Plasma kallikrein inhibitors for treating childhood hereditary angioedema attacks and uses thereof

Abstract

To provide a novel regimen for treating or preventing hereditary angioedema (HAE) attacks using an antibody capable of binding to and inhibiting the active form of human plasma kallikrein (pKal).SOLUTION: Provided is a method for treating or preventing hereditary angioedema (HAE) attacks, comprising administering an antibody comprising the same complementarity-determining region (CDR) as DX-2930 during a first treatment period, the antibody being administered multiple times at approximately 150 mg every two weeks or every four weeks to a human subject during the first treatment period, and the human subject having, being suspected of having, or being at risk for HAE, and being a pediatric subject between the ages of 2 and 12.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] Related Applications This application claims the benefit under 35 U.S.C. §119(e) of U.S. Provisional Application No. 62 / 960,333, filed January 13, 2020, which is incorporated herein by reference in its entirety. [Background technology]

[0002] Plasma kallikrein, a serine protease component of the contact system, is a promising drug target for various inflammatory, cardiovascular, infectious (sepsis), and neoplastic diseases (Sainz IM et al., Thromb Haemost 98, 77-83, 2007). The contact system is activated by factor XIIa upon exposure to foreign or negatively charged surfaces or by prolylcarboxypeptidase on the surface of endothelial cells (Sainz IM et al., Thromb Haemost 98, 77-83, 2007). Activation of plasma kallikrein amplifies intrinsic coagulation through its feedback activation of factor XII and enhances inflammation through the production of the proinflammatory nonapeptide bradykinin. As the major kininogenase in the circulation, plasma kallikrein is critically involved in the production of bradykinin in the vascular system. A genetic deficiency in the C1 inhibitor protein (C1-INH), the major natural inhibitor of plasma kallikrein, causes hereditary angioedema (HAE). Patients with HAE suffer from acute attacks of painful edema, often caused by unknown triggers (Zuraw BLet et al., N Engl J Med 359, 1027-1036, 2008). Summary of the Invention

[0003] Provided herein are regimens for treating, reducing the rate of, or preventing hereditary angioedema (HAE) attacks using antibodies capable of binding to and inhibiting the active form of human plasma kallikrein (pKal), for example, antibodies having the same complementarity-determining regions (CDRs) as DX-2930 (SHP643, also known as lanadelumab).

[0004] Aspects of the present disclosure relate to a method for treating hereditary angioedema (HAE) attacks or reducing the rate of HAE attacks, comprising administering to a human subject in need thereof an antibody comprising the same complementarity-determining region (CDR) as DX-2930 for a first treatment period; during the first treatment period, the antibody is administered to the human subject multiple times at about 150 mg or 300 mg every two weeks or every four weeks; and the human subject has, is suspected of having, or is at risk for HAE and is a pediatric subject between the ages of 2 and 12. In some embodiments, the human subject is between the ages of 2 and 6, and the antibody is administered at about 150 mg every four weeks. In some embodiments, the human subject is between the ages of 6 and 12, and the antibody is administered at about 150 mg every two weeks. In some embodiments, the human subject is between the ages of 2 and 6, and the antibody is administered at about 300 mg every four weeks. In some embodiments, the human subject is between 6 and 12 years of age and the antibody is administered at about 300 mg every two weeks.

[0005] In some embodiments, the antibody is a full-length antibody or an antigen-binding fragment thereof. In some examples, the antibody comprises a heavy chain variable region set forth in SEQ ID NO:3 and / or a light chain variable region set forth in SEQ ID NO:4. In some examples, the antibody comprises a heavy chain set forth in SEQ ID NO:1 and a light chain set forth in SEQ ID NO:2. In some embodiments, the antibody is DX-2930 (also known as lanadelumab). In some embodiments, the antibody is TAKHZYRO® (lanadelumab-flyo). In some embodiments, the antibody is administered subcutaneously.

[0006] In any of the methods described herein, the antibody may be formulated into a pharmaceutical composition comprising a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical composition comprises sodium phosphate, citric acid, histidine, sodium chloride, and polysorbate 80. In one example, the sodium phosphate is at a concentration of about 30 mM, the citric acid is at a concentration of about 19 mM, the histidine is at a concentration of about 50 mM, the sodium chloride is at a concentration of about 90 mM, and the polysorbate 80 is about 0.01%.

[0007] In some embodiments, the human subject has HAE type I or II. In some embodiments, the human subject has an HAE attack rate of at least 1 HAE attack per 12 weeks prior to the first treatment period.

[0008] In some embodiments, the method further comprises administering the antibody to the subject for a second treatment period after the first treatment period.

[0009] The details of one or more embodiments of the invention are set forth in the description below. Other features or advantages of the invention will be apparent from the following drawings and detailed description of certain embodiments, and from the appended claims. [Brief explanation of the drawings]

[0010] [Figure 1] 1 shows an exemplary dosing regimen, including a screening period, an observation period, a first treatment period ("Treatment Period A"), a second treatment period ("Treatment Period B"), and a follow-up period. During the first and second treatment periods, patients between 2 and less than 6 years of age receive 150 mg of lanadelumab every 4 weeks for 26 weeks, whereas patients between 6 and less than 21 years of age receive 150 mg of lanadelumab every 2 weeks for 26 weeks. DETAILED DESCRIPTION OF THE INVENTION

[0011] definition For convenience, before further description of the present invention, certain terms employed in the specification, examples, and appended claims are defined here. Other terms are defined as they appear herein.

[0012] Unless the context clearly indicates otherwise, the singular forms "a," "an," and "the" include plural referents.

[0013] As used herein, the term "about" refers to + / - 5% of a particular value. For example, about 150 mg of antibody includes any amount of antibody between 142.5 mg and 157.5 mg.

[0014] The term "antibody" refers to an immunoglobulin molecule that can specifically bind to a target, such as a carbohydrate, polynucleotide, lipid, or polypeptide, through at least one antigen recognition site located in the variable region of the immunoglobulin molecule. Antibodies are composed of heavy chain immunoglobulin variable domains (V H ) at least one heavy (H) chain, a light chain immunoglobulin variable domain (V L For example, an antibody may comprise at least one light chain comprising a heavy (H) chain variable region (referred to herein as V H or HV) and light (L) chain variable region (V L or abbreviated LV). In another example, the antibody comprises two heavy (H) chain variable regions and two light (L) chain variable regions.

[0015] As used herein, the term "antibody" encompasses not only intact (i.e., full-length) polyclonal or monoclonal antibodies, but also antigen-binding fragments thereof (e.g., Fab, Fab', F(ab'), Fv, etc.), single-chain (scFv), domain antibody (dAb) fragments (de Wildt et al., Euro. J. Immunol. (1996) 26(3):629-639), any variants thereof, fusion proteins comprising antibody portions, humanized antibodies, chimeric antibodies, diabodies, linear antibodies, single-chain antibodies, multispecific antibodies (e.g., bispecific antibodies), and any other modified configurations of immunoglobulin molecules containing an antigen recognition site of the required specificity, including glycosylation variants of antibodies, amino acid sequence variants of antibodies, and covalently modified antibodies. Antibodies include antibodies of any class, such as IgD, IgE, IgG, IgA, or IgM (or subclasses thereof), and antibodies need not be of any particular class. Immunoglobulins can be assigned to different classes depending on the antibody amino acid sequence of the constant domain of their heavy chains. There are five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, some of which can be further divided into subclasses (isotypes), e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The heavy chain constant domains corresponding to the different classes of immunoglobulins are called alpha, delta, epsilon, gamma, and mu, respectively. The subunit structures and three-dimensional configurations of the different classes of immunoglobulins are well known. Antibodies can be from any source, but primates (human and non-human primates) and primatized antibodies are preferred.

[0016] V H and / or V LA region may comprise all or part of the amino acid sequence of a naturally occurring variable domain. For example, the sequence may lack one, two, or more N- or C-terminal amino acids, internal amino acids, may contain one or more inserted or additional terminal amino acids, or may contain other modifications. In one embodiment, a polypeptide comprising an immunoglobulin variable domain sequence may associate with another immunoglobulin variable domain sequence to form a structure that preferentially interacts with an antigen-binding site, e.g., plasma kallikrein.

[0017] V H and V L The regions can be further subdivided into regions of hypervariability termed "complementarity-determining regions" ("CDRs"), interspersed with more conserved regions termed "framework regions" ("FRs"). The extent of the framework regions and CDRs has been defined (see Kabat, E.A., et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, USDapartment of Health and Human Services, NIH Publication No. 91-3242, and Chothia, C. et al. (1987) J. Mol. Biol. 196:901-917). The Kabat definition is used herein. Each VH and VL is typically composed of three CDRs and four FRs, arranged from the amino terminus to the carboxy terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.

[0018] V H or V LIn addition to the variable region, the heavy or light chain of the antibody may further comprise all or a portion of a heavy or light chain constant region. In one embodiment, an antibody is a tetramer consisting of two immunoglobulin heavy chains and two immunoglobulin light chains, which are interconnected, for example, by disulfide bonds. In IgG, the heavy chain constant region comprises three immunoglobulin domains, CH1, CH2, and CH3. The light chain constant region comprises a CL domain. The variable regions of the heavy and light chains contain binding domains that interact with antigens. The constant region of an antibody typically mediates binding of the antibody to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (C1q) of the classical complement system. The light chain of the immunoglobulin may be of the kappa or lambda type. In one embodiment, the antibody is glycosylated. The antibody may be functional for antibody-dependent cellular cytotoxicity and / or complement-mediated cytotoxicity.

[0019] One or more regions of an antibody can be human or effectively human. For example, one or more variable regions can be human or effectively human. For example, one or more of the CDRs can be human, e.g., HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and / or LC CDR3. Each of the light chain (LC) and / or heavy chain (HC) CDRs can be human. The HC CDR3 can be human. One or more framework regions, e.g., FR1, FR2, FR3, and / or FR4 of the HC and / or LC, can be human, e.g., the Fc region can be human. In one embodiment, all framework regions are human, e.g., derived from a human somatic cell, e.g., a hematopoietic cell that produces immunoglobulins or a non-hematopoietic cell. In one embodiment, the human sequences are germline sequences, e.g., encoded by a germline nucleic acid. In one embodiment, framework (FR) residues of a selected Fab can be converted to the amino acid type of the corresponding residues in the most similar primate germline gene, particularly a human germline gene. One or more of the constant regions can be human or effectively human. For example, at least 70, 75, 80, 85, 90, 92, 95, 98, or 100% of the immunoglobulin variable domains, constant regions, constant domains (CH1, CH2, CH3, and / or CL1), or the entire antibody can be human or effectively human.

[0020] Antibodies can be encoded by immunoglobulin genes or segments thereof. Exemplary human immunoglobulin genes include the kappa, lambda, alpha (IgA1 and IgA2), gamma (IgG1, IgG2, IgG3, IgG4), delta, epsilon, and mu constant region genes, as well as many immunoglobulin variable region genes. Full-length immunoglobulin "light chains" (approximately 25 KDa or approximately 214 amino acids) are encoded by an NH2-terminal variable region gene (approximately 110 amino acids) and a COOH-terminal kappa or lambda constant region gene. Full-length immunoglobulin "heavy chains" (approximately 50 KDa or approximately 446 amino acids) are similarly encoded by a variable region gene (approximately 116 amino acids) and one of the other aforementioned constant region genes, such as gamma (encoding approximately 330 amino acids). The length of the human HC varies considerably, with the HC CDR3 varying from approximately 3 amino acid residues to more than 35 amino acid residues.

[0021] The term "antigen-binding fragment" of a full-length antibody refers to one or more fragments of a full-length antibody that retain the ability to specifically bind to a target of interest. Examples of functional binding fragments encompassed by the term "antigen-binding fragment" of a full-length antibody include: (i) Fab fragments, i.e., V L , V H , C L (ii) a F(ab')2 fragment, i.e., a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; (iii) a V H and an Fd fragment consisting of the CH1 domain, (iv) a V of a single arm of an antibody L and V H Fv fragment consisting of domains, (v) V H (vi) isolated complementarity-determining regions (CDRs); and (vi) isolated complementarity-determining regions (CDRs). In addition, the two domains V of the Fv fragment are also included. L and V HAlthough the V are encoded by separate genes, they can be joined by a synthetic linker that allows them to be made as a single protein chain using recombinant methods, and the V L and V H The domains pair to form monovalent molecules known as single-chain Fvs (scFvs). See, e.g., U.S. Patent Nos. 5,260,203, 4,946,778, and 4,881,175; Bird et al. (1988) Science 242:423-426; and Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883. Antibody fragments can be obtained using any suitable technique, including conventional techniques known to those skilled in the art.

[0022] The term "monospecific antibody" refers to an antibody that displays a single binding specificity and affinity for a particular target, e.g., epitope. This term includes "monoclonal antibody" or "monoclonal antibody composition," which, as used herein, refer to a preparation of antibodies or fragments thereof of single molecular composition, regardless of how the antibody was produced. An antibody may be "germlined" by reverting one or more non-germline amino acids in the framework regions to the corresponding germline amino acid of the antibody, so long as binding characteristics are substantially retained.

[0023] Inhibition constant (K i ) is a measure of inhibitor potency. It is the concentration of inhibitor required to reduce enzyme activity by half and is independent of enzyme or substrate concentration. Apparent K i (K i,app ) is obtained at various substrate concentrations by measuring the inhibitory effect of various concentrations of an inhibitor (e.g., an inhibitory binding protein) on the extent of a reaction (e.g., enzyme activity). Fitting the change in the pseudo-first-order rate constant as a function of inhibitor concentration to the Morrison equation (Equation 1) gives the apparent K i This gives an estimate of the value of K i is K i,app It is obtained from the y-intercept extracted from linear regression analysis of a plot of α versus substrate concentration.

number

[0024] As used herein, "binding affinity" refers to the apparent association constant or K A K A is the dissociation constant (K D ) is the reciprocal of the . The binding antibody has a specific target molecule, e.g., plasma kallikrein, and a specific binding affinity of at least 10 5 , 10 6 , 10 7 , 10 8 , 10 9 , 10 10 , and 10 11 M -1 The higher affinity binding of a binding antibody to a first target than to a second target can be expressed as a higher K A (or a number K D ) for binding to the first target A is high (or the number K D In such cases, the binding antibody has specificity for a first target (e.g., a protein or mimetic thereof in a first conformation) over a second target (e.g., the same protein or mimetic thereof in a second conformation, or a second protein). The difference in binding affinity (e.g., specificity or other comparative difference) is at least 1.5, 2, 3, 4, 5, 10, 15, 20, 30, 40, 50, 70, 80, 90, 100, 500, 1000, 10,000, or 10 5 It could be double.

