoglobin level in the individual as compared to a baseline level.

[0155] Embodiment 36, Use of a bispecific antibody for the manufacture of a medicament for increasing the hemoglobin level in an individual diagnosed with hemophilia A, wherein the bispecific antibody binds to a) blood coagulation factor IX and/or activated blood coagulation factor IX, and b) blood coagulation factor X, wherein the bispecific antibody comprises

(i) a first antibody H chain comprising a variable region comprising complementarity determining regions (CDRs) 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 3, 4 and 5, respectively;

(ii) a second antibody H chain comprising a variable region comprising CDRs 1,

2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 8, 9, and 10, respectively; and

(iii) identical first and second antibody L chains, each L chain comprising a variable region comprising CDRs 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 13, 14 and 15, respectively, wherein the medicament increases the hemoglobin level in the individual as compared to a baseline level.

[0156] Embodiment 37, A bispecific antibody for use in the treatment of hemophilia A in an individual, wherein the bispecific antibody binds to a) blood coagulation factor IX and/or activated blood coagulation factor IX, and b) blood coagulation factor X, wherein the bispecific antibody comprises (i) a first antibody H chain comprising a variable region comprising complementarity determining regions (CDRs) 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 3, 4 and 5, respectively;

(ii) a second antibody H chain comprising a variable region comprising CDRs 1,

2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 8, 9, and 10, respectively; and

(iii) identical first and second antibody L chains, each L chain comprising a variable region comprising CDRs 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 13, 14 and 15, respectively; and wherein the treatment increases the hemoglobin level in the individual as compared to a baseline level.

[0157] A bispecific antibody for use in increasing the hemoglobin level in an individual diagnosed with hemophilia A, wherein the bispecific antibody binds to a) blood coagulation factor IX and/or activated blood coagulation factor IX, and b) blood coagulation factor X, wherein the bispecific antibody comprises

(i) a first antibody H chain comprising a variable region comprising complementarity determining regions (CDRs) 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 3, 4 and 5, respectively;

(ii) a second antibody H chain comprising a variable region comprising CDRs 1,

2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 8, 9, and 10, respectively; and

(iii) identical first and second antibody L chains, each L chain comprising a variable region comprising CDRs 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 13, 14 and 15, respectively, wherein the use increases the hemoglobin level in the individual as compared to a baseline level.

EXAMPLES

[0158] The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the present invention and are not intended to limit the scope of what the inventors regard as their invention nor are they intended to represent that the experiments below are all or the only experiments performed. Efforts have been made to ensure accuracy with respect to numbers used (e.g., amounts, temperature, etc.), but some experimental errors and deviations should be accounted for.

Example 1: Emicizumab prophylaxis treatment of patients with hemophilia A led to improvement in hemoglobin levels.

[0159] This Example describes the assessment of the change from baseline in hemoglobin and associated red blood cell (RBC) indices in participants (>12 years) receiving emicizumab prophylaxis for severe hemophilia A.

Methods

[0160] A post-hoc analysis was performed on the data collected from the HAVEN 1 (NCT02622321), HAVEN 3 (NCT02847637), HAVEN 4 (NCT03020160), and STASEY (NCT03191799) clinical trials. In these trials, a total of N=501 adolescent and adult participants (>12 years) with severe hemophilia A initially received Emicizumab prophylaxis at 3 mg/kg/wk subcutaneously for 4 weeks before being randomized to 1.5mg/kg emicizumab per week, 3.0mg/kg emicizumab per two-week period, or 6.0mg/kg emicizumab per four-week period administered via subcutaneous injection as described in the clinical trial protocols. FIGs. 1A-1D provide schematic overviews of each trial. Clinical trial protocols can be found at: https://clinicaltrials.gov/study/NCT02622321 (HAVEN 1), https://clinicaltrials.gov/study/NCT02847637 (HAVEN 3), https://clinicaltrials.gov/study/NCT03020160 (HAVEN 4), and https://clinicaltrials.gov/study/NCT03191799 (STASEY), each of which is hereby incorporated by reference in their entireties.

