Plasma kallikrein inhibitors and their use in 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 of 35 U.S.C. § 119(e) of U.S. Provisional Patent Application No. 62 / 725,216, filed Aug. 30, 2018, and U.S. Provisional Patent Application No. 62 / 808,612, filed Feb. 21, 2019, each of which is incorporated herein by reference in its entirety.

Background Art

[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 I.M. et al., Thromb Haemost 98, 77-83, 2007). The contact system is activated by factor XIIa upon exposure to foreign substances or charged surfaces or by prolyl carboxypeptidase on the endothelial cell surface (Sainz I.M. et al., Thromb Haemost 98, 77-83, 2007). Activation of plasma kallikrein amplifies endogenous coagulation via its feedback activation of factor XII and enhances inflammation via the production of bradykinin, an inflammatory promoting nonapeptide. As the major kininogenase in circulation, plasma kallikrein contributes significantly to bradykinin production in blood vessels. Genetic deficiency of 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 triggered by unknown stimuli (Zuraw B.L. et al., N Engl J Med 359, 1027-1036, 2008).

Summary of the Invention

[0003] This specification provides a treatment plan for treating hereditary angioedema (HAE) attacks, reducing the incidence of HAE attacks, or preventing HAE attacks, using an antibody capable of binding to and inhibiting active human plasma kallikrein (pKal), such as an antibody having the same complementarity-determining region (CDR) as DX-2930 (also known as SHP643, lanadermab).

[0004] In some embodiments, the Disclosure provides a method for treating or reducing the incidence of hereditary angioedema (HAE) attacks, comprising administering (e.g., subcutaneously) one 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 approximately 300 mg every two weeks during a first treatment period. In some embodiments, the subject is a woman who has, is suspected of having, or is at risk of having HAE; is under 18 years of age or between 40 and 65 years of age; and / or has previously experienced at least one laryngeal HAE attack.

[0005] In some embodiments, the Disclosure provides a method for treating or reducing the incidence of hereditary angioedema (HAE) attacks, comprising administering (e.g., subcutaneously) one 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 approximately 150 mg every four weeks, approximately 300 mg every four weeks, or approximately 300 mg every two weeks. In some embodiments, the subject is an adolescent aged 12–18 years.

[0006] Any method described herein may further include administering the antibody to the subject in a second treatment period after a first treatment period. In some embodiments, the first dose of the second treatment period is approximately two weeks after the last dose of the first treatment period. In some embodiments, the second treatment period includes one or more doses of the antibody at approximately 300 mg each. In some embodiments, the second treatment period includes multiple doses of the antibody at approximately 300 mg each, every two weeks.

[0007] Any method described herein may further include (a) administering the antibody to a human subject in a single dose of approximately 300 mg after a first treatment period; and (b) if the subject experiences an HAE attack after (a), administering the antibody to the subject in one or more doses of approximately 300 mg. In some embodiments, in step (b), the subject is administered the antibody in multiple doses of approximately 300 mg every two weeks. In some embodiments, the initial dose in step (b) is within one week after an HAE attack. In some embodiments, the single dose in (a) and the initial 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 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, subjects treated with any of the methods described herein, including the use of any of the anti-pKal antibodies described herein (e.g., DX-2930), have received one or more HAE treatments prior to the first dose of the anti-pKal antibody. Such prior HAE treatments may include C1 inhibitors (e.g., C1-INH), plasma kallikrein inhibitors (e.g., ecalantide), bradykinin receptor antagonists (e.g., icatibant), androgens (e.g., danazol), antifibrinolytic agents (e.g., tranexamic acid), or combinations thereof. In such subjects, a tapering period may be established to gradually transition from the prior HAE treatment to the anti-pKal antibody treatment described herein. In some examples, the tapering period is approximately 2 to 4 weeks. The prior HAE treatment may be terminated either before the first dose of the antibody to the subject or within 3 weeks after the first dose of the antibody. Alternatively, the subject may be directly transitioned from any of the pre-HAE treatments to the anti-pKal antibody treatment described herein.

[0010] In some embodiments, subjects have not received HAE treatment prior to the first dose of the anti-pKal antibody. In some embodiments, subjects have not received prior HAE treatment for at least two weeks prior to the first dose of the antibody.

[0011] In some embodiments, the subjects have not received long-term prophylaxis to HAE, or HAE treatment including angiotensin-converting enzyme (ACE) inhibitors, estrogen-containing drug therapy, or androgens, prior to, 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 includes a heavy chain variable region indicated by SEQ ID NO: 3 and / or a light chain variable region indicated by SEQ ID NO: 4. In some examples, the antibody includes a heavy chain indicated by SEQ ID NO: 1 and a light chain indicated by SEQ ID NO: 2.

[0013] In any of the methods described herein, the antibody can be formulated into a pharmaceutical composition comprising a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical composition comprises sodium phosphate, citrate, histidine, sodium chloride, and polysorbate 80. In one example, the concentration of sodium phosphate is about 30 mM, citrate is about 19 mM, histidine is about 50 mM, sodium chloride is about 90 mM, and polysorbate 80 is about 0.01%.

[0014] Details of one or more embodiments of the present invention are shown in the following description. Other features or advantages of the present invention will become apparent from the following drawings and detailed descriptions of some embodiments, and also from the appended claims. [Brief explanation of the drawing]

[0015] [Figure 1]Figures 1A–1C include Poisson regression plots of HAE attacks observed by researchers during the treatment period (days 0–182) for patients, based on the number of HAE attacks during the acclimatization period. Figure 1A: 1–<2 HAE attacks per month during the acclimatization period. Figure 1B: 2–<3 HAE attacks per month during the acclimatization period. Figure 1C: ≥3 HAE attacks per month during the acclimatization period. [Figure 2A] Figures 2A-2B include diagrams showing the HAE attack rates in patients who previously received long-term prophylaxis with a C1 inhibitor (C1-INH). Figure 2A: Past (3-month) mean (standard deviation), baseline, and monthly HAE attack rates during lanadermab treatment (days 0-182). Figure 2B: Reduction in HAE attack rates in each HAE patient in the indicated lanadermab treatment group. [Figure 2B] Same as above. [Figure 3-1] Figures 3A–3C include plots of monthly HAE seizure rates in adolescent subjects. Figure 3A shows a plot of estimated least squares mean (LS) versus placebo for monthly seizure frequency in adolescent patients, within a 95% confidence interval. Figure 3B: Plot of monthly HAE seizure frequency versus baseline for rollover and non-rollover adolescent subjects during the lanadelmab treatment period. Figure 3C shows a plot of estimated least squares mean (vs. placebo) for monthly seizure rate in adolescent patients, within a 95% confidence interval, for each of the indicated lanadelmab treatment groups. [Figure 3-2] Same as above. [Figure 4-1] Figures 4A-4E plot the percentage reduction in HAE attack rate from placebo, within the 95% confidence interval, for each of the demographic groups indicated. Figure 4A: Age; Figure 4B: Sex; Figure 4C: Weight; Figure 4D: HAE type; Figure 4E: History of laryngeal attacks. For each group indicated, the bar graphs, from left to right, correspond to 150 mg every 4 weeks, 300 mg every 4 weeks, and 300 mg every 2 weeks. The "n" below the plot indicates the number of subjects in each group. [Figure 4-2] Same as above. [Figure 4-3] Same as above. [Figure 5]Figure 5 shows a forest plot of the incidence ratio of HAE attacks confirmed by researchers, based on the demographic data described. [Modes for carrying out the invention]

[0016] definition For convenience, before describing the present invention, certain terms used 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 otherwise clearly indicated by the context.

[0018] Where used herein, the term "approximately" refers to a range of + / - 5% of a given value. For example, approximately 300 mg of antibody contains any amount between 285 mg and 315 mg of antibody.

[0019] The term "antibody" refers to an immunoglobulin molecule that can specifically bind to targets such as carbohydrates, polynucleotides, lipids, and polypeptides through at least one antigen recognition site located in the variable domain of the immunoglobulin molecule. Antibodies are heavy chain immunoglobulins with variable domains (V H ) containing at least one heavy (H) chain, light chain immunoglobulin variable domain (V L The antibody may contain at least one or both light chains, including the heavy (H) chain variable region (V in this specification). For example, the antibody may contain the heavy (H) chain variable region (V in this specification). H (or abbreviated as HV) and light (L) chain variable region (V in this specification) L It may include (or abbreviated as LV). In another example, the antibody contains two heavy (H) chain variable regions and two light (L) chain variable regions.

[0020] As used herein, the term “antibody” encompasses not only intact (i.e., full-length) polyclonal or monoclonal antibodies, but also their antigen-binding fragments (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), any mutant thereof, fusion proteins containing the antibody moiety, humanized antibodies, chimeric antibodies, diabodies, linear antibodies, single-chain antibodies, polyspecific antibodies (e.g., bispecific antibodies), and any other modified conformation of an immunoglobulin molecule containing an antigen-recognition site having the required specificity, including glycosylated variants of antibodies, amino acid sequence variants of antibodies, and covalently modified antibodies. Antibodies include any class of antibody, such as IgD, IgE, IgG, IgA, or IgM (or their subclasses), but an antibody does not have to be an antibody of any particular class. Depending on the antibody amino acid sequence of the heavy chain constant domain, immunoglobulins can be assigned to various 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 different classes of immunoglobulin are called alpha, delta, epsilon, gamma, and mu, respectively. The subunit structures and three-dimensional configurations of different classes of immunoglobulins are well known. Antibodies can originate from any source, but primate antibodies (human and non-human primates) and primate-modified antibodies are preferred.

[0021] V H and / or V LThe region may include all or part of the amino acid sequence of a natural variable domain. For example, the sequence may lack 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 include other modifications. In one embodiment, a polypeptide comprising an immunoglobulin variable domain sequence can associate with another immunoglobulin variable domain sequence to form a structure that preferentially interacts with an antigen-binding site, such as plasma kallikrein.

[0022] V H and V L The region can be further subdivided into hypervariable regions called "complementary determining regions" ("CDRs") interspersed with more conserved regions called "framework regions" ("FRs"). The ranges covered by the framework regions and CDRs are defined (see Kabat, E.A., et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department 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 typically consists of three CDRs and four FRs and is arranged in the following order from the amino terminus to the carboxy terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.

[0023] V H or V LIn addition to the constant region, the heavy or light chain of the antibody may further include all or part of the heavy or light chain constant region. In one embodiment, the antibody is a tetramer of two heavy immunoglobulin chains and two light immunoglobulin chains, which are interconnected, for example, by disulfide bonds. In IgG, the heavy chain constant region includes three immunoglobulin domains, CH1, CH2, and CH3. The light chain constant region includes a CL domain. The variable regions of the heavy and light chains contain binding domains that interact with the antigen. The constant region of the antibody generally mediates the 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 kappa or lambda type. In one embodiment, the antibody is glycosylated. The antibody may be functional against antibody-dependent cell-mediated cytotoxicity and / or complement-mediated cytotoxicity.

[0024] One or more regions of an antibody may be human or effectively human. For example, one or more variable regions may be human or effectively human. For example, one or more CDRs, e.g., HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and / or LC CDR3, may be human. Each of the light chain (LC) and / or heavy chain (HC) CDRs may be human. HC CDR3 may be human. One or more framework regions, e.g., FR1, FR2, FR3, and / or FR4 of HC and / or LC, may be human. For example, the Fc region may be human. In one embodiment, all framework regions are human and derived, for example, from human somatic cells, e.g., hematopoietic cells or non-hematopoietic cells that produce immunoglobulins. In one embodiment, the human sequence is a germline sequence and is encoded, for example, by germline nucleic acid. In one embodiment, selected framework (FR) residues of Fab can be converted to the amino acid types of the corresponding residues in the most similar primate germline genes, particularly human germline genes. One or more constant regions may be human or effectively human. For example, at least 70, 75, 80, 85, 90, 92, 95, 98, or 100% of the immunoglobulin variable domain, constant region, constant domain (CH1, CH2, CH3, and / or CL1), or the entire antibody may be human or effectively human.