[0025] Binding affinity can be determined by a variety of methods, including equilibrium dialysis, equilibrium binding, gel filtration, ELISA, surface plasmon resonance, or spectroscopy (e.g., using a fluorescence assay). Exemplary conditions for assessing binding affinity are in HBS-P buffer (10 mM HEPES pH 7.4, 150 mM NaCl, 0.005% (v / v) Surfactant P20). These techniques can be used to measure the concentration of bound and free binding protein as a function of binding protein (or target) concentration. The concentration of bound binding protein ([Bound]) is related to the concentration of free binding protein ([Free]) and the concentration of binding sites of the binding protein on the target by the following equation, where (N) is the number of binding sites per target molecule: [Bound]=N·[Free] / ((1 / KA)+[Free])

[0026] However, K A It is not always necessary to precisely determine the K because sometimes it is possible to obtain a quantitative measure of affinity (e.g., determined using methods such as ELISA or FACS analysis) and A This is because it is sufficient to obtain a qualitative measure of affinity, or to obtain an estimate of affinity (e.g., by activity in a functional assay, e.g., an in vitro or in vivo assay), which can be used for comparison, such as to determine whether a higher affinity is proportional to (e.g., 2-fold higher), or to obtain a qualitative measure of affinity, or to obtain an estimate of affinity (e.g., by activity in a functional assay, e.g., an in vitro or in vivo assay).

[0027] The term "binding antibody" (or "binding protein," as used interchangeably herein) refers to an antibody that can interact with a target molecule. The term "target molecule" is used interchangeably with "ligand." A "plasma kallikrein-binding antibody" refers to an antibody that can interact with (e.g., bind to) plasma kallikrein, and particularly includes antibodies that preferentially or specifically interact with and / or inhibit plasma kallikrein. An antibody inhibits plasma kallikrein if it causes a decrease in the activity of plasma kallikrein compared to the activity of plasma kallikrein in the absence of the antibody and under the same conditions.

[0028] A "conservative amino acid substitution" is one in which an amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues with similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), β-branched side chains (e.g., threonine, valine, isoleucine), and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine).

[0029] Amino acid residues in one or more framework and / or CDRs of the binding protein can comprise one or more mutations (e.g., substitutions (e.g., conservative substitutions or non-essential amino acid substitutions), insertions, or deletions) compared to the binding proteins described herein. A plasma kallikrein binding protein can have mutations (e.g., substitutions (e.g., conservative substitutions or non-essential amino acid substitutions), insertions, or deletions) (e.g., at least 1, 2, 3, or 4, and / or fewer than 15, 12, 10, 9, 8, 7, 6, 5, 4, 3, or 2 mutations) compared to the binding proteins described herein, e.g., mutations that do not substantially affect protein function. The mutations can be present in framework regions, CDRs, and / or constant regions. In some embodiments, the mutations are present in framework regions. In some embodiments, the mutations are present in CDRs. In some embodiments, the mutations are present in constant regions. Whether a particular substitution will be tolerated, i.e., will not adversely affect biological properties such as binding activity, can be predicted, for example, by assessing whether the mutation is conservative, or by the method of Bowie, et al. Science (1990) 247:1306-1310.

[0030] An "effectively human" immunoglobulin variable region is an immunoglobulin variable region that contains a sufficient number of human framework amino acid positions so that the immunoglobulin variable region does not elicit an immunogenic response in normal humans. An "effectively human" antibody is an antibody that contains a sufficient number of human amino acid positions so that the antibody does not elicit an immunogenic response in normal humans.

[0031] "Epitope" refers to a site on a target compound that is bound by a binding protein (e.g., an antibody, such as a Fab or full-length antibody). When the target compound is a protein, the site may be composed entirely of amino acid constituents, entirely of chemical modifications of amino acids of the protein (e.g., glycosyl moieties), or a combination thereof. Overlapping epitopes contain at least one common amino acid residue, glycosyl group, phosphate group, sulfate group, or other molecular feature.

[0032] A "humanized" immunoglobulin variable region is an immunoglobulin variable region that has been modified to contain a sufficient number of human framework amino acid positions so that the immunoglobulin variable region does not elicit an immunogenic response in normal humans. Examples of descriptions of "humanized" immunoglobulins include U.S. Patent No. 6,407,213 and U.S. Patent No. 5,693,762.

[0033] An "isolated" antibody refers to an antibody that is at least 90% removed from at least one component of the natural sample from which it is obtained. An antibody can be "at least" somewhat pure if the species or population of species of interest is at least 5, 10, 25, 50, 75, 80, 90, 92, 95, 98, or 99% pure on a weight / weight basis.

[0034] The methods described herein include administering multiple doses of an antibody to a human subject in need thereof. The terms "patient," "subject," or "host" may be used interchangeably. The subject may have received a previous treatment for HAE, such as a treatment comprising an antibody described herein. In some embodiments, the subject is a pediatric subject (e.g., an infant, child, or adolescent subject). In some embodiments, the human subject is a pediatric subject under the age of 12. In some embodiments, the human subject is a pediatric subject between the ages of 2 and 12. In some embodiments, the human subject is a pediatric subject between the ages of 2 and 12. In some embodiments, the human subject is a pediatric subject between the ages of 2 and 6. In some embodiments, the human subject is a pediatric subject between the ages of 2 and 6. In some embodiments, the human subject is a pediatric subject between the ages of 6 and 12. In some embodiments, the human subject is a pediatric subject between the ages of 6 and 12.

[0035] The terms "prekallikrein" and "pre-plasma kallikrein" are used interchangeably herein and refer to the zymogen form of active plasma kallikrein, also known as prekallikrein.

[0036] As used herein, the term "substantially identical" (or "substantially homologous") is used herein to refer to a first amino acid or nucleic acid sequence that contains a sufficient number of identical or equivalent amino acid residues or nucleotides (e.g., having similar side chains, e.g., conservative amino acid substitutions) as a second amino acid or nucleic acid sequence such that the first and second amino acid or nucleic acid sequences have (or encode proteins having) a similar activity, e.g., binding activity, binding preference, or biological activity. In the case of antibodies, the second antibody has the same specificity and at least 50%, at least 25%, or at least 10% of the affinity for the same antigen.

[0037] Statistical significance can be determined by any method known in the art. Exemplary statistical tests include the Student's t-test, the nonparametric Mann-Whitney U test, and the nonparametric Wilcoxon statistical test. Some statistically significant relationships have a P value of less than 0.05 or 0.02. Certain binding proteins may exhibit statistically significant (e.g., P value < 0.05 or 0.02) differences, such as differences in specificity or binding. For example, terms such as "induce," "inhibit," "enhance," "elevate," "increase," and "decrease," which represent distinct qualitative or quantitative differences between two states, may refer to differences, such as statistically significant differences, between two states.

[0038] A "therapeutically effective dose" preferably modulates a measurable parameter, e.g., plasma kallikrein activity, by a statistically significant degree or by at least about 20%, more preferably at least about 40%, even more preferably at least about 60%, and even more preferably at least about 80% compared to untreated subjects. The ability of a compound to modulate a measurable parameter, e.g., a disease-related parameter, can be assessed in animal model systems predictive of efficacy in human disorders and conditions. Alternatively, this property of a composition can be assessed by examining the compound's ability to modulate the parameter in vitro.

[0039] The term "treating," as used herein, refers to the application or administration of a composition containing one or more active agents to a subject who has, has symptoms of, is suspected of, is predisposed to, or is at risk for having HAE, with the intent to cure, heal, alleviate, mitigate, alter, repair, ameliorate, improve, or affect the disease, symptoms of the disease, or predisposition to the disease. "Prophylactic treatment," also known as "preventive treatment," refers to treatment that aims to protect a person from or reduce the risk of disease to which they have been or may be exposed. In some embodiments, the treatment methods described herein aim to prevent the occurrence and / or recurrence of HAE.

[0040] The term "preventing" a disease in a subject refers to subjecting the subject to a drug treatment, e.g., administering a drug, such that at least one symptom of the disease is prevented, i.e., administered prior to clinical manifestation of an undesirable condition (e.g., a disease or other undesirable pathology in a host animal), so as to protect the host from developing the undesirable condition. "Preventing" a disease is sometimes also referred to as "prophylaxis" or "prophylactic treatment."

[0041] A "prophylactically effective amount" means an amount effective, at dosages and for periods of time necessary, to achieve the desired prophylactic result. Typically, since a prophylactic dose is used in subjects prior to or at an earlier stage of disease, the prophylactically effective amount will be less than the therapeutically effective amount.

[0042] Antibodies that bind to plasma kallikrein (pKal) Plasma kallikrein-binding antibodies (anti-pKal antibodies) for use in the methods described herein may be full-length (e.g., IgG (including IgG1, IgG2, IgG3, IgG4), IgM, IgA (including IgA1, IgA2), IgD, and IgE) or may comprise only an antigen-binding fragment (e.g., Fab, F(ab')2, or scFv fragment). The binding antibody may comprise two heavy-chain immunoglobulins and two light-chain immunoglobulins, or may be a single-chain antibody. Plasma kallikrein-binding antibodies may be recombinant proteins such as humanized antibodies, CDR-grafted antibodies, chimeric antibodies, deimmunized antibodies, or in vitro-generated antibodies, and may optionally comprise constant regions derived from human germline immunoglobulin sequences. In one embodiment, the plasma kallikrein-binding antibodies are monoclonal antibodies.

[0043] In one aspect, the disclosure features an antibody (e.g., an isolated antibody) that binds to plasma kallikrein (e.g., human plasma kallikrein and / or mouse kallikrein) and includes at least one immunoglobulin variable region. For example, the antibody includes a heavy chain (HC) immunoglobulin variable domain sequence and / or a light chain (LC) immunoglobulin variable domain sequence. In one embodiment, the antibody binds to and inhibits plasma kallikrein, e.g., human plasma kallikrein and / or mouse kallikrein.

[0044] In some embodiments, the antibodies described herein have the same CDR sequences as DX-2930 (also known as SHP643 or lanadelumab), e.g., the heavy chain CDR sequences set forth as SEQ ID NOs:5-7 and the light chain CDR sequences set forth as SEQ ID NOs:8-10. In some embodiments, the antibodies comprise the same CDR sequences as DX-2930 and an LC immunoglobulin variable domain sequence that is at least 85, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% identical (e.g., in entirety or in framework regions) to an LC variable domain described herein. In some embodiments, the antibodies comprise the same CDR sequences as DX-2930 and an HC immunoglobulin variable domain sequence that is at least 85, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% identical (e.g., in entirety or in framework regions) to an HC variable domain described herein. In some embodiments, the antibody comprises the same CDR sequences as DX-2930 and an LC sequence that is at least 85, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% identical (e.g., entirely or in framework regions) to an LC sequence described herein. In some embodiments, the antibody comprises the same CDR sequences as DX-2930 and an HC sequence that is at least 85, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% identical (e.g., entirely or in framework regions) to an HC sequence described herein.

[0045] The plasma kallikrein binding protein may be an isolated antibody (e.g., at least 70, 80, 90, 95, or 99% free of other proteins). In some embodiments, the plasma kallikrein binding antibody or composition thereof is inactive or partially active (e.g., has a K of 5000 nM or greater) compared to the plasma kallikrein binding antibody. i,appThe plasma kallikrein-binding antibody is isolated from antibody cleavage fragments (e.g., DX-2930) that bind to plasma kallikrein at a specific site. For example, the plasma kallikrein-binding antibody is at least 70% free of such antibody cleavage fragments. In other embodiments, the binding antibody is at least 80%, at least 90%, at least 95%, at least 99%, or even 100% free of inactive or partially active antibody cleavage fragments.

[0046] The plasma kallikrein binding antibody may additionally inhibit plasma kallikrein, eg, human plasma kallikrein.

[0047] In some embodiments, the plasma kallikrein binding antibody does not bind to prekallikrein (e.g., human prekallikrein and / or mouse prekallikrein), but binds to the activated form of plasma kallikrein (e.g., human plasma kallikrein and / or mouse kallikrein).

[0048] In certain embodiments, the antibody binds to or near the active site of the catalytic domain of plasma kallikrein or a fragment thereof, or binds to an epitope that overlaps with the active site of plasma kallikrein.

[0049] The antibody has a binding affinity of at least 10 to plasma kallikrein, e.g., human plasma kallikrein. 5 , 10 6 , 10 7 , 10 8 , 10 9 , 10 10 , and 10 11 M -1 In one embodiment, the antibody can bind with a binding affinity of 1×10 -3 , 5×10 -4 s -1 , or 1 × 10 -4 s -1 slower than K off In one embodiment, the antibody binds to human plasma kallikrein at a concentration of 1 x 10 2 , 1×10 3 , or 5 × 10 3 M -1 s-1 Faster than K on In one embodiment, the antibody binds to plasma kallikrein but does not bind to tissue kallikrein and / or plasma prekallikrein (e.g., the antibody binds to tissue kallikrein and / or plasma prekallikrein less efficiently than it binds to plasma kallikrein (e.g., 5-, 10-, 50-, 100-, or 1000-fold less efficiently than a negative control, or does not bind at all).

[0050] In one embodiment, the antibody is, e.g., 10 -5 , 10 -6 , 10 -7 , 10 -8 , 10 -9 , and 10 -10 K less than M i The antibody inhibits human plasma kallikrein activity with an IC of less than, for example, 100 nM, 10 nM, 1, 0.5, or 0.2 nM. 50 For example, the antibody may modulate plasma kallikrein activity and the production of factor XIIa (e.g., derived from factor XII) and / or bradykinin (e.g., derived from high molecular weight kininogen (HMWK)). The antibody may inhibit plasma kallikrein activity and / or the production of factor XIIa (e.g., derived from factor XII) and / or bradykinin (e.g., derived from high molecular weight kininogen (HMWK)). The affinity of the antibody for human plasma kallikrein may be a K of less than 100 nM, less than 10 nM, less than 5 nM, less than 1 nM, or less than 0.5 nM. D In one embodiment, the antibody inhibits plasma kallikrein but not tissue kallikrein (e.g., the antibody inhibits tissue kallikrein less effectively than it inhibits plasma kallikrein (e.g., 5-, 10-, 50-, 100-, or 1000-fold less, or not at all, compared to a negative control).

[0051] In some embodiments, the antibody has an apparent inhibition constant (K) of less than 1000, 500, 100, 5, 1, 0.5, or 0.2 nM. i,app )

[0052] Plasma kallikrein binding antibodies may have their HC and LC variable domain sequences contained in a single polypeptide (eg, scFv) or in different polypeptides (eg, IgG or Fab).

