Plasma kallikrein inhibitors and their use for treating hereditary angioedema attacks

Abstract

To provide methods for treating hereditary angioedema (HAE) attacks or reducing the rate of HAE attacks.SOLUTION: Provided is a composition comprising an antibody capable of binding to and inhibiting active human plasma kallikrein (pKal), the composition being administered such that the antibody is administered to a human subject multiple times at about 300 mg every two weeks during a first treatment period, the human subject having, being suspected of having, or being at risk for HAE, the human subject (i) being female; (ii) being under 18 years of age or between 40 and 65 years of age; (iii) having had at least one previous attack of laryngeal HAE; (iv) having had 1-2, 2-3, or 4 or more attacks of HAE in the four weeks prior to the first administration of the first treatment period; and / or (v) having been treated with a C1-inhibitor prior to the first treatment period.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] This application claims the benefit under 35 U.S.C. §119(e) of U.S. Provisional Application No. 62 / 725,216, filed August 30, 2018, and U.S. Provisional Application No. 62 / 808,612, filed February 21, 2019, each of which is incorporated by reference in its entirety herein. [Background technology]

[0002] Plasma kallikrein is a serine protease component of the contact system and a potential drug target for different inflammatory, cardiovascular, infectious (sepsis) and tumor diseases (Sainz IM et al., Thromb Haemost 98, 77-83, 2007). The contact system is activated either by factor XIIa upon exposure to foreign bodies or negatively charged surfaces, or by prolylcarboxypeptidase on the endothelial cell surface (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 potentiates inflammation through the production of the proinflammatory nonapeptide bradykinin. As the major kininogenase in the circulation, plasma kallikrein contributes significantly to bradykinin production in blood vessels. A genetic deficiency in the C1-inhibitor protein (C1-INH), the major natural inhibitor of plasma kallikrein, leads to hereditary angioedema (HAE). HAE patients suffer from acute attacks of painful edema, often precipitated by unknown triggers (Zuraw BLet al., N Engl J Med 359, 1027-1036, 2008). Summary of the Invention

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

[0004] In some aspects, the disclosure provides a method for treating hereditary angioedema (HAE) attacks or reducing the rate of HAE attacks, comprising administering (e.g., subcutaneously) any of the antibodies described herein (e.g., DX-2930) to a human subject in need thereof. In some embodiments, the antibody is administered to the subject in multiple doses of about 300 mg every two weeks in a first treatment period. In some embodiments, the subject is a female who has, is suspected of having, or is at risk for HAE; is under the age of 18 or between 40 and 65 years of age; and / or has previously experienced at least one laryngeal HAE attack.

[0005] In some aspects, the disclosure provides a method for treating hereditary angioedema (HAE) attacks or reducing the rate of HAE attacks, comprising administering (e.g., subcutaneously) any of the antibodies described herein (e.g., DX-2930) to a human subject in need thereof. In some embodiments, the antibody is administered to the subject at about 150 mg every 4 weeks, about 300 mg every 4 weeks, or about 300 mg every 2 weeks. In some embodiments, the subject is an adolescent between 12 and 18 years of age.

[0006] Any of the methods described herein may further comprise administering the antibody to the subject in a second treatment period after the first treatment period. In some embodiments, the first dose of the second treatment period is about 2 weeks after the last dose of the first treatment period. In some embodiments, the second treatment period comprises one or more doses of the antibody at about 300 mg. In some embodiments, the second treatment period comprises multiple doses of the antibody at about 300 mg every 2 weeks.

[0007] Any of the methods described herein may further comprise: (a) administering the antibody to the human subject in a single dose of about 300 mg after the first treatment period; and (b) if the subject experiences an HAE attack after (a), further administering the antibody to the subject in one or more doses of about 300 mg. In some embodiments, in step (b), the subject is administered the antibody in multiple doses of about 300 mg every two weeks. In some embodiments, the first dose in step (b) is within one week after the HAE attack. In some embodiments, the single dose in (a) and the first dose in (b) are separated by at least 10 days.

[0008] In any of the methods described herein, the human subject may have type I or type II HAE. For example, the subject may have experienced at least two HAE attacks per year prior to the first treatment period. In some embodiments, the subject has had at least one HAE attack in the four weeks prior to the first dose of the first treatment period, or at least two HAE attacks in the eight weeks prior to the first dose of the first treatment period.

[0009] In some embodiments, a subject treated by any of the methods described herein, including the use of any of the anti-pKal antibodies described herein (e.g., DX-2930), has undergone one or more HAE treatments prior to the first dose of anti-pKal antibody. Such prior HAE treatments may include C1-inhibitors (e.g., C1-INH), plasma kallikrein inhibitors (e.g., ecallantide), bradykinin receptor antagonists (e.g., icatibant), androgens (e.g., danazol), antifibrinolytics (e.g., tranexamic acid), or combinations thereof. Such subjects may be gradually transitioned from the prior HAE treatment to the anti-pKal antibody treatment described herein, with a tapering period. In some instances, the tapering period is about 2-4 weeks. The prior HAE treatment may be terminated either prior to the subject's first dose of the antibody or within 3 weeks after the first dose of the antibody. Alternatively, subjects may be transitioned directly from any prior HAE treatment to the anti-pKal antibody treatment described herein.

[0010] In some embodiments, the subject has not had a prior HAE treatment prior to the first dose of the anti-pKal antibody, hi some embodiments, the subject has not had a prior HAE treatment for at least 2 weeks prior to the first dose of the antibody.

[0011] In some embodiments, the subject has not received long-term prophylaxis for HAE or HAE treatment including angiotensin-converting enzyme (ACE) inhibitors, estrogen-containing medications, or androgens prior to the first treatment period, during the first treatment period, and / or during the second treatment period.

[0012] 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.

[0013] In any of the methods described herein, the antibody can be formulated into a pharmaceutical composition comprising a pharma- ceutically 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%.

[0014] The details of one or more embodiments of the invention are set forth in the description below. Other features and advantages of the invention will be apparent from the following drawings and detailed description of certain embodiments, and also from the appended claims. [Brief description of the drawings]

[0015] [Figure 1]Figures 1A-1C contain Poisson regression plots of investigator-confirmed HAE attacks during the treatment period (days 0-182) for patients based on the number of HAE attacks during the run-in period. Figure 1A: 1 to <2 HAE attacks per month during the run-in period. Figure 1B: 2 to <3 HAE attacks per month during the run-in period. Figure 1C: ≥3 HAE attacks per month during the run-in period. [Figure 2A] Figures 2A-2B contain diagrams showing HAE attack rates in patients previously treated with long-term prophylaxis with C1-inhibitor (C1-INH). Figure 2A: Historical (3-month) mean (standard deviation), HAE attack rates per month at baseline and during lanadelumab treatment (days 0-182). Figure 2B: Reduction in HAE attack rates in each of the HAE patients in the indicated lanadelumab treatment groups. [Figure 2B] Same as above. [Figure 3-1] Figures 3A-3C include plots of monthly HAE attack rates in adolescent subjects. Figure 3A shows a plot of the least squares mean (LS) estimated monthly attack frequency for adolescent patients versus placebo, with 95% confidence intervals. Figure 3B: A plot of monthly HAE attack frequency during lanadelumab treatment versus baseline for rollover and non-rollover adolescent subjects. Figure 3C shows a plot of the least squares mean estimated monthly attack rate for adolescent patients (versus placebo), with 95% confidence intervals, in each of the indicated lanadelumab treatment groups. [Figure 3-2] Same as above. [Figure 4-1] Figures 4A-4E show plots of the percentage reduction in HAE attack rate from placebo with 95% confidence intervals for each of the indicated demographics. Figure 4A: Age; Figure 4B: Gender; Figure 4C: Weight; Figure 4D: HAE type; Figure 4E: History of laryngeal attacks. For each of the indicated groups, the bars correspond, from left to right, to 150 mg every 4 weeks, 300 mg every 4 weeks, and 300 mg every 2 weeks. The "n" below the plots refers to the number of subjects in each group. [Figure 4-2] Same as above. [Figure 4-3] Same as above. [Diagram 5]FIG. 5 shows a forest plot of incidence rate ratios of the number of investigator-confirmed HAE attacks based on the demographics indicated. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0016] definition For convenience, prior to description of the invention, certain terms employed in the specification, examples and appended claims are defined herein. Other terms are defined as they appear herein.

[0017] The singular forms "a," "an," and "the" include plural references unless the context clearly indicates otherwise.

[0018] As used herein, the term "about" refers to + / - 5% of a particular value. For example, about 300 mg of antibody includes any amount of antibody between 285 mg and 315 mg.

[0019] The term "antibody" refers to an immunoglobulin molecule capable of specifically binding 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 made up of heavy chain immunoglobulin variable domains (V H At least one heavy (H) chain containing a light chain immunoglobulin variable domain (V L For example, an antibody can include at least one light chain comprising a heavy (H) chain variable region (referred to herein as V H or HV) and the light (L) chain variable region (herein referred to as V L or LV). In another example, the antibody comprises two heavy (H) chain variable regions and two light (L) chain variable regions.

[0020] As used herein, the term "antibody" includes not only intact (i.e., full-length) polyclonal or monoclonal antibodies, but also antigen-binding fragments thereof (Fab, Fab', F(ab')2, Fv, etc.), single chain (scFv), domain antibody (dAb) fragments (de Wildt et.al., Euro. J. Immunol. (1996) 26(3):629-639), mutants of any of these, 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 configuration of an immunoglobulin molecule that contains an antigen recognition site with 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), but an antibody need not be of any particular class. Depending on the antibody amino acid sequence of the constant domain of its heavy chain, immunoglobulins can be assigned to different classes. 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, although primate (human and non-human primate) and primatized antibodies are preferred.

[0021] V H and / or V LA region may comprise all or a portion of the amino acid sequence of a naturally occurring variable domain. For example, the sequence may be deleted of one, two or more N- or C-terminal amino acids, internal amino acids, may include one or more insertions or additional terminal amino acids, or may contain other alterations. In one embodiment, a polypeptide comprising an immunoglobulin variable domain sequence is capable of associating with another immunoglobulin variable domain sequence to form a structure that preferentially interacts with an antigen binding site, e.g., plasma kallikrein.

[0022] V H and V L The regions can be further subdivided into regions of hypervariability, termed "complementarity determining regions" ("CDRs"), interspersed with regions of greater conservedness, 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, USDepartment 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 amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.

[0023] V H Or V LIn addition to the domains, 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, the antibody is a tetramer of two heavy and two light immunoglobulin chains, the heavy and light immunoglobulin chains being 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 regions of the antibody generally mediate 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.

[0024] One or more regions of the antibody can be human or effectively human. For example, one or more of the 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 of the framework regions can be human, e.g., FR1, FR2, FR3 and / or FR4 of the HC and / or LC. For example, the Fc region can be human. In one embodiment, the framework regions are all 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, the framework (FR) residues of the selected Fab can be converted to the amino acid type of the corresponding residue 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 domain, the constant region, the constant domains (CH1, CH2, CH3 and / or CL1) or the entire antibody can be human or effectively human.

[0025] Antibodies may be encoded by immunoglobulin genes or segments thereof. Exemplary human immunoglobulin genes include 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" (about 25 KDa or about 214 amino acids) are encoded by a variable region gene at the NH2-terminus (about 110 amino acids) and a kappa or lambda constant region gene at the COOH-terminus. Full-length immunoglobulin "heavy chains" (about 50 KDa or about 446 amino acids) are similarly encoded by a variable region gene (about 116 amino acids) and one of the other constant region genes mentioned above, such as gamma (encoding about 330 amino acids). The length of the human HC varies considerably, as the HC CDR3 varies from about 3 amino acid residues to more than 35 amino acid residues.