[0025] Antibodies can 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. The full-length immunoglobulin "light chain" (approximately 25 kDa or 214 amino acids) is encoded by a variable region gene at the NH2-terminus (approximately 110 amino acids) and a kappa or lambda constant region gene at the COOH-terminus. The full-length immunoglobulin "heavy chain" (approximately 50 kDa or 446 amino acids) is similarly encoded by a variable region gene (approximately 116 amino acids) and one of the other constant region genes mentioned above, such as gamma (encoding approximately 330 amino acids). The length of human HC varies considerably, as HC CDR3 ranges from approximately 3 amino acid residues to over 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 that are included in the term "antigen-binding fragment" of a full-length antibody and retain functionality include (i) Fab fragments, i.e., V L , V H , C L (ii) a monovalent fragment consisting of the CH1 domain; (ii) a bivalent fragment containing the F(ab')2 fragment, i.e., two Fab fragments linked by disulfide bridges in the hinge region; (iii) V H and Fd fragment consisting of CH1 domain; (iv) V of single arm of antibody L and V H Fv fragment consisting of domains, (v)V H (vi) The dAb fragment consisting of domains (Ward et al., (1989) Nature 341:544-546); and (vi) the isolated complementarity-determining region (CDR). Furthermore, the two domains of the Fv fragment, V L and V H These are encoded by individual genes, but these are V L and V HThey can be linked using recombinant methods by synthetic linkers, which allow the regions to pair and be prepared as a single protein chain forming a monovalent molecule known as a single-chain Fv (scFv). See, for example, U.S. Patents No. 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 the prior art known to those skilled in the art.

[0027] The term “monocretinative antibody” refers to an antibody that exhibits single-binding specificity and affinity for a specific target, such as an epitope. This term includes “monoclonal antibody” or “monoclonal antibody composition,” which, as used herein, refers to a single-molecule composition of an antibody or a preparation of a fragment thereof, regardless of how the antibody was produced. An antibody is “germ-serumized” by converting one or more non-germ-series amino acids in its framework region back to the corresponding germ-series amino acids of the antibody, provided that its binding properties are substantially preserved.

[0028] Inhibition constant (K i ) provides a measure of inhibitory activity; this is the concentration of the inhibitor required to halve the enzyme activity, and is independent of the 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 inhibitors (e.g., inhibitory binding proteins) on the degree of reaction (e.g., enzyme activity); and the apparent K is obtained by fitting the change in the pseudo-first-order rate constant as a function of the inhibitor concentration to Morrison's equation (Equation 1). i An estimated value of K is obtained. i is, K i,app The y-intercept of the plot against substrate concentration is obtained by deriving it from linear regression analysis.

number

[0029] As used herein, “binding affinity” refers to the apparent association constant or K A It refers to K. A The dissociation constant (K D This is the reciprocal of ). The binding antibody may have binding affinities of at least 105, 106, 107, 108, 109, 1010, and 1011M-1 to a specific target molecule, such as plasma kallikrein. Higher affinity binding of the binding antibody to the first target compared to the second target indicates K for binding to the first target. A K regarding binding to the second target A (or numerical value K) D ) is higher than (or numerical value K) D This can be indicated by (a small value). In such cases, the binding antibody has specificity for the first target (e.g., the protein with the first three-dimensional structure or its mimetic) compared to the second target (e.g., the same protein with the second three-dimensional structure or its mimetic); or the second protein. The difference in binding affinity (e.g., with respect to specificity or other comparisons) is at least 1.5, 2, 3, 4, 5, 10, 15, 20, 30, 40, 50, 70, 80, 90, 100, 500, 1000, 10,000 or 10 5 It could be double.

[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 fluorescence assays). Exemplary conditions for evaluating binding affinity are in HBS-P buffer (10 mM HEPES pH 7.4, 150 mM NaCl, 0.005% (v / v) surfactant P20). Using these techniques, the concentrations of bound and free binding proteins can be measured as a function of the 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 at the target, where (N) is the number of binding sites per target molecule by the following formula: [Bound]=N·[Free] / ((1 / KA)+[Free]).

[0031] K A While it is not always necessary to make an accurate determination, it is sometimes determined using methods such as ELISA or FACS analysis, and K A Since it is sufficient to obtain a quantitative measurement of affinity proportional to the activity, it can therefore be used for comparison, for example, to determine whether the affinity is higher, for example, twice as high, in order to obtain a quantitative measurement of affinity or to obtain an estimate of affinity by using a functional assay, for example, activity in an in vitro or in vivo assay.

[0032] The term “conjugated antibody” (or as used interchangeably with “conjugated protein” herein) refers to an antibody capable of interacting with a target molecule. The term “target molecule” is used interchangeably with “ligand.” “Plasma kallikrein-conjugated antibody” refers to an antibody capable of interacting with (e.g., binding to) plasma kallikrein, and includes antibodies that preferentially or specifically interact with and / or inhibit plasma kallikrein. An antibody inhibits plasma kallikrein if it causes a decrease in the activity of plasma kallikrein compared to the activity of plasma kallikrein in the absence of the antibody and under the same conditions.

[0033] A "conservative amino acid substitution" is a substitution in which an amino acid residue is replaced by an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains are defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), amino acids with acidic side chains (e.g., aspartic acid, glutamic acid), amino acids with non-loading side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), amino acids with nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), amino acids with beta-branched side chains (e.g., threonine, valine, isoleucine), and amino acids with aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine).

[0034] One or more framework and / or CDR amino acid residues of a binding protein may contain 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. Plasma kallikrein-binding proteins may 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 mutations, and / or fewer than 15, 12, 10, 9, 8, 7, 6, 5, 4, 3, or 2 mutations), such as mutations that do not substantially affect protein function. Mutations may be located in the framework region, CDR, and / or constant region. In some embodiments, mutations are located in the framework region. In some embodiments, mutations are located in the CDR. In some embodiments, mutations are located in the constant region. Whether a particular substitution is permissible, that is, whether it has a detrimental effect on biological properties such as binding activity, can be predicted, for example, by evaluating whether the mutation is conservative, or by the method described in 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 so as not to elicit an immunogenic response in a normal human. A "substantially human" antibody is one that contains a sufficient number of human amino acid positions so as not to elicit an immunogenic response in a normal human.

[0036] An "epitope" refers to a site on a target compound to which a binding protein (e.g., an antibody such as Fab or a full-length antibody) binds. If the target compound is a protein, this site may consist entirely of amino acid components, entirely of chemical modifications of amino acids in the protein (e.g., glycosyl moieties), or a combination thereof. Overlapping epitopes contain at least one common amino acid residue, glycosyl group, phosphate group, sulfate group, or other molecular characteristics.

[0037] A “humanized” immunoglobulin variable region is an immunoglobulin variable region that has been modified to include a sufficient number of human framework amino acid positions so that it does not elicit an immunogenic response in normal humans. Descriptions of “humanized” immunoglobulins include, for example, U.S. Patent Applications No. 6,407,213 and No. 5,693,762.

[0038] An “isolated” antibody refers to an antibody from which at least 90% of at least one component of the natural sample from which the isolated antibody can be obtained has been removed. An antibody can be “at least” of a certain 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-to-weight basis.

[0039] The methods described herein involve administering multiple doses of an antibody to a human subject in need. The terms “patient,” “subject,” or “host” may be used interchangeably. A subject may be a subject that has undergone prior treatment for HAE, such as treatment containing the antibodies described herein. In some embodiments, the subject is a pediatric subject (e.g., an infant, child, or adolescent subject). In some embodiments, the human subject is an adolescent under 18 years of age. In some embodiments, the human subject is an adolescent between 12 and 18 years of age. In some embodiments, the subject is between 40 and under 65 years of age.

[0040] In some embodiments, the human subject is defined by sex. For example, 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, a human subject is defined by having or not having a history 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 the antibody described herein. In some embodiments, the subject has not experienced a laryngeal attack prior to administration of the antibody described herein.

[0043] The terms “prekallikrein” and “preplasma kallikrein” are used interchangeably herein and refer to the enzyme precursor form of active plasma kallikrein, also known as prekallikrein.

[0044] As used herein, the term “substantially identical” (or “substantially homologous”) means, in this specification, a first amino acid or nucleic acid sequence containing a sufficient number of amino acid residues or nucleotides identical or equivalent to (e.g., having similar side chains, e.g., conserved amino acid substitutions) to the second amino acid or nucleic acid sequence, such that the first and second amino acid or nucleic acid sequences have similar activity, e.g., binding activity, binding preference, or biological activity (or encode proteins having such activity). In the case of an antibody, the second antibody has the same specificity and has 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 some 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 p-values ​​less than 0.05 or 0.02. Certain binding proteins may show statistically significant differences in, for example, specificity or binding (e.g., p-value < 0.05 or 0.02). Terms such as “inducing,” “inhibiting,” “enhancing,” “increasing,” “increasing,” and “decreasing” may refer to differences, such as statistically significant differences between two situations, indicating distinguishable qualitative or quantitative differences between two situations.

[0046] The "therapeutically effective dose" preferably adjusts a measurable parameter, such as plasma kallikrein activity, by a statistically significant amount, or by 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 an untreated control group. The compound's ability to adjust a measurable parameter, such as a disease-related parameter, can be evaluated in animal model systems that predict efficacy in human disorders and conditions. Alternatively, this property of a composition can be evaluated by examining the compound's ability to adjust parameters in vitro.

[0047] Where used herein, the term “to treat” refers to the application or administration of a composition comprising one or more active substances to a person who has HAE, has HAE symptoms, is suspected of having HAE, or is predisposed to or at risk of HAE, with the aim of curing, resolving, reducing, mitigating, modifying, treating, relieving, improving, or influencing the disease, symptoms of the disease, or predisposition to the disease. “Prophylactic treatment,” also known as “preventive treatment,” refers to treatment aimed at protecting a person from or reducing the risk of a disease to which they have been or may be exposed. In some embodiments, the treatment methods described herein are aimed at preventing the onset and / or recurrence of HAE.

[0048] The term "preventing" a disease in a subject refers to administering a pharmaceutical treatment, such as a drug, to the subject in such a way that at least one symptom of the disease is prevented; that is, it is administered before the onset of clinical symptoms of the undesirable condition (e.g., the disease or another undesirable condition in the host animal) in order to protect the host from the onset of the undesirable condition. "Preventing" a disease may also be called "prevention" or "preventive treatment."

[0049] The “prophylactic effective dose” refers to the dosage and amount effective over the required time to achieve the desired prophylactic outcome. Typically, the prophylactic effective dose is less than the therapeutic effective dose, as prophylactic doses are used in subjects before or earlier in the course of the disease.

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

[0051] In one embodiment, 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 contains at least one immunoglobulin variable region. For example, the antibody contains 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, for example, heavy chain CDR sequences shown as SEQ ID NOs. 5-7 and light chain CDR sequences shown as SEQ ID NOs. 8-10. In some embodiments, the antibody includes the same CDR sequences as DX-2930 and LC immunoglobulin variable domain sequences that are at least 85, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100% identical (e.g., in the whole or in the framework region) to the LC variable domains described herein. In some embodiments, the antibody includes the same CDR sequences as DX-2930 and HC immunoglobulin variable domain sequences that are at least 85, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100% identical (e.g., in the whole or in the framework region) to the HC variable domains described herein. In some embodiments, the antibody includes an LC sequence that is identical (e.g., in whole or in framework regions) 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 the same CDR sequence as DX-2930 and the LC sequences described herein. In some embodiments, the antibody includes an HC sequence that is identical (e.g., in whole or in framework regions) 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 the same CDR sequence as DX-2930 and the HC sequences described herein.