[0053] In one embodiment, the HC and LC variable domain sequences are components of the same polypeptide chain. In another embodiment, the HC and LC variable domain sequences are components of different polypeptide chains. For example, the antibody is an IgG, e.g., IgG1, IgG2, IgG3, or IgG4. The antibody can be a soluble Fab. In other implementations, the antibody includes a Fab2', scFv, minibody, scFv::Fc fusion, Fab::HSA fusion, HSA::Fab fusion, Fab::HSA::Fab fusion, or other molecule containing the antigen-binding site of one of the binding proteins herein. The VH and VL regions of these Fabs can be provided as an IgG, Fab, Fab2, Fab2', scFv, PEGylated Fab, PEGylated scFv, PEGylated Fab2, VH::CH1::HSA+LC, HSA::VH::CH1+LC, LC::HSA+VH::CH1, HSA::LC+VH::CH1, or other suitable construct.

[0054] In one embodiment, the antibody is a human or humanized antibody, or is non-immunogenic in humans. For example, the antibody comprises one or more human antibody framework regions, such as all human framework regions, or framework regions that are at least 85, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to a human framework region. In one embodiment, the antibody comprises a human Fc domain, or an Fc domain that is at least 95, 96, 97, 98, or 99% identical to a human Fc domain.

[0055] In one embodiment, the antibody is a primate or primatized antibody, or is non-immunogenic in humans. For example, the antibody comprises one or more primate antibody framework regions, e.g., all primate framework regions, or framework regions at least 85, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to a primate framework region. In one embodiment, the antibody comprises a primate Fc domain, or an Fc domain at least 95, 96, 97, 98, or 99% identical to a primate Fc domain. "Primates" include humans (Homo sapiens), chimpanzees (Pan troglodytes and Pan paniscus (bonobo)), gorillas (Gorilla gorilla), gibbons, monkeys, lemurs, aye-ayes (Daubentonia madagascariensis), and tarsiers. In some embodiments, the affinity of the primate antibody for human plasma kallikrein is less than 1000, 500, 100, 10, 5, 1, or 0.5 nM, e.g., less than 10 nM, less than 1 nM, or less than 0.5 nM. D It is characterized by:

[0056] In certain embodiments, the antibody does not include sequences from a mouse or rabbit (eg, is not a mouse or rabbit antibody).

[0057] In some embodiments, the antibody used in the methods described herein can be DX-2930 or a functional variant thereof as described herein.

[0058] In one example, a functional variant of DX-2930 contains the same complementarity determining regions (CDRs) as DX-2930. In another example, a functional variant of DX-2930 contains the same VDRs as DX-2930. H and V L Compared to the FR of V H or V LThe functional variants may contain one or more mutations (e.g., conservative substitutions) in any of the FRs of DX-2930. Preferably, such mutations do not occur in residues predicted to interact with one or more of the CDRs (which can be determined by routine techniques). In other embodiments, the functional variants described herein contain one or more (e.g., 1, 2, or 3) mutations in one or more of the CDR regions of DX-2930. Preferably, such functional variants retain the same regions / residues involved in antigen binding as the parent. In yet other embodiments, the functional variants of DX-2930 contain one or more mutations in the V of DX-2930. H V comprising an amino acid sequence that is at least 85% (e.g., 90%, 92%, 94%, 95%, 96%, 97%, 98%, or 99%) identical to the amino acid sequence of H Chain, and / or DX-2930 V L V having an amino acid sequence that is at least 85% (e.g., 90%, 92%, 94%, 95%, 96%, 97%, 98%, or 99%) identical to the amino acid sequence of L These variants can bind to activated plasma kallikrein and preferably do not bind to prekallikrein.

[0059] The "percent identity" of two amino acid sequences is determined using the algorithm of Karlin and Altschul Proc. Natl. Acad. Sci. USA 87:2264-68, 1990, as modified in Karlin and Altschul Proc. Natl. Acad. Sci. USA 90:5873-77, 1993. Such an algorithm has been incorporated into the NBLAST and XBLAST programs (version 2.0) of Altschul, et al. J. Mol. Biol. 215:403-10, 1990. BLAST protein searches can be performed using the XBLAST program (score = 50, word length = 3) to obtain amino acid sequences homologous to the protein molecule of interest. When gaps exist between the two sequences, Gapped BLAST can be utilized as described in Altschul et al., Nucleic Acids Res. 25(17):3389-3402, 1997. When utilizing BLAST and Gapped BLAST programs, the default parameters of the respective programs (e.g., XBLAST and NBLAST) can be used.

[0060] In some embodiments, the antibody used in the methods and compositions described herein may be the DX-2930 antibody (also known as "lanadelumab"). The complete and variable sequences of the heavy and light chains of DX-2930 are shown below, with the signal sequence in italics. The CDRs are in bold and underlined. DX-2930 heavy chain amino acid sequence (451 amino acids, 49439.02 Da) [ka] DX-2930 light chain amino acid sequence (213 amino acids, 23419.08 Da) [ka] DX-2930 heavy chain variable domain amino acid sequence [ka] DX-2930 light chain variable domain amino acid sequence [ka]

[0061] [Table 1]

[0062] Antibody preparation The antibodies described herein (e.g., DX-2930) can be produced by any method known in the art (see, e.g., Harlow and Lane, (1988) Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, New York, and Greenfield, (2013) Antibodies: A Laboratory Manual, Second edition, Cold Spring Harbor Laboratory Press).

[0063] The sequence encoding the antibody of interest, e.g., DX-2930, may be maintained in a vector within a host cell, which can be propagated and frozen for later use. Alternatively, the polynucleotide sequence may be used in genetic engineering to "humanize" the antibody or improve its affinity (affinity maturation) or other properties. For example, if the antibody is to be used in clinical trials and treatments in humans, the constant region may be engineered to more closely resemble human constant regions to avoid an immune response. It may be desirable to genetically engineer an antibody sequence to obtain higher affinity for the target antigen and greater efficacy in inhibiting the activity of PKal. It will be apparent to those skilled in the art that one or more polynucleotide changes can be made to an antibody while maintaining its binding specificity for the target antigen.

[0064] In other embodiments, fully human antibodies can be obtained by using commercially available mice engineered to express specific human immunoglobulin proteins. Transgenic animals designed to generate more desirable (e.g., fully human antibodies) or more robust immune responses may also be used to generate humanized or human antibodies. Examples of such technology are Xenomouse® from Amgen, Inc. (Fremont, Calif.), and HuMAb-Mouse® and TC Mouse™ from Medarex, Inc. (Princeton, NJ). In another alternative, antibodies may be produced recombinantly by phage display or yeast technology. See, e.g., U.S. Patent Nos. 5,565,332; 5,580,717; 5,733,743; and 6,265,150; and Winter et al. (1994) Annu. Rev. Immunol. 12:433-455. Alternatively, phage display technology (McCafferty et al., (1990) Nature 348:552-553) can be used to produce human antibodies and antibody fragments in vitro, from immunoglobulin variable (V) domain gene repertoires from unimmunized donors.

[0065] Antigen-binding fragments of intact antibodies (full-length antibodies) can be prepared by conventional methods, for example, F(ab')2 fragments can be generated by pepsin digestion of the antibody molecule, and Fab fragments can be generated by reducing the disulfide bridges of the F(ab')2 fragment.

[0066] Genetically engineered antibodies, such as humanized antibodies, chimeric antibodies, single-chain antibodies, and bispecific antibodies, can be produced, for example, through conventional recombinant techniques. In one example, DNA encoding a monoclonal antibody specific to a target antigen can be readily isolated or synthesized. The DNA can be placed into one or more expression vectors and then transfected into host cells, such as E. coli cells, monkey COS cells, Chinese hamster ovary (CHO) cells, or myeloma cells, that do not otherwise produce immunoglobulin proteins, to obtain the synthesis of monoclonal antibodies in the recombinant host cells. See, for example, PCT Publication No. WO 87 / 04462. The DNA can then be modified, for example, by substituting coding sequences for human heavy and light chain constant domains for the homologous murine sequences (Morrison et al., (1984) Proc. Nat. Acad. Sci. 81:6851), or by covalently linking all or part of the coding sequence for a non-immunoglobulin polypeptide to the immunoglobulin coding sequence. In this manner, genetically engineered antibodies such as "chimeric" or "hybrid" antibodies can be prepared that have the binding specificity of a target antigen.

[0067] Techniques developed for the production of "chimeric antibodies" are well known in the art. See, e.g., Morrison et al. (1984) Proc. Natl. Acad. Sci. USA 81, 6851; Neuberger et al. (1984) Nature 312, 604; and Takeda et al. (1984) Nature 314:452.

[0068] Methods for constructing humanized antibodies are also well known in the art. See, e.g., Queen et al., Proc. Natl. Acad. Sci. USA, 86:10029-10033 (1989). In one example, the V of a parent non-human antibody is H and V LThe variable regions of the parent V are subjected to three-dimensional molecular modeling analysis according to methods known in the art. Framework amino acid residues predicted to be important for the formation of the correct CDR structure are then identified using the same molecular modeling analysis. In parallel, the parent V H and V L Using the sequence as a search query, a human V antibody having an amino acid sequence that is homologous to the amino acid sequence of the parent non-human antibody is identified. H and V L The human V chains are then identified from any antibody gene database. H and V L The donor gene is selected.

[0069] The CDR regions in the selected human donor gene can be replaced with CDR regions from the parent non-human antibody or a functional variant thereof. If necessary, residues in the framework regions of the parent chain that are predicted to be important for interactions with the CDR regions (see above) can be used to replace the corresponding residues in the human donor gene.

[0070] Single-chain antibodies can be prepared by recombinantly linking a nucleotide sequence encoding a heavy-chain variable region with a nucleotide sequence encoding a light-chain variable region. Preferably, a flexible linker is incorporated between the two variable regions. Alternatively, techniques described for producing single-chain antibodies (U.S. Pat. Nos. 4,946,778 and 4,704,692) can be adapted to generate phage or yeast scFv libraries, and PKal-specific scFv clones can be identified from the libraries using routine procedures. Positive clones can be subjected to further screening to identify clones that inhibit PKal activity.

[0071] Some antibodies, e.g., Fabs, can be produced in bacterial cells, e.g., E. coli cells (see, e.g., Nadkarni, A. et al., 2007 Protein Expr Purif 52(1):219-29). For example, if the Fab is encoded by a sequence in a phage display vector that contains a suppressible stop codon between the display entity and the bacteriophage protein (or fragment thereof), the nucleic acid of the vector can be introduced into bacterial cells that cannot suppress the stop codon. In this case, the Fab is not fused to the gene III protein and is secreted into the periplasm and / or medium.

[0072] Antibodies can also be produced in eukaryotic cells. In one embodiment, the antibody (e.g., scFv) is expressed in yeast cells such as Pichia (see, e.g., Powers et al., 2001, J. Immunol. Methods. 251:123-35; Schoonooghe S. et al., 2009 BMC Biotechnol. 9:70; Abdel-Salam, H.A. et al., 2001 Appl Microbiol Biotechnol 56(1-2):157-64; Takahashi K. et al., 2000 Biosci Biotechnol Biochem 64(10):2138-44; Edqvist, J. et al., 1991 J Biotechnol 20(3):291-300), Hanseula, or Saccharomyces.Those skilled in the art will be familiar with, for example, oxygen conditions (see, e.g., Baumann K., et al. 2010 BMC Syst. Biol. 4:141), osmolarity (see, e.g., Dragosits, M. et al., 2010 BMC Genomics 11:207), temperature (see, e.g., Dragosits, M. et al., 2009 J Proteome Res. 8(3):1380-92), fermentation conditions (see, e.g., Ning, D. et al. 2005 J. Biochem. and Mol. Biol. 38(3):294-299), yeast strains (see, e.g., Kozyr, AV et al. 2004 Mol Biol (Mosk) 38(6):1067-75; Horwitz, AH. et al., 1988 Proc Natl Acad Sci USA 85(22):8678-82; Bowdish, K. et al. 1991 J Biol Chem 266(18):11901-8), overexpression of proteins to enhance antibody production (see, e.g., Gasser, B. et al., 2006 Biotechnol. Bioeng. 94(2):353-61), acidity level of the culture (see, e.g., Kobayashi H., et al., 1997 FEMS Microbiol Lett 152(2):235-42), and substrate and / or ion concentrations (see, e.g., Ko JH. et al., 2996 Appl Biochem Biotechnol 60(1):41-8). In addition, yeast systems can be used to produce antibodies with extended half-lives (see, eg, Smith, BJ. et al. 2001 Bioconjug Chem 12(5):750-756).

[0073] In a preferred embodiment, the antibody is produced in mammalian cells. Preferred mammalian host cells for expressing a clonal antibody or its antigen-binding fragment include Chinese hamster ovary (CHO) cells (dhfr-CHO cells described in Urlaub and Chasin, 1980, Proc. Natl. Acad. Sci. USA 77:4216-4220, which are used with a DHFR selection marker, e.g., as described in Kaufman and Sharp, 1982, Mol. Biol. 159:601 621), lymphoid cell lines, e.g., NS0 myeloma cells and SP2 cells, COS cells, HEK293T cells (J. Immunol. Methods (2004) 289(1-2):65-80), and cells from transgenic animals, e.g., transgenic mammals. For example, the cells are mammary epithelial cells.

[0074] In some embodiments, plasma kallikrein binding antibodies are produced in plants or cell-free systems (see, eg, Galeffi, P., et al., 2006 J Transl Med 4:39).

[0075] In addition to the nucleic acid sequences encoding the various immunoglobulin domains, recombinant expression vectors may carry additional sequences, such as sequences that regulate replication of the vector in host cells (e.g., origins of replication) and selectable marker genes. The selectable marker gene can facilitate selection of host cells into which the vector has been introduced (see, e.g., U.S. Pat. Nos. 4,399,216, 4,634,665, and 5,179,017). For example, the selectable marker gene typically confers resistance to drugs, such as G418, hygromycin, or methotrexate, on the host cells into which the vector has been introduced. Preferred selectable marker genes include the dihydrofolate reductase (DHFR) gene (dhfr with methotrexate selection / propagation). - host cell), and the neo gene (for G418 selection).

[0076] In an exemplary system for recombinant expression of an antibody or antigen-binding portion thereof, a recombinant expression vector encoding both the antibody heavy chain and the antibody light chain is transfected with dhfr - The antibody heavy and light chain genes are introduced into CHO cells. Within the recombinant expression vector, the antibody heavy and light chain genes are each operably linked to an enhancer / promoter regulatory element (e.g., derived from SV40, CMV, adenovirus, etc., e.g., a CMV enhancer / AdMLP promoter regulatory element or an SV40 enhancer / AdMLP promoter regulatory element) to drive high levels of gene transcription. The recombinant expression vector also contains a DHFR gene, which allows for selection of CHO cells transfected with the vector using methotrexate selection / amplification. Selected transformant host cells are cultured to allow expression of the antibody heavy and light chains, and intact antibody is recovered from the culture medium. Standard molecular biology techniques are used to prepare the recombinant expression vector, transfect the host cells, select for transformants, culture the host cells, and recover the antibody from the culture medium. For example, some antibodies can be isolated by affinity chromatography using a protein A- or protein G-bound matrix.