[0161] In the current post-hoc analysis, biomarkers such as annualized bleed rates, hemoglobin (g/L), red blood cell counts (10¹²/L), hematocrit (fraction of 1), erythrocyte mean corpuscular volume (fL), and red blood cell distribution width (fraction of 1) were assessed as described in each protocol. Baseline levels of each biomarker were assessed prior to administration of the first dose of emicizumab or placebo. Experimental levels of each biomarker were collected at multiple time points over 73-78 weeks. Experimental biomarker levels were then compared to baseline to analyze change from baseline, as shown in Table 1 below. [0162] Hemoglobin change from baseline was evaluated 1) in the pooled population (N=497) and 2) in subgroups according to i. prior treatment regimen, ii. inhibitor status, and iii. baseline hemoglobin level. Baseline hemoglobin levels were stratified into the following subgroups:

• 80 g/L < hemoglobin < 100 g/L

• 100 g/L < hemoglobin < 120 g/L

• 120 g/L < hemoglobin < 140 g/L

• 140 g/L < hemoglobin < 180 g/L

[0163] Change from baseline in associated RBC indices was assessed.

Results

[0164] Improvement in mean hemoglobin was observed across the pooled population of patients with hemophilia A. In the pooled population, mean hemoglobin (standard deviation [SD], g/L) was 145.3 [15.8] at baseline and 151.3 [13.2] at Weeks 73-78 (Table 1). Mean hemoglobin increased from baseline to Weeks 73-78 for all subgroups (baseline hemoglobin level, prior treatment regimen, and baseline inhibitor status; Table 1). These improvements were continually observed over time, especially for those participants who displayed low baseline hemoglobin (FIG. 2).

[0165] The greatest improvement in hemoglobin was observed in the subgroups of patients with inhibitors and among patients with the lowest hemoglobin levels at baseline. The subgroup of patients with inhibitors included patients who were previously managed with bypass agents (i.e., aPCC and rFVIIa) prior to trial enrollment and treatment with emicizumab. Patients with inhibitors had lower baseline hemoglobin than those without inhibitors, and experienced a greater increase in hemoglobin (mean [SD], g/L) by Weeks 73-78 (8.1 [15.9] for inhibitor patients vs 2.9 [10.9] for noninhibitor patients).

[0166] Among the additional subgroups of patients, the mean hemoglobin increased from baseline to Weeks 73-78 for all subgroups (baseline hemoglobin level, prior treatment regimen, and baseline inhibitor status; Table 1). Table 1. Change in hemoglobin level and other laboratory measurements from baseline to Weeks 73-78 by participant subgroup.

Data for Red Blood Cell Count (10¹²/L) and Red Cell Distribution Width (fraction of 1) are sourced from the HAVEN 1 study only. CBU, chromogenic Bethesda unit; F, factor; HgB, hemoglobin; NE, not estimable; RBC, red blood cell; SD, standard deviation.

[0167] Additional laboratory measurements are presented in Table 1 above. Further analysis of participant subpopulations identified that annualized bleed rate (ABR; at Weeks 27-52) and hemoglobin (at Week 52) were negatively correlated (r=-0.48) among participants with low baseline hemoglobin (<120g/L). [0168] These data complement the previously observed reduction in ABR for participants with hemophilia A receiving emicizumab; all subgroups studied experienced hemoglobin increases. [0169] The present disclosure is not intended to be limited in scope to the particular disclosed embodiments, which are provided, for example, to illustrate various aspects of the disclosure. Various modifications to the compositions and methods described will become apparent from the description and teachings herein. Such variations may be practiced without departing from the true scope and spirit of the disclosure and are intended to fall within the scope of the present disclosure.

SEQUENCE TABLE

Claims

1. A method comprising:

(a) administering a bispecific antibody that binds to blood coagulation factor IX and/or activated blood coagulation factor IX, and binds to blood coagulation factor X in a treatment regimen to an individual diagnosed with hemophilia A, wherein the bispecific antibody comprises

(i) a first antibody H chain comprising a variable region comprising complementarity determining regions (CDRs) 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 3, 4 and 5, respectively;

(ii) a second antibody H chain comprising a variable region comprising CDRs 1,

2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 8, 9, and 10, respectively; and

(iii) identical first and second antibody L chains, each L chain comprising a variable region comprising CDRs 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 13, 14 and 15, respectively; and

(b) increasing the hemoglobin level in the individual as compared to a baseline level.