[0053] Plasma kallikrein-binding protein can be an isolated antibody (e.g., containing at least 70, 80, 90, 95, or 99% of other proteins). In some embodiments, the plasma kallikrein-binding antibody or its composition is inactive or partially active (e.g., with a K content of 5000 nM or higher) compared to the plasma kallikrein-binding antibody. i,app The antibody cleavage fragments (e.g., DX-2930) that bind to plasma kallikrein are isolated. For example, the plasma kallikrein-conjugated antibody contains at least 70% of such antibody cleavage fragments; in other embodiments, the conjugated antibody contains at least 80%, at least 90%, at least 95%, at least 99%, or even 100% of antibody cleavage fragments that are inactive or partially conjugated.

[0054] Plasma kallikrein-binding antibodies can further inhibit plasma kallikrein, such as human plasma kallikrein.

[0055] In some embodiments, the plasma kallikrein-conjugating antibody does not bind to prekallikrein (e.g., human prekallikrein and / or mouse prekallikrein), but binds to the active form of plasma kallikrein (e.g., 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 fragments thereof, or to an epitope that overlaps with the active site of plasma kallikrein.

[0057] This antibody is at least 10 5 , 10 6 , 10 7 , 10 8 , 10 9 , 10 10 and 10 11 M -1 With binding affinity, it can bind to plasma kallikrein, for example, human plasma kallikrein. In one embodiment, the antibody is 1 × 10⁶ -3 , 5×10 -4 s -1 or 1 × 10 -4 s -1 Slower than K off It binds to human plasma kallikrein. In one embodiment, the antibody is 1 × 10⁶ 2 , 1 x 10 3 or 5 x 10 3 M -1 s -1 Faster K onIt binds to human plasma kallikrein. In one embodiment, the antibody binds to plasma kallikrein but not to tissue kallikrein and / or plasma prekallikrein (for example, the antibody binds to tissue kallikrein and / or plasma prekallikrein with lower efficacy than it would if it bound to plasma kallikrein (for example, 1 / 5, 10, 50, 100 or 1 / 1000 or none 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 This antibody inhibits human plasma kallikrein activity at Ki levels less than M. For example, IC levels of less than 100 nM, 10 nM, 1, 0.5, or 0.2 nM are effective. 50 This antibody may have the following properties. For example, this antibody may modulate plasma kallikrein activity and the production of factor XIIa (e.g., from factor XII) and / or bradykinin (e.g., from high molecular weight kininogen (HMWK)). This antibody may inhibit plasma kallikrein activity and / or the production of factor XIIa (e.g., from factor XII) and / or bradykinin (e.g., from high molecular weight kininogen (HMWK)). The affinity of this antibody for human plasma kallikrein is less than 100 nm, less than 10 nM, less than 5 nM, less than 1 nM, and less than 0.5 nM of K. D The antibody may be characterized in that, in one embodiment, it inhibits plasma kallikrein but not tissue kallikrein (for example, the antibody inhibits tissue kallikrein with lower efficacy than it would inhibit plasma kallikrein (for example, 5-, 10-, 50-, 100- or 1 / 1000th or none compared to a negative control)).

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

[0060] Plasma kallikrein-conjugated antibodies may be contained within a single polypeptide (e.g., scFv) or may have their HC and LC variable domain sequences on different polypeptides (e.g., 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 IgG, e.g., IgG1, IgG2, IgG3, or IgG4. The antibody may be a soluble Fab. In other embodiments, the antibody comprises Fab2', scFv, minibody, scFv::Fc fusion, Fab::HSA fusion, HSA::Fab fusion, Fab::HSA::Fab fusion, or other molecules containing an antigen combination site of one of the binding proteins described herein. The VH and VL regions of these Fabs may 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 human or a humanized antibody, or is non-immunogenic in humans. For example, the antibody includes one or more human antibody framework regions, such as all human framework regions or framework regions that are at least 85, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to human framework regions. In one embodiment, the antibody includes 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 primate-like antibody, or is non-immunogenic in humans. For example, the antibody includes 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, or 99% identical to primate framework regions. In one embodiment, the antibody includes 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" include 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 to human plasma kallikrein is less than 1000, 500, 100, 10, 5, 1, and 0.5 nM, for example, less than 10 nM, less than 1 nM, and less than 0.5 nM of K D It is characterized by the following.

[0065] In certain embodiments, the antibody does not contain sequences derived from mice or rabbits (for example, it is not a mouse or rabbit antibody).

[0066] In some embodiments, the antibodies used in the methods described herein may be DX-2930 or functional variants thereof as described herein.

[0067] In one example, a functional mutant of DX-2930 contains the same complementarity-determining region (CDR) as DX-2930. In another example, a functional mutant of DX-2930 contains the same V H and V L Compared to the case of V H or V LOne or more mutations (e.g., conservative substitutions) may be contained in any of the FRs. Preferably, such mutations do not occur in residues that are expected to interact with one or more CDRs that can be determined by conventional 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 antigen-binding regions / residues as the parent. In other embodiments, the functional variant of DX-2930 is V H V contains 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 V of DX-2930 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 V. L These variants may contain chains. These variants can bind to the active form of plasma kallikrein, and preferably do not bind to prekallikrein.

[0068] The "percent identicality" 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 described 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 searching can be performed using the XBLAST program, score=50, word length=3, to obtain amino acid sequence homology for the target protein molecule. If a gap exists between two sequences, Gapped BLAST can be used, as described in Altschul et al., Nucleic Acids Res. 25(17):3389-3402, 1997. When using the BLAST and Gapped BLAST programs, the initial settings parameters of the individual programs (e.g., XBLAST and NBLAST) can be used.

[0069] In some embodiments, the antibody used in the methods and compositions described herein may be DX-2930 antibody. The full weight and light chain lengths and variable sequences for DX-2930 are provided below, with signal sequences in italics. CDR is in bold and underlined.

[0070] [ka]

[0071] [Table 1]

[0072] Antibody preparation The antibodies described herein (e.g., DX-2930) can be prepared by any method known in the art. See, for example, 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 target antibody, for example DX-2930, can be maintained in a vector within host cells, which can then be grown in the host cells and frozen for later use. Alternatively, this polynucleotide sequence may be used for genetic engineering to "humanize" the antibody or to improve its affinity (affinity maturation) or other characteristics. For example, the constant region may be manipulated to more closely resemble the human constant region in order to avoid an immune response when the antibody is used in clinical trials and treatments in humans. Genetic engineering of the antibody sequence may be desired to obtain higher affinity for the target antigen and greater efficacy in inhibiting PKal activity. It will be apparent to those skilled in the art that one or more polynucleotide changes can be made to an antibody while still maintaining its binding specificity to the target antigen.

[0074] In other embodiments, complete human antibodies can be obtained by using commercially available mice that have been modified to express specific human immunoglobulin proteins. Transgenic animals designed to produce a more desirable (e.g., completely human antibodies) or more robust immune response may also be used for humanization or the production of human antibodies. Examples of such techniques include Xenomouse® from Amgen, Inc. (Fremont, Calif.) and HuMAb-Mouse® and TC Mouse® from Medarex, Inc. (Princeton, NJ). In alternative methods, antibodies can be produced recombinantly by phage display or yeast techniques. See, for example, U.S. Patent Nos. 5,565,332; 5,580,717; 5,733,743; and 6,265,150; and Winter et al., (1994) Annu. Rev. Immunol. 12:433-455. Alternatively, phage display technology (McCafferty et al., (1990) Nature 348:552-553) can be used to produce human antibodies and antibody fragments in vitro from a repertoire of immunoglobulin variable (V) domain genes from non-immune donors.

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

[0076] Genetically modified antibodies, such as humanized antibodies, chimeric antibodies, single-chain antibodies, and bispecific antibodies, can be produced, for example, through conventional recombinant techniques. For example, DNA encoding a monoclonal antibody specific to a target antigen can be readily isolated or synthesized. The DNA may be placed in one or more expression vectors, which are then transferred into host cells that do not otherwise express immunoglobulin proteins, such as Escherichia coli (E. coli) cells, monkey COS cells, Chinese hamster ovary (CHO) cells, or myeloma cells, to obtain synthesis of monoclonal antibodies in recombinant host cells. See, for example, International Publication No. 87 / 04462. The DNA can then be modified, for example, by substituting the coding sequences of human heavy and light chain constant domains for homologous mouse sequences, as described by Morrison et al., (1984) Proc. Nat. Acad. Sci. 81:6851, or by covalently linking all or part of the coding sequence of a non-immunoglobulin polypeptide to the immunoglobulin coding sequence. In this way, genetically modified antibodies with binding specificity to target antigens, such as "chimeric" or "hybrid" antibodies, can be prepared.

[0077] The techniques developed for producing "chimeric antibodies" are well known in this field. See, for example, 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 this field. See, for example, Queen et al., Proc. Natl. Acad. Sci. USA, 86:10029-10033 (1989). In one example, the non-human antibody of the parent is used. H and V LThe variable region is subjected to three-dimensional molecular modeling analysis according to methods known in the art. Next, the same molecular modeling analysis is used to identify framework amino acid residues that are expected to be important for the formation of the accurate CDR structure. In parallel, parent V is used as the search query. H and V L Using the sequence, from some antibody gene database, a human V antibody with an amino acid sequence homologous to the amino acid sequence of the parent's non-human antibody is identified. H and V L Identify the chain. Next, human V H and V L Select the acceptor gene.

[0079] The CDR region within the selected human acceptor gene can be replaced with a CDR region from a parental non-human antibody or a functional variant thereof. If necessary, residues within the parental chain framework region that are expected to be important in the interaction with the CDR region (see above) can be used to replace the corresponding residue in the human acceptor gene.

[0080] Single-chain antibodies can be prepared via recombinant techniques by linking nucleotide sequences encoding a heavy-chain variable region and a light-chain variable region. Preferably, a flexible linker is incorporated between these two variable regions. Alternatively, techniques described for the production of single-chain antibodies (U.S. Patents No. 4,946,778 and 4,704,692) can be adapted for the preparation of phage or yeast scFv libraries, and PKal-specific scFv clones can be identified from the library according to standard procedures. Positive clones can be subjected to further screening to identify clones that inhibit PKal activity.

[0081] Some antibodies, such as Fab, can be produced in bacterial cells, such as Escherichia coli (E. coli) cells (see, e.g., Nadkarni, A. et al., 2007 Protein Expr Purif 52(1):219-29). For example, if Fab is encoded by a sequence in a phage display vector that includes an inhibitory stop codon between the display portion and a bacteriophage protein (or a fragment thereof), the vector nucleic acid can be transferred into bacterial cells that cannot inhibit the stop codon. In this case, Fab is not fused to the gene III protein and is secreted into the periplasm and / or culture medium.

[0082] Antibodies can also be produced in eukaryotic cells. In one embodiment, this antibody (e.g., scFv's) is expressed in yeast cells such as Pichia (e.g., Powers et al., 2001, J.Immunol.Methods.251:123-35; Schoonooghe S. et al., 2009 BMC Biotechnol.9:70; Abdel-Salam, HA. et al., 2001 Appl Microbiol Biotechnol 56(1-2):157-64; Takahashi K. et al., 2000 Biosci Biotechnol Biochem 64(10):2138-44; Edqvist, J. et al., 1991 J Biotechnol 20(3):291-300), Hanseula, or Saccharomyces.Those skilled in the art will know, for example, oxygen conditions (see, e.g., Baumann K., et al. 2010 BMC Syst. Biol. 4:141), molar osmotic pressure (see, e.g., Dragosits, M. et al., 2010 BMC Genomics 11:207), temperature (see, e.g., Dragosits, M. et al., 2009 J Proteome Res. 8(3):1380-92), fermentation conditions (see, e.g., Ning, D. et al. 2005 J. Biochem. and Mol. Biol. 38(3):294-299), and yeast strains (see, e.g., Kozyr, AV et al. 2004 Mol Biol(Mosk) 38(6):1067-75; Horwitz, AH. et al., 1988 Proc Natl Acad Sci USA 85(22):8678-82; Bowdish, K. et al. Antibody production in yeast can be optimized by optimizing factors such as overexpression of proteins to enhance antibody production (see, for example, Gasser, B. et al., 2006 Biotechol. Bioeng. 94(2):353-61), the acidity level of the culture (see, for example, Kobayashi H., et al., 1997 FEMS Microbiol Lett 152(2):235-42), and the concentration of substrates and / or ions (see, for example, Ko JH. et al., 2996 Appl Biochem Biotechnol 60(1):41-8). Furthermore, the yeast system can be used to produce antibodies with longer half-lives (see, for example, Smith, BJ. et al. 2001 Bioconjug Chem 12(5):750-756).