[0077] In the case of antibodies containing an Fc domain, the antibody production system can produce antibodies in which the Fc region is glycosylated. For example, the Fc domain of an IgG molecule is glycosylated at asparagine 297 in the CH2 domain. This asparagine is a site for modification with biantennary oligosaccharides. This glycosylation has been demonstrated to be necessary for effector functions mediated by Fcγ receptors and complement C1q (Burton and Woof, 1992, Adv. Immunol. 51:1-84; Jefferis et al., 1998, Immunol. Rev. 163:59-76). In one embodiment, the Fc domain is produced in a mammalian expression system that properly glycosylates the residue corresponding to asparagine 297. The Fc domain may also contain other eukaryotic post-translational modifications.

[0078] Antibodies can also be produced by transgenic animals. For example, U.S. Patent No. 5,849,992 describes a method for expressing antibodies in the mammary gland of transgenic mammals. A transgene is constructed that includes a milk-specific promoter, a nucleic acid encoding an antibody of interest, and a signal sequence for secretion. The milk produced by such transgenic mammalian females contains the antibody of interest secreted therein. The antibody can be purified from the milk or used directly for some applications.

[0079] Pharmaceutical Composition An antibody described herein (e.g., DX-2930) can be present in a composition, e.g., a pharmaceutically acceptable composition or pharmaceutical composition. An antibody described herein (e.g., DX-2930) can be formulated with a pharmaceutically acceptable carrier. In some embodiments, 150 mg of DX-2930 antibody, optionally with a pharmaceutically acceptable carrier, is present in a composition, e.g., a pharmaceutically acceptable composition or pharmaceutical composition. In some embodiments, 300 mg of DX-2930 antibody, optionally with a pharmaceutically acceptable carrier, is present in a composition, e.g., a pharmaceutically acceptable composition or pharmaceutical composition.

[0080] A "pharmaceutically acceptable carrier" includes any and all physiologically compatible solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, etc. Preferably, the carrier is suitable for subcutaneous, intravenous, intramuscular, parenteral, spinal, or epidermal administration (e.g., by injection or infusion), although carriers suitable for inhaled and intranasal administration are also contemplated.

[0081] The pharmaceutically acceptable carrier in the pharmaceutical compositions described herein may include one or more of a buffering agent, an amino acid, and an osmolality adjusting agent. Any suitable buffering agent or combination of buffering agents may be used in the pharmaceutical compositions described herein to maintain or help maintain the proper pH of the composition. Non-limiting examples of buffering agents include sodium phosphate, potassium phosphate, citric acid, sodium succinate, histidine, Tris, and sodium acetate. In some embodiments, the buffering agent may be at a concentration of about 5-100 mM, 5-50 mM, 10-50 mM, 15-50 mM, or about 15-40 mM. For example, the one or more buffering agents may be at a concentration of about 15 mM, 16 mM, 17 mM, 18 mM, 19 mM, 20 mM, 21 mM, 22 mM, 23 mM, 24 mM, 25 mM, 26 mM, 27 mM, 28 mM, 29 mM, 30 mM, 31 mM, 32 mM, 33 mM, 35 mM, 36 mM, 37 mM, 38 mM, 39 mM, or about 40 mM. In some examples, the pharmaceutically acceptable carrier comprises sodium phosphate and citric acid, which may be at a concentration of about 30 mM and about 19 mM, respectively.

[0082] In some embodiments, a pharmaceutically acceptable carrier comprises one or more amino acids, which can reduce antibody aggregation and / or increase antibody stability during storage prior to administration. Exemplary amino acids for use in making the pharmaceutical compositions described herein include, but are not limited to, alanine, arginine, asparagine, aspartic acid, glycine, histidine, lysine, proline, or serine. In some examples, the concentration of an amino acid in a pharmaceutical composition can be about 5-100 mM, 10-90 mM, 20-80 mM, 30-70 mM, 40-60 mM, or about 45-55 mM. In some examples, the concentration of an amino acid (e.g., histidine) can be about 40 mM, 41 mM, 42 mM, 43 mM, 44 mM, 45 mM, 46 mM, 47 mM, 48 mM, 49 mM, 50 mM, 51 mM, 52 mM, 53 mM, 54 mM, 55 mM, 56 mM, 57 mM, 58 mM, 59 mM, or about 60 mM. In one example, the pharmaceutical composition comprises histidine at a concentration of about 50 nM.

[0083] Any suitable osmolality adjuster may be used to prepare the pharmaceutical compositions described herein. In some embodiments, the osmolality adjuster is a salt or an amino acid. Examples of suitable salts include, but are not limited to, sodium chloride, sodium succinate, sodium sulfate, potassium chloride, magnesium chloride, magnesium sulfate, and calcium chloride. In some embodiments, the osmolality adjuster in the pharmaceutical composition may be at a concentration of about 10-150 mM, 50-150 mM, 50-100 mM, 75-100 mM, or about 85-95 mM. In some embodiments, the osmolality modifier may be at a concentration of about 80 mM, 81 mM, 82 mM, 83 mM, 84 mM, 85 mM, 86 mM, 87 mM, 88 mM, 89 mM, 90 mM, 91 mM, 92 mM, 93 mM, 94 mM, 95 mM, 96 mM, 97 mM, 98 mM, 99 mM, or about 100 mM. In one example, the osmolality modifier may be sodium chloride, which may be at a concentration of about 90 mM.

[0084] The pharmaceutically acceptable carrier in the pharmaceutical compositions described herein may further comprise one or more pharmaceutically acceptable excipients. Generally, pharmaceutically acceptable excipients are pharmacologically inactive substances. Non-limiting examples of excipients include lactose, glycerol, xylitol, sorbitol, mannitol, maltose, inositol, trehalose, glucose, bovine serum albumin (BSA), dextran, polyvinyl acetate (PVA), hydroxypropylmethylcellulose (HPMC), polyethyleneimine (PEI), gelatin, polyvinylpyrrolidone (PVP), hydroxyethylcellulose (HEC), polyethylene glycol (PEG), ethylene glycol, glycerol, dimethyl sulfoxide (DMSO), dimethylformamide (DM F), polyoxyethylene sorbitan monolaurate (Tween-20), polyoxyethylene sorbitan monooleate (Tween-80), sodium dodecyl sulfate (SDS), polysorbate, polyoxyethylene copolymer, potassium phosphate, sodium acetate, ammonium sulfate, magnesium sulfate, sodium sulfate, trimethylamine N-oxide, betaine, zinc ions, copper ions, calcium ions, manganese ions, magnesium ions, CHAPS, sucrose monolaurate, and 2-O-beta-mannoglycerate. In some embodiments, the pharmaceutically acceptable carrier comprises between about 0.001%-0.1%, 0.001%-0.05%, 0.005-0.1%, 0.005%-0.05%, 0.008%-0.05%, 0.008%-0.03%, or about 0.009%-0.02% of an excipient. In some embodiments, the excipient is about 0.005%, 0.006%, 0.007%, 0.008%, 0.009%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, or about 0.1%. In some embodiments, the excipient is polyoxyethylene sorbitan monooleate (Tween-80). In one example, the pharmaceutically acceptable carrier contains 0.01% Tween-80.

[0085] In some examples, the pharmaceutical compositions described herein comprise an anti-pKal antibody (e.g., DX-2930) also described herein and one or more of sodium phosphate (e.g., sodium phosphate dibasic dihydrate), citric acid (e.g., citric acid monohydrate), histidine (e.g., L-histidine), sodium chloride, and polysorbate 80. For example, the pharmaceutical composition may comprise an antibody, sodium phosphate, citric acid, histidine, sodium chloride, and polysorbate 80. In some examples, the antibody is formulated in about 30 mM sodium phosphate, about 19 mM citric acid, about 50 mM histidine, about 90 mM sodium chloride, and about 0.01% polysorbate 80. The concentration of the antibody (e.g., DX-2930) in the composition may be about 150 mg / mL. In one example, the composition comprises or consists of about 150 mg of DX-2930, about 30 mM sodium phosphate dibasic dihydrate, about 19 mM (e.g., 19.6 mM) citric acid monohydrate, about 50 mM L-histidine, about 90 mM sodium chloride, and about 0.01% polysorbate 80 per mL of solution.

[0086] A "pharmaceutically acceptable salt" is a salt that retains the desired biological activity of a compound and does not impart any undesired toxicological effects. (See, e.g., Berge, SM, et al., 1977, J. Pharm. Sci. 66:1-19.) Examples of such salts include acid addition salts and base addition salts. Acid addition salts include those derived from non-toxic inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, and phosphorous acid, as well as those derived from non-toxic organic acids such as aliphatic mono- and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxyalkanoic acids, aromatic acids, and aliphatic and aromatic sulfonic acids. Base addition salts include those derived from alkaline earth metals such as sodium, potassium, magnesium, and calcium, and those derived from non-toxic organic amines such as N,N'-dibenzylethylenediamine, N-methylglucamine, chloroprocaine, choline, diethanolamine, ethylenediamine, and procaine.

[0087] The compositions may be in a variety of forms. These include, for example, liquid, semi-solid, and solid dosage forms, such as liquid solutions (e.g., injections and infusions), dispersions or suspensions, tablets, pills, powders, liposomes, and suppositories. The form may vary depending on the intended mode of administration and therapeutic use. Many compositions are in the form of injections or infusions, such as compositions similar to those used to administer antibodies to humans. An exemplary mode of administration is parenteral (e.g., intravenous, subcutaneous, intraperitoneal, intramuscular). In one embodiment, the plasma kallikrein binding protein is administered by intravenous infusion or injection. In another embodiment, the plasma kallikrein binding protein is administered by intramuscular injection. In another embodiment, the plasma kallikrein binding protein is administered by subcutaneous injection. In another preferred embodiment, the plasma kallikrein binding protein is administered by intraperitoneal injection.

[0088] The phrases "parenteral administration" and "administered parenterally," as used herein, refer to modes of administration other than enteral and topical administration, usually by injection, including, but not limited to, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injection and infusion. In some embodiments, the antibody is administered subcutaneously.

[0089] The composition can be formulated as a solution, microemulsion, dispersion, liposome, or other ordered structure suitable for high drug concentration.Sterile injectable solutions can be prepared by incorporating the required amount of binding protein with one or a combination of the above-listed ingredients in an appropriate solvent, and then optionally sterilizing by filtration.Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle containing the basic dispersion medium and the other required ingredients listed above.For sterile powders for preparing sterile injectable solutions, the preferred preparation method is vacuum drying and freeze-drying, which produces a powder of the active ingredient plus any desired additional ingredients from its solution that has been previously sterile-filtered.The proper fluidity of the solution can be maintained, for example, by using a coating such as lecithin, by maintaining the required particle size in the case of dispersions, and by using surfactants.Prolonged absorption of injectable compositions can be achieved by including an agent that delays absorption, such as monostearate salts and gelatin, in the composition.

[0090] The antibodies described herein (e.g., DX-2930) can be administered by a variety of methods, including intravenous injection, subcutaneous injection, or infusion. For example, in some therapeutic applications, the antibodies are administered by intravenous infusion at a rate of less than 30, 20, 10, 5, or 1 mg / min to provide a dose of about 1-100 mg / m. 2 or 7 to 25 mg / m 2A dose of 100 mg / kg or more can be achieved. The route and / or mode of administration will vary depending on the desired results. In certain embodiments, the active compound may be prepared with a carrier that protects the compound against rapid release, such as a controlled-release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid can be used. Many methods are available for preparing such formulations. See, for example, *Sustained and Controlled Release Drug Delivery Systems*, JR Robinson, ed., 1978, Marcel Dekker, Inc., New York.

[0091] The pharmaceutical composition can be administered using a medical device. For example, in one embodiment, the pharmaceutical compositions disclosed herein can be administered in conjunction with a device, such as a needleless hypodermic injection device, a pump, or an implant.

[0092] In certain embodiments, the antibodies described herein (e.g., DX-2930) can be formulated to ensure adequate distribution in vivo. For example, the blood-brain barrier (BBB) ​​excludes many highly hydrophilic compounds. To ensure that the therapeutic compounds disclosed herein cross the BBB (if desired), they can be formulated, for example, in liposomes. For methods of manufacturing liposomes, see, e.g., U.S. Patent Nos. 4,522,811; 5,374,548; and 5,399,331. Liposomes may contain one or more moieties that are selectively transported into specific cells or organs, thereby enhancing targeted drug delivery (see, e.g., V. V. Ranade, 1989, J. Clin. Pharmacol. 29:685).

[0093] Dosage regimens are adjusted to provide the optimum desired response (e.g., therapeutic response). For example, a single bolus may be administered, several divided doses may be administered over time, or the dose may be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation. It is particularly advantageous to formulate parenteral compositions in unit dosage forms for ease of administration and uniformity of dosage. As used herein, unit dosage form refers to physically discrete units suitable as unitary dosages for the subject to be treated, each unit containing a predetermined amount of active compound calculated to produce the desired therapeutic effect in combination with the required pharmaceutical carrier. The specifications for the unit dosage forms are dictated by and directly depend on (a) the inherent characteristics of the active compound and the particular therapeutic effect to be achieved, and (b) the limitations inherent in the art of formulating such active compounds to suit the therapeutic susceptibility of individuals.

[0094] An exemplary, non-limiting range for a therapeutically or prophylactically effective amount of an antibody described herein (e.g., DX-2930) is about 150 mg. In some embodiments, a therapeutically or prophylactically effective amount of an antibody described herein (e.g., DX-2930) is about 300 mg. As will be appreciated by those of skill in the art, the therapeutically or prophylactically effective amount of an antibody for pediatric subjects may be lower than for adult subjects. In some embodiments, the effective amount administered to a pediatric subject is a fixed dose or a weight-based dose. In some embodiments, an effective amount of less than about 150 mg is administered to a pediatric subject. In some embodiments, a therapeutically or prophylactically effective amount of an antibody is administered every two weeks or every four weeks during a first treatment period. In some embodiments, an antibody may be administered to a subject during a second treatment period. In some embodiments, the therapeutically or prophylactically effective amount of an antibody during a first treatment period is the same as the therapeutically or prophylactically effective amount of an antibody during a second treatment period. In some embodiments, the therapeutically or prophylactically effective amount of an antibody during a first treatment period is 150 mg, and the therapeutically or prophylactically effective amount of an antibody during a second treatment period is 150 mg. In some embodiments, the therapeutically or prophylactically effective amount of the antibody during a first treatment period is 300 mg, and the therapeutically or prophylactically effective amount of the antibody during a second treatment period is 300 mg. In some embodiments, the therapeutically or prophylactically effective amount of the antibody during a first treatment period is 150 mg, and the therapeutically or prophylactically effective amount of the antibody during a second treatment period is 300 mg. In some embodiments, the therapeutically or prophylactically effective amount of the antibody during a first treatment period is 300 mg, and the therapeutically or prophylactically effective amount of the antibody during a second treatment period is 150 mg. In some embodiments, the therapeutically or prophylactically effective amount of the antibody during a first treatment period is different from the therapeutically or prophylactically effective amount of the antibody during a second treatment period. In some embodiments, the therapeutically or prophylactically effective amount of the antibody during a first treatment period is the same as the therapeutically or prophylactically effective amount of the antibody during a second treatment period.