[0026] 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 binding fragments encompassed within the term "antigen-binding fragment" of a full-length antibody and that retain functionality include: (i) a Fab fragment, i.e., a V L , V H , C L and the CH1 domain; (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 fragment of a single arm of an antibody. L and V H (v) an Fv fragment consisting of a V H (vi) isolated complementarity determining regions (CDRs); and (vi) dAb fragments consisting of two domains, the V and VD domains (Ward et al., (1989) Nature 341:544-546). L and V H are encoded by individual genes, which are collectively referred to as V L and V HThe regions can be linked using recombinant methods by synthetic linkers that allow them to be produced as a single protein chain that pairs to form a monovalent molecule known as a single chain Fv (scFv). See, for example, 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.

[0027] The term "monospecific antibody" refers to an antibody that displays a single binding specificity and affinity for a particular target, e.g., epitope. The term includes "monoclonal antibody" or "monoclonal antibody composition," which as used herein refers to a preparation of antibodies or fragments thereof of single molecular composition, regardless of how the antibody was made. 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 the binding characteristics are substantially retained.

[0028] Inhibition constant (K i ) provides a measure of inhibitory 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 different substrate concentrations by measuring the inhibitory effect of different concentrations of an inhibitor (e.g., an inhibitory binding protein) on the extent of a reaction (e.g., enzyme activity); the apparent K is determined by fitting the change in the pseudo-first-order rate constant as a function of inhibitor concentration to the Morrison equation (Equation 1): i This gives an estimate of the value of K i is K i,app The y-intercept of the plot versus substrate concentration is derived from linear regression analysis.

number

[0029] As used herein, "binding affinity" refers to the apparent association constant or K A Refers to. A is the dissociation constant (K D ) is the reciprocal of the K for binding to the first target. A binding antibody can have a binding affinity for a particular target molecule, e.g., plasma kallikrein, of, for example, at least 10, 10, 10, 10, 10, and 10 M. A higher affinity binding of a binding antibody to a first target compared to a second target is indicative of a higher K for binding to the first target. A is the K for binding of the second target A (or number K D ) higher than (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) compared to 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., for specificity or other comparison) may be 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.

[0030] 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). Using these techniques, the concentration of bound and free binding protein can be measured 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 for the binding protein on the target, where (N) is the number of binding sites per target molecule according to the following formula: [Bound]=N·[Free] / ((1 / KA)+[Free]).

[0031] K A It is not necessary to make an exact determination of K, which is sometimes determined using methods such as ELISA or FACS analysis. A It is sufficient to obtain a quantitative measure of affinity that is proportional to the affinity of the target protein, and thus, for example, a functional assay, such as activity in an in vitro or in vivo assay, can be used for comparison, such as determining whether the affinity is higher, e.g., 2-fold higher, to obtain a quantitative measure of affinity or to obtain an inference of affinity.

[0032] The term "binding antibody" (or used interchangeably herein with "binding protein") 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 when compared to the activity of plasma kallikrein in the absence of the antibody and under the same conditions.

[0033] 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 having 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), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine).

[0034] One or more framework and / or CDR amino acid residues of the binding protein can include one or more mutations (e.g., substitutions (e.g., conservative substitutions or substitutions of non-essential amino acids), insertions or deletions) compared to the binding proteins described herein. The plasma kallikrein binding protein can have mutations (e.g., substitutions (e.g., conservative substitutions or substitutions of non-essential amino acids), 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 the framework regions, CDRs and / or constant regions. In some embodiments, the mutations are present in the framework regions. In some embodiments, the mutations are present in the CDRs. In some embodiments, the mutations are present in the constant regions. Whether a particular substitution is tolerated, i.e., does not have a deleterious effect on 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. (1990) Science 247:1306-1310.

[0035] An "effectively human" immunoglobulin variable region is one that contains a sufficient number of human framework amino acid positions such that the immunoglobulin variable region does not elicit an immunogenic response in normal humans. A "substantially human" antibody is one that contains a sufficient number of human amino acid positions such that the antibody does not elicit an immunogenic response in normal humans.

[0036] "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 the protein's amino acids (e.g., glycosyl moieties), or a combination thereof. Overlapping epitopes include at least one common amino acid residue, glycosyl group, phosphate group, sulfate group, or other molecular feature.

[0037] 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. Descriptions of "humanized" immunoglobulins include, for example, U.S. Patent Applications 6,407,213 and 5,693,762.

[0038] An "isolated" antibody refers to an antibody that is removed from at least 90% of at least one component of the natural sample from which it may be obtained. An antibody may be "at least" of a certain degree of purity 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-by-weight basis.

[0039] 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 be a subject who has undergone prior 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, a child, or an adolescent subject). In some embodiments, the human subject is an adolescent less than 18 years of age. In some embodiments, the human subject is an adolescent between 12-18 years of age. In some embodiments, the subject is less than 40-65 years of age.

[0040] In some embodiments, the human subject is defined by gender, e.g., in some embodiments, the subject is female.

[0041] In some embodiments, the human subject is defined by body weight. In some embodiments, the human subject weighs less than 50 kg. In some embodiments, the human subject weighs between 50 kg and 75 kg. In some embodiments, the human subject weighs between 75 kg and 100 kg. In some embodiments, the human subject weighs 100 kg or more.

[0042] In some embodiments, the human subject is defined by a history or lack of laryngeal attacks. In some embodiments, the subject has experienced at least one (e.g., 1, 2, 3, 4, 5 or more) laryngeal attack (i.e., laryngeal HAE attack) prior to administration of an antibody described herein. In some embodiments, the subject has not experienced a laryngeal attack prior to administration of an antibody described herein.

[0043] 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.

[0044] 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 (e.g., having similar side chains, e.g., conservative amino acid substitutions) amino acid residues or nucleotides to 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 compared to the same antigen.

[0045] Statistical significance can be determined by any method known in the art. Representative statistical tests include the Student's T-test, the Mann Whitney U nonparametric test, and the Wilcoxon nonparametric statistical test. Some statistically significant relationships have a P value of less than 0.05 or 0.02. Certain binding proteins can show statistically significant differences, for example, in specificity or binding (e.g., P value < 0.05 or 0.02). For example, the terms "induce," "inhibit," "enhance," "elevate," "increase," "decrease," and the like, which indicate a discernible qualitative or quantitative difference between two situations, can refer to a difference, for example, a statistically significant difference between two situations.

[0046] A "therapeutically effective dose" preferably modulates a measurable parameter, such as plasma kallikrein activity, to a statistically significant extent, or at least about 20%, more preferably at least about 40%, even more preferably at least about 60% and also more preferably at least about 80% compared to untreated subjects. The ability of a compound to modulate a measurable parameter, such as a disease-related parameter, can be evaluated in an animal model system predictive of efficacy in human disorders and conditions. Alternatively, this property of a composition can be evaluated by examining the ability of the compound to modulate the parameter in vitro.

[0047] The term "treating" as used herein refers to the application or administration of a composition comprising one or more active agents to a subject having HAE, having symptoms of HAE, suspected of having HAE, or having a predisposition to or risk for HAE, with the intent to cure, heal, alleviate, alleviate, alter, treat, ameliorate, improve, or affect the disease, symptoms of the disease, or predisposition to the disease. "Prophylactic treatment," also known as "preventative treatment," refers to treatment that is intended to protect a person from or reduce the risk of a disease to which the person has been or may be exposed. In some embodiments, the treatment methods described herein are intended to prevent the onset and / or recurrence of HAE.

[0048] The term "preventing" a disease in a subject refers to administering a pharmaceutical treatment, e.g., a drug, to a subject such that at least one symptom of the disease is prevented, i.e., administered prior to the onset of clinical symptoms of an undesirable condition (e.g., a disease or other undesirable condition in a host animal) such that the host is protected from the onset of the undesirable condition. "Preventing" a disease may also be referred to as "prophylaxis" or "prophylactic treatment."

[0049] A "prophylactically effective amount" refers to 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.

[0050] Antibody binding to plasma kallikrein (pKal) Plasma kallikrein binding antibodies (anti-pKal antibodies) for use in the methods described herein can be full length (e.g., IgG (including IgG1, IgG2, IgG3, IgG4), IgM, IgA (including IgA1, IgA2), IgD and IgE) or can comprise only antigen-binding fragments (e.g., Fab, F(ab')2 or scFv fragments. The binding antibody can comprise two heavy chain immunoglobulins and two light chain immunoglobulins, or can be a single chain antibody. The plasma kallikrein binding antibody can be a recombinant protein, such as a humanized, CDR-grafted, chimeric, deimmunized or in vitro generated antibody, and can optionally comprise a constant region derived from human germline immunoglobulin sequences. In one embodiment, the plasma kallikrein binding antibody is a monoclonal antibody.

[0051] 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 comprises at least one immunoglobulin variable region. For example, the antibody comprises 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.

[0052] In some embodiments, the antibodies described herein have the same CDR sequences as DX-2930, 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 whole 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 whole 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., in whole 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., in whole or in framework regions) to an HC sequence described herein.

[0053] The plasma kallikrein binding protein can 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,app The plasma kallikrein-binding antibody is isolated from antibody cleavage fragments (e.g., DX-2930) that bind to plasma kallikrein at a specific concentration. 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 antibody cleavage fragments that are inactive or partial.

[0054] The plasma kallikrein binding antibody may further inhibit plasma kallikrein, for example, human plasma kallikrein.

[0055] In some embodiments, the plasma kallikrein binding antibody does not bind to prekallikrein (eg, human prekallikrein and / or mouse prekallikrein), but binds to the active form of plasma kallikrein (eg, human plasma kallikrein and / or mouse kallikrein).

[0056] 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.

[0057] The antibody is at least 10 5 , 10 6 , 10 7 , 10 8 , 10 9 , 10 10 and 10 11 M -1 In one embodiment, the antibody can bind to plasma kallikrein, e.g., human plasma kallikrein, 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 1×10 2 , 1×10 3 Or 5 x 10 3 M -1 s -1 Faster K onIn 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 effectively than it binds to plasma kallikrein (e.g., 5, 10, 50, 100, or 1000-fold less, or not at all, when compared to a negative control).

[0058] In one embodiment, the antibody is, for example, 10 -5 , 10 -6 , 10 -7 , 10 -8 , 10 -9 and 10 -10 The antibodies inhibit human plasma kallikrein activity with a Ki of less than 1 M. The antibodies have an IC of less than 100 nM, 10 nM, 1, 0.5, or 0.2 nM, for example. 50 For example, the antibody may modulate plasma kallikrein activity and production of factor XIIa (e.g., from factor XII) and / or bradykinin (e.g., from high molecular weight kininogen (HMWK)). The antibody may inhibit plasma kallikrein activity and / or production of factor XIIa (e.g., from factor XII) and / or bradykinin (e.g., from high molecular weight kininogen (HMWK)). The affinity of the antibody for human plasma kallikrein may be less than 100 nm, less than 10 nM, less than 5 nM, less than 1 nM, 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, when compared to a negative control).

[0059] In some embodiments, the antibodies have an apparent inhibition constant (K i,app ) is less than 1000, 500, 100, 5, 1, 0.5 or 0.2 nM.

[0060] Plasma kallikrein binding antibodies may have their HC and LC variable domain sequences contained in a single polypeptide (eg, scFv) or may be on different polypeptides (eg, IgG or Fab).

[0061] 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 comprises a Fab2', scFv, minibody, scFv::Fc fusion, Fab::HSA fusion, HSA::Fab fusion, Fab::HSA::Fab fusion, or other molecule that comprises the antigen combining site of one of the binding proteins herein. The VH and VL regions of these Fabs can be provided as 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 constructs.