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

[0084] In some embodiments, plasma kallikrein-conjugated antibodies are produced in plants or cell-free systems (see, for example, Galeffi, P., et al., 2006 J Transl Med 4:39).

[0085] In addition to nucleic acid sequences encoding diversified immunoglobulin domains, recombinant expression vectors may contain further sequences, such as sequences that control vector replication in host cells (e.g., origins of replication) and selectable marker genes. Selectable marker genes facilitate the selection of host cells into which the vector has been introduced (see, for example, U.S. Patents 4,399,216, 4,634,665, and 5,179,017). Typically, for example, selectable marker genes confer resistance to drugs, such as G418, hygromycin, or methotrexate, in host cells into which the vector has been introduced. Preferred selectable marker genes include the dihydrofolate reductase (DHFR) gene (dhfr selected / amplified by methotrexate). - Examples include (for use in host cells) and neo genes (for G418 selection).

[0086] In an exemplary system for the recombinant expression of an antibody or its antigen-binding moiety, dhfr is expressed by calcium phosphate-mediated gene transfer. - A recombinant expression vector encoding both the antibody heavy chain and antibody light chain is introduced into CHO cells. Within the recombinant expression vector, the antibody heavy chain and light chain genes are manipulably ligated to enhancer / promoter regulatory elements (e.g., derived from SV40, CMV, adenovirus, etc., e.g., a CMV enhancer / AdMLP promoter regulatory element or an SV40 enhancer / AdMLP promoter regulatory element) respectively, driving high transcription levels of the genes. The recombinant expression vector also contains the DHFR gene, which allows selection of CHO cells to which the vector has been transfected using methotrexate selection / amplification. Selected transformant host cells are cultured, intact antibodies are recovered from the culture medium, and antibody heavy chain and light chain expression is enabled. Using standard molecular biology techniques, the recombinant expression vector is prepared, transfected into host cells, transformants are selected, host cells are cultured, and antibodies are recovered 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 containing an Fc domain, the antibody production system can produce antibodies in which the Fc region is glycosylated. For example, the Fc domain of the IgG molecule is glycosylated with asparagine 297 in the CH2 domain. This asparagine is a site for modification by a branched oligosaccharide. It has been shown that this glycosylation is necessary for the effector function mediated by the Fcγ receptor 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 appropriately 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, U.S. Patent No. 5,849,992 describes a method for expressing antibodies in the mammary glands of a transgenic mammal. A transgene is constructed comprising a milk-specific promoter and a nucleic acid encoding the antibody of interest, and a signal sequence for secretion. The milk produced by such a female transgenic mammal contains the antibody of interest secreted therein. This antibody can be purified from the milk or, for certain applications, can be used directly.

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

[0090] Pharmaceutically acceptable carriers include any physiologically compatible solvents, dispersions, coatings, antimicrobial and antifungal agents, isotonic agents and absorption retarders. Preferably, carriers are suitable for subcutaneous, intravenous, intramuscular, parenteral, spinal, or epidermal administration (e.g., by injection or infusion), but carriers suitable for inhalation and intranasal administration are also intended.

[0091] The pharmaceutically acceptable carriers in the pharmaceutical compositions described herein may comprise one or more of the following: buffers, amino acids, and osmotic regulators. Any suitable buffer or combination of buffers may be used in the pharmaceutical compositions described herein to maintain or help maintain the appropriate pH of the composition. Non-limiting examples of buffers include sodium phosphate, potassium phosphate, citrate, sodium succinate, histidine, Tris, and sodium acetate. In some embodiments, the buffer may be at concentrations of about 5–100 mM, 5–50 mM, 10–50 mM, 15–50 mM, or about 15–40 mM. For example, one or more buffers may be present in concentrations of approximately 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 approximately 40 mM. In some cases, pharmaceutically acceptable carriers may include sodium phosphate and citrate, which may be present in concentrations of approximately 30 mM and approximately 19 mM, respectively.

[0092] In some embodiments, pharmaceutically acceptable carriers include one or more amino acids that can reduce antibody aggregation during pre-administration storage and / or improve antibody stability. Representative amino acids for use in the preparation of 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 amino acids 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) may be approximately 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 approximately 60 mM. In one example, the pharmaceutical composition contains histidine at a concentration of approximately 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 concentrations 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 concentrations 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. For example, the osmotic pressure modifier could be sodium chloride, which may be at a concentration of approximately 90 mM.

[0094] A pharmaceutically acceptable carrier in a pharmaceutical composition described herein may further comprise one or more pharmaceutically acceptable excipients. Generally, pharmaceutically acceptable excipients are pharmacologically inert 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, dimethyl sulfoxide (DMSO), and dimethylformamide (DMSO). Examples of excipients include polyoxyethylene sorbitan monolaurate (Tween-20), polyoxyethylene sorbitan monooleate (Tween-80), sodium dodecyl sulfate (SDS), polysorbate, polyoxyethylene copolymer, potassium phosphate, sodium acetate, ammonium sulfate, magnesium sulfate, sodium sulfate, trimethylamine N-oxide, betaine, zinc ions, copper ions, calcium ions, manganese ions, magnesium ions, CHAPS, sucrose monolaurate, and 2-O-beta-mannoglycerate. In some embodiments, pharmaceutically acceptable carriers contain excipients in amounts of about 0.001% to 0.1%, 0.001% to 0.05%, 0.005% to 0.1%, 0.005% to 0.05%, 0.008% to 0.05%, 0.008% to 0.03%, or about 0.009% to 0.02%. In some embodiments, the excipient is present in amounts of approximately 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 approximately 0.1%. In some embodiments, the excipient is polyoxyethylene sorbitan monooleate (Tween-80). In one example, the pharmaceutically acceptable carrier contains 0.01% Tween-80.

[0095] In some cases, the pharmaceutical compositions described herein include an anti-pKal antibody (e.g., DX-2930) and one or more of the following: sodium phosphate (e.g., dibasic sodium phosphate dihydrate), citrate (e.g., citrate monohydrate), histidine (e.g., L-histidine), sodium chloride, and polysorbate 80. For example, a pharmaceutical composition may include the antibody, sodium phosphate, citrate, histidine, sodium chloride, and polysorbate 80. In some cases, the antibody is formulated in about 30 mM sodium phosphate, about 19 mM citrate, 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 approximately 150 mg DX-2930 / 1 mL solution, approximately 30 mM dibasic sodium phosphate dihydrate, approximately 19 mM (e.g., 19.6 mM) citric acid monohydrate, approximately 50 mM L-histidine, approximately 90 mM sodium chloride, and approximately 0.01% polysorbate 80. In another example, the composition comprises or consists of approximately 300 mg DX-2930 / 1 mL solution, approximately 30 mM dibasic sodium phosphate dihydrate, approximately 19 mM (e.g., 19.6 mM) citric acid monohydrate, approximately 50 mM L-histidine, approximately 90 mM sodium chloride, and approximately 0.01% polysorbate 80.

[0096] A pharmaceutically acceptable salt is one that retains the desired biological activity of the compound without conferring any undesirable toxic 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, and phosphorous acid, as well as salts derived from non-toxic organic acids such as aliphatic mono- and dicarboxylic acids, phenyl-substituted alkanos, hydroxyalkanoics, aromatic acids, and aliphatic and aromatic sulfonic acids. Base addition salts include salts derived from alkaline earth metals such as sodium, potassium, magnesium, and calcium, as well as salts derived from non-toxic organic amines such as N,N'-dibenzylethylenediamine, N-methylglucamine, chloroprocaine, choline, diethanolamine, ethylenediamine, and procaine.

[0097] This composition may be in various forms. These include, for example, liquid, semi-solid, and solid dosage forms, such as liquid solutions (e.g., injectable and drip-able solutions), dispersions or suspensions, tablets, pills, powders, liposomes, and suppositories. The form may depend on the intended method of administration and therapeutic application. Many compositions are in the form of injectable or drip-able solutions, such as compositions similar to those used for administering antibodies to humans. Exemplary methods of administration are parenteral (e.g., intravenous, subcutaneous, intraperitoneal, intramuscular). In one embodiment, plasma kallikrein-binding protein is administered by intravenous drip or injection. In another embodiment, plasma kallikrein-binding protein is administered by intramuscular injection. In yet another embodiment, plasma kallikrein-binding protein is administered by subcutaneous injection. In yet another preferred embodiment, plasma kallikrein-binding protein is administered by intraperitoneal injection.

[0098] When used herein, the phrases “parenteral administration” and “administered parenterally” mean a method of administration other than intestinal and local administration, usually by injection, including, but not limited to, intravenous, intramuscular, intra-arterial, subarachnoid, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subepidermal, intra-articular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injections and infusions. In some embodiments, the antibody is administered subcutaneously.

[0099] The composition may be formulated as a solution, microemulsion, dispersion, liposome, or other ordered structure suitable for high drug concentrations. Sterile injectable solutions may be prepared by incorporating the required amount of binding protein in a suitable solvent along with one or a combination of the components listed above, and subsequently by filtration sterilization as necessary. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle containing a basic dispersion medium and other required components from the components listed above. For sterile powders for the preparation of sterile injectable solutions, preferred preparation methods are vacuum drying and freeze-drying, which yield a powder of the active component + any further desired components from a pre-filtered solution. Proper fluidity of the solution can be maintained by the use of a coating such as lecithin, by maintaining the required particle size in the case of a dispersion, and by the use of a surfactant. Sustained absorption of the injectable composition may be achieved by including absorption-delaying substances in the composition, such as monostearate and gelatin.

[0100] The antibodies described herein (e.g., DX-2930) may be administered by various methods, including intravenous injection, subcutaneous injection, or intravenous infusion. For example, in some therapeutic applications, the dose is approximately 1 to 100 mg / m². 2 Or 7-25 mg / m² 2The antibody can be administered by intravenous infusion at a rate of less than 30, 20, 10, 5, or 1 mg / min to reach the desired dose. The route and / or method of administration varies depending on the desired outcome. In certain embodiments, the active compound may be prepared with a carrier that protects the compound from rapid release, such as a controlled-release formulation including implants and microcapsule-encapsulated delivery systems. Biodegradable, biocompatible polymers, such as ethylene vinyl acetate, polyacid anhydride, polyglycolic acid, collagen, polyoltoesters, and polylactic acid, can be used. Many methods are available for preparing such formulations. See, for example, Sustained and Controlled Release Drug Delivery Systems, JR Robinson, ed., 1978, Marcel Dekker, Inc., New York.

[0101] Pharmaceutical compositions may be administered using medical devices. For example, in one embodiment, the pharmaceutical compositions disclosed herein may be administered by a device, such as a needleless subcutaneous injection device, a pump, or an indwelling.