[0095] kit An antibody described herein (e.g., DX-2930) can be provided in a kit, e.g., as a component of the kit. For example, the kit includes (a) a DX-2930 antibody, e.g., a composition (e.g., a pharmaceutical composition) comprising the antibody, and optionally (b) informational material. The informational material can be descriptive, instructional, marketing, or other material regarding the methods described herein and / or use of (e.g., for) the antibodies described herein (e.g., DX-2930). In some embodiments, the kit includes one or more doses of DX-2930. In some embodiments, the one or more doses is 150 mg. In some embodiments, the one or more doses is 300 mg.

[0096] The informational material of the kit is not limited in its form. In one embodiment, the informational material can include information regarding the manufacture of the compound, the molecular weight of the compound, concentration, expiration date, batch or manufacturing site information, etc. In one embodiment, the informational material relates to the use of the antibody for the treatment, prevention, or diagnosis of disorders and conditions, e.g., diseases or conditions associated with plasma kallikrein.

[0097] In one embodiment, the informational material can include instructions for administering an antibody described herein (e.g., DX-2930) in a manner suitable for practicing a method described herein, e.g., at an appropriate dose, dosage form, mode of administration, or dosing schedule (e.g., a dose, dosage form, dosing schedule, or mode of administration described herein). In another embodiment, the informational material can include instructions for administering an antibody described herein (e.g., DX-2930) to a suitable subject, e.g., a human, e.g., a human having or at risk for a disease or condition associated with plasma kallikrein. For example, the material can include instructions for administering an antibody described herein (e.g., DX-2930), e.g., according to a dosing schedule described herein, to a patient having a disorder or condition described herein, e.g., a disease associated with plasma kallikrein. The informational material of the kit is not limited in its form. Often, the informational material, e.g., instructions, is provided in printed form, but can also be in other formats, such as computer-readable material.

[0098] The antibodies described herein (e.g., DX-2930) can be provided in any form, for example, liquid, dried, or lyophilized. Preferably, the antibodies are substantially pure and / or sterile. When the antibodies are provided in a liquid solution, the liquid solution is preferably an aqueous solution, with a sterile aqueous solution being preferred. When the antibodies are provided in a dry form, reconstitution is generally by the addition of a suitable solvent. The solvent, e.g., sterile water or buffer, can optionally be provided in the kit.

[0099] The kit can include one or more containers for a composition containing an antibody described herein (e.g., DX-2930). In some embodiments, the kit contains separate containers, dividers, or compartments for the composition and informational material. For example, the composition can be contained in a bottle, vial, or syringe, and the informational material can be contained with the container. In other embodiments, the separate elements of the kit are contained in a single, undivided container. For example, the composition is contained in a bottle, vial, or syringe, which has informational material in the form of a label attached thereto. In some embodiments, the kit includes multiple (e.g., a pack) individual containers, each containing one or more unit dosage forms (e.g., dosage forms described herein) of an antibody described herein (e.g., DX-2930). For example, the kit includes multiple syringes, ampoules, foil packets, or blister packs, each containing a single unit dose of an antibody described herein (e.g., DX-2930). The containers of the kit can be airtight, waterproof (e.g., impervious to changes in moisture and evaporation), and / or light-tight.

[0100] The kit optionally includes a device suitable for administering the composition, e.g., a syringe, or any such delivery device. In one embodiment, the device is an implantable device that dispenses a metered dose of the antibody. The disclosure also features methods of providing a kit, e.g., by combining the components described herein.

[0101] treatment In some aspects, the disclosure provides for the use of an antibody described herein (eg, DX-2930) in the treatment of HAE.

[0102] (i) Hereditary angioedema Hereditary angioedema (HAE) is also known as "Quincke's edema," C1 esterase inhibitor deficiency, C1 inhibitor deficiency, and hereditary angioedema (HANE). HAE is characterized by unpredictable, recurrent attacks of severe subcutaneous or submucosal swelling (angioedema), which can affect, for example, the limbs, face, genitals, gastrointestinal tract, and respiratory tract (Zuraw, 2008). Symptoms of HAE include, for example, swelling of the arms, legs, lips, eyes, tongue, and / or throat; swelling of the throat (larynx), airway obstruction, which may be accompanied by sudden hoarseness and / or death from asphyxiation (Bork et al., 2012; Bork et al., 2000). Approximately 50% of all HAE patients will experience a laryngeal attack during their lifetime, and there is no way to predict which patients are at risk for a laryngeal attack (Bork et al., 2003; Bork et al., 2006). Symptoms of HAE include recurrent episodes of abdominal cramps without apparent cause; and / or intestinal swelling, which can be severe and can cause abdominal cramps, vomiting, dehydration, diarrhea, pain, shock, and / or intestinal symptoms resembling an abdominal emergency, potentially leading to unnecessary surgery (Zuraw, 2008). Swelling can last up to five days or more. Approximately one-third of individuals with HAE develop a non-itchy rash called erythema marginatum during attacks. Most patients suffer from multiple attacks per year.

[0103] HAE is a rare disorder and its exact prevalence is unknown, but current estimates range from 1 per 10,000 to 1 per 150,000, with most authors agreeing that 1 per 50,000 is probably the closest estimate (Bygum, 2009; Goring et al., 1998; Lei et al., 2011; Nordenfelt et al., 2014; Roche et al., 2005).

[0104] Plasma kallikrein plays an important role in the pathogenesis of HAE attacks (Davis, 2006; Kaplan and Joseph, 2010). In normal physiology, C1-INH regulates not only plasma kallikrein but also the activity of various other proteases, such as C1r, C1s, factor XIa, and factor XIIa. Plasma kallikrein regulates the release of bradykinin from high-molecular-weight kininogen (HMWK). Deficiency of C1-INH in HAE results in uncontrolled plasma kallikrein activity, leading to excessive production of bradykinin. Bradykinin is a vasodilator that is thought to be involved in the characteristic HAE symptoms of localized swelling, inflammation, and pain (Craig et al., 2012; Zuraw et al., 2013).

[0105] Airway swelling can be life-threatening and can cause death in some patients. Mortality rates are estimated to be 15-33%. HAE results in approximately 15,000-30,000 emergency department visits per year.

[0106] Trauma or stress, such as dental procedures, illness (e.g., viral illnesses such as colds and influenza), menstruation, and surgery, can trigger attacks of angioedema. To prevent acute attacks of HAE, patients may attempt to avoid specific stimuli that previously triggered attacks. However, attacks often occur without any known trigger. Symptoms of HAE typically first appear in childhood and worsen during adolescence. On average, untreated individuals experience attacks every 1–2 weeks, with most episodes lasting approximately 3–4 days (ghr.nlm.nih.gov / condition / hereditary-angioedema). The frequency and duration of attacks vary widely among people with hereditary angioedema, even within the same family.

[0107] There are three types of HAE, known as type I, type II, and type III, all of which can be treated by the methods described herein. HAE is estimated to affect 1 in 50,000 people, with type I accounting for approximately 85% of cases, type II accounting for approximately 15% of cases, and type III being extremely rare. Type III is the most recently described form and was originally thought to occur only in women, but families with affected males have been identified.

[0108] HAE is inherited in an autosomal dominant pattern, meaning that affected individuals may have inherited a mutation from one affected parent. De novo mutations can also occur in genes, so HAE can occur in individuals with no family history of the disorder. It is estimated that 20-25% of cases are due to de novo spontaneous mutations.

[0109] Mutations in the SERPING1 gene cause hereditary angioedema types I and II. The SERPING1 gene directs the production of the C1 inhibitor protein, which is important for controlling inflammation. C1 inhibitor blocks the activity of certain proteins that promote inflammation. Mutations that cause hereditary angioedema type I result in reduced levels of C1 inhibitor in the blood. In contrast, mutations that cause type II result in the production of abnormally functioning C1 inhibitor. Approximately 85% of patients have type I HAE, which is characterized by very low production of functionally normal C1-INH protein, while the remaining approximately 15% have type II HAE, which produces normal or elevated levels of functionally impaired C1-INH (Zuraw, 2008). Without adequate levels of functional C1 inhibitor, excessive amounts of bradykinin are produced from high molecular weight kininogen (HMWK), increasing vascular leakage mediated by bradykinin binding to B2 receptors (B2-R) on the surface of endothelial cells (Zuraw, 2008). Bradykinin promotes inflammation by increasing fluid leakage from blood vessel walls into body tissues. Excessive fluid accumulation in body tissues leads to episodes of swelling seen in individuals with hereditary angioedema types I and II.

[0110] Mutations in the F12 gene have been associated with some cases of hereditary angioedema type III. The F12 gene provides the instructions for making clotting factor XII. In addition to playing a key role in blood clotting (clotting), factor XII is also an important stimulator of inflammation and is involved in the production of bradykinin. Specific mutations in the F12 gene result in the production of increased activity of factor XII. As a result, more bradykinin is produced, making blood vessel walls leakier and leading to episodes of swelling. The cause of other cases of hereditary angioedema type III remains unknown. Mutations in one or more as-yet-unidentified genes may be involved in the disorder in these cases.

[0111] Although HAE may present similarly to other forms of angioedema caused by allergies or other medical conditions, the causes and treatment are quite different. When hereditary angioedema is misdiagnosed as an allergy, it is most commonly treated with antihistamines, steroids, and / or epinephrine. While epinephrine may be used for life-threatening reactions, these are typically ineffective in HAE. Misdiagnosis can lead to unnecessary exploratory surgery in patients with abdominal swelling, and some HAE patients have their abdominal pain incorrectly diagnosed as psychogenic.

[0112] Like adults, children with HAE can suffer from recurrent, debilitating attacks. Symptoms can appear very early in childhood, with upper airway angioedema reported in patients with HAE as young as 3 years of age (Bork et al., 2003). In one case study of 49 pediatric HAE patients, 23 had suffered at least one episode of airway angioedema by age 18 (Farkas, 2010). Because the disease typically worsens after puberty, there is a significant unmet medical need for children with HAE, particularly adolescents (Bennett and Craig, 2015; Zuraw, 2008).

[0113] C1 inhibitor therapy, and other therapies for HAE, are described in Kaplan, AP, J Allergy Clin Immunol, 2010, 126(5):918-925.

[0114] Acute treatment of an HAE attack is administered to stop the progression of edema as quickly as possible. C1 inhibitor concentrate from donor blood, administered intravenously, is one acute treatment. However, this treatment is not available in many countries. In emergency situations when C1 inhibitor concentrate is unavailable, fresh frozen plasma (FFP) can be used as an alternative because it also contains C1 inhibitor.

[0115] Purified C1 inhibitor derived from human blood has been used in Europe since 1979. Several C1 inhibitor treatments are currently available in the United States, and two C1 inhibitor products are currently available in Canada. Pasteurized Verinert P (CSL Behring) was approved by the FDA for acute attacks in 2009. Nanofiltered Synlyse (ViroPharma) was approved by the FDA for prophylaxis in 2008. Rhucin (Pharming) is an investigational recombinant C1 inhibitor that does not carry the risk of infectious disease transmission from human bloodborne pathogens.

[0116] Treatment for acute HAE attacks may also include medication and / or intravenous fluids for pain relief.

[0117] Other treatment modalities can stimulate C1 inhibitor synthesis or reduce C1 inhibitor consumption. Androgen medications such as danazol can reduce the frequency and severity of attacks by stimulating C1 inhibitor production.

[0118] Helicobacter pylori can trigger abdominal attacks. Antibiotics that treat H. pylori may reduce abdominal attacks.

[0119] Newer treatments attack the contact cascade. Ecallantide (KALBITOR®, DX-88, Dyax) inhibits plasma kallikrein and is approved in the United States. Icatibant (FIRAZYR®, Shire) inhibits the bradykinin B2 receptor and is approved in Europe and the United States.

[0120] Diagnosis of HAE may depend, for example, on family history and / or blood tests. Laboratory findings associated with types I, II, and III HAE are described, for example, in Kaplan, AP, J Allergy Clin Immunol, 2010, 126(5):918-925. In type I HAE, C1 inhibitor levels are decreased, as are C4 levels, while C1q levels are normal. In type II HAE, C1 inhibitor levels are normal or increased, but C1 inhibitor function is abnormal. C4 levels are decreased, and C1q levels are normal. In type III, C1 inhibitor, C4, and C1q levels may all be normal.

[0121] Symptoms of HAE can be assessed, for example, using a questionnaire (e.g., a questionnaire completed by the patient, clinician, or family member). Such questionnaires are known in the art and include, for example, visual analog scales. See, e.g., McMillan, CV et al. Patient. 2012;5(2):113-26. In some embodiments, the subject has type I HAE or type II HAE. Type I HAE or type II HAE may be diagnosed using any method known in the art, for example, by a clinical history consistent with HAE (e.g., episodes of non-pruritic swelling of the subcutaneous or mucous membranes) or diagnostic testing (e.g., evaluation of C1-INH function tests and C4 levels).

[0122] (ii) Treatment of HAE with anti-PKal antibodies The present disclosure provides methods of treating hereditary angioedema (HAE) (e.g., ameliorating, stabilizing, or eliminating one or more symptoms) by administering an antibody described herein (e.g., a therapeutically effective amount of an antibody described herein) to a subject having or suspected of having HAE, e.g., according to a dosing schedule described herein. Additionally provided are methods of treating HAE by administering an antibody described herein (e.g., a therapeutically effective amount of an antibody described herein), e.g., according to a dosing schedule described herein, or in combination with a second therapy, e.g., in combination with another agent (e.g., one described herein). The present disclosure also provides methods of preventing HAE or symptoms thereof by administering an antibody described herein (e.g., a prophylactically effective amount of an antibody described herein) to a subject at risk of developing HAE (e.g., a subject who has a family member with HAE or a genetic predisposition thereto), e.g., according to a dosing schedule described herein. In some examples, the subject may be a human patient who does not have symptoms of HAE at the time of treatment. In some embodiments, the subject is a human patient with type I HAE or type II HAE. In some embodiments, the subject is a human patient who has experienced at least two (eg, 2, 3, 4, 5, or more) attacks of HAE in the year prior to treatment.