[0062] 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, 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.

[0063] 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, such as all primate framework regions or framework regions that are at least 85, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% identical to a primate framework region. In one embodiment, the antibody comprises a primate Fc domain or an Fc domain that is at least 95, 96, 97, 98, or 99% identical to a primate Fc domain. "Primates" includes humans (Homo sapiens), chimpanzees (Pan troglodytes and Pan paniscus (bonobo)), gorillas (Gorilla gorilla), gibbons, monkeys, lemurs, aye-ayes (Daubentonia madagascariensis), and tarsiers.

[0064] In some embodiments, the affinity of the primate antibody for human plasma kallikrein is less than 1000, 500, 100, 10, 5, 1, 0.5 nM, such as less than 10 nM, less than 1 nM, and less than 0.5 nM. D It is characterized by:

[0065] In certain embodiments, the antibody does not contain sequences derived from a mouse or rabbit (eg, is not a mouse or rabbit antibody).

[0066] In some embodiments, the antibody used in the methods described herein can be DX-2930 or a functional variant thereof as described herein.

[0067] In one example, the functional variant of DX-2930 contains the same complementarity determining regions (CDRs) as DX-2930. In another example, the functional variant of DX-2930 contains the V H and V L Compared to the case of V H Or V LPreferably, such mutations do not occur at residues predicted to interact with one or more of the CDRs, as can be determined by routine techniques. In other embodiments, the functional variants described herein contain one or more mutations (e.g., 1, 2, or 3) within one or more CDR regions of DX-2930. Preferably, such functional variants retain the same regions / residues that contribute to antigen binding as the parent. In yet other embodiments, the functional variants of DX-2930 contain one or more mutations (e.g., 1, 2, or 3) within one or more CDR regions 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 has 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 the active form of plasma kallikrein and preferably do not bind to prekallikrein.

[0068] 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, modified as in Karlin and Altschul Proc. Natl. Acad. Sci. USA 90:5873-77, 1993. Such an algorithm is 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 with the XBLAST program, score=50, wordlength=3, to obtain amino acid sequences homologous to protein molecules 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.

[0069] In some embodiments, the antibody used in the methods and compositions described herein can be a DX-2930 antibody. The full length heavy and light chain and variable sequences for DX-2930 are provided below, with the signal sequence in italics. The CDRs are in bold and underlined.

[0070] [ka]

[0071] [Table 1]

[0072] 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).

[0073] The sequence encoding the antibody of interest, e.g., DX-2930, can be maintained in a vector in a host cell, which can then be propagated and frozen for later use. In the alternative, the polynucleotide sequence can be used for genetic engineering to "humanize" the antibody or to improve the affinity (affinity maturation) or other characteristics of the antibody. For example, the constant region can be engineered to be more similar to human constant regions to avoid immune reactions when the antibody is used in human clinical trials and treatments. It may be desirable to genetically engineer the antibody sequence to obtain higher affinity for the target antigen and greater efficacy in inhibiting the activity of PKal. It will be apparent to one of skill in the art that one or more polynucleotide changes can be made to an antibody and still maintain its binding specificity for the target antigen.

[0074] In other embodiments, fully human antibodies can be obtained by using commercially available mice that have been modified to express specific human immunoglobulin proteins. For the production of humanized or human antibodies, transgenic animals designed to generate a more desirable (e.g., fully human antibodies) or more robust immune response can also be used. 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 can be recombinantly produced by phage display or yeast technology. See, e.g., U.S. Pat. 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.

[0075] Antigen-binding fragments of intact antibodies (full-length antibodies) can be prepared via conventional methods, for example, F(ab')2 fragments can be produced by pepsin digestion of the antibody molecule and Fab fragments can be generated by reducing the disulfide bridges of the F(ab')2 fragment.

[0076] 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 for a target antigen can be readily isolated or synthesized. The DNA can be placed into one or more expression vectors, which are then introduced into host cells, such as Escherichia coli (E. coli) cells, monkey COS cells, Chinese hamster ovary (CHO) cells or myeloma cells, that do not otherwise express immunoglobulin proteins, to obtain the synthesis of monoclonal antibodies in the recombinant host cells. See, for example, WO 87 / 04462. The DNA can then be modified, for example, by substituting the coding sequences for human heavy and light chain constant domains in place of 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.

[0077] 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).

[0078] 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 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. The same molecular modeling analysis is then used to identify framework amino acid residues that are predicted to be important for the formation of accurate CDR structures. In parallel, the parent V is used as a search query. H and V L The sequence is used to identify human VH1 antibodies from any antibody gene database that have amino acid sequences that are homologous to the amino acid sequence of the parent non-human antibody. H and V L Next, identify the human V H and V L An acceptor gene is selected.

[0079] The CDR regions in the selected human acceptor 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 predicted to be important in interactions with the CDR regions (see above) can be used to substitute for the corresponding residues in the human acceptor gene.

[0080] Single chain antibodies can be prepared via recombinant techniques by linking a nucleotide sequence encoding a heavy chain variable region and a nucleotide sequence encoding a light chain variable region. Preferably, a flexible linker is incorporated between the two variable regions. Alternatively, techniques described for the production of 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 according to conventional procedures. Positive clones can be subjected to further screening to identify clones that inhibit PKal activity.

[0081] Some antibodies, e.g., Fabs, can be made 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 moiety and a bacteriophage protein (or a fragment thereof), the vector nucleic acid can be transferred into a bacterial cell 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.

[0082] Antibodies may also be produced in eukaryotic cells. In one embodiment, the antibodies (e.g., scFv's) are 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.The skilled artisan will be familiar with the various aspects of the present invention, including, for example, oxygen conditions (see, for example, Baumann K., et al. 2010 BMC Syst. Biol. 4:141), osmolarity (see, for example, Dragosits, M. et al., 2010 BMC Genomics 11:207), temperature (see, for example, Dragosits, M. et al., 2009 J Proteome Res. 8(3):1380-92), fermentation conditions (see, for example, Ning, D. et al. 2005 J. Biochem. and Mol. Biol. 38(3):294-299), yeast strains (see, for example, 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. Antibody production in yeast can be optimized by optimizing the acidity of the culture (see, e.g., Gasser, B. 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 Biotechol. 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), substrate and / or ion concentration (see, e.g., Ko JH. et al. 2996 Appl Biochem Biotechnol 60(1):41-8). Additionally, yeast systems can be used to produce antibodies with longer half-lives (see, e.g., Smith, BJ. et al. 2001 Bioconjug Chem 12(5):750-756).

[0083] In a preferred embodiment, the antibody is produced in a mammalian cell. Preferred mammalian host cells for expressing the clonal antibody or antigen-binding fragment thereof include Chinese Hamster Ovary (CHO) cells (including dhfr-CHO cells as described in Urlaub and Chasin, 1980, Proc. Natl. Acad. Sci. USA 77:4216-4220, used with the DHFR selectable marker as described in Kaufman and Sharp, 1982, Mol. Biol. 159:601 621), lymphoid cell lines such as NS0 myeloma cells and SP2 cells, COS cells, HEK293T cells (J. Immunol. Methods (2004) 289(1-2):65-80) and cells from transgenic animals, such as transgenic mammals. For example, the cell is a mammalian epithelial cell.

[0084] 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).

[0085] In addition to the nucleic acid sequence encoding the diversified immunoglobulin domain, the recombinant expression vector may carry additional sequences, such as sequences that control replication of the vector in a host cell (e.g., origins of replication) and selectable marker genes. The selectable marker gene facilitates selection of a host cell 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, typically the selectable marker gene confers resistance to drugs, such as G418, hygromycin or methotrexate, in a host cell into which the vector has been introduced. Preferred selectable marker genes include the dihydrofolate reductase (DHFR) gene (dhfr gene with methotrexate selection / amplification). - The promoter and promoter region include the ribosome-specific nucleotide sequence (for use in a host cell) and the neo gene (for G418 selection).

[0086] In an exemplary system for recombinant expression of an antibody or antigen-binding portion thereof, dhfr expression is achieved by calcium phosphate-mediated gene transfer. - A recombinant expression vector encoding both the antibody heavy chain and the antibody light chain is introduced into the 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., CMV enhancer / AdMLP promoter regulatory element or SV40 enhancer / AdMLP promoter regulatory element, e.g., derived from SV40, CMV, adenovirus, etc.) to drive high transcription levels of the genes. The recombinant expression vector also carries a DHFR gene, which allows for selection of CHO cells transfected with the vector using methotrexate selection / amplification. The selected transformant host cells are cultured, and intact antibody is recovered from the medium, allowing expression of the antibody heavy and light chains. 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 coupling matrix.

[0087] For antibodies that include an Fc domain, the antibody production system may 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 the site for modification with biantennary oligosaccharides. It has been shown that this glycosylation is required 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 include other eukaryotic post-translational modifications.

[0088] Antibodies can also be produced by transgenic animals. For example, US Patent No. 5,849,992 describes a method for expressing an antibody in the mammary gland of a transgenic mammal. A transgene is constructed that contains a milk-specific promoter and a nucleic acid encoding the antibody of interest and a signal sequence for secretion. The milk produced by females of such transgenic mammals contains the antibody of interest secreted therein. The antibody can be purified from the milk or, for some applications, can be used directly.

[0089] Pharmaceutical Compositions The antibodies described herein (e.g., DX-2930) can be present in a composition, e.g., a pharma- ceutically acceptable composition or pharmaceutical composition. The antibodies described herein (e.g., DX-2930) can be formulated with a pharma- ceutically acceptable carrier. In some embodiments, 150 mg or 300 mg of DX-2930 antibody is present in a composition, e.g., a pharma- ceutically acceptable composition or pharmaceutical composition, optionally with a pharma- ceutically acceptable carrier.

[0090] Pharmaceutically acceptable carriers include any and all 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.

[0091] The pharma- ceutically 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 a suitable 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 can 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 pharma- ceutically acceptable carrier comprises sodium phosphate and citric acid, which can be at a concentration of about 30 mM and about 19 mM, respectively.

[0092] In some embodiments, the pharma- ceutically acceptable carrier includes one or more amino acids that may reduce aggregation of the antibody during storage prior to administration and / or improve the stability of the antibody. 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 the amino acid in the pharmaceutical composition may 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 contains histidine at a concentration of about 50 mM.

[0093] Any suitable osmotic modifier may be used to prepare the pharmaceutical compositions described herein. In some embodiments, the osmotic modifier 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 osmotic modifier 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 osmotic 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 osmolarity modifier can be sodium chloride, which can be at a concentration of about 90 mM.

[0094] The pharma- ceutically acceptable carrier in the pharmaceutical compositions described herein may further include one or more pharma- ceutically acceptable excipients. In general, pharma- ceutically 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), hydroxypropyl methylcellulose (HPMC), polyethyleneimine (PEI), gelatin, polyvinylpyrrolidone (PVP), hydroxyethylcellulose (HEC), polyethylene glycol (PEG), ethylene glycol, glycerol, dimethylsulfoxide (DMSO), dimethylformamide (DMSO), and the like. 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 ion, copper ion, calcium ion, manganese ion, magnesium ion, CHAPS, sucrose monolaurate, and 2-O-beta-mannoglycerate. In some embodiments, the pharma- ceutically acceptable carrier comprises 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 the 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 pharma- ceutically acceptable carrier contains 0.01% Tween-80.