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

[0103] The administration regimen is adjusted to provide the optimal 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 gradually reduced or increased as indicated by the emergency situation of the treatment. It is particularly advantageous to formulate parenteral compositions in unit dose forms to facilitate administration and ensure uniformity of the dose. When used herein, a unit dose form refers to a physically distinct unit suitable as a unit dose for the subject being treated; each unit contains a predetermined amount of the active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specifications for unit dose forms may depend on and be directly influenced by (a) the specific characteristics of the active compound and the particular therapeutic effect to be achieved and (b) the inherent limitations in the field of the art of compounding such active compounds for treating the susceptibility of the individual.

[0104] The exemplary non-limiting range for the therapeutic or prophylactic effective dose of the antibody described herein (e.g., DX-2930) is approximately 150 mg or 300 mg. As will be understood by those skilled in the art, the therapeutic or prophylactic effective dose of the antibody may be lower for pediatric subjects than for adult subjects. In some embodiments, the effective dose administered to pediatric subjects is a fixed dose or a dose based on body weight. In some embodiments, an effective dose of less than approximately 150 mg or 300 mg is administered to pediatric subjects. In some embodiments, the therapeutic or prophylactic effective dose of the antibody is administered every two weeks or every four weeks during a first treatment period. In some embodiments, the antibody may be administered to subjects during a second treatment period. In some embodiments, the therapeutic or prophylactic effective dose of the antibody in the first treatment period is different from the therapeutic or prophylactic effective dose of the antibody in the second treatment period. In some embodiments, the therapeutic or prophylactic effective dose of the antibody in the first treatment period is 150 mg, and the therapeutic or prophylactic effective dose of the antibody in the second treatment period is 300 mg. In some embodiments, the therapeutic or prophylactic effective dose of antibody in the first treatment period is the same as the therapeutic or prophylactic effective dose of antibody in the second treatment period. In one example, the therapeutic or prophylactic effective dose of antibody in both the first and second treatment periods is 300 mg.

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

[0106] kit The antibody described herein (e.g., DX-2930) may be provided in a kit, for example, as a component of the kit. For example, this kit includes (a) the DX-2930 antibody, for example, a composition containing the antibody (e.g., a pharmaceutical composition), and optionally (b) informational material. The informational material may be explanatory, instructive, marketing, or other material relating to the method described herein and / or the use of the antibody described herein (e.g., DX-2930), for example, the method described herein. In some embodiments, this kit includes one or more doses of DX-2930. In some embodiments, one or more doses are 150 mg or 300 mg.

[0107] The informational materials of this kit are not limited to their form. In one embodiment, the informational materials may include information on the preparation of the compound, the molecular weight, concentration, expiration date, batch or place of manufacture information, etc. In one embodiment, the informational materials may relate to the use of antibodies to treat, prevent or diagnose disorders and conditions, such as plasma kallikrein-related diseases or conditions.

[0108] In one embodiment, the information material may include instructions for administering the antibody described herein (e.g., DX-2930) in a manner appropriate to carry out the method described herein, for example, in an appropriate dose, dosage form, administration method or administration schedule (e.g., the dose, dosage form, administration schedule or administration method described herein). In another embodiment, the information material may include instructions for administering the antibody described herein (e.g., DX-2930) to an appropriate subject, for example, a human, for example, a human having or at risk of having plasma kallikrein-related disease or condition. For example, this material may include instructions for administering the antibody described herein (e.g., DX-2930) to a patient having a disorder or condition described herein, for example, plasma kallikrein-related disease, in accordance with the administration schedule described herein. The information material of this kit is not limited to its form. Often, the information material, for example, instructions, is provided in printed form, but it may also be in other forms, such as computer-readable material.

[0109] The antibodies described herein (e.g., DX-2930) may be provided in any form, such as liquid, dry, or lyophilized. 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, and a sterile aqueous solution is preferred. If the antibodies are provided in a dry form, reconstitution is generally by adding a suitable solvent. The solvent, such as sterile water or buffer, may optionally be provided in the kit.

[0110] This kit may include one or more containers for a composition containing the antibody described herein (e.g., DX-2930). In some embodiments, the kit includes separate containers, dividers, or compartments for the composition and informational materials. For example, the composition may be contained in a bottle, vial, or syringe, and the informational materials may be contained in conjunction with the container. In other embodiments, the individual elements of the kit are contained in a single, undivided container. For example, the composition is contained in a bottle, vial, or syringe, with informational materials attached therein in the form of a label. In some embodiments, the kit includes a plurality of individual containers (e.g., packs), each containing one or more unit dose formulations (e.g., dosage forms described herein) of the antibody described herein (e.g., DX-2930). For example, the kit includes a plurality of syringes, ampoules, foil packets, or blister packs, each containing a single unit dose of the antibody described herein (e.g., DX-2930). The container for this kit may be airtight, water-resistant (e.g., impermeable to changes in moisture or evaporation), and / or light-shielding.

[0111] The kit optionally includes a device suitable for administering the composition, such as a syringe or any such delivery device. In one embodiment, the device is an embedded device for dispensing a fixed dose of the antibody. The disclosure also features a method of providing the kit by combining, for example, the components described herein.

[0112] treatment In some embodiments, this disclosure provides the use of antibodies 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 angioedema (HANE). HAE is characterized by unpredictable, recurrent episodes of severe subcutaneous or submucosal swelling (angioedema), which can affect, for example, the limbs, face, genitals, gastrointestinal tract, and airways (Zuraw, 2008). Symptoms of HAE include, for example, swelling of the arms, legs, lips, eyes, tongue, and / or throat; swelling of the throat (larynx), which may include sudden hoarseness, and / or airway obstruction that can lead to death by asphyxiation (Bork et al., 2012; Bork et al., 2000). Approximately 50% of all HAE patients experience a laryngeal attack during their lifetime, and there is no way to predict whether a patient is at risk of a laryngeal attack (Bork et al., 2003; Bork et al., 2006). HAE symptoms include abdominal colic with no apparent cause; and / or recurrent episodes of bowel swelling that can be severe and lead to bowel symptoms similar to an abdominal emergency, such as abdominal colic, vomiting, dehydration, diarrhea, pain, shock, and / or unnecessary surgery (Zuraw, 2008). The swelling can last for up to 5 days or more. About one-third of individuals with HAE develop a non-itchy rash called erythema marginata during attacks. Most patients experience multiple attacks per year.

[0114] HAEs are rare disorders, and their exact prevalence is unknown, but current estimates range from 1 in 10,000 to 1 in 150,000 people, with many authors agreeing that 1 in 50,000 people 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 a crucial role in the pathogenesis 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, resulting in overproduction of bradykinin. Bradykinin is a vasodilator thought to be responsible for the characteristic HAE symptoms of localized swelling, inflammation, and pain (Craig et al., 2012; Zuraw et al., 2013).

[0116] Airway swelling can be life-threatening, and in some patients, it can lead to death. The mortality rate is estimated to be between 15 and 33%. HAEs result in approximately 15,000 to 30,000 emergency transports per year.

[0117] Trauma or stress, such as dental procedures, illnesses (e.g., viral illnesses like the common cold and influenza), menstruation, and surgery, can trigger angioedema attacks. To prevent acute HAE attacks, patients can try to avoid certain stimuli that have previously triggered attacks. However, attacks often occur without known triggers. 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 approximately 3-4 days (ghr.nlm.nih.gov / condition / hereditary-angioedema). The frequency and duration of attacks vary considerably among individuals with hereditary angioedema and even within the same family.

[0118] There are three types of HAE, known as type I, type II, and type III, all of which can be treated by the methods described herein. HAE is estimated to occur in 1 in 50,000 people, with type I accounting for about 85 percent of cases, type II accounting for about 15 percent, and type III being extremely rare. Type III is the most recently described form and was initially thought to occur only in women, but families with affected men have been identified.

[0119] HAE is inherited in an autosomal dominant pattern, meaning that affected individuals can inherit a mutation from one affected parent. New mutations can also occur within the gene, and therefore HAE can occur even in individuals with no family history of the disorder. It is estimated that 20–25% of cases are due to new, spontaneous mutations.

[0120] Mutations in the SERPING1 gene cause type I and type II hereditary angioedema. The SERPING1 gene provides instructions for the production of the C1 inhibitor protein, which is crucial for controlling inflammation. The C1 inhibitor blocks the activity of certain proteins that promote inflammation. Mutations that cause type I hereditary angioedema lead to decreased levels of C1 inhibitor in the blood. In contrast, mutations that cause type II result in the production of abnormally functioning C1 inhibitors. Approximately 85% of patients have type I HAE, characterized by very low production of functionally normal C1-INH protein, while the remaining approximately 15% of patients have type II HAE, resulting in normal or high levels of functionally defective C1-INH (Zuraw, 2008). If functional C1 inhibitor levels are not adequate, an excess of bradykinin is produced from high molecular weight kininogen (HMWK), leading to 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 into body tissues through the blood vessel walls. Excessive fluid accumulation in body tissues causes episodes of swelling seen in individuals with type I and type II hereditary angioedema.

[0121] Mutations in the F12 gene are associated with several causes of type III hereditary angioedema. The F12 gene provides instructions for the production of coagulation factor XII. In addition to playing a crucial role in blood coagulation, factor XII is also a major stimulant 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, making the blood vessel walls more prone to leakage, which leads to episodes of swelling. The causes of other cases of type III hereditary angioedema remain unknown. Mutations in one or more unidentified genes may be responsible for the impairment in these cases.

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

[0123] Similar to adults, children with HAE can experience recurrent and debilitating attacks. Symptoms can appear as early as 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 a significant unmet medical need among children with HAE, particularly in adolescence, because the disease generally worsens after adolescence (Bennett and Craig, 2015; Zuraw, 2008).

[0124] Treatment of HAEs with C1 inhibitors and other therapies is described in Kaplan, AP, J Allergy Clin Immunol, 2010, 126(5):918-925.

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

[0126] Purified C1 inhibitors derived from human blood have 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 Behring P (CSL Behring) was approved by the FDA in 2009 for acute exacerbations. Nanofiltered ViroPharma was approved by the FDA in 2008 for prophylactic use. Rhucin (Pharming) is a recombinant C1 inhibitor under investigation that does not carry the risk of transmission of infectious diseases from human blood-derived pathogens.

[0127] Treatment for acute HAE attacks may include pain medication and / or intravenous fluid administration.

[0128] Other therapeutic modalities may stimulate the synthesis of C1 inhibitors or reduce their consumption. Androgens such as danazol may reduce the frequency and severity of seizures by stimulating C1 inhibitor production.

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

[0130] Newer treatments target the contact cascade. Ecalantide (KALBITOR®, DX-88, Dyax) inhibits plasma kallikrein and is approved in the United States. Icatibant (Firazyr®, Shire) inhibits bradykinin B2 receptors and is approved in Europe and the United States.

[0131] The 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; 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] The symptoms of HAE can be assessed using, for example, questionnaires, such as those 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 a clinical history consistent with HAE (e.g., subcutaneous or mucosal, non-inflammatory swelling episodes) or by diagnostic tests (e.g., C1-INH function tests and C4 level assessment).

[0133] (ii) Treatment of HAE with anti-PKal antibody This disclosure provides a method for treating hereditary angioedema (HAE) (e.g., relieving, stabilizing, or eliminating one or more symptoms) by administering an antibody described herein (e.g., a therapeutically effective dose of the antibody described herein) to a subject who has or is suspected of having HAE, for example, according to the administration schedule described herein. Furthermore, a method is provided for treating HAE by administering an antibody described herein (e.g., a therapeutically effective dose of the antibody described herein) to a subject who has or is suspected of having HAE, for example, according to the administration schedule described herein, or in combination with a second treatment, for example, one other agent described herein. This disclosure also provides a method for preventing HAE or its symptoms by administering an antibody described herein (e.g., a prophylactically effective dose of the 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 to it), for example, according to the administration schedule described herein. In some examples, the subject may be a human patient who does not have HAE symptoms at the time of treatment. In some embodiments, the subject is a human patient who has HAE type I or HAE type II. In some embodiments, the subjects are human patients who have experienced at least two HAE attacks per year prior to treatment (e.g., two, three, four, or five or more).