[0123] In some embodiments, the subject is a pediatric subject under the age of 12. In some embodiments, the subject is a pediatric subject between the ages of 2 and 12. In some embodiments, the subject is a pediatric subject between the ages of 2 and 6. In some embodiments, the subject is a pediatric subject between the ages of 6 and 12. ...

[0124] In some embodiments, any of a subgroup of human patients may be administered an anti-pKal antibody (e.g., DX-2930) at about 150 mg every two weeks. In some embodiments, any of a subgroup of human patients may be administered an anti-pKal antibody (e.g., DX-2930) at about 150 mg every four weeks. In some embodiments, the subjects are pediatric subjects aged 2 to 6 years, and the subjects are administered an antibody at about 150 mg every four weeks. In some embodiments, the subjects are pediatric subjects aged 6 to 12 years, and the subjects are administered an antibody at about 150 mg every two weeks. In some embodiments, human patients are administered an anti-pKal antibody (e.g., DX-2930) at about 150 mg every two weeks, and once the subject has been seizure-free for at least six months, the dosing of the antibody is reduced to about 150 mg every four weeks.

[0125] In some embodiments, any subgroup of human patients may receive an anti-pKal antibody (e.g., DX-2930) at about 300 mg every two weeks. In some embodiments, any subgroup of human patients may receive an anti-pKal antibody (e.g., DX-2930) at about 300 mg every four weeks.

[0126] Treatment includes administering an amount effective to alleviate, relieve, alter, repair, ameliorate, improve, or affect the disorder, the symptoms of the disorder, or the predisposition to the disorder. Treatment can also delay the onset, e.g., prevent the onset, or prevent the worsening of the disease or condition.

[0127] Methods for administering the DX-2930 antibody are also described in the "Pharmaceutical Compositions" section. The appropriate dosage of the antibody used may depend on the age and weight of the subject and the particular drug used. The antibody can be used, for example, as a competitive agent to inhibit or reduce undesirable interactions between plasma kallikrein and its substrate (e.g., factor XII or HMWK). The dose of the antibody can be sufficient to block 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.9% of plasma kallikrein activity in the patient, particularly at the disease site. In some embodiments, 150 mg of the antibody is administered to the subject every two weeks or every four weeks. In some embodiments, the antibody is administered to the subject for a first treatment period comprising administering 150 mg of the antibody every two weeks or every four weeks. In some embodiments, 300 mg of the antibody is administered to the subject every two weeks or every four weeks. In some embodiments, the antibody is administered to the subject in a first treatment period comprising administering 300 mg of the antibody every two weeks or every four weeks. In some embodiments, the first treatment period is 26 weeks. In some embodiments, the first treatment period is 52 weeks. In some embodiments, the subject is administered 13 times in the first treatment period. In some embodiments, the subject is administered 14 times in the first treatment period. In some embodiments, the subject is administered 26 times in the first treatment period. In some embodiments, the subject is administered 27 times in the first treatment period. In some embodiments, the antibody is administered to the subject in a second treatment period following the first treatment period.

[0128] In one embodiment, the antibody is used to inhibit the activity of plasma kallikrein (e.g., inhibit at least one activity of plasma kallikrein, e.g., reduce the production of factor XIIa and / or bradykinin), e.g., in vivo. The binding protein can be used by itself or can be conjugated to an agent, e.g., a cytotoxic drug, a cytotoxic enzyme, or a radioisotope.

[0129] Antibodies can be used directly in vivo to eliminate antigen-expressing cells via natural complement-dependent cytotoxicity (CDC) or antibody-dependent cellular cytotoxicity (ADCC). The antibodies described herein can comprise a complement-binding effector domain, such as the Fc portion from IgG1, -2, or -3, or the corresponding portion of IgM that binds complement. In one embodiment, a population of target cells is treated ex vivo with an antibody described herein and appropriate effector cells. This treatment can be supplemented by the addition of complement or serum containing complement. Furthermore, phagocytosis of target cells coated with the antibodies described herein can be improved by binding of complement proteins. In another embodiment, target cells coated with an antibody containing a complement-binding effector domain are lysed by complement.

[0130] Methods for administering the DX-2930 antibody are described in the "Pharmaceutical Compositions" section. The appropriate dosage of the molecule used will depend on the age and weight of the subject and the particular drug used. The antibody can be used, for example, as a competitive agent to inhibit or reduce undesired interactions between natural or pathological agents and plasma kallikrein.

[0131] A therapeutically effective amount of the antibodies described herein can be administered to a subject having, suspected of having, or at risk for HAE to thereby treat the disorder (e.g., ameliorate or improve symptoms or features of the disorder, slow, stabilize, and / or halt progression of the disease).

[0132] The antibodies described herein can be administered in a therapeutically effective amount, which is an amount that, when administered to a subject in single or multiple doses, is effective in treating the subject, e.g., in curing, alleviating, reducing, or ameliorating at least one symptom of a disorder in the subject to an extent greater than would be expected in the absence of such treatment.

[0133] Dosage regimens can be adjusted to provide the optimum desired response (e.g., therapeutic response). For example, a single bolus may be administered, or several divided doses may be administered over time, or the dose may be proportionally reduced or increased based on the exigencies of the therapeutic situation. In other examples, several doses may be administered over time followed by a bolus, or the dose may be proportionally reduced or increased based on the exigencies of the therapeutic situation. In other examples, the dose may be divided into several doses and administered over time. For ease of administration and uniformity of dosage, it is particularly advantageous to formulate parenteral compositions into unit dosage forms. As used herein, unit dosage form refers to physically discrete units suitable as unitary dosages for the subject to be treated, each unit containing a predetermined amount of active compound calculated to produce the desired therapeutic effect in combination with the required pharmaceutical carrier.

[0134] In some embodiments, the antibodies described herein are administered in a dosing regimen during a first treatment period. In some embodiments, the antibody is administered multiple times during the first treatment period. During this period, a therapeutically or prophylactically effective amount of the antibody (e.g., DX-2930) may be about 150 mg administered weekly, every 2 weeks, every 3 weeks, every 4 weeks, every 5 weeks, every 6 weeks, every 7 weeks, every 8 weeks, or more. In some embodiments, a therapeutically or prophylactically effective amount of the antibody (e.g., DX-2930) may be about 150 mg administered weekly, every 2 weeks, every 3 weeks, every 4 weeks, every 5 weeks, every 6 weeks, every 7 weeks, every 8 weeks, or more to subjects under the age of 12. In some embodiments, a therapeutically or prophylactically effective amount of the antibody (e.g., DX-2930) during the first treatment period may be about 300 mg administered weekly, every 2 weeks, every 3 weeks, every 4 weeks, every 5 weeks, every 6 weeks, every 7 weeks, every 8 weeks, or more. In some embodiments, a therapeutically or prophylactically effective amount of an antibody (e.g., DX-2930) can be about 300 mg administered to a subject under the age of 12 every week, every 2 weeks, every 3 weeks, every 4 weeks, every 5 weeks, every 6 weeks, every 7 weeks, every 8 weeks or more.

[0135] In some embodiments, a therapeutically or prophylactically effective amount of an antibody (e.g., DX-2930) can be about 150 mg administered every week, every 2 weeks, every 3 weeks, every 4 weeks, every 5 weeks, every 6 weeks, every 7 weeks, every 8 weeks or more to a subject between the ages of 2 and under 6. In some examples, a therapeutically or prophylactically effective amount of an antibody (e.g., DX-2930) can be about 150 mg administered every 4 weeks to a subject between the ages of 2 and under 6.

[0136] In some embodiments, a therapeutically or prophylactically effective amount of an antibody (e.g., DX-2930) can be about 150 mg administered every week, every two weeks, every three weeks, every four weeks, every five weeks, every six weeks, every seven weeks, every eight weeks or more to a subject between the ages of 6 and under 12. In some embodiments, a therapeutically or prophylactically effective amount of an antibody (e.g., DX-2930) can be about 150 mg administered every two weeks to a subject between the ages of 6 and under 12.

[0137] In some embodiments, a therapeutically or prophylactically effective amount of an antibody (e.g., DX-2930) can be about 150 mg and is administered every two weeks or every four weeks. In some embodiments, a therapeutically or prophylactically effective amount of an antibody (e.g., DX-2930) can be 150 mg and is administered to a subject every two weeks. In some embodiments, a therapeutically or prophylactically effective amount of an antibody (e.g., DX-2930) can be 150 mg and is administered to a subject every four weeks. In some embodiments, a therapeutically or prophylactically effective amount is administered at least two times, at least three times, at least four times, at least five times, at least six times, at least seven times, at least eight times, at least nine times, at least ten times, at least eleven times, at least 12 times, at least thirteen times, or more.

[0138] In some embodiments, a therapeutically or prophylactically effective amount of an antibody (e.g., DX-2930) can be about 300 mg administered every week, every 2 weeks, every 3 weeks, every 4 weeks, every 5 weeks, every 6 weeks, every 7 weeks, every 8 weeks or more to a subject between the ages of 2 and under 6. In some examples, a therapeutically or prophylactically effective amount of an antibody (e.g., DX-2930) can be about 300 mg administered every 4 weeks to a subject between the ages of 2 and under 6.

[0139] In some embodiments, a therapeutically or prophylactically effective amount of an antibody (e.g., DX-2930) can be about 300 mg administered every week, every two weeks, every three weeks, every four weeks, every five weeks, every six weeks, every seven weeks, every eight weeks or more to a subject between the ages of 6 and under 12. In some embodiments, a therapeutically or prophylactically effective amount of an antibody (e.g., DX-2930) can be about 300 mg administered every two weeks to a subject between the ages of 6 and under 12.

[0140] In some embodiments, a therapeutically or prophylactically effective amount of an antibody (e.g., DX-2930) can be about 300 mg and is administered every two weeks or every four weeks. In some embodiments, a therapeutically or prophylactically effective amount of an antibody (e.g., DX-2930) can be 300 mg and is administered to a subject every two weeks. In some embodiments, a therapeutically or prophylactically effective amount of an antibody (e.g., DX-2930) can be 300 mg and is administered to a subject every four weeks. In some embodiments, a therapeutically or prophylactically effective amount is administered at least two times, at least three times, at least four times, at least five times, at least six times, at least seven times, at least eight times, at least nine times, at least ten times, at least eleven times, at least 12 times, at least thirteen times, or more.

[0141] In some embodiments, the first treatment period is 26 weeks. In some embodiments, the therapeutically or prophylactically effective amount is 150 mg or 300 mg and is administered to a subject every two weeks (e.g., every two weeks for 26 weeks, resulting in a total of 13 deliveries). In some embodiments, the therapeutically or prophylactically effective amount is 150 mg or 300 mg and is administered to a subject every two weeks (e.g., every two weeks for 26 weeks and follow-up, resulting in a total of 13 deliveries). In some embodiments, the therapeutically or prophylactically effective amount is 150 mg or 300 mg and is administered to a subject every four weeks (e.g., every four weeks for 26 weeks, resulting in a total of 7 deliveries). In some embodiments, the therapeutically or prophylactically effective amount is 150 mg or 300 mg and is administered to a subject every two weeks (e.g., every two weeks for 26 weeks, resulting in a total of 13 deliveries). In some embodiments, the therapeutically or prophylactically effective amount is 150 mg or 300 mg and is administered to a subject every four weeks (e.g., every four weeks for 26 weeks, resulting in a total of 7 deliveries).

[0142] In one example, the first treatment period is 26 weeks, and the antibody is administered on days 0, 28, 56, 84, 112, 140, and 168. In another example, the first treatment period is 26 weeks, and the antibody is administered on days 0, 14, 28, 42, 56, 70, 84, 98, 112, 126, 140, 154, and 168. Those skilled in the art will understand that the recited treatment schedules allow for a time frame of ±4 days (e.g., ±3 days, ±2 days, or ±1 day). For example, doses given on days 10-18 would be included in the day 14 dose described above.

[0143] In some embodiments, the first treatment period is 52 weeks. In some embodiments, the therapeutically or prophylactically effective amount is 150 mg and is administered to a subject every two weeks (e.g., every two weeks for 52 weeks, resulting in a total of 26 deliveries). In some embodiments, the first treatment period is 52 weeks. In some embodiments, the therapeutically or prophylactically effective amount is 300 mg and is administered to a subject every two weeks (e.g., every two weeks for 52 weeks, resulting in a total of 26 deliveries). In some embodiments, the therapeutically or prophylactically effective amount is 150 mg and is administered to a subject every four weeks (e.g., every four weeks for 52 weeks, resulting in a total of 13 deliveries). In some embodiments, the therapeutically or prophylactically effective amount is 300 mg and is administered to a subject every four weeks (e.g., every four weeks for 52 weeks, resulting in a total of 13 deliveries).

[0144] In one example, the first treatment period is 52 weeks, and the antibody is administered on days 0, 28, 56, 84, 112, 140, 168, 196, 224, 252, 280, 308, and 336. In another example, the first treatment period is 52 weeks, and the antibody is administered on days 0, 14, 28, 42, 56, 70, 84, 98, 112, 126, 140, 154, 168, 182, 196, 210, 224, 238, 252, 266, 280, 294, 308, 322, 336, and 350. Those skilled in the art will understand that the recited treatment schedules allow for a time frame of ±3 days (e.g., ±3 days, ±2 days, or ±1 day). For example, doses given on days 10-18 would be included in the day 14 dose described above.

[0145] In some embodiments, the therapeutically or prophylactically effective amount is administered in a dosing regimen during a second treatment period following a first treatment period. In some embodiments, the therapeutically or prophylactically effective amount is different between the first and second treatment periods. In some embodiments, the therapeutically or prophylactically effective amount during the second treatment period is about 150 mg. During this period, the antibody may be administered in multiple doses of about 150 mg, such as 150 mg every two weeks or 150 mg every four weeks. In some embodiments, the therapeutically or prophylactically effective amount during the second treatment period is about 300 mg. During this period, the antibody may be administered in multiple doses of about 300 mg, such as 300 mg every two weeks or 300 mg every four weeks. In some embodiments, the dosing frequency may be reduced if the subject has been seizure-free for at least six months. For example, a subject administered 150 mg or 300 mg of the antibody every two weeks may be administered about 150 mg of the antibody every four weeks if the subject has been seizure-free for at least six months. In some embodiments, multiple doses of antibody are administered at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times, at least 11 times, at least 12 times, or at least 13 times during the second treatment period.