[0095] In some examples, the pharmaceutical compositions described herein include an anti-pKal antibody also described herein (e.g., DX-2930) and one or more of sodium phosphate (e.g., dibasic sodium phosphate dihydrate), citric acid (e.g., citric acid monohydrate), histidine (e.g., L-histidine), sodium chloride, and polysorbate 80. For example, the pharmaceutical composition may include 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 or 300 mg / mL. In one example, the composition comprises or consists of about 150 mg DX-2930 / 1 mL solution, about 30 mM dibasic sodium phosphate 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. In another example, the composition comprises or consists of about 300 mg DX-2930 / 1 mL solution, about 30 mM dibasic sodium phosphate 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.

[0096] Pharmaceutically acceptable salts are salts that retain the desired biological activity of the compounds and do 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 salts derived from non-toxic inorganic acids, such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, phosphorous acid, and the like, as well as salts derived from non-toxic organic acids, such as aliphatic mono- and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxyalkanoic acids, aromatic acids, aliphatic and aromatic sulfonic acids, and the like. Base addition salts include salts derived from alkaline earth metals, such as sodium, potassium, magnesium, calcium, and the like, as well as salts derived from non-toxic organic amines, such as N,N'-dibenzylethylenediamine, N-methylglucamine, chloroprocaine, choline, diethanolamine, ethylenediamine, procaine, and the like.

[0097] 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., injectable and infusible solutions), dispersions or suspensions, tablets, pills, powders, liposomes and suppositories. The form may depend on the intended mode of administration and therapeutic application. Many compositions are in the form of injectable or infusible solutions, such as compositions similar to those used for administering 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.

[0098] 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.

[0099] 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 in a suitable solvent with one or a combination of the above listed ingredients, followed by filtration sterilization as required. In general, dispersions are prepared by incorporating the active compound into a sterile vehicle containing a basic dispersion medium and the required other ingredients from the above listed ingredients. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred preparation method is vacuum drying and freeze-drying, which produces a powder of the active ingredient plus any additional desired ingredients from the solution previously sterilized by filtration. The proper fluidity of the solution can be maintained, for example, by the use of a coating such as lecithin, by maintaining the required particle size in the case of dispersion, and by the use of surfactants. Prolonged absorption of injectable compositions can be achieved by including in the composition an agent that delays absorption, for example, monostearate salts and gelatin.

[0100] 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, for some therapeutic applications, the antibody may be administered at a dose of about 1-100 mg / m 2 Or 7 to 25 mg / m 2The antibody can be administered by intravenous infusion at a rate of less than 30, 20, 10, 5 or 1 mg / min to reach a dose of 100 mg / min. The route and / or mode of administration varies depending on the desired results. In certain embodiments, the active compound can be prepared with a carrier that will protect the compound against rapid release, such as a controlled release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters and polylactic acid. Many methods are available for the preparation of such formulations. See, for example, Sustained and Controlled Release Drug Delivery Systems, JR Robinson, ed., 1978, Marcel Dekker, Inc., New York.

[0101] The pharmaceutical composition may be administered using a medical device. For example, in one embodiment, the pharmaceutical compositions disclosed herein may be administered with a device, such as a needleless hypodermic injection device, a pump, or an indwelling.

[0102] In certain embodiments, the antibodies described herein (e.g., DX-2930) can be formulated to ensure proper distribution in vivo. For example, the blood-brain barrier (BBB) ​​excludes many highly hydrophilic compounds. To ensure that the therapeutic compounds disclosed herein (if necessary) cross the BBB, they can be formulated, for example, in liposomes. For methods of producing liposomes, see, for example, U.S. Pat. Nos. 4,522,811; 5,374,548; and 5,399,331. Liposomes can contain one or more moieties that selectively transport to specific cells or organs, thus enhancing targeted drug delivery (see, for example, VV Ranade, 1989, J. Clin. Pharmacol. 29:685).

[0103] The dosage regimen is 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 tapered or gradually 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. Unit dosage form, as used herein, refers to a physically discrete unit suitable as a unitary dosage for the subject to be treated; each unit contains a predetermined amount of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specifications for the unit dosage form are governed by and directly depend on (a) the unique characteristics of the active compound and the particular therapeutic effect to be achieved, and (b) the inherent limitations in the art of formulating such active compounds for the treatment of susceptibility in an individual.

[0104] 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 or 300 mg. As will be appreciated by one of skill in the art, the therapeutically or prophylactically effective amount of an antibody may be lower for a pediatric subject than for an adult subject. 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 or 300 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 in a first treatment period. In some embodiments, the antibody may be administered to a subject in a second treatment period. In some embodiments, the therapeutically or prophylactically effective amount of an antibody in a first treatment period is different from the therapeutically or prophylactically effective amount of an antibody in a second treatment period. In some embodiments, a therapeutically or prophylactically effective amount of an antibody in a first treatment period is 150 mg and a therapeutically or prophylactically effective amount of an antibody in a second treatment period is 300 mg. In some embodiments, the therapeutically or prophylactically effective amount of the antibody in the first treatment period is the same as the therapeutically or prophylactically effective amount of the antibody in the second treatment period. In one example, the therapeutically or prophylactically effective amount of the antibody in the first treatment period and the second treatment period is 300 mg.

[0105] In some embodiments, an exemplary non-limiting range for a therapeutically or prophylactically effective amount of an antibody described herein (e.g., DX-2930) is about 300 mg. In some embodiments, a therapeutically or prophylactically effective amount of an antibody is administered in a single dose. If the subject experiences an HAE attack, the antibody may be further administered to the subject in multiple doses, such as administering a dose of about 300 mg every two weeks.

[0106] kit An antibody described herein (e.g., DX-2930) can be provided in a kit, e.g., as a component of a 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 relating to the methods described herein and / or use of the antibody described herein (e.g., DX-2930), e.g., for the methods described herein. In some embodiments, the kit includes one or more doses of DX-2930. In some embodiments, the one or more doses is 150 mg or 300 mg.

[0107] The informational material of the kit is not limited in its form. In one embodiment, the informational material may include information regarding the preparation of the compound, the molecular weight of the compound, concentration, expiration date, batch or place of manufacture information, etc. In one embodiment, the informational material relates to using the antibody to treat, prevent or diagnose disorders and conditions, e.g., plasma kallikrein-associated diseases or conditions.

[0108] 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 the methods described herein, e.g., at an appropriate dose, dosage form, mode of administration, or administration schedule (e.g., at a dose, dosage form, administration 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 plasma kallikrein-associated disease or condition. For example, the material can include instructions for administering an antibody described herein (e.g., DX-2930) to a patient having a disorder or condition described herein, e.g., a plasma kallikrein-associated disease, e.g., according to an administration schedule described herein. The informational material of the kit is not limited in its form. In many cases, the informational material, e.g., instructions, is provided in printed form, but can also be in other formats, such as computer-readable material.

[0109] The antibodies described herein (e.g., DX-2930) may be provided in any form, such as liquid, dried or lyophilized form. The antibodies are preferably substantially pure and / or sterile. If the antibodies are provided in a liquid solution, the liquid solution is preferably an aqueous solution, with a sterile aqueous solution being preferred. If the antibodies are provided in a dry form, reconstitution is generally by the addition of a suitable solvent. A solvent, such as sterile water or buffer, may optionally be provided in the kit.

[0110] The kit may include one or more containers for the 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 may be contained in a bottle, vial, or syringe, and the informational material may be contained in association with the container. In other embodiments, the separate elements of the kit are contained within a single undivided container. For example, the composition is contained in a bottle, vial, or syringe that has attached thereto the informational material in the form of a label. In some embodiments, the kit includes a plurality (e.g., a pack) of individual containers, each of which contains 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 a plurality of syringes, ampoules, foil packets, or blister packs, each of which contains a single unit dose of an antibody described herein (e.g., DX-2930). The containers of the kits can be air tight, waterproof (eg, impermeable to changes in moisture or evaporation), and / or light-tight.

[0111] The kit optionally includes a device suitable for administration of the composition, such as 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, such as by combining the components described herein.

[0112] treatment In some aspects, the disclosure provides for the use of an antibody described herein (e.g., DX-2930) in the treatment of HAE.

[0113] (i) Hereditary angioedema Hereditary angioedema (HAE) is also known as "Quincke's edema", C1 esterase inhibitor deficiency, C1 inhibitor deficiency, and hereditary angioneurotic edema (HANE). HAE is characterized by unpredictable, recurrent attacks of severe subcutaneous or submucosal swelling (angioedema), which can affect, for example, the extremities, face, genitals, gastrointestinal tract, and airway (Zuraw, 2008). Symptoms of HAE include, for example, swelling in the arms, legs, lips, eyes, tongue, and / or throat; swelling of the throat (larynx), airway obstruction that may include sudden hoarseness, and / or death by suffocation (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 that a patient is at risk for a laryngeal attack (Bork et al., 2003; Bork et al., 2006). HAE symptoms include repeated episodes of abdominal cramps without obvious cause; and / or intestinal swelling that can be severe and lead to intestinal symptoms similar to abdominal cramps, vomiting, dehydration, diarrhea, pain, shock, and / or unnecessary surgery (Zuraw, 2008). The swelling can last up to five days or more. Approximately one-third of individuals with this HAE develop a non-itchy rash called erythema marginatum during attacks. Most patients experience multiple attacks per year.

[0114] HAE is a rare disorder and its exact prevalence is unknown, although 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).

[0115] Plasma kallikrein plays an important role in the pathology of HAE attacks (Davis, 2006; Kaplan and Joseph, 2010). In normal physiology, C1-INH regulates the activity of plasma kallikrein as well as various other proteases, such as Clr, Cls, factor XIa, and factor XIIa. Plasma kallikrein regulates the release of bradykinin from high molecular weight kininogen (HMWK). Deficiency of C1-INH in HAE leads to uncontrolled plasma kallikrein activity, which leads to overproduction of bradykinin. Bradykinin is a vasodilator that is believed to be responsible for the characteristic HAE symptoms of local swelling, inflammation, and pain (Craig et al., 2012; Zuraw et al., 2013).

[0116] Airway swelling can be life-threatening and causes death in some patients. Mortality is estimated at 15-33%. HAE results in approximately 15,000-30,000 emergency room visits annually.

[0117] 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 try to avoid certain stimuli that have previously caused attacks. However, in many cases, attacks occur without a known trigger. Typically, HAE symptoms first appear in childhood and worsen during adolescence. On average, untreated individuals experience attacks every 1-2 weeks, with most episodes lasting about 3-4 days (ghr.nlm.nih.gov / condition / hereditary-angioedema). The frequency and duration of attacks vary widely among individuals with hereditary angioedema and even among members of the same family.

[0118] There are three types of HAE, known as types I, II, and 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 percent of cases, type II accounting for approximately 15 percent of cases, and type III being very rare. Type III is the most recently described form, and was initially thought to occur only in women, but families with affected males have been identified.

[0119] HAE is inherited in an autosomal dominant pattern, meaning that affected individuals may inherit a mutation from one affected parent. De novo mutations in the gene can also occur, so HAE can occur in individuals with no prior family history of the disorder. It is estimated that 20-25% of cases are due to de novo spontaneous mutations.

[0120] Mutations in the SERPING1 gene cause type I and type II hereditary angioedema. The SERPING1 gene provides instructions for producing the C1 inhibitor protein, which is important in controlling inflammation. C1 inhibitor blocks the activity of certain proteins that promote inflammation. Mutations that cause type I hereditary angioedema lead to reduced levels of C1 inhibitor in the blood. In contrast, mutations that cause type II result in the production of an 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% of patients have type II HAE, which results in normal or high levels of functionally defective C1-INH (Zuraw, 2008). Without adequate levels of functional C1 inhibitor, excessive amounts of bradykinin are generated from high molecular weight kininogen (HMWK), resulting in increased 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 through blood vessel walls into body tissues. Excessive accumulation of fluid in body tissues causes episodes of swelling seen in individuals with Type I and Type II hereditary angioedema.