[0134] In some embodiments, the subjects are female. In some embodiments, the subjects are children. In some embodiments, the subjects are adolescents under 18 years of age. In some embodiments, the subjects are adolescents between 12 and 18 years of age. In some embodiments, the subjects are between 40 and 65 years of age.

[0135] In some embodiments, the subject may be defined by sex. 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, an anti-pKal antibody (e.g., DX-2930) of approximately 300 mg may be administered to any of the human patient subgroups every two weeks. In other embodiments, such human patients may be administered approximately 150 mg of the antibody every two or four weeks. In yet another embodiment, such human patients may be administered approximately 300 mg of the antibody every four weeks.

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

[0139] Treatment involves administering an effective dose to reduce, alleviate, modify, treat, induce remission, improve, or influence a disorder, its symptoms, or predisposition to the disorder. This treatment may also delay the onset of the disease or condition, for example, by preventing its onset or exacerbation.

[0140] The method of administering the DX-2930 antibody is also described in "Pharmaceutical Compositions". The appropriate dose of the antibody used may depend on the age and weight of the subject and the specific drug used. This antibody can be used as a competitor to inhibit or reduce undesirable interactions between plasma kallikrein and its substrates (e.g., factor XII or HMWK). The antibody dose 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 this antibody is administered every two weeks or every four weeks. In some embodiments, this antibody is administered to the subject in a first treatment period, which includes administration of 150 mg or 300 mg of this antibody every two weeks or every four weeks. In some embodiments, this antibody is administered to the subject in a second treatment period following the first treatment period. In some embodiments, 300 mg of this antibody is administered as a single dose. If the subject experiences an HAE attack after the single dose, the antibody may be administered at a dose of 300 mg every two weeks.

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

[0142] This antibody can be used directly in vivo to eliminate antigen-expressing cells via innate complement-dependent cytotoxicity (CDC) or antibody-dependent cytotoxicity (ADCC). The antibodies described herein may include a complement-binding effector domain, such as an Fc portion from IgG1, -2, or -3, or a corresponding portion of IgM that binds to 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, the phagocytosis of target cells coated with the antibodies described herein can be improved by the binding of complement proteins. In another embodiment, target cells coated with the antibody containing the complement-binding effector domain are lysed with complement.

[0143] The method of administering the DX-2930 antibody is described in "Pharmaceutical Compositions." The appropriate dosage of the molecule used depends on the age and weight of the subject and the specific drug being used. This antibody may be used as a competitor to inhibit or reduce undesirable interactions, for example, between natural or pathological substances and plasma kallikrein.

[0144] A therapeutically effective dose of any antibody described herein may be administered to a subject who has, is suspected of having, or is at risk of having HAE, thereby treating the disorder (e.g., alleviating or improving the symptoms or characteristics of the disorder, slowing, stabilizing, and / or halting disease progression).

[0145] The antibodies described herein may be administered in therapeutically effective doses. A therapeutically effective dose of an antibody is an amount effective, in a single dose or multiple doses administered to a subject, to treat the subject, for example, to cure, reduce, alleviate or improve at least one symptom of a disorder in the subject to a degree exceeding what would be expected if such treatment were not performed.

[0146] The drug regimen can be adjusted to provide the optimal desired response (e.g., therapeutic response). For example, a single bolus may be administered, multiple divided doses may be administered over time, or the dose may be gradually reduced or increased as indicated by the emergency of the treatment situation. In other examples, multiple doses may be administered over time after a bolus, or the dose may be gradually reduced or increased as indicated by the emergency of the treatment situation. In other examples, the dose may be divided into multiple doses and administered over time. For ease of administration and for dose uniformity, it is particularly advantageous to formulate parenteral compositions in unit dose forms. When used herein, unit dose form refers to a physically separate unit appropriate as a unit dose for the subject being treated; each unit contains a predetermined amount of the active compound calculated to produce the desired therapeutic effect in relation to the required pharmaceutical carrier.

[0147] In some embodiments, the antibody described herein is administered in a drug regimen during a first treatment period. In some embodiments, the antibody is administered in multiple doses during the first treatment period. During this period, the therapeutic or prophylactic effective dose of the antibody (e.g., DX-2930) may be about 150 mg or 300 mg and is administered weekly, every two weeks, every three weeks, every four weeks, every five weeks, every six weeks, every seven weeks, or every eight weeks or more. In some embodiments, the therapeutic or prophylactic effective dose of the antibody (e.g., DX-2930) may be about 300 mg and is administered to female subjects weekly, every two weeks, every three weeks, every four weeks, every five weeks, every six weeks, every seven weeks, or every eight weeks or more. In some embodiments, the therapeutic or prophylactic effective dose of the antibody (e.g., DX-2930) may be about 300 mg and is administered to subjects under 18 years of age weekly, every two weeks, every three weeks, every four weeks, every five weeks, every six weeks, every seven weeks, or every eight weeks or more. In some embodiments, the therapeutic or prophylactic effective dose of this antibody (e.g., DX-2930) may be approximately 300 mg, administered weekly, every two weeks, every three weeks, every four weeks, every five weeks, every six weeks, every seven weeks, or every eight weeks or more to subjects aged 40-65 years.

[0148] In some embodiments, the therapeutic or prophylactic effective dose of this antibody (e.g., DX-2930) may be approximately 300 mg, administered weekly, every two weeks, every three weeks, every four weeks, every five weeks, every six weeks, every seven weeks, or every eight weeks or more to subjects aged 65 years or older. In a specific example, the antibody is administered to subjects at a dose of approximately 300 mg every two weeks. In another specific example, the antibody is administered to subjects at a dose of approximately 300 mg every four weeks.

[0149] In some embodiments, the therapeutic or prophylactic effective dose of this antibody (e.g., DX-2930) may be approximately 300 mg, administered weekly, every two weeks, every three weeks, every four weeks, every five weeks, every six weeks, every seven weeks, or every eight weeks or more to subjects who have experienced at least one laryngeal HAE attack. In a specific example, the antibody is administered to subjects at a dose of approximately 300 mg every two weeks. In another specific example, the antibody is administered to subjects at a dose of approximately 300 mg every four weeks.

[0150] In some embodiments, the therapeutic or prophylactic effective dose of this antibody (e.g., DX-2930) may be approximately 150 mg or 300 mg, administered weekly, every two weeks, every three weeks, every four weeks, every five weeks, every six weeks, every seven weeks, or every eight weeks or more to subjects under 18 years of age. In a specific example, the antibody is administered to a subject at a dose of approximately 300 mg every two weeks. In another specific example, the antibody is administered to a subject at a dose of approximately 300 mg every four weeks.

[0151] In some embodiments, the therapeutic or prophylactic effective dose of the antibody (e.g., DX-2930) may be approximately 150 mg or 300 mg, administered every two weeks or every four weeks. In some embodiments, the therapeutic or prophylactic effective dose of the antibody (e.g., DX-2930) may be 300 mg, administered to the subject every two weeks. In some embodiments, the therapeutic or prophylactic effective dose of the antibody (e.g., DX-2930) may be 300 mg, administered to the subject every four weeks. In some embodiments, the therapeutic or prophylactic effective dose of the antibody (e.g., DX-2930) may be 150 mg, administered to the subject every four weeks. In some embodiments, the therapeutic or prophylactic effective dose is administered at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least twelve, and at least thirteen times or more. In some embodiments, the first treatment period is 26 weeks. In some embodiments, the therapeutic or prophylactic effective dose is 150 mg, administered to the subject every four weeks (e.g., every four weeks for 26 weeks, resulting in a total of seven doses delivered). In some embodiments, the therapeutic or prophylactic effective dose is 300 mg, administered to the subject every two weeks (e.g., every two weeks for 26 weeks, resulting in a total of 13 doses delivered). In some embodiments, the therapeutic or prophylactic effective dose is 300 mg, administered to the subject every four weeks (e.g., every four weeks for 26 weeks, resulting in a total of seven doses 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 treatment schedule described allows for a margin 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 described above.

[0153] In some embodiments, the therapeutic or prophylactic effective dose is administered in a dosing regimen during a second treatment period following a first treatment period. In some embodiments, the therapeutic or prophylactic effective dose differs between the first and second treatment periods. In some embodiments, the therapeutic or prophylactic effective dose for the second treatment period is approximately 300 mg. During this period, the antibody may be administered in multiple doses of approximately 300 mg, such as 300 mg every two weeks. In some embodiments, during the second treatment period, multiple doses of the antibody are administered at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 times. In some embodiments, the second treatment period is 26 weeks. In some embodiments, the antibody is administered at doses of approximately 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 is administered approximately two weeks after the last dose of the first treatment period.

[0154] In any of the embodiments described herein, the timing of administration of the antibody is approximate and may include the three days before and the three days after a specified day (for example, administration every two weeks may include administration on day 11, day 12, day 13, day 14, day 15, day 16, or day 17).

[0155] In some embodiments, the antibody described herein is administered as a single dose of approximately 300 mg to a subject who has previously received prior HAE treatment (first treatment), such as multi-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 approximately 300 mg every two weeks for an appropriate period, e.g., 26 weeks. In some embodiments, the first of multiple doses is administered within one week of an HAE attack (e.g., within day 1, 2, 3, 4, 5, 6, or 7 of the HAE attack). In some embodiments, the antibody is administered at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least twelve, or at least thirteen times or more.

[0156] Pre-HAE treatment may include the same antibody described herein (e.g., DX-2930). In some embodiments, pre-HAE treatment may include multiple doses of DX-2930 administered every two weeks or every four weeks. In some embodiments, DX-2930 is administered to the subject (e.g., subcutaneously) at doses of 150 mg every four weeks, 300 mg every two weeks, or 300 mg every four weeks. In one example, the subject was pre-treated with the antibody every two weeks or every four weeks for 26 weeks prior to a single dose of the antibody. In some embodiments, the multiple doses of the antibody in pre-treatment are administered at least two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, or thirteen times. In some embodiments, the antibody was pre-treated on days 0, 28, 56, 84, 112, 140, and 168. In some embodiments, a single dose of approximately 300 mg of this antibody is administered approximately two weeks after the last dose of pre-treatment. In one example, the single dose for 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 approximate and includes the three days before and the three days after the specified day (for example, administration every two weeks includes administration on day 11, day 12, day 13, day 14, day 15, day 16, or day 17).

[0158] In some embodiments, subjects may be evaluated to establish a baseline HAE seizure rate before administering an antibody according to any of the methods described herein. Such an evaluation period may be called a “break-in period.” In some embodiments, the baseline HAE seizure rate must match or exceed a minimum number of HAE seizures over a given period. In one example, a subject experiences at least one HAE seizure during a 4-week break-in period prior to the first dose of the antibody. In another example, a subject experiences less than 1 to 2 seizures per month during a 4-week break-in period prior to the first dose of the antibody. In yet another example, a subject experiences less than 2 to 3 seizures per month during a 4-week break-in period prior to the first dose of the antibody. In yet another example, a subject experiences 3 or more seizures per month during a 4-week break-in period prior to the first dose of the antibody. In yet another example, a subject experiences at least two HAE seizures during an 8-week break-in period prior to the first dose of the antibody. In another example, the subjects experienced at least one HAE attack per month on average.

[0159] In some embodiments, the therapeutic or prophylactic effective dose of this antibody (e.g., DX-2930) may be approximately 150 mg or 300 mg, administered every 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, or 8 weeks or more to subjects who have experienced fewer than 1-2 HAE attacks per month during the acclimatization period before the first dose of this antibody. In some embodiments, the therapeutic or prophylactic effective dose of this antibody (e.g., DX-2930) may be approximately 150 mg or 300 mg, administered every 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, or 8 weeks or more to subjects who have experienced fewer than 2-3 HAE attacks per month during the acclimatization period before the first dose of this antibody. In some embodiments, the therapeutic or prophylactic effective dose of this antibody (e.g., DX-2930) may be approximately 150 mg or 300 mg, administered every 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, or 8 weeks or more to subjects who have experienced more than 3 HAE attacks per month during the adjustment period prior to the initial administration of this antibody.