2. A method of treating hemophilia A comprising: administering a bispecific antibody that binds to blood coagulation factor IX and/or activated blood coagulation factor IX, and binds to blood coagulation factor X, to an individual diagnosed with hemophilia A, wherein the bispecific antibody comprises

(i) a first antibody H chain comprising a variable region comprising complementarity determining regions (CDRs) 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 3, 4 and 5, respectively;

(ii) a second antibody H chain comprising a variable region comprising CDRs 1,

2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 8, 9, and 10, respectively; and

(iii) identical first and second antibody L chains, each L chain comprising a variable region comprising CDRs 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 13, 14 and 15, respectively; and wherein the hemoglobin level in the individual is increased as compared to a baseline level.

3. A method of increasing the hemoglobin level in an individual diagnosed with hemophilia A, comprising administering a bispecific antibody that binds to blood coagulation factor IX and/or activated blood coagulation factor IX, and binds to blood coagulation factor X, to an individual diagnosed with hemophilia A, wherein the bispecific antibody comprises

(i) a first antibody H chain comprising a variable region comprising complementarity determining regions (CDRs) 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 3, 4 and 5, respectively;

(ii) a second antibody H chain comprising a variable region comprising CDRs 1,

2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 8, 9, and 10, respectively; and

(iii) identical first and second antibody L chains, each L chain comprising a variable region comprising CDRs 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 13, 14 and 15, respectively; and wherein the hemoglobin level in the individual is increased as compared to a baseline level.

4. The method of claim 1, 2 or 3, wherein the first antibody H chain variable region comprises the amino acid sequence set forth in SEQ ID NO:2.

5. The method of claim 1, 2, 3 or 4, wherein the second antibody H chain variable region comprises the amino acid sequence set forth in SEQ ID NO:7.

6. The method of any one of claims 1 to 5, wherein the identical first and second antibody L chain variable regions each comprises the amino acid sequences set forth in SEQ ID NO: 12.

7. The method of any one of claims 1 to 6, wherein the amino acid sequence of the first antibody full-length H chain is set forth in SEQ ID NO:1.

8. The method of any one of claims 1 to 7, wherein the amino acid sequence of the second antibody full-length H chain is set forth in SEQ ID NO:6.

9. The method of any one of claims 1 to 8, wherein the amino acid sequence of the identical first and second antibody full-length L chains each is set forth in SEQ ID NOT E

10. The method of any one of claims 1 to 9, wherein the bispecific antibody is administered at an initial dose of 3 mg/kg every week for one or more weeks.

11. The method of claim 10, wherein the bispecific antibody is administered at an initial dose of 3 mg/kg every week for four weeks.

12. The method of any one of claims 1 to 11, wherein the bispecific antibody is administered at a maintenance dose of 1.5 mg/kg every week for at least 12 weeks, at least 24 weeks or at least 52 weeks.

13. The method of any one of claims 1 to 11, wherein the bispecific antibody is administered at a maintenance dose of 3 mg/kg every two weeks for at least 12 weeks, at least 24 weeks or at least 52 weeks.

14. The method of any one of claims 1 to 11, wherein the bispecific antibody is administered at a maintenance dose of 6 mg/kg every 4 weeks for at least 12 weeks, at least 24 weeks or at least 52 weeks.

15. The method of any one of claims 1 to 14, wherein the treatment is an episodic treatment or a prophylactic treatment.

16. The method of any one of claims 1 to 14, wherein the treatment comprises both an episodic treatment and a prophylactic treatment.

17. The method of any one of claims 1 to 16, wherein the individual has Factor VIII inhibitors in the bloodstream.

18. The method of any one of claims 1 to 16, wherein the individual does not have Factor VIII inhibitors in the bloodstream.

19. The method of any one of claims 1 to 18, wherein the individual is at least 12 years of age.

20. The method of any one of claims 1 to 19, wherein the baseline level of hemoglobin is between 80 and 100 g/L.

21. The method of any one of claims 1 to 19, wherein the baseline level of hemoglobin is between 100 and 120 g/L.

22. The method of any one of claims 1 to 19, wherein the baseline level of hemoglobin is between 120 and 140 g/L.

23. The method of any one of claims 1 to 19, wherein the baseline level of hemoglobin is between 140 and 220 g/L.

24. The method of any one of claims 1 to 23, wherein the hemoglobin level increases by 5 g/L, 10 g/L or 15 g/L as compared to a baseline level.

25. The method of any one of claims 1 to 23, wherein the hemoglobin level increases by 20 g/L, 25 g/L or 30 g/L as compared to a baseline level.