[0121] Mutations in the F12 gene have been linked to some causes of hereditary angioedema type III. The F12 gene provides the instructions for producing clotting factor XII. In addition to playing a crucial role in blood clotting (clotting), factor XII is also a critical stimulator of inflammation and is involved in bradykinin production. Certain mutations in the F12 gene result in the production of factor XII with increased activity. As a result, more bradykinin is produced and blood vessel walls become leakier, which leads 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 responsible for the disorder in these cases.

[0122] Although HAE may present with similar symptoms to other forms of angioedema due to allergies or other medical conditions, it has significantly different causes and treatments. When hereditary angioedema is misdiagnosed as an allergy, it is most often treated with antihistamines, steroids, and / or epinephrine, and although epinephrine may be used for life-threatening reactions, this treatment is generally ineffective in HAE. Misdiagnosis has also led to unnecessary exploratory surgery in patients with abdominal swelling, and abdominal pain has been misdiagnosed as psychosomatic in some HAE patients.

[0123] Similar to adults, children with HAE can experience recurrent and debilitating attacks. Symptoms can appear very early in childhood, with upper airway angioedema reported in HAE patients as young as 3 years of age (Bork et al., 2003). In one case study of 49 pediatric HAE patients, 23 had at least one episode of airway angioedema by age 18 (Farkas, 2010). There is significant unmet medical need among children with HAE, especially adolescents, as the disease commonly worsens after adolescence (Bennett and Craig, 2015; Zuraw, 2008).

[0124] C1 inhibitor therapy as well as other treatments for HAE are described in Kaplan, AP, J Allergy Clin Immunol, 2010, 126(5):918-925.

[0125] Acute treatment of HAE attacks is provided to halt the progression of edema as quickly as possible. C1 inhibitor concentrate from donor blood administered intravenously is one emergency treatment; however, this procedure is not available in many countries. In emergency situations where C1 inhibitor concentrate is not available, fresh frozen plasma (FFP) can be used as an alternative, as it also contains C1 inhibitor.

[0126] 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 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 pose a risk of infectious disease transmission by human blood-borne pathogens.

[0127] Treatment of an acute HAE attack may also include medication and / or IV fluids for pain relief.

[0128] Other treatment modalities may stimulate the synthesis of C1 inhibitor or reduce C1 inhibitor consumption. Androgen medications such as danazol may reduce the frequency and severity of attacks by stimulating C1 inhibitor production.

[0129] Helicobacter pylori can trigger abdominal attacks. Antibiotics to treat H. pylori reduce abdominal attacks.

[0130] Newer treatments attack the contact cascade: ecallantide (KALBITOR®, DX-88, Dyax) inhibits plasma kallikrein and is approved in the United States. Icatibant (Firadzyl®, Shire) inhibits the bradykinin B2 receptor and is approved in Europe and the United States.

[0131] Diagnosis of HAE may depend, for example, on family history and / or blood tests. Research findings related to 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 elevated; however, 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.

[0132] Symptoms of HAE can be assessed using, for example, questionnaires, such as questionnaires completed by the patient, physician, or family. Such questionnaires are known in the art and include, for example, visual analog scales. See, for example, 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 can be diagnosed using any method known in the art, such as by clinical history (e.g., subcutaneous or mucosal, non-itchy swelling episodes) consistent with HAE or by diagnostic testing (e.g., C1-INH function test and C4 level evaluation).

[0133] (ii) Treatment of HAE with anti-PKal antibody 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, methods of treating HAE are provided 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 treatment, e.g., one other agent 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 with a family history of HAE or a genetic predisposition thereto), e.g., according to a dosing schedule described herein. In some examples, the subject can be a human patient who does not have HAE symptoms at the time of treatment. In some embodiments, the subject is a human patient with HAE type I or HAE type II. 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 per year prior to treatment.

[0134] In some embodiments, the subject is a female. In some embodiments, the subject is a pediatric subject. In some embodiments, the subject is an adolescent under 18 years of age. In some embodiments, the subject is an adolescent between 12 and 18 years of age. In some embodiments, the subject is between 40 and 65 years of age.

[0135] In some embodiments, the subject may be defined by gender, for example, in some embodiments, the subject is female.

[0136] In some embodiments, the human subject is defined by body weight. In some embodiments, the human subject weighs less than 50 kg. In some embodiments, the human subject weighs between 50 kg and 75 kg. In some embodiments, the human subject weighs between 75 kg and 100 kg. In some embodiments, the human subject weighs 100 kg or more.

[0137] In some embodiments, any of the human patient subgroups may be administered about 300 mg of an anti-pKal antibody (e.g., DX-2930) every two weeks. In other examples, such human patients may be administered about 150 mg of the antibody every two or four weeks. In yet other examples, such human patients may be administered about 300 mg of the antibody every four weeks.

[0138] In some embodiments, the human subject is defined by a history or lack of a laryngeal attack. In some embodiments, the subject has experienced at least one (e.g., 1, 2, 3, 4, 5 or more) laryngeal attack (i.e., laryngeal HAE attack) prior to administration of an antibody described herein. In some embodiments, the subject has not experienced a laryngeal attack prior to administration of an antibody described herein.

[0139] Treatment includes administering an amount effective to relieve, alleviate, alter, cure, ameliorate, improve, or affect the disorder, the symptoms of the disorder, or the predisposition to the disorder. The treatment may also delay the onset of the disease or condition, e.g., prevent its development or prevent its worsening.

[0140] Methods for administering the DX-2930 antibody are also described in "Pharmaceutical Compositions." The appropriate dosage of the antibody used may depend on the age and weight of the subject and the particular drug used. The antibody may be used as a competitor, for example, to inhibit or reduce undesirable interactions between plasma kallikrein and its substrate (e.g., factor XII or HMWK). The dose of the antibody may be sufficient to block 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% 99% or 99.9% of the activity of plasma kallikrein in the patient, particularly at the disease site. In some embodiments, 150 mg or 300 mg of the antibody is administered every two weeks or every four weeks. In some embodiments, the antibody is administered to the subject for a first treatment period comprising administration of 150 mg or 300 mg of the antibody every two weeks or every four weeks. In some embodiments, the antibody is administered to the subject for a second treatment period following the first treatment period. In some embodiments, the antibody is administered in a single dose of 300 mg. If the subject experiences an HAE attack after a single dose, the antibody may be administered at 300 mg every two weeks.

[0141] In one embodiment, the antibodies are used, for example, to inhibit the activity of plasma kallikrein in vivo (e.g., inhibit at least one activity of plasma kallikrein, e.g., reduce factor XIIa and / or bradykinin production). The binding proteins can be used alone or conjugated to an agent, such as, for example, a cytotoxic drug, a cytotoxic enzyme, or a radioisotope.

[0142] The 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 contain a complement-binding effector domain, such as an 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 the antibodies described herein and appropriate effector cells. This treatment can be supplemented by the addition of complement or complement-containing serum. 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 the antibodies containing a complement-binding effector domain are lysed by complement.

[0143] Methods for administering the DX-2930 antibody are described in "Pharmaceutical Compositions." 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 as a competitor, for example, to inhibit or reduce undesirable interactions between natural or pathological substances and plasma kallikrein.

[0144] A therapeutically effective amount of the antibodies described herein can be administered to a subject having, suspected of having, or at risk of having HAE to thereby treat the disorder (e.g., ameliorate or ameliorate symptoms or characteristics of the disorder, slow, stabilize, and / or halt disease progression).

[0145] The antibodies described herein may be administered in a therapeutically effective amount, which is an amount that, upon administration to a subject in a single dose or multiple doses, is effective to treat the subject, e.g., cure, alleviate, ameliorate, or ameliorate at least one symptom of a disorder in the subject to an extent greater than would be expected in the absence of such treatment.

[0146] Dosage regimens can be adjusted to provide the optimum desired response (e.g., therapeutic response). For example, a single bolus can be administered, multiple divided doses can be administered over time, or the dose can be tapered or increased as indicated by the exigencies of the therapeutic situation. In other examples, a bolus can be administered followed by multiple doses over time, or the dose can be tapered or increased as indicated by the exigencies of the therapeutic situation. In other examples, the dose can be divided into multiple doses and administered over time. For ease of administration and uniformity of dosage, it is particularly advantageous to formulate parenteral compositions in unit dosage form. Unit dosage form, as used herein, refers to a physically discrete unit suitable as a unitary dosage for the subject to be treated; each unit contains a predetermined amount of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier.

[0147] In some embodiments, the antibody described herein is administered in a dosing regimen during the first treatment period. In some embodiments, the antibody is administered in multiple doses 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 or 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. In some embodiments, a therapeutically or prophylactically effective amount of the antibody (e.g., DX-2930) may 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 female subject. In some embodiments, a therapeutically or prophylactically effective amount of the antibody (e.g., DX-2930) may 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 who is under the age of 18. In some embodiments, a therapeutically or prophylactically effective amount of the antibody (e.g., DX-2930) can 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 to a subject between the ages of 40 and 65.

[0148] In some embodiments, a therapeutically or prophylactically effective amount of the antibody (e.g., DX-2930) 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, to a subject aged 65 years or older. In a particular example, the antibody is administered to a subject at about 300 mg every 2 weeks. In another particular example, the antibody is administered to a subject at about 300 mg every 4 weeks.

[0149] In some embodiments, a therapeutically or prophylactically effective amount of the antibody (e.g., DX-2930) may be about 300 mg and is 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 a subject who has experienced at least one attack of laryngeal HAE. In a specific example, the antibody is given to a subject at about 300 mg every 2 weeks. In another specific example, the antibody is given to a subject at about 300 mg every 4 weeks.

[0150] In some embodiments, a therapeutically or prophylactically effective amount of the antibody (e.g., DX-2930) may be about 150 mg or 300 mg and is administered to a subject under the age of 18 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. In a specific example, the antibody is administered to a subject at about 300 mg every two weeks. In another specific example, the antibody is administered to a subject at about 300 mg every four weeks.

[0151] In some embodiments, a therapeutically or prophylactically effective amount of the antibody (e.g., DX-2930) may be about 150 mg or 300 mg and is administered every 2 weeks or every 4 weeks. In some embodiments, a therapeutically or prophylactically effective amount of the antibody (e.g., DX-2930) may be 300 mg and is administered to a subject every 2 weeks. In some embodiments, a therapeutically or prophylactically effective amount of the antibody (e.g., DX-2930) may be 300 mg and is administered to a subject every 4 weeks. In some embodiments, a therapeutically or prophylactically effective amount of the antibody (e.g., DX-2930) may be 150 mg and is administered to a subject every 4 weeks. In some embodiments, a therapeutically or prophylactically effective amount is 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, at least 13 times or more. In some embodiments, the first treatment period is 26 weeks. In some embodiments, the therapeutically or prophylactically effective amount is 150 mg and is administered to the subject every 4 weeks (e.g., every 4 weeks for 26 weeks, resulting in a total of 7 doses being delivered). In some embodiments, the therapeutically or prophylactically effective amount is 300 mg and is administered to the subject every 2 weeks (e.g., every 2 weeks for 26 weeks, resulting in a total of 13 doses being delivered). In some embodiments, the therapeutically or prophylactically effective amount is 300 mg and is administered to the subject every 4 weeks (e.g., every 4 weeks for 26 weeks, resulting in a total of 7 doses being delivered).

[0152] 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. It will be understood by those skilled in the art that the described treatment schedule allows for a window of ±4 days (e.g., ±3 days, ±2 days, or ±1 day). For example, doses administered on days 10-18 are included in the dose for day 14 above.