[0160] In some embodiments, administration of the antibody according to any of the methods described herein results in a reduction in the mean HAE seizure rate in subjects. In some embodiments, the percentage reduction in the mean HAE seizure rate after administration of the antibody according to any of the methods described herein can be determined by comparing it to the HAE seizure rate in subjects that did not receive the antibody (e.g., subjects who received a placebo). In some embodiments, the percentage reduction in the mean HAE seizure rate may be at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95% compared to the HAE seizure rate in subjects who did not receive the antibody.

[0161] Any of the subjects described herein may have previously received pretreatment for HAE, such as prophylactic or therapeutic treatment for HAE. Aspects of this disclosure also provide a method for administering an antibody described herein (e.g., DX-2930) to a subject that has previously received one or more pretreatments for HAE. In some embodiments, the pretreatment for HAE is a treatment comprising an antibody described herein (e.g., DX-2930). In some embodiments, the subject was pre-treated with multiple doses of DX-2930 every two weeks or every four weeks. In some embodiments, the subject was pre-treated with 150 mg of DX-2930 every two weeks. In some embodiments, the subject was pre-treated with 300 mg of DX-2930 every two weeks. In some embodiments, the subject was pre-treated with 300 mg of DX-2930 every four weeks. In some embodiments, multiple doses of the pretreatment antibody are administered at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 times.

[0162] In some embodiments, the subject has received one or more prior treatments for HAE, such as long-term prophylactic treatment, which may include any of the HAE therapeutic agents known in the art. Typical anti-HAE agents include, but are not limited to, C1 inhibitors (e.g., SynRise®, Berinart®, or Ruconest®), plasma kallikrein inhibitors (e.g., Kalbitor®), bradykinin receptor inhibitors (e.g., Filazyr®), attenuated androgens (e.g., Danazol), and antifibrinolytic agents (e.g., traexamic acid). In some embodiments, the subject had received treatment with a C1 inhibitor prior to the first treatment period. In some examples, the subject may undergo a dose-tasting period before receiving anti-pKal antibody treatment as described herein. The dose tapering period refers to the period prior to anti-pKal antibody treatment, during which the dose, frequency, or both of the anti-HAE agent is gradually reduced so that subjects receiving prior HAE treatment (e.g., C1-INH, oral androgens, and / or oral antifibrinolytic agents) can gradually transition from the prior HAE treatment to the anti-pKal antibody treatment described herein. In some embodiments, dose tapering includes a progressive or stepwise method of reducing the dose of the prior treatment and / or the frequency in which the prior treatment is administered. The tapering period may last 2 to 4 weeks and may vary depending on the factors of the individual patent. In some examples, the prior treatment is completed before the start of anti-pKal antibody treatment. In other examples, the prior treatment may be completed within an appropriate time frame (e.g., 2, 3, or 4 weeks) after the subject has received the initial dose of the anti-pKal antibody.

[0163] Alternatively, subjects who have received prior HAE treatment may be directly transitioned to anti-pKal antibody treatment as described herein without a dose-tasking period.

[0164] In some embodiments, the therapeutic or prophylactic effective dose of this 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, or every 8 weeks or more to subjects who have received one or more prior treatments for HAE.

[0165] In other embodiments, the subject receives no pretreatment of HAE before the initial treatment, no initial treatment period, and / or no additional single-dose or multi-dose treatments (second treatment period) as described herein. In some embodiments, the subject receives no treatment other than the antibodies described herein during the first treatment period and / or the second treatment period. In some embodiments, the subject receives no pretreatment of HAE for at least two weeks (e.g., at least two, three, four, or five weeks) before, during, or after the initial treatment or the first treatment period. In some embodiments, the subject does not receive long-term prophylaxis of HAE (e.g., C1 inhibitors, weakened androgens, antifibrinolytic agents) for at least two weeks before, during, or after the initial treatment or the first treatment period. In some embodiments, subjects do not receive HAE treatment including angiotensin-converting enzyme (ACE) inhibitors for at least 4 weeks prior to, during, and / or during the first treatment period or the first treatment period. In some embodiments, subjects do not receive estrogen-containing drug therapy for at least 4 weeks prior to, during, and / or during the first treatment period or the first treatment period. In some embodiments, subjects do not receive androgens (e.g., stanozolol, danazol, oxandrolone, methyltestosterone, testosterone) for at least 2 weeks prior to, during, and / or during the first treatment period or the first treatment period.

[0166] Any of the methods described herein may further include monitoring the patient for adverse effects (e.g., elevated creatine phosphatase levels) and / or the level of antibody-mediated pKal inhibition (e.g., serum or plasma antibody concentration or pKal activity level) before, during, or after the procedure. If one or more adverse effects are observed, the antibody dose may be reduced or the procedure may be terminated. If the inhibition level falls below the minimum therapeutic level, a further dose of the antibody may be administered to the patient. Patients may also be evaluated for: antibody production 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)); priority of subcutaneous administration of the antibody (e.g., D-2930) compared to other injectable agents; and 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 a series of treatments (e.g., after the initial dose) to evaluate the effectiveness of the treatment. If the plasma or serum concentration of the antibody is lower than about 80 nM, an additional dose may be required, which may be the same as or higher than the initial 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 the subject treated with the antibody. Such assays may include synthetic substrate assays or Western blot assays for measuring cleaved kininogen as described herein.

[0168] Alternatively, plasma or serum levels of creatine kinase and / or one or more coagulation parameters (e.g., activated partial thromboplastin time (aPTT), prothrombin time (PT), hemorrhagic events) can be monitored during the treatment. If plasma or serum levels of creatine kinase are observed to increase during the treatment, the antibody dose may be reduced or the treatment may be terminated. Similarly, if one or more coagulation parameters are found to be significantly affected during the treatment, the antibody dose may be changed or the treatment may be terminated.

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

[0170] (iii) Combination therapy The antibodies described herein (e.g., DX-2930) may be administered in combination with one or more other therapeutic agents to treat diseases or conditions related to plasma kallikrein activity, for example, the diseases or conditions described herein. For example, the antibodies described herein (e.g., DX-2930) may be used therapeutically or prophylactically (e.g., before, during, or after a series of procedures) with another anti-plasma kallikrein Fab or IgG (e.g., another Fab or IgG described herein), another plasma kallikrein inhibitor, peptide inhibitor, small molecule inhibitor, or surgical procedure. Examples of plasma kallikrein inhibitors that may be used in combination therapy with plasma kallikrein conjugated to the antibodies described herein include, for example, the plasma kallikrein inhibitors described in International Publication No. 95 / 21601 or International Publication No. 2003 / 103475.

[0171] One or more plasma kallikrein inhibitors can be used in combination with the antibodies described herein (e.g., DX-2930). For example, the combination may reduce the required inhibitor dose, thereby mitigating 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 used concurrently with a second anti-HAE therapeutic agent such as ecalantide, a C1 esterase inhibitor (e.g., SynRise®), 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. The drugs or therapeutic agents may 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 performed at different times. The time between the administration of one drug and another may be in minutes, hours, days, or weeks. The use of plasma kallikrein-conjugated antibodies as described herein may also be used to reduce the dose of another therapeutic agent, for example, to mitigate side effects associated with the other drug being administered. Thus, the combination may involve administering the second drug at a dose at least 10, 20, 30, or 50% lower than when used in the absence of the plasma kallikrein-conjugated antibody. In some embodiments, the subject may be administered a C1-inhibitor as a loading IV dose or SC dose simultaneously with the initial dose of the anti-pKal antibody (e.g., DX-2930) as described herein. Next, the patient can continue anti-pKal antibody treatment (without further administration of C1 inhibitors).

[0174] Combination therapy may include administering medications that reduce the side effects of other therapeutic agents. These medications may be those that reduce the side effects of treatment for plasma kallikrein-related diseases.

[0175] (iv) Assays for evaluating treatment regimens Assay methods for evaluating the effectiveness of any of the treatment methods described herein are also within the scope of this disclosure. In some embodiments, the plasma or serum concentration of one or more HAE-related biomarkers (e.g., 2-chain HMWK) may be measured before and / or during a series of treatments (e.g., after the first dose) to evaluate the effectiveness of the treatment. In some embodiments, the plasma or serum concentration (level) of one or more HAE-related biomarkers obtained at a time after dose administration is compared to the concentration of the biomarker in a sample obtained at an earlier time after dose administration or before the first dose administration. In some embodiments, the biomarker is 2-HMWK.

[0176] The level of a biomarker can be measured by detecting the biomarker in a plasma or serum sample obtained from a subject using an antibody that specifically detects the biomarker, for example, by an immunoassay such as a Western blot assay or ELISA. In some embodiments, the level of 2-HWMK in a plasma or serum sample obtained from a subject is evaluated by an 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 obvious to those skilled in the art.

[0177] Without further effort, those skilled in the art can utilize the present invention to the greatest extent possible based on the above. Accordingly, the following specific embodiments should be construed as merely illustrative and not limiting the remainder of this disclosure. All publications cited herein are incorporated by reference with respect to the purposes or subjects referred to herein. [Examples]

[0178] Example 1: Efficacy and safety of DX-2930 treatment in a human patient subpopulation. Lanadermab is a sterile, preservative-free solution for injection with a pH of 6.0. The active ingredient, antibody DX-2930, is formulated using the following official components: 30 mM dibasic sodium phosphate dihydrate, 19.6 mM citrate monohydrate, 50 mM L-histidine, 90 mM sodium chloride, and 0.01% polysorbate 80. Each vial contains a nominal concentration of 150 mg of the active ingredient DX-2930 in 1 mL of solution. The test product is administered by subcutaneous (SC) injection into the upper arm in a blinded manner.

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

[0180] Patients aged ≥12 years with baseline HAE type I / II and ≥1 seizure / month were randomized in a 2:2:2:3 ratio to receive lanadelmab 150 mg every 4 weeks (q4wks), 300 mg every 4 weeks, 300 mg every 2 weeks, or placebo. A preliminary analysis was planned for subpopulations with an appropriate number of patients for Poisson regression.

[0181] The following primary and secondary efficacy endpoints were evaluated from day 14 to day 182. The primary endpoints of this study were the number of HAE attacks and the mean HAE attack rate. The secondary endpoints were, in order of priority: 1. Number of HAE attacks requiring emergency treatment 2. Number of moderate to severe HAE attacks.

[0182] Exploratory efficacy endpoints 1. The time from the 14th day until the first seizure occurs, i.e., the period of time from the 14th day until the first seizure occurs in the subject, during which the subject was seizure-free. 2. The number of high-pathological HAE seizures per week; a high-pathological HAE seizure is defined as any seizure having at least one of the following characteristics: a severe seizure requiring hospitalization (except hospitalization for <24 hours of observation), a hemodynamically significant seizure (systolic blood pressure <90, requiring IV fluid replacement, or accompanied by syncope or near-syncope), or a laryngeal seizure.

[0183] Clinical Tests Clinical tests were performed on patients included in the clinical trial, including general safety parameters (hematology, coagulation, urinalysis, and serological chemistry), serology, pregnancy tests, C1-INH function assays, C4 assays, C1q assays, PK samples, plasma anti-drug antibody tests, and PD samples. All clinical tests were performed using established and validated methods.