26. The method of any one of claims 1 to 23, wherein the hemoglobin level increases by 5 - 10 g/L, 10-15 g/L, 15-20 g/L, 20-25 g/L, or 25-30 g/L as compared to a baseline level.

27. The method of any one of claims 1 to 26, wherein the hemoglobin level is 80-100 g/L, 100-120 g/L, or 120-140 g/L at least four weeks after administration of the initial dose.

28. The method of any one of claims 1 to 26, wherein the hemoglobin level is 100-120 g/L, 120-140 g/L or 140-160 g/L at least twelve weeks after administration of the initial dose.

29. The method of any one of claims 1 to 26, wherein the hemoglobin level is 100-120 g/L, 120-140 g/L or 140-160 g/L at least twenty-four weeks after administration of the initial dose.

30. The method of any one of claims 1 to 29, wherein the hematocrit level is further measured in percentage.

31. The method of any one of claims 1 to 30, wherein erythrocyte mean corpuscular volume levels are further measured in 10¹²/L.

32. The method of any one of claims 1 to 31, wherein red blood cell count is further measured in percentage.

33. The method of any one of claims 1 to 32, wherein red cell distribution width levels are further measured in percentage.

34. The method of any one of claims 1 to 33, wherein the antibody is emicizumab.

35. Use of a bispecific antibody for the manufacture of a medicament for the treatment of hemophilia A in an individual, wherein the bispecific antibody binds to a) blood coagulation factor IX and/or activated blood coagulation factor IX, and b) blood coagulation factor X, wherein the bispecific antibody comprises

(i) a first antibody H chain comprising a variable region comprising complementarity determining regions (CDRs) 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 3, 4 and 5, respectively;

(ii) a second antibody H chain comprising a variable region comprising CDRs 1,

2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 8, 9, and 10, respectively; and

(iii) identical first and second antibody L chains, each L chain comprising a variable region comprising CDRs 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 13, 14 and 15, respectively; and wherein the treatment increases the hemoglobin level in the individual as compared to a baseline level.

36. Use of a bispecific antibody for the manufacture of a medicament for increasing the hemoglobin level in an individual diagnosed with hemophilia A, wherein the bispecific antibody binds to a) blood coagulation factor IX and/or activated blood coagulation factor IX, and b) blood coagulation factor X, wherein the bispecific antibody comprises

(i) a first antibody H chain comprising a variable region comprising complementarity determining regions (CDRs) 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 3, 4 and 5, respectively; (ii) a second antibody H chain comprising a variable region comprising CDRs 1,

2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 8, 9, and 10, respectively; and

(iii) identical first and second antibody L chains, each L chain comprising a variable region comprising CDRs 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 13, 14 and 15, respectively, wherein the medicament increases the hemoglobin level in the individual as compared to a baseline level.

37. A bispecific antibody for use in the treatment of hemophilia A in an individual, wherein the bispecific antibody binds to a) blood coagulation factor IX and/or activated blood coagulation factor IX, and b) blood coagulation factor X, wherein the bispecific antibody comprises

(i) a first antibody H chain comprising a variable region comprising complementarity determining regions (CDRs) 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 3, 4 and 5, respectively;

(ii) a second antibody H chain comprising a variable region comprising CDRs 1,

2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 8, 9, and 10, respectively; and

(iii) identical first and second antibody L chains, each L chain comprising a variable region comprising CDRs 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 13, 14 and 15, respectively; and wherein the treatment increases the hemoglobin level in the individual as compared to a baseline level.

38. A bispecific antibody for use in increasing the hemoglobin level in an individual diagnosed with hemophilia A, wherein the bispecific antibody binds to a) blood coagulation factor IX and/or activated blood coagulation factor IX, and b) blood coagulation factor X, wherein the bispecific antibody comprises (i) a first antibody H chain comprising a variable region comprising complementarity determining regions (CDRs) 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 3, 4 and 5, respectively;

(ii) a second antibody H chain comprising a variable region comprising CDRs 1,

2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 8, 9, and 10, respectively; and

(iii) identical first and second antibody L chains, each L chain comprising a variable region comprising CDRs 1, 2, and 3 that comprise the amino acid sequences set forth in SEQ ID NOs: 13, 14 and 15, respectively, wherein the use increases the hemoglobin level in the individual as compared to a baseline level.