[0153] In some embodiments, the therapeutically or prophylactically effective amount is administered in a dosing regimen during a second treatment period after a first treatment period. In some embodiments, the therapeutically or prophylactically effective amount is different during the first and second treatment periods. 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 administered every two weeks. In some embodiments, during the second treatment period, multiple doses of the 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, at least 13 times. In some embodiments, the second treatment period is 26 weeks. In some embodiments, the antibody is administered at a dose of about 300 mg every two weeks for 26 weeks (e.g., 13 doses are delivered). In some embodiments, the single initial dose of the second treatment period occurs about two weeks after the last administration of the first treatment period.

[0154] In any of the embodiments described herein, the timing of administration of the antibody is approximately and can include 3 days before and 3 days after a specified date (e.g., administration every two weeks would include administration on days 11, 12, 13, 14, 15, 16, or 17).

[0155] In some embodiments, the antibodies described herein are administered in a single dose of about 300 mg to a subject who has undergone a prior HAE treatment (first treatment), such as a multiple dose treatment with the same anti-pKal antibody described herein (e.g., DX-2930). If the subject experiences an HAE attack after the single dose, the subject can be treated with multiple doses of the antibody at about 300 mg every two weeks for an appropriate period of time, e.g., 26 weeks. In some embodiments, the first of the multiple doses is administered within one week of an HAE attack (e.g., within 1, 2, 3, 4, 5, 6, or 7 days of an HAE attack). In some embodiments, the antibodies 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, at least 13 times, or more.

[0156] The prior HAE treatment may include the same antibody (e.g., DX-2930) described herein. In some embodiments, the prior HAE treatment may include multiple doses of DX-2930 every 2 weeks or every 4 weeks. In some embodiments, DX-2930 is administered (e.g., subcutaneously) to the subject at 150 mg every 4 weeks, 300 mg every 2 weeks, or 300 mg every 4 weeks. In one example, the subject was pre-administered with the antibody every 2 weeks or every 4 weeks for 26 weeks prior to administration of a single dose of the antibody. In some embodiments, the pre-treatment multiple doses of the 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. In some embodiments, the antibody was pre-administered on days 0, 28, 56, 84, 112, 140, and 168. In some embodiments, a single dose of about 300 mg of the antibody is administered about 2 weeks after the last dose of the prior treatment. In one example, the single dose of the second treatment period is administered on day 182 of the first treatment period.

[0157] In any of the embodiments described herein, the timing of administration of the antibody is approximately, including 3 days before and 3 days after the specified date (e.g., administration every 2 weeks would include administration on days 11, 12, 13, 14, 15, 16, or 17).

[0158] In some embodiments, a subject may be evaluated to establish a baseline HAE attack rate prior to administration of 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 HAE attack must meet or exceed a minimum number of HAE attacks in a given period. In one example, the subject experiences at least one HAE attack in a four week run-in period prior to the first administration of the antibody. In another example, the subject experiences less than one to two attacks per month in a four week run-in period prior to the first administration of the antibody. In another example, the subject experiences less than two to three attacks per month in a four week run-in period prior to the first administration of the antibody. In another example, the subject experiences three or more attacks per month in a four week run-in period prior to the first administration of the antibody. In another example, the subject experiences at least two HAE attacks in an eight week run-in period prior to the first administration of the antibody. In yet another example, the subject experiences an average of at least one attack of HAE per month.

[0159] In some embodiments, a therapeutically or prophylactically effective amount of the antibody (e.g., DX-2930) may be about 150 mg or 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 less than 1-2 attacks of HAE per month during a run-in period before the first administration of the antibody. In some embodiments, a therapeutically or prophylactically effective amount of the antibody (e.g., DX-2930) may be about 150 mg or 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 less than 2-3 attacks of HAE per month during a run-in period before the first administration of the antibody. In some embodiments, a therapeutically or prophylactically effective amount of the antibody (e.g., DX-2930) may be about 150 mg or 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 more than 3 attacks of HAE per month during the run-in period prior to the first administration of the antibody.

[0160] In some embodiments, administration of the antibody according to any of the methods described herein results in a reduction in the average HAE attack rate in a subject. In some embodiments, the percent reduction in the average HAE attack rate following administration of the antibody according to any of the methods described herein can be determined relative 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 can 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% relative to the HAE attack rate in subjects who did not receive the antibody.

[0161] Any of the subjects described herein may have received a prior treatment for HAE, such as a preventative or therapeutic treatment for HAE. Aspects of the present disclosure also provide a method of administering an antibody described herein (e.g., DX-2930) to a subject who has received one or more prior treatments for HAE. In some embodiments, the prior treatment for HAE is a treatment comprising an antibody described herein (e.g., DX-2930). In some embodiments, the subject has previously been administered multiple doses of DX-2930 every two weeks or every four weeks. In some embodiments, the subject has previously been administered 150 mg of DX-2930 every two weeks. In some embodiments, the subject has previously been administered 300 mg of DX-2930 every two weeks. In some embodiments, the subject has previously been administered 300 mg of DX-2930 every four weeks. In some embodiments, multiple doses of the pretreatment 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.

[0162] In some embodiments, the subject has undergone one or more prior treatments for HAE, such as long-term preventative treatments, which may include any of the therapeutic agents for HAE known in the art. Exemplary anti-HAE agents include, but are not limited to, C1-inhibitors (e.g., Synlyse®, Verinert®, 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 has undergone treatment with C1-inhibitors prior to the first treatment period. In some instances, the subject may undergo a dose tapering period prior to undergoing the anti-pKal antibody treatment described herein. A dose tapering period refers to a period prior to anti-pKal antibody treatment during which the dosage, frequency, or both of the anti-HAE agent is gradually reduced such that a subject receiving prior HAE treatment (e.g., C1-INH, oral androgens, and / or oral antifibrinolytic agents) may be gradually transitioned from the prior HAE treatment to the anti-pKal antibody treatment described herein. In some embodiments, dose tapering involves a gradual or stepwise method of decreasing the dosage of the prior treatment and / or the frequency with which the prior treatment is administered. The tapering period may last from 2 to 4 weeks and may vary based on individual patent factors. In some instances, the prior treatment is terminated prior to the initiation of anti-pKal antibody treatment. In other instances, the prior treatment may be terminated within an appropriate time frame (e.g., 2 weeks, 3 weeks, or 4 weeks) after the subject is administered the first dose of anti-pKal antibody.

[0163] Alternatively, subjects receiving prior HAE treatment may be transitioned directly to the anti-pKal antibody treatment described herein without a dose tapering period.

[0164] In some embodiments, a therapeutically or prophylactically effective amount of the antibody (e.g., DX-2930) may be about 150 mg or 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 undergone one or more prior treatments for HAE.

[0165] In other embodiments, the subject does not receive any prior treatment for HAE prior to the first treatment, the first treatment period, and / or the additional single and multiple dose treatments (second treatment period) described herein. In some embodiments, the subject does not receive any treatment other than the antibodies described herein during the first treatment period and / or during the second treatment period. In some embodiments, the subject does not receive any prior treatment for HAE for at least 2 weeks (e.g., at least 2, 3, 4, 5 weeks or more) prior to the first treatment or first treatment period, during the first treatment or first treatment period, and / or during the second treatment period. In some embodiments, the subject does not receive long-term prevention of HAE (e.g., C1 inhibitor, attenuated androgen, antifibrinolytic agent) for at least 2 weeks prior to the first treatment or first treatment period, during the first treatment period, and / or during the second treatment period. In some embodiments, the subject does not receive an HAE treatment including an angiotensin-converting enzyme (ACE) inhibitor for at least 4 weeks before the initial treatment or first treatment period, during the first treatment period, and / or during the second treatment period. In some embodiments, the subject does not receive an estrogen-containing medication for at least 4 weeks before the initial treatment or first treatment period, during the first treatment period, and / or during the second treatment period. In some embodiments, the subject does not receive an androgen (e.g., stanozolol, danazol, oxandrolone, methyltestosterone, testosterone) for at least 2 weeks before the initial treatment or first treatment period, during the first treatment period, and / or during the second treatment period.

[0166] Any of the methods described herein may further include monitoring the patient for side effects (e.g., elevated creatine phosphatase levels) and / or levels of inhibition of pKal by the antibody (e.g., serum or plasma concentration of the antibody or pKal activity levels) before, during, or after treatment or during a course of treatment. If one or more adverse effects are observed, the dose of the antibody may be reduced or treatment may be terminated. If the level of inhibition falls below the minimum therapeutic level, an additional dose of the antibody may be administered to the patient. Patients may also be assessed for production of antibodies against the administered antibody; activity of C1-inhibitor, C4 and / or C1q; quality of life; incidence of any 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., D-2930) over other injectable agents, quality of life (e.g., Angioedema-Quality of Life (AE-QOL), EuroQoL Group 5-dimension report).

[0167] In some embodiments, the plasma or serum concentration of the antibody (e.g., DX-2930) may be measured during the course of treatment (e.g., after the first dose) to assess the effectiveness of the treatment. If the plasma or serum concentration of the antibody is lower than about 80 nM, a booster dose may be required, which may be the same or higher than the first dose. 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 an immunoassay or 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 an assay may include a synthetic substrate assay or a Western blot assay for measuring cleaved kininogen as described herein.

[0168] 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 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 altered or treatment may be terminated.

[0169] In some embodiments, the optimal dosage (e.g., optimal prophylactic or therapeutic dosage) of the antibody (e.g., DX-2930) can be determined as follows: The antibody is administered to a subject in need of treatment at an initial dose. The plasma concentration of the antibody in the subject is measured. If the plasma concentration is lower than 80 nM, the dose of the antibody is increased in subsequent administrations. The dose of the antibody that maintains an antibody plasma concentration above about 80 nM can be selected as the optimal dose for the subject. The subject's creatine phosphokinase levels can be monitored during the course of treatment, and the optimal dose for the subject can be further adjusted based on the creatine phosphokinase level, for example, if an increase in creatine phosphokinase is observed during treatment, the dose of the antibody can be reduced.

[0170] (iii) Combination therapy The antibodies described herein (e.g., DX-2930) can be administered in combination with one or more other therapeutic agents to treat a disease or condition associated with plasma kallikrein activity, such as a disease or condition described herein. For example, the antibodies described herein (e.g., DX-2930) can be used therapeutically or prophylactically (e.g., before, during, or after a course of treatment) 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. Examples of plasma kallikrein inhibitors that can be used in combination therapy of plasma kallikrein binding to the antibodies described herein include, for example, plasma kallikrein inhibitors described in WO 95 / 21601 or WO 2003 / 103475.

[0171] One or more plasma kallikrein inhibitors can be used in combination with an antibody described herein (e.g., DX-2930), e.g., by combination, a lower dose of the inhibitor is required, which can result in reduced side effects.

[0172] The antibodies described herein (e.g., DX-2930) may be administered in combination with one or more current therapeutic agents for treating HAE. For example, the DX-2930 antibody may be co-administered with a second anti-HAE therapeutic agent, such as ecallantide, a C1 esterase inhibitor (e.g., Synlize™), aprotinin (TRASYLOL®), and / or a bradykinin B2 receptor inhibitor (e.g., icatibant (Firazyr®)).