[0184] result Overall, 125 patients were treated with either lanadermab (n=84) or placebo (n=41). The mean HAE attack rate was determined for all patients and therefore for all patient subgroups. Using the mean number of HAE attacks, the percentage reduction in the mean HAE attack rate in patients treated with DX-2930 compared to patients treated with placebo was determined. The HAE attack rate was consistently reduced with DX-2930 compared to placebo in all patients and patient subgroups. However, as shown in Table 2, in some patient subgroups, a more significant percentage reduction, i.e., a more therapeutically effective reduction, was observed when 300 mg of DX-2930 was administered every two weeks compared to 300 mg of DX-2930 every four weeks (or 150 mg of DX-2930 every four weeks) compared to placebo treatment. Specifically, in patients under 18 years of age who received 300 mg of DX-2930 every four weeks, the HAE seizure rate decreased by 20.5% compared to placebo; in patients under 18 years of age who received 300 mg of DX-2930 every two weeks, a further reduction of approximately 42 percentage points in the seizure rate was observed (62.3%) (Figure 3A). In patients aged 40 to <65 years who received 300 mg of DX-2930 every four weeks, the HAE seizure rate decreased by 71.5% compared to placebo; and in patients aged 40 to <65 years who received 300 mg of DX-2930 every two weeks, the incidence rate decreased further by approximately 18 percentage points (89.8%). In female patients administered 300 mg of DX-2930 every four weeks, the rate of HAE attacks decreased by 69.6% compared to placebo; in female patients administered 300 mg of DX-2930 every two weeks, a further reduction of approximately 16 percentage points was observed (85.8%). In patients with a history of laryngeal attacks who were administered 300 mg of DX-2930 every four weeks, the rate of HAE attacks decreased by 64.2% compared to placebo; in patients with a history of laryngeal attacks who were administered 300 mg of DX-2930 every two weeks, a further reduction of less than 21 percentage points was observed (85.7%).

[0185] [Table 2]

[0186] All HAE type I / II patients treated with lanadelmab 300 mg q2wks or q4wks experienced a clinically meaningful and sustained reduction in HAE attack rates compared to placebo. However, certain subgroups of patients, such as women, patients <18 years or 40–65 years of age, and patients with a history of at least one laryngeal attack, showed better therapeutic efficacy with 300 mg every two weeks.

[0187] Patients were further stratified based on HAE attacks during the adjustment period, and the effectiveness of each lanadermab treatment regimen was evaluated in these patient subgroups. As shown in Tables 3–5 and Figures 1A–1C, each lanadermab treatment regimen resulted in a significant reduction in HAE attack rates compared to placebo in all subgroups.

[0188] [Table 3]

[0189] [Table 4]

[0190] [Table 5]

[0191] In patients using only C1 inhibitors (C1-INH) for long-term prophylaxis, baseline seizure rates were elevated during protocol-based C1-INH discontinuation compared to historical rates (over the past 3 months) (Figure 2A). Seizure rates during lanadelmab treatment were lower than historical rates. Seizure rates decreased by an average of 68.8%, 59.3%, and 82.1% during lanadelmab 150 mg q4wks, 300 mg q4wks, and 300 mg q2wks, respectively, compared to historical seizure rates during long-term prophylaxis.

[0192] Using a Poisson regression model, lanadelmab demonstrated consistent therapeutic effects compared to placebo in patients who received C1-INH alone for prophylaxis and in patients who did not receive long-term prophylaxis (Figure 2B). In patients who received C1-INH alone for long-term prophylaxis before lanadelmab administration, the mean seizure rate was significantly reduced by 73.6%, 71.6%, and 82.5% in the lanadelmab 150 mg q4wks, 300 mg q4wks, and 300 mg q2wks regimens, respectively, compared to placebo (P<0.001 for all comparisons).

[0193] Based on age, sex, weight, type of HAE (e.g., type I or type II), and history of laryngeal attacks, the percentage reduction in HAE attack rates in each drug regimen was evaluated for each subgroup of subjects receiving lanadelumab compared to placebo. Figures 4A-4E and 5.

[0194] Lanadermab significantly suppressed pKal activation, as demonstrated by its effect on cHMWK levels. A fixed-dose regimen of 300 mg every two weeks resulted in optimal clinical responses across a wide body weight range in both adolescents and adults.

[0195] Example 2: Efficacy and safety of DX-2930 (lanadermab) treatment in human adolescent patients In this Phase 3 and Open-Label Long-Term (OLE) trial, the efficacy and safety of lanadermab, a monoclonal antibody targeting plasma kallikrein, were investigated in adolescent patients with HAE who had C1 inhibitor deficiency.

[0196] For the Phase 3 trial, patients aged ≥12 years with physician-confirmed seizures ≥1 per 4 weeks were randomized to receive placebo or lanadelmab every 4 weeks at a dose of 150 mg (150 mg q4w), 300 mg q4w, or 300 mg q2w. In the Phase 3 trial, 10 out of 125 patients (8%) were adolescent (≥12 to <18 years). Prior to the start of the Phase 3 trial, 60.0% of patients had received C1-INH simply for long-term prophylaxis.

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

[0198] In the open-label long-term trial, non-rollover participants will receive an open-label dose of 300 mg lanadelmab subcutaneously on day 0 and continue to receive open-label 300 mg lanadelmab subcutaneously every two weeks for the treatment period according to the planned dosing regimen. A total of 26 doses will be administered, with the final dose given at the visit on day 350.

[0199] In rollover patients, 62.5% received C1-INH alone before the start of the open-label long-term trial. In non-rollover adolescent patients, 61.6% received long-term prophylactic treatment (C1-INH alone or C1-INH and oral therapy) before the start of the trial (primarily C1-INH alone; 46.2%). Monthly seizure rates (MAR) and other treatment-induced adverse events (TEAEs) were recorded.

[0200] Three adolescent subjects experienced 13 non-serious adverse events (TEAEs) during treatment. In an open-label long-term trial, 21 out of 212 patients (9.9%) were adolescent. Rollover patients (n=8) and non-rollover patients (n=13) had mean (SD) monthly seizure rates of 1.65 (1.158) and 1.54 (0.971) at baseline, respectively, and 0.35 (0.635) and 0.07 (0.166) during the treatment period, representing mean (SD) percentage changes of -84.371 (18.9415) and -94.893 (10.5230), respectively. Nine patients experienced 65 non-serious lanadermab-related TEAEs.

[0201] The results from this trial are provided in Table 6 below. Lanadermab was found to be effective and safe in reducing MAR in adolescent patients with HAE.

[0202] [Table 6]

[0203] In the Phase 3 trial, patients treated with lanadelmab 300 mg q4wks (n=3; 0.436[0.253]) or lanadelmab 300 mg q2wks (n=2; 0.207[0.148]) showed lower least squares mean (SE) HAE seizure rates from day 0 to day 182 compared to patients given placebo (n=4; 0.548[0.224]). This could not be estimated in the 150 mg q4wks treatment arm as it included only one adolescent patient. The estimated least squares mean monthly seizure rate ratio (vs. placebo) was favorable for lanadelmab treatment, particularly the 300 mg q2wks dose regimen, at 95% CI (Figure 3C). In an open-label long-term trial, the mean (SD) percentage change from baseline in the mean monthly seizure rate was 84.37 (18.94) (n=8; usual dosing stage) in rollover patients and -94.89 (10.52) (n=13; Figure 3B) in non-rollover patients.

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

[0205] In both the Phase 3 trial and its OLE, there were no deaths or trial discontinuations due to TEAEs.

[0206]

Table 7

[0207] In conclusion, lanadermab administration demonstrated good tolerability and reduced the monthly seizure rate in adolescents in phase 3 and open-label long-term studies.

[0208] Other Embodiments All properties disclosed herein can be combined in any combination. Each property disclosed herein can be replaced by an alternative property that serves the same, equivalent, or similar purpose. Thus, unless otherwise clearly stated, each disclosed property is merely an example of a comprehensive set of equivalent or similar properties.

[0209] From the above description, those skilled in the art can easily identify the basic features of the present invention and make various modifications and alterations to adapt it to various uses and conditions without departing from its spirit and scope. Accordingly, other embodiments are also within the scope of the claims.

[0210] equivalent While several embodiments of the present invention have been described and illustrated herein, those skilled in the art will readily recall a variety of other means and / or structures for performing the functions described herein and / or obtaining the results and / or one or more advantages, and each of such variations and / or modifications will be considered to fall within the scope of the embodiments of the present 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 illustrative, and that actual parameters, dimensions, materials and / or configurations will depend on the specific application in which the teachings of the present invention are used. Those skilled in the art will be able to recognize many equivalents to the specific embodiments of the present invention described herein or to verify them by mere ordinary experimentation. Thus, it will be understood that the embodiments described herein are presented merely as examples, and that embodiments of the present invention may be carried out in ways other than those specifically described and claimed, within the scope of the appended claims and their equivalents. Embodiments of this disclosure cover each individual property, system, article, material, kit and / or method described herein. Furthermore, any combination of two or more such properties, systems, articles, materials, kits and / or methods is included within the scope of the invention of this disclosure, provided that such properties, systems, articles, materials, kits and / or methods are not mutually inconsistent.

[0211] All definitions defined and used herein should be understood to take precedence over dictionary definitions, definitions in documents incorporated by reference, and / or the common meaning of the terms defined.

[0212] Where used herein and in the claims, the indefinite articles "a" and "an" should be understood to mean "at least one" unless the opposite is clearly indicated.

[0213] When used herein and in the claims, the phrase “and / or” should be understood to mean “either or both” of the elements thus linked, i.e., elements that exist in some cases together and in other cases separately. Any multiple elements listed with “and / or” should be interpreted in the same form, i.e., “one or more” of the elements thus linked. Other elements may exist, at their discretion and with or without relation to the specifically identified elements, other than those specifically identified by the “and / or” clause. Thus, as a non-limiting example, when used in conjunction with an open-ended word such as “including,” in one embodiment it may refer to A only (including, at their discretion, elements other than B); in another embodiment it may refer to B only (including, at their discretion, elements other than A); and in yet another embodiment it may refer to both A and B (including, at their discretion, other elements).

[0214] Where used herein and in the 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 interpreted as inclusive, that is, including not just a number of elements or at least one of the items in the list, but also multiple and, by any choice, further unlisted items. Only terms that are clearly indicated to be the opposite, such as “one of” or “exactly one,” or, when used in the claims, “consisting of,” refer to including exactly one element from a number of elements or list. In general, the term “or” will be interpreted only, when used herein, to refer to an exclusive substitute (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”

[0215] Where used herein and in the claims, the phrase “at least one” when referring to a list of one or more elements should be understood to mean at least one element selected from any one or more elements in the element list, but not necessarily including at least one of every element specifically enumerated in the element list, and not excluding any combination of elements in the element list. This definition also allows for the optional presence of elements other than those specifically identified in the list of elements referred to by the phrase “at least one,” regardless of whether they are related to the specifically identified elements. Therefore, as a non-limiting example, "at least one of A and B (or equivalently, "at least one of A or B" or equivalently, "at least one of A and / or B") may, in one embodiment, mean at least one that includes multiple A and B (and optionally include elements other than B); in another embodiment, mean at least one that includes multiple B and A (and optionally include elements other than A); and in yet another embodiment, mean at least one that includes multiple A and B (and optionally include other elements), etc.

[0216] Unless otherwise clearly indicated, it should be understood that in any method asserted herein, including multiple steps or actions, the order of the steps or actions of the Method is not necessarily limited to the order in which the steps or actions of the Method are enumerated.

[0217] In the claims and in the above specification, all transitional clauses such as “comprising,” “including,” “carrying,” “having,” “containing,” “involving,” “holding,” and “composed of” should be understood to be open-ended, meaning that they include but are not limited. As shown in Section 2111.03 of the U.S. Patent and Trademark Office's Manual of Patent Examining Procedures, only the transitional clauses “consisting of” and “consisting essentially of” are closed or semi-closed transitional clauses, respectively.

Claims

1