[0146] In some embodiments, the second treatment period is 52 weeks. In some embodiments, the antibody is administered at a dose of about 150 mg or 300 mg every two weeks for 52 weeks (e.g., resulting in 26 deliveries). In some embodiments, the antibody is administered at a dose of about 150 mg or 300 mg every four weeks for 26 weeks (e.g., resulting in 13 deliveries). In some embodiments, the second treatment period is 26 weeks. In some embodiments, the antibody is administered at a dose of about 150 mg or 300 mg every two weeks for 26 weeks (e.g., resulting in 13 deliveries). In some embodiments, the antibody is administered at a dose of about 150 mg or 300 mg every four weeks for 26 weeks (e.g., resulting in 7 deliveries). In some embodiments, the antibody is administered at a dose of about 150 mg or 300 mg every two weeks for 26 weeks (e.g., resulting in 13 deliveries). In some embodiments, the first single dose of the second treatment period is administered about two weeks after the last dose of the first treatment period.

[0147] In any of the embodiments described herein, the timing of antibody administration is approximate and may include three days before and three days after the indicated day (e.g., administration every two weeks would include administration on day 11, 12, 13, 14, 15, 16, or 17).

[0148] The prior HAE treatment may include the same antibody (e.g., DX-2930) as described herein. In some embodiments, the prior HAE treatment may include multiple doses of DX-2930 every two weeks or every four weeks. In some embodiments, DX-2930 is administered to the subject (e.g., subcutaneously) at 150 mg every two weeks or 150 mg every four weeks. In some embodiments, DX-2930 is administered to the subject (e.g., subcutaneously) at 300 mg every two weeks or 300 mg every four weeks. In one example, the subject was previously administered the antibody every two weeks or every four weeks for 26 weeks prior to the single administration of the antibody. In some embodiments, the multiple doses of antibody in the prior treatment are administered at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least 11, at least 12, or at least 13 times. In some embodiments, the antibody was previously administered on days 0, 28, 56, 84, 112, 140, and 168. In some embodiments, a single dose of about 150 mg of antibody is administered about two weeks after the last dose of the previous treatment. In some embodiments, a single dose of about 300 mg of antibody is administered about two weeks after the last dose of the previous treatment. In one example, a single dose of the second treatment period is administered on day 182 of the first treatment period.

[0149] In some embodiments, a subject may be evaluated to establish a baseline rate of HAE attacks prior to administering an antibody according to any of the methods described herein. Such an evaluation period may be referred to as a "run-in period." In some embodiments, the baseline rate of HAE attacks must meet or exceed a minimum number of HAE attacks within a given period. In one example, the subject experiences at least one HAE attack during the four-week run-in period before first administering the antibody. In another example, the subject experiences between one and fewer than two attacks per month during the four-week run-in period before first administering the antibody. In another example, the subject experiences between two and fewer than three attacks per month during the four-week run-in period before first administering the antibody. In another example, the subject experiences three or more attacks per month during the four-week run-in period before first administering the antibody. In another example, the subject experiences at least two HAE attacks during the eight-week run-in period before first administering the antibody. In yet another example, the subject experiences an average of at least one HAE attack per month.

[0150] In some embodiments, a therapeutically or prophylactically effective amount of an antibody (e.g., DX-2930) can be about 150 mg and is administered to a subject who has experienced a past HAE attack rate of at least one HAE attack per 12 weeks during the period before the antibody is first administered. In some embodiments, a therapeutically or prophylactically effective amount of an antibody (e.g., DX-2930) can be about 150 mg and is administered every 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, or more during the period before the antibody is first administered. In some embodiments, a therapeutically or prophylactically effective amount of an antibody (e.g., DX-2930) can be about 300 mg and is administered to a subject who has experienced a past HAE attack rate of at least one HAE attack per 12 weeks during the period before the antibody is first administered. In some embodiments, a therapeutically or prophylactically effective amount of an antibody (e.g., DX-2930) can be about 300 mg administered every 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks or more to a subject who has experienced at least one HAE attack per 12 weeks during the period prior to first administration of the antibody.

[0151] In some embodiments, administration of an antibody according to any of the methods described herein reduces the average HAE attack rate in a subject. In some embodiments, the percent reduction in the average HAE attack rate after administration of an antibody according to any of the methods described herein may be determined compared to the HAE attack rate in subjects who did not receive the antibody (e.g., subjects who received a placebo). In some embodiments, the percent reduction in the average HAE attack rate may be at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95% compared to the HAE attack rate in subjects who did not receive the antibody (e.g., subjects who received a placebo).

[0152] Any of the subjects described herein may have received a previous treatment for HAE, such as a prophylactic or therapeutic treatment for HAE. Aspects of the present disclosure also provide methods of administering an antibody described herein (e.g., DX-2930) to a subject who has received one or more previous treatments for HAE. In some embodiments, the previous treatment for HAE is a treatment comprising an antibody described herein (e.g., DX-2930). In some embodiments, the subject previously received multiple doses of DX-2930 every two weeks or every four weeks. In some embodiments, the subject previously received DX-2930 at 150 mg every two weeks. In some embodiments, the subject previously received DX-2930 at 150 mg every four weeks. In some embodiments, the subject previously received DX-2930 at 300 mg every two weeks. In some embodiments, the subject previously received DX-2930 at 300 mg every four weeks. In some embodiments, the multiple antibody doses of the prior treatment are administered at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times, at least 11 times, at least 12 times, or at least 13 times.

[0153] In some embodiments, the subject has received one or more prior therapies for HAE, such as long-term preventative treatment, which may include any of the HAE treatments known in the art. Exemplary anti-HAE agents include, but are not limited to, C1 inhibitors (e.g., Cinryze®, Berinert®, or Ruconest®), plasma kallikrein inhibitors (e.g., Kalbitor®), bradykinin receptor inhibitors (e.g., Firazyr®), attenuated androgens (e.g., danazol), and antifibrinolytic agents (e.g., traexamic acid). In some embodiments, the subject received treatment with a C1 inhibitor prior to the first treatment period. In some instances, the subject may undergo a tapering period before receiving the anti-pKal antibody treatment described herein. A tapering period refers to a period prior to anti-pKal antibody treatment during which a subject receiving an anti-HAE treatment (e.g., C1-INH, oral androgen, and / or oral antifibrinolytic agent) gradually reduces the dosage, frequency, or both of the anti-HAE agent to allow the subject to gradually transition from the previous HAE treatment to the anti-pKal antibody treatment described herein. In some embodiments, tapering involves a gradual or stepwise method of reducing the dosage of the previous treatment and / or the frequency with which the previous treatment is administered. The tapering period may last 2-4 weeks, which may vary depending on individual patient factors. In some instances, the previous treatment is terminated before anti-pKal antibody treatment begins. In other instances, the previous treatment may be terminated within an appropriate time frame (e.g., 2, 3, or 4 weeks) after the subject receives the first dose of anti-pKal antibody.

[0154] Alternatively, subjects undergoing a previous HAE treatment may be transitioned directly to the anti-pKal antibody treatment described herein without a tapering period.

[0155] In some embodiments, a therapeutically or prophylactically effective amount of an antibody (e.g., DX-2930) can be about 150 mg or 300 mg and is administered every 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks or more to a subject who has received one or more prior treatments for HAE.

[0156] In other embodiments, the subject has not received any prior treatment for HAE prior to the initial treatment, first treatment period, and / or subsequent single and multiple dose treatments (second treatment period) described herein. In some embodiments, the subject has not received any treatment other than the antibodies described herein during the first treatment period and / or second treatment period. In some embodiments, the subject has not received any prior treatment for HAE for at least two weeks (e.g., at least two, three, four, five, or more weeks) prior to the initial treatment or first treatment period, or during the initial treatment or first treatment period and / or second treatment period. In some embodiments, the subject has not received long-term prophylaxis for HAE (e.g., C1 inhibitor, attenuated androgen, antifibrinolytic agent) for at least two weeks prior to the initial treatment or first treatment period, or during the first treatment period and / or second treatment period. In some embodiments, the subject has not received HAE treatment including an angiotensin-converting enzyme (ACE) inhibitor for at least four weeks prior to the initial treatment or first treatment period, during the first treatment period, and / or second treatment period. In some embodiments, the subject has not received estrogen-containing medication for at least 4 weeks prior to the initial treatment or first treatment period, during the first treatment period, and / or during the second treatment period. In some embodiments, the subject has not received androgens (e.g., stanozolol, danazol, oxandrolone, methyltestosterone, testosterone) for at least 2 weeks prior to the initial treatment or first treatment period, during the first treatment period, and / or during the second treatment period.

[0157] Any of the methods described herein may further include monitoring the patient for side effects (e.g., elevated creatine phosphatase levels) and / or the level of inhibition of pKal by the antibody (e.g., serum or plasma concentration of the antibody or pKal activity level) before, after, or during the course of treatment. If one or more adverse effects are observed, the dose (dosage or dosing frequency) of the antibody may be reduced or treatment may be discontinued. If the level of inhibition is below the minimum therapeutic level, an additional dose of the antibody may be administered to the patient. Patients may also be assessed for antibody production against the administered antibody; activity of C1 inhibitor, C4, and / or C1q; quality of life; incidence of HAE attacks, health-related quality of life, anxiety and / or depression (e.g., Hospital Anxiety and Depression Scale (HADS)), work productivity (e.g., Work Productivity and Activity Impairment Questionnaire (WPAI)), preference for subcutaneous administration of the antibody (e.g., DX-2930) over other injectable agents, quality of life (e.g., Angioedema Quality of Life (AE-QOL), EuroQoL Group 5-item report).

[0158] In some embodiments, the plasma or serum concentration of an antibody (e.g., DX-2930) may be measured during the course of treatment (e.g., after an initial dose) to assess the effectiveness of treatment. If the plasma or serum concentration of the antibody is less than about 80 nM, a follow-up dose equal to or greater than the initial dose may be required. The plasma or serum concentration of the antibody may be measured by determining the protein level of the antibody in a plasma or serum sample obtained from the subject, for example, by immunoassay or mass spectrometry (MS) assay. The plasma or serum concentration of the antibody may also be measured by determining the inhibition level of pKal in a plasma or serum sample obtained from a subject treated with the antibody. Such assays may include synthetic substrate assays or Western blot assays for measuring cleaved kininogen, such as those described herein.

[0159] Alternatively or additionally, plasma or serum levels of creatine kinase and / or one or more coagulation parameters (e.g., activated partial thromboplastin time (aPTT), prothrombin time (PT), bleeding events) can be monitored during the course of treatment. If plasma or serum levels of creatine kinase are found to increase during treatment, the dosage (dosage or dosing frequency) of the antibody may be reduced or treatment may be terminated. Similarly, if one or more coagulation parameters are found to be significantly affected during treatment, the dosage of the antibody may be modified or treatment may be terminated.

[0160] In some embodiments, the optimal dosage (e.g., optimal prophylactic dosage or optimal therapeutic dosage) of an antibody (e.g., DX-2930) may be determined as follows: An initial dose of the antibody is given to a subject in need of treatment. The plasma concentration of the antibody in the subject is measured. If the plasma concentration is less than 80 nM, the dose of the antibody is increased in subsequent administrations. The antibody dosage that maintains the antibody plasma concentration above about 80 nM can be selected as the optimal dosage for the subject. The subject's creatine phosphokinase levels can be monitored during the course of treatment, and the subject's optimal dosage can be further adjusted based on the creatine phosphokinase level. For example, if an increase in creatine phosphokinase is observed during treatment, the antibody dosage may be reduced.

[0161] (iii) Combination therapy An antibody described herein (e.g., DX-2930) can be administered in combination with one or more other therapies to treat a disease or condition associated with plasma kallikrein activity, e.g., a disease or condition described herein. For example, an antibody described herein (e.g., DX-2930) can be used therapeutically or prophylactically with another anti-plasma kallikrein Fab or IgG (e.g., another Fab or IgG described herein), another plasma kallikrein inhibitor, a peptide inhibitor, a small molecule inhibitor, or surgery (e.g., before, during, or after a course of treatment). Examples of plasma kallikrein inhibitors that can be used in combination therapy with a plasma kallikrein-binding antibody described herein include, for example, the plasma kallikrein inhibitors described in PCT Publication Nos. WO1995 / 21601 or WO2003 / 103475.

[0162] One or more plasma kallikrein inhibitors can be used in combination with an antibody described herein (e.g., DX-2930), e.g., by doing so, a lower dose of the inhibitor may be required, thereby reducing side effects.

[0163] The antibodies described herein (e.g., DX-2930) can be administered in combination with one or more current therapies for treating HAE. For example, the DX-2930 antibody can be used with a second anti-HAE therapeutic agent, such as ecallantide, a C1 esterase inhibitor (e.g., CINRYZE™), aprotinin (TRASYLOL®), and / or a bradykinin B2 receptor inhibitor (e.g., icatibant (FIRAZYR®)).

[0164] The term "combination" refers to the use of two or more agents or therapies to treat the same patient, where the use or effects of the agents or therapies overlap in time. The agents or therapies can be administered simultaneously (e.g., as a single formulation administered to the patient or as two separate formulations administered simultaneously) or sequentially in any order. Sequential administration is administration given at different times. The time between administration of one agent and administration of another agent can be minutes, hours, days, or weeks. The use of plasma kallikrein-binding antibodies described herein can also be used to reduce the dosage of another therapy, for example, to reduce side effects associated with the other agent being administered. Thus, a combination can include administering the second agent at a dosage at least 10, 20, 30, or 50% less than would be used in the absence of the plasma kallikrein-binding antibody. In some embodiments, a subject may receive C1 inhibitor as an intravenous (IV) or subcutaneous (SC) loading dose concurrently with the initial administration of an anti-pKal antibody (e.g., DX-2930) described herein. Thereafter, the subject can continue anti-pKal antibody treatment (without further administration of C1 inhibitor).

[0165] Combination therapy can include the administration of an agent that reduces the side effects of the other therapy, which can be an agent that reduces the side effects of treating a disease associated with plasma kallikrein.

[0166] (iv) Assays for evaluating treatment regimens Also within the scope of the present disclosure are assay methods for assessing the effectiveness of any of the treatment methods described herein. In some embodiments, to assess the effectiveness of a treatment, the plasma or serum concentration of one or more biomarkers associated with HAE (e.g., double-chain HMWK) may be measured before treatment and / or during the course of treatment (e.g., after the first dose). In some embodiments, the plasma or serum concentration (level) of one or more biomarkers associated with HAE obtained at a time point after administration of the dose is compared to the concentration of the biomarker in a sample obtained at an earlier time point after administration of the dose or before administration of the first dose. In some embodiments, the biomarker is 2-HMWK.