[0173] The term "combination" refers to the use of two or more drugs or therapeutic agents to treat the same patient, where the use or action of the drugs or therapeutic agents overlaps in time. Drugs or therapeutic agents 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 that occurs at different times. The time between administration of one drug and another drug can be minutes, hours, days, or weeks. The use of plasma kallikrein binding antibodies described herein can also be used to lower the dosage of another therapeutic agent, for example, to reduce side effects associated with another drug being administered. Thus, a combination can include administering the second drug at a dosage that is at least 10, 20, 30, or 50% lower than would be used in the absence of the plasma kallikrein binding antibody. In some embodiments, a subject can be administered C1-inhibitor as a loading IV or SC dose concurrently with the first administration of an anti-pKal antibody (e.g., DX-2930) described herein. The subject can then continue anti-pKal antibody treatment (without further administration of C1-inhibitor).

[0174] The combination therapy may include administering an agent that reduces the side effects of the other therapeutic agent. The agent may be an agent that reduces the side effects of treating a plasma kallikrein-associated disease.

[0175] (iv) Assays for evaluating treatment regimens Assay methods for assessing the effectiveness of any of the treatment methods described herein are also within the scope of this disclosure. In some embodiments, plasma or serum concentrations of one or more biomarkers associated with HAE (e.g., 2-chain HMWK) may be measured before and / or during a course of treatment (e.g., after the first dose) to assess the effectiveness of the treatment. 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.

[0176] The level of the biomarker can be measured by detecting the biomarker in a plasma or serum sample obtained from the subject, for example by an immunoassay such as a 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 the subject is assessed by immunoassay. Antibodies for use in immunoassays for the detection of 2-HWMK are known in the art, and the selection of such antibodies for use in the methods described herein will be apparent to one of ordinary skill in the art.

[0177] Without further elaboration, it is believed that one skilled in the art can, based on the above, utilize the present invention to its fullest extent. Accordingly, the following specific embodiments are 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 purposes or subject matter discussed herein. EXAMPLES

[0178] Example 1: Efficacy and Safety of DX-2930 Treatment in Human Patient Subpopulations Lanadelumab is a sterile, preservative-free solution for injection at pH 6.0. The active ingredient, antibody DX-2930, is formulated with the following nominal components: 30 mM dibasic sodium phosphate dihydrate, 19.6 mM citric acid monohydrate, 50 mM L-histidine, 90 mM sodium chloride, 0.01% polysorbate 80. Each vial contains a nominal concentration of 150 mg DX-2930 active ingredient in 1 mL solution. The study product is administered by subcutaneous (SC) injection in the upper arm in a blinded fashion.

[0179] The placebo consisted of an inactive formulation of the test product: 30 mM dibasic sodium phosphate dihydrate, 19.6 mM citric acid monohydrate, 50 mM L-histidine, 90 mM sodium chloride, pH 6.0, with 0.01% polysorbate 80. Subjects randomized to the placebo treatment arm received a placebo dose, and subjects randomized to the 300 mg or 150 mg DX-2930 treatment arms every 4 weeks received a placebo dose between doses of DX-2930.

[0180] Patients ≥12 years of age with HAE type I / II and ≥1 attack / month at baseline were randomized 2:2:2:3 to lanadelumab 150 mg every 4 weeks (q4wks), 300 mg every 4 weeks, 300 mg every 2 weeks, or placebo. Exploratory analyses were planned for subpopulations with adequate numbers of patients for Poisson regression.

[0181] The following primary and secondary efficacy outcomes were assessed from Day 14 to Day 182. The primary outcomes of the study were the number of HAE attacks and the mean HAE attack rate. Secondary outcomes included, ranked as follows: 1. Number of HAE attacks requiring emergency treatment 2. Moderate to severe number of HAE attacks.

[0182] Exploratory efficacy endpoints 1. Time to first seizure occurrence after day 14, i.e., the seizure-free duration that a subject remains until the first seizure occurs after day 14. 2. Number of high morbidity HAE attacks per week; a high morbidity HAE attack is defined as any attack with at least one of the following characteristics: severe attack requiring hospitalization (excluding hospitalization for observation <24 hours), hemodynamically significant attack (systolic blood pressure <90, requiring IV hydration, or involving syncope or near syncope) or laryngeal attack.

[0183] Lab Test Patients included in the clinical trial were subjected to laboratory tests including general safety parameters (hematology, coagulation, urinalysis and serum chemistry), serology, pregnancy test, C1-INH functional assay, C4 assay, C1q assay, PK samples, plasma anti-drug antibody tests and PD samples. All laboratory tests are performed using established and validated methods.

[0184] result Overall, 125 patients were treated with lanadelumab (n=84) or placebo (n=41). The average HAE attack rate was determined for all patients and therefore for all patient subgroups. The average number of HAE attacks was used to determine the percent reduction in the average HAE attack rate for patients who received DX-2930 compared to those who received placebo. HAE attack rates were consistently reduced by DX-2930 compared to placebo in all patients and patient subgroups. However, as shown in Table 2, a larger percent reduction, i.e., a more therapeutically effective reduction, was observed for some patient subgroups when 300 mg of DX-2930 was administered every 2 weeks compared to 300 mg of DX-2930 every 4 weeks (or 150 mg of DX-2930 every 4 weeks). Specifically, patients <18 years of age receiving 300 mg DX-2930 every 4 weeks experienced a 20.5% reduction in HAE attack rates compared to placebo; patients <18 years of age receiving 300 mg DX-2930 every 2 weeks experienced an additional attack rate reduction of approximately 42 percentage points (62.3%) (Figure 3A). Patients 40 to <65 years of age receiving 300 mg DX-2930 every 4 weeks experienced a 71.5% reduction in HAE attack rates compared to placebo; patients 40 to <65 years of age receiving 300 mg DX-2930 every 2 weeks experienced an additional reduction in incidence of approximately 18 percentage points (89.8%). Female patients receiving 300 mg of DX-2930 every 4 weeks had a 69.6% reduction in HAE attack rates compared to placebo; those receiving 300 mg of DX-2930 every 2 weeks had an additional reduction of approximately 16 percentage points (85.8%). Patients with a history of laryngeal attacks receiving 300 mg of DX-2930 every 4 weeks had a 64.2% reduction in HAE attack rates compared to placebo; those with a history of laryngeal attacks receiving 300 mg of DX-2930 every 2 weeks had an additional reduction of approximately 21 percentage points (85.7%).

[0185] [Table 2]

[0186] All HAE type I / II patients treated with lanadelumab 300 mg q2wks or q4wks experienced a clinically meaningful and sustained reduction in HAE attack rates compared to placebo, although certain subpopulations of patients, such as women, patients aged <18 years or 40-65 years, and patients with a history of at least one laryngeal attack, showed better treatment efficacy with 300 mg every 2 weeks.

[0187] Patients were also stratified based on HAE attacks during the run-in period to evaluate the efficacy of each of the lanadelumab treatment regimens in these patient subgroups. As shown in Tables 3-5 and Figures 1A-1C, each of the lanadelumab treatment regimens resulted in a significant reduction in the rate of HAE attacks when compared to placebo in all subgroups.

[0188] [Table 3]

[0189] [Table 4]

[0190] [Table 5]

[0191] In patients who used only C1-inhibitor (C1-INH) for long-term prophylaxis, baseline attack rates increased during protocol-based C1-INH interruption compared with historical rates (during the last 3 months) (Figure 2A). Attack rates during lanadelumab treatment were lower than historical rates. Attack rates decreased by a mean of 68.8%, 59.3%, and 82.1% during treatment with lanadelumab 150 mg q4wks, 300 mg q4wks, and 300 mg q2wks, respectively, compared with historical rates during long-term prophylaxis.

[0192] A consistent treatment effect of lanadelumab was observed in patients who used C1-INH for prophylaxis alone and in patients who did not use long-term prophylaxis when compared with placebo using a Poisson regression model (Figure 2B). In patients who used C1-INH alone for long-term prophylaxis before lanadelumab, the mean attack rate was significantly reduced by 73.6%, 71.6%, and 82.5% in the lanadelumab 150 mg q4wks, 300 mg q4wks, and 300 mg q2wks regimens, respectively, compared with placebo (P<0.001 for all comparisons).

[0193] The percentage reduction in HAE attack rate in subjects receiving lanadelumab at each dosing regimen compared to placebo was assessed for subgroups of subjects based on age, sex, weight, type of HAE (e.g., type I or type II), and history of laryngeal attacks (Figures 4A-4E and 5).

[0194] Lanadelumab significantly inhibited pKal activation, as indicated by its effect on cHMWK levels. Optimal clinical responses were observed in adolescents and adults across a wide weight range at a fixed dose regimen of 300 mg every 2 weeks.

[0195] Example 2: Efficacy and Safety of DX-2930 (Lanadelumab) Treatment in Human Adolescent Patients This phase 3, open-label, long-term (OLE) study investigated the efficacy and safety of lanadelumab, a monoclonal antibody targeting plasma kallikrein, in adolescent patients with HAE and C1 inhibitor deficiency.

[0196] For the phase 3 study, patients ≥12 years of age with physician-confirmed attacks ≥1 per 4 weeks were randomized to placebo or lanadelumab 150 mg every 4 weeks (150 mg q4w), 300 mg q4w, or 300 mg q2w. In the phase 3 study, 10 of 125 patients (8%) were adolescents (≥12 to <18 years of age). Prior to the start of the phase 3 study, 60.0% of patients were receiving C1-INH solely for long-term prophylaxis.

[0197] Generally, rollover subjects in the open-label extension study were treated with lanadelumab according to the treatment regimen of the Phase 3 study (i.e., 150 mg every 4 weeks, 300 mg every 4 weeks, 300 mg every 2 weeks). In the open-label extension study, subjects receive a single open-label dose of 300 mg lanadelumab subcutaneously on day 0. Subjects did not receive additional lanadelumab doses until their first reported, physician-confirmed HAE attack. Once the rollover subject reports their first HAE attack, they receive a second open-label dose of lanadelumab as soon as possible, with at least 10 days between the first and second open-label doses. After the second dose, rollover subjects continue to receive repeat subcutaneous doses of open-label 300 mg lanadelumab every 2 weeks for the remainder of the treatment period according to planned dosing. The treatment period lasts for 350 days from the date of the first open-label administration.

[0198] Non-rollover subjects in the open-label long-term study will receive an open-label dose of 300 mg lanadelumab subcutaneously on Day 0 and will continue to receive open-label 300 mg lanadelumab subcutaneously every 2 weeks for the treatment period according to planned dosing. A total of 26 doses will be administered, with the final dose being administered at the Day 350 visit.

[0199] Among rollover patients, 62.5% received C1-INH alone before the start of the open-label long-term study. Among non-rollover adolescent patients, 61.6% received long-term prophylaxis (C1-INH alone or C1-INH and oral therapy) before the start of the study (mainly C1-INH alone; 46.2%). Monthly attack rates (MAR) and other treatment-emergent adverse events (TEAEs) were recorded.

[0200] Three adolescent subjects experienced 13 non-serious treatment-emergent adverse events (TEAEs). In the open-label, extended-term study, 21 / 212 patients (9.9%) were adolescents. Rollover (n=8) and non-rollover (n=13) patients had mean (SD) monthly attack rates of 1.65 (1.158) and 1.54 (0.971) at baseline and 0.35 (0.635) and 0.07 (0.166), respectively, for a mean (SD) percent change of -84.371 (18.9415) and -94.893 (10.5230), respectively. Nine patients experienced 65 non-serious lanadelumab-related TEAEs.

[0201] Results from this study are provided below in Table 6. Lanadelumab was found to be effective and safe in reducing MAR in adolescent patients with HAE.

[0202] [Table 6]

[0203] In the phase 3 study, lower least squares mean (SE) HAE attack rates were observed on days 0–182 in patients treated with lanadelumab 300 mg q4wks (n=3; 0.436 [0.253]) or lanadelumab 300 mg q2wks (n=2; 0.207 [0.148]) compared with patients receiving placebo (n=4; 0.548 [0.224]). This was not estimated for the 150 mg q4wks treatment arm because it included only one adolescent patient. The least squares mean estimates of monthly attack rate ratios (vs. placebo), with 95% CI, favored treatment with lanadelumab, especially the 300 mg q2wks dose regimen (Figure 3C). In the open-label long-term study, the mean (SD) percent change from baseline in monthly mean seizure rate was 84.37 (18.94) in rollover patients (n = 8; usual dosing phase) and -94.89 (10.52) in non-rollover patients (n = 13; Figure 3B).

[0204] In the phase 3 study, there were 13 non-serious lanadelumab-related TEAEs in three adolescent patients (Table 7). The most common TEAEs occurring in >1 patient during treatment with lanadelumab were injection site pain (3 patients) and redness (2 patients). In the open-label long-term study, there were 65 non-serious lanadelumab-related TEAEs in nine patients over a mean follow-up time of 0.63 years. The most common TEAEs occurring in >1 patient were injection site pain (9 patients), viral upper respiratory tract infection (3 patients), influenza (2 patients), streptococcal pharyngitis (2 patients), upper respiratory tract infection (2 patients), abdominal pain (2 patients), and headache (2 patients). Overall, the most common TEAE associated with lanadelumab administration recorded in >1 patient was injection site pain (3 patients in the phase 3 study and 8 patients in the OLE study; Table 7). These were similar to those identified in the overall Phase 3 study population.

[0205] There were no deaths or discontinuations due to TEAEs in both the Phase 3 study and its OLE.

[0206] [Table 7] HAE: hereditary angioedema; m = number of events; TEAE = treatment-emergent adverse events; q2wks = every 2 weeks; q4wks = every 4 weeks. TEAEs are presented during the treatment period (Days 0-182) for Phase 3 studies. *There were no serious or severe TEAEs associated with lanadelumab in the Phase 3 study. A serious TEAE was defined as any TEAE that resulted in death, a life-threatening experience, an unplanned hospitalization, persistent / significant disability / incapacity, a major medical event, or a congenital anomaly / birth defect. A severe TEAE was one that was classified by the investigator as severe (Grade 3, resulting in significant limitation of activity, usually requiring some assistance, requiring medical intervention / treatment, and / or likely hospitalization) or life-threatening (Grade 4, extreme limitation of activity requiring significant assistance, requiring significant medical intervention / treatment, and / or likely hospitalization / hospice care).

[0207] In conclusion, lanadelumab was well tolerated and reduced monthly attack rates in adolescents in phase 3 and open-label long-term studies.

[0208] Other embodiments All features disclosed herein may be combined in any combination. Each feature disclosed herein may be replaced by alternative features serving the same, equivalent or similar purpose. Thus, unless expressly stated otherwise, each feature disclosed is merely an example of a generic series of equivalent or similar features.

[0209] From the above description, those skilled in the art can easily ascertain the basic 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. Therefore, other embodiments are also within the scope of the claims.

[0210] equivalent While several embodiments of the invention have been described and illustrated herein, those skilled in the art will readily envision various other means and / or structures for performing the functions described herein and / or obtaining the results and / or one or more advantages, and each such variation and / or modification is deemed to be within the scope of the embodiments of the invention described herein. More generally, those skilled in the art will readily recognize that all parameters, dimensions, materials and / or configurations described herein are intended to be exemplary, and that the actual parameters, dimensions, materials and / or configurations will depend on the particular application(s) for which the teachings of the 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 embodiments of the invention described herein. Thus, it is understood that the foregoing embodiments are presented by way of example only, and that within the scope of the appended claims and equivalents thereto, embodiments of the invention may be practiced other than as specifically described and claimed. The embodiments of the present disclosure are directed to each individual feature, system, article, material, kit and / or method described herein. Furthermore, any combination of two or more such features, systems, articles, materials, kits, and / or methods is included within the inventive scope of the present disclosure, if such features, systems, articles, materials, kits, and / or methods are not mutually inconsistent.

[0211] All definitions, as defined and used herein, should be understood to supersede any dictionary definitions, definitions in documents incorporated by reference, and / or ordinary meanings of the defined terms.

[0212] 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."

[0213] The phrase "and / or" as used herein in the specification and claims should be understood to mean "either or both" of the elements so connected, i.e., elements that are present conjointly in some cases and separately 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 connected. Other elements, whether in association with the specifically identified elements or not, may optionally be present other than the elements specifically identified by the "and / or" clause. Thus, as a non-limiting example, a reference to "A and / or B", when used in conjunction with an open-ended term such as "comprising", may in one embodiment refer to only A (optionally including elements other than B); in another embodiment, refer to only B (optionally including elements other than A); or in yet another embodiment, refer to both A and B (optionally including other elements).

[0214] 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 to be inclusive, i.e., including not only at least one of a number or list of elements, but also a plurality, and optionally further unlisted items. Only terms clearly indicated to the contrary, such as "only one of" or "exactly one of," or, when used in the claims, "consisting of," refer to the inclusion of exactly one element of a number or list of elements. In general, the term "or" will only be construed to refer to exclusive alternatives (i.e., "one or the other but not both) when preceded by an exclusive term such as "either," "one of," "only one of," or "exactly one of." "Consisting essentially of," when used in the claims, has its ordinary meaning as used in the field of patent law.

[0215] As used herein in the specification and claims, the phrase "at least one" with 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 does not exclude any combination of elements in the list of elements. This definition also allows that elements other than those specifically identified in the list of elements to which the phrase "at least one" refers may optionally be present, whether or not related to the specifically identified elements. 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") can refer, in one embodiment, to at least one, optionally including more than one, A, B being absent (and optionally including elements other than B); in another embodiment, to at least one, optionally including more than one, B, A being absent (and optionally including elements other than A); in yet another embodiment, to at least one, optionally including more than one, A, and at least one, optionally including more than one, B (and optionally including other elements), etc.

[0216] It should also be understood that, unless expressly indicated to the contrary, in any method claimed herein that includes more than one step or act, the order of the method steps or acts is not necessarily limited to the order in which the method steps or acts are recited.

[0217] In the claims, as well as in the above specification, transitional phrases such as "comprising," "including," "carrying," "having," "containing," "involving," "holding," "composed of," and the like, are all to be understood to be open-ended, i.e., to mean including but not limited to. Only the transitional phrases "consisting of" and "consisting essentially of" are closed or semi-closed transitional phrases, respectively, as set forth in the U.S. Patent Office Manual of Patent Examining Procedures, Section 2111.03.

Claims

1. 1. A composition for use in a method of treating hereditary angioedema (HAE) attacks or reducing the rate of HAE attacks, said method comprising: administering to a human subject in need thereof an antibody comprising heavy chain complementarity determining regions (CDRs) set forth as SEQ ID NOs:5-7 and light chain CDRs set forth as SEQ ID NOs:8-10 during a first treatment period; during said first treatment period, said antibody is administered to said human subject in multiple doses at about 300 mg every two weeks; the human subject is afflicted with, suspected of being afflicted with, or at risk of being afflicted with HAE; (i) is female; (ii) is between 40 and 65 years old; (iii) has had 1-2, 2-3, or more than 3 attacks of HAE in the 4 weeks prior to the first dose of the first treatment period; and / or (iv) has received treatment with C1-inhibitor prior to the first treatment period; composition.

2. A composition for use in a method for treating hereditary angioedema (HAE) attacks or reducing the rate of HAE attacks, the method comprising: administering to a human subject in need thereof an antibody comprising heavy chain complementarity determining regions (CDRs) set forth as SEQ ID NOs:5-7 and light chain CDRs set forth as SEQ ID NOs:8-10; 12. The method of claim 11, wherein the human subject (i) an adolescent between 12 and 18 years of age; and / or (ii) had 2-3 or more than 3 attacks of HAE in the 4 weeks prior to the first dose of the antibody; the antibody is administered to the human subject at about 150 mg every 4 weeks, about 300 mg every 4 weeks, or about 300 mg every 2 weeks; composition.

3. The composition described in claim 1 or claim 2, wherein the antibody is a full-length antibody or an antigen-binding fragment thereof.

4. The composition described in any one of claims 1 to 3, 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.

5. A composition described in any one of claims 1 to 4, wherein the antibody comprises a heavy chain represented by sequence number 1 and a light chain represented by sequence number 2.

6. The composition described in any one of claims 1 to 5, wherein the antibody is formulated in a pharmaceutical composition comprising a pharma- ceutically acceptable carrier.

7. The composition described in claim 6, wherein the pharmaceutical composition comprises sodium phosphate, citric acid, histidine, sodium chloride and polysorbate 80.

8. The composition of claim 7, 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 a concentration of about 0.01%.

9. The composition of claim 1, wherein the antibody is administered subcutaneously.

10. The composition described in any one of claims 1 to 9, wherein the human subject has type I or type II HAE.

11. A composition described in any one of claims 1 to 10, wherein the human subject has experienced at least two HAE attacks per year prior to the first treatment period.

12. The composition described in any one of claims 1 to 11, wherein the human subject has undergone one or more prior HAE treatments prior to the first treatment period.

13. The composition of claim 12, wherein the prior HAE treatment comprises a C1-inhibitor, a plasma kallikrein inhibitor, a bradykinin receptor antagonist, an androgen, an antifibrinolytic drug, or a combination thereof.

14. The composition of claim 13, wherein the prior HAE treatment comprises C1-INH, ecallantide, icatibant, danazol, tranexamic acid, or a combination thereof.

15. A composition described in any one of claims 12 to 14, wherein the method includes a tapering period of the one or more prior HAE treatments.

16. The composition of claim 15, wherein the tapering period is about 2 to 4 weeks.

17. A composition described in any one of claims 12 to 16, wherein the one or more prior HAE treatments are completed either before the first dose of the antibody or within three weeks after the first dose of the antibody.

18. The composition described in any one of claims 1 to 11, wherein the human subject has not undergone prior HAE treatment.

19. The composition described in claim 18, wherein the human subject has not undergone prior HAE treatment for at least two weeks prior to the first dose of the antibody.

20. The composition described in any one of claims 1 to 19, wherein the human subject has had at least one HAE attack in the 4 weeks prior to the first dose of the first treatment period or at least two HAE attacks in the 8 weeks prior to the first dose of the first treatment period.

21. The composition described in any one of claims 1 to 20, wherein the method further comprises administering the antibody to the subject during a second treatment period after the first treatment period.

22. The composition of claim 21, wherein the first dose of the second treatment period is approximately 2 weeks after the last administration of the first treatment period.

23. The composition described in claim 21 or claim 22, wherein the second treatment period comprises one or more administrations of the antibody at about 300 mg.

24. The composition of claim 23, wherein the second treatment period comprises multiple doses of the antibody at about 300 mg every two weeks.

25. The composition described in any one of claims 1 to 24, wherein the human subject has not received long-term prophylaxis for HAE or HAE treatment including angiotensin-converting enzyme (ACE) inhibitors, estrogen-containing medications or androgens prior to, during, and / or during the first treatment period.

26. The method of claim 20, (a) after the first treatment period, administering the antibody in a single dose of about 300 mg to a human subject in need thereof; (b) if the subject experiences an HAE attack after (a), further administering the antibody to the subject in one or more doses of about 300 mg; The composition of any one of claims 1 and 3-25, further comprising:

27. The composition of claim 26, wherein in step (b), the subject is administered the antibody in multiple doses of about 300 mg every two weeks.

28. The composition described in claim 27, wherein the initial dose of step (b) is within one week after the HAE attack.

29. The composition of any one of claims 26 to 28, wherein the single dose of (a) and the first dose of (b) are separated by at least 10 days.

Images