[0167] The level of a biomarker can be measured by detecting the biomarker in a plasma or serum sample obtained from a subject, for example, by immunoassay such as Western blot assay or ELISA, using an antibody that specifically detects the biomarker. In some embodiments, the level of 2-HWMK in a plasma or serum sample obtained from a subject is assessed by immunoassay. Antibodies for use in immunoassays to detect 2-HWMK are known to those skilled in the art, and the selection of such antibodies for use in the methods described herein will be clear to those skilled in the art.

[0168] Without further elaboration, it is believed that one skilled in the art can utilize the present invention to its fullest extent based on the preceding description. The following specific embodiments are, therefore, to be construed as merely illustrative, and not limitative of the remainder of the disclosure in any way. All publications cited herein are incorporated by reference for the purpose or subject matter discussed herein. [Example]

[0169] Example: Efficacy and Safety of Lanadelumab (DX-2930) Treatment in Human Pediatric Patients There is an unmet need for a safe, effective, and convenient long-term preventive therapy for pediatric patients under 12 years of age with hereditary angioedema (HAE). Lanadelumab (DX-2930), a fully human monoclonal antibody targeting plasma kallikrein, is approved for the routine prevention of recurrent HAE attacks in patients aged 12 years and older. We describe an open-label, multicenter, phase 3 study to investigate the safety, pharmacokinetics (PK), and pharmacodynamics (PD) of lanadelumab for the prevention of HAE attacks in pediatric patients aged 2 to under 12 years with type I or type II HAE.

[0170] Lanadelumab is a preservative-free sterile solution for injection (pH 6.0). The active ingredient, antibody DX-2930, is formulated using the following compendial ingredients: 30 mM sodium phosphate dibasic dihydrate, 19.6 mM citric acid monohydrate, 50 mM L-histidine, 90 mM sodium chloride, and 0.01% polysorbate 80. Each vial contains a nominal concentration of 150 mg of DX-2930 active ingredient in 1 mL of solution ("Test Product"). The Test Product is administered by subcutaneous (SC) injection into the upper arm in a blinded fashion.

[0171] The placebo will consist of an inactive formulation of the test product: 30 mM sodium phosphate dibasic dihydrate, 19.6 mM citric acid monohydrate, 50 mM L-histidine, 90 mM sodium chloride (pH 6.0) with 0.01% polysorbate 80. Placebo administration will occur for subjects randomized to the placebo treatment group and between doses of DX-2930 for subjects randomized to 150 mg of DX-2930 every 4 weeks.

[0172] Patients will be enrolled who are between 2 and 12 years old at the time of screening, have type I / II HAE, and have a documented attack of at least one per 12 weeks. There will be at least five patients in each age group (2 to 6 years old and 6 to 12 years old), and we plan to enroll at least 20 patients. Exploratory analyses will be conducted on subgroups with sufficient numbers of patients for Poisson regression. The inclusion and exclusion criteria are listed below.

[0173] Selection criteria included: Children (male or female) aged between 2 and 12 years at the time of screening. A documented diagnosis of HAE (Type I or Type II) based on both: Documented clinical history consistent with HAE (SC without urticaria or episodes of non-pruritic swelling of mucous membranes), Diagnostic test results obtained during screening from a sponsor-approved central laboratory confirming functional C1-INH levels are less than 40 percent (%) of normal. Participants with functional C1 esterase inhibitor (C1-INH) levels between 40 and 50% of normal levels may be enrolled if their complement 4 (C4) levels are also below the normal range. Participants may be retested during the baseline observation period, with prior sponsor approval, if the investigator believes the results are inconsistent with clinical history or confounded by recent complement 1 (C1) inhibitor use. Historical baseline HAE attack rate of at least one attack per 3 months. Note: Additionally, participants who experienced ≥ 1.0 angioedema attacks per 3 months during the 12-week baseline observation period and remained eligible according to the inclusion criteria will enter the lanadelumab treatment period. · Agree to comply with the schedule of treatments, evaluations, and procedures prescribed in the protocol. · Before any study-specific procedures are performed, the nature of the study is explained to them and they have a parent / legal guardian who is able to provide written informed consent for their child to participate in the study (with assent from the child, if appropriate). Women of childbearing potential must agree to abstain for the duration of the study, from screening through 70 days after the final study visit, or agree to comply with the applicable contraceptive requirements of this protocol.

[0174] Exclusion criteria included: · Concurrent diagnosis of another form of chronic, recurrent angioedema, such as acquired angioedema (AAE), HAE with normal C1-INH, idiopathic angioedema, or recurrent angioedema associated with urticaria. · Receipt of an investigational drug or exposure to an investigational device within 4 weeks prior to screening. · Pregnant or breastfeeding. - Androgen therapy (e.g., stanozolol, danazol, oxandrolone, methyltestosterone, and testosterone) was initiated within 2 weeks prior to the observation period. · Exposure to angiotensin-converting enzyme (ACE) inhibitors or estrogen-containing medications with systemic absorption (e.g., oral contraceptives or hormone replacement therapy) within 4 weeks prior to screening. Any active infectious illness or fever defined by oral temperature >38 degrees Celsius (°C) (100.4 degrees Fahrenheit [°F]), tympanic temperature >38.5°C (101.3°F), axillary temperature >38°C (100.4°F), or rectal / core temperature >38.5°C (101.3°F) within 24 hours prior to the first dose of study drug in Treatment Period A. Any unresolved HAE attack prior to the first dose of study drug in Treatment Period A. · Having any of the following liver function test abnormalities: alanine aminotransferase (ALT) more than 3 times the upper limit of normal (ULN), or aspartate aminotransferase (AST) more than 3 times the ULN, or total bilirubin more than 2 times the ULN (unless the elevated bilirubin is the result of Gilbert syndrome). · Have any condition (any surgical or medical condition) that, in the opinion of the investigator or sponsor, may compromise their safety or compliance, prevent the successful conduct of the study, or interfere with the interpretation of the results (e.g., a significant pre-existing illness or other major addiction that may confound the investigator's interpretation of the study results). The participant has a known hypersensitivity to the investigational product or any of its components.

[0175] Following a 4- to 12-week observation period (or less than 12 weeks if an HAE attack occurs; the "run-in period"), patients receive subcutaneous treatment with lanadelumab, as shown in Figure 1. Patients aged 2 to less than 6 years at the time of informed consent receive 150 mg every 4 weeks (q4weeks), and patients aged 6 to less than 12 years at the time of informed consent receive 150 mg every 2 weeks (q2wks). An interim analysis will occur after 26 weeks of treatment, and a final analysis will occur after a total of 52 weeks of treatment and 4 weeks of follow-up. If patients remain seizure-free for 6 months, the dosage of 150 mg every 2 weeks may be changed to 150 mg every 4 weeks. The primary endpoints of the study include safety and pharmacokinetic (PK) measures of lanadelumab treatment, including adverse events, vital signs (e.g., blood pressure, heart rate, temperature, and respiratory rate), clinical laboratories (e.g., hematology, clinical chemistry, coagulation), measurements of lanadelumab plasma concentrations, and other PK parameters in plasma. Secondary endpoints include clinical outcome measures, such as the number of HAE attacks during the treatment period; the number of HAE attacks requiring the use of acute therapy; the number of moderate or severe HAE attacks; time to first attack; characteristics of the HAE attacks (e.g., duration, severity, location of attacks, rescue medications used); achievement of an attack-free state during the evaluation period; pharmacodynamic (PD) measures, including plasma kallikrein activity (e.g., as measured via cleaved high-molecular-weight kininogen levels); and the immunogenicity of lanadelumab in the pediatric population studied (e.g., neutralizing and non-neutralizing anti-drug antibodies). Additional exploratory endpoints include quality of life assessments (e.g., Pediatric Quality of Life Generic Core Scale (PedsQL), PedsQL-PIM, and EQ-5D-Y questionnaire); exploratory pharmacodynamic endpoints, and the effects of lanadelumab on C1-INH, C4, HK1, and C1q.

[0176] Other embodiments All features disclosed herein may be combined in any combination. Each feature disclosed herein may be replaced by an alternative feature serving the same, equivalent, or similar purpose. Thus, unless expressly stated otherwise, each feature disclosed is only one example of a generic series of equivalent or similar features.

[0177] From the foregoing description, those skilled in the art can readily ascertain the essential features of the present invention, and can make various changes and modifications to the present invention to adapt it to various uses and conditions without departing from the spirit and scope thereof. Accordingly, other embodiments are also within the scope of the following claims.

[0178] equivalent While several inventive embodiments have been described and illustrated herein, those skilled in the art will readily envision various other means and / or structures for performing the functions and / or obtaining the results and / or one or more advantages described herein. Each such variation and / or modification is deemed to be within the scope of the inventive embodiments described herein. More generally, those skilled in the art will readily appreciate that all parameters, dimensions, materials, and configurations described herein are meant to be exemplary, and that the actual parameters, dimensions, materials, and / or configurations will depend on the specific application(s) for which the teachings of the present invention are used. Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific inventive embodiments described herein. Accordingly, it should be understood that the foregoing embodiments are presented by way of example only and that, within the scope of the appended claims and their equivalents, embodiments of the present disclosure are directed to each individual feature, system, article, material, kit, and / or method described herein. Additionally, any combination of two or more of such features, systems, articles, materials, kits, and / or methods, if such features, systems, articles, materials, kits, and / or methods are not mutually inconsistent, is within the inventive scope of the present disclosure.

[0179] All definitions, as defined and used herein, should be understood to supersede dictionary definitions and definitions in documents incorporated by reference and / or ordinary meanings of the defined terms.

[0180] The indefinite articles "a" and "an," as used herein in the specification and claims, unless clearly indicated to the contrary, should be understood to mean "at least one."

[0181] The phrase "and / or," as used herein in the specification and claims, should be understood to mean "either or both" of the elements so conjoined, i.e., elements present conjunctively in some cases and disjunctively in other cases. Multiple elements listed with "and / or" should be construed in the same manner, i.e., "one or more" of the elements so conjoined. Other elements may optionally be present other than the elements specifically identified by the "and / or" clause, whether related to those elements specifically identified or not. Thus, as a non-limiting example, a reference to "A and / or B," when used in conjunction with open-ended language such as "comprising," can refer, in one embodiment, to A only (optionally including elements other than B); in another embodiment, to B only (optionally including elements other than A); in yet another embodiment, to both A and B (optionally including other elements); and so forth.

[0182] As used herein in the specification and claims, "or" should be understood to have the same meaning as "and / or" as defined above. For example, when separating items in a list, "or" or "and / or" should be construed as inclusive, i.e., including at least one of any number or list of elements, but also including more than one element, optionally including additional unlisted items. Only terms clearly indicated otherwise, such as "only one of," "exactly one of," or, when used in the claims, "consisting of," refer to the inclusion of exactly one of any number or list of elements. In general, the term "or," when used herein with exclusive terms such as "either," "one of," "only one of," or "exactly one of," should only be construed as indicating exclusive alternatives (i.e., one or the other, but not both). "Consisting essentially of," when used in the claims, shall have its ordinary meaning as used in the field of patent law.

[0183] As used herein in the specification and claims, the phrase "at least one" in reference to a list of one or more elements should be understood to mean at least one element selected from any one or more of the elements in the list of elements, but not necessarily including at least one of each and every element specifically listed in the list of elements, and not excluding any combinations of elements in the list of elements. This definition also allows for elements other than those specifically identified in the list of elements to which the phrase "at least one" refers, may optionally be present, whether related to those specifically identified elements or not. Thus, as a non-limiting example, "at least one of A and B" (or, equivalently, "at least one of A or B," or, equivalently, "at least one of A and / or B") may refer to, in one embodiment, at least one A, optionally including two or more As, and no Bs (optionally including elements other than B); in another embodiment, at least one B, optionally including two or more Bs, and no As (optionally including elements other than A); in yet another embodiment, at least one A, optionally including two or more As, and at least one B, optionally including two or more Bs (optionally including other elements); etc.

[0184] Also, unless expressly stated to the contrary, it should be understood that in any method claimed herein including more than one step or action, the order of the method steps or actions is not necessarily limited to the order in which the method steps or actions are described.

[0185] In the claims and the above specification, all transitional phrases such as "comprising," "including," "carrying," "having," "containing," "involving," "holding," "composed of," and the like, are to be understood to mean open-ended, i.e., "including, but not limited to." Only the transitional phrases "consisting of" and "consisting essentially of" shall be closed or semi-closed transitional phrases, respectively, as set forth in the U.S. Patent and Trademark Office's United States Manual of Patent Examining Procedures, Section 2111.03.

Claims

1. 1. A method for treating or reducing the rate of hereditary angioedema (HAE) attacks, comprising: administering to a human subject in need thereof an antibody comprising the same complementarity-determining regions (CDRs) as DX-2930 during a first treatment period; during the first treatment period, the antibody is administered to the human subject multiple times at about 150 mg every two weeks or every four weeks; the human subject has, is suspected of having, or is at risk for HAE, and is a pediatric subject between the ages of 2 and 12; The method.

2. 10. The method of claim 1, wherein the human subject is between 2 and less than 6 years of age and the antibody is administered at about 150 mg every 4 weeks.

3. 10. The method of claim 1, wherein the human subject is between 6 and less than 12 years of age and the antibody is administered at about 150 mg every two weeks.

4. The method of any one of claims 1 to 3, wherein the antibody is a full-length antibody or an antigen-binding fragment thereof.

5. The method of any one of claims 1 to 4, wherein the antibody comprises a heavy chain variable region represented by SEQ ID NO:3 and / or a light chain variable region represented by SEQ ID NO:

4.

6. The method of any one of claims 1 to 5, wherein the antibody comprises a heavy chain represented by SEQ ID NO: 1 and a light chain represented by SEQ ID NO:

2.

7. The method of any one of claims 1 to 6, wherein the antibody is formulated into a pharmaceutical composition comprising a pharmaceutically acceptable carrier.

8. 8. The method of claim 7, wherein the pharmaceutical composition comprises sodium phosphate, citric acid, histidine, sodium chloride, and polysorbate 80.

9. 9. The method of claim 8, wherein the sodium phosphate is at a concentration of about 30 mM, the citric acid is at a concentration of about 19 mM, the histidine is at a concentration of about 50 mM, the sodium chloride is at a concentration of about 90 mM, and the polysorbate 80 is at about 0.01%.

10. The method of any one of claims 1 to 9, wherein the antibody is administered subcutaneously.

11. The method of any one of claims 1 to 10, wherein the human subject has type I or type II HAE.

12. 12. The method of any one of claims 1 to 11, wherein the human subject has an HAE attack rate of at least 1 HAE attack per 12 weeks prior to the first treatment period.

13. 13. The method of any one of claims 1 to 12, wherein the method further comprises administering the antibody to the subject for a second therapeutic period after the first therapeutic period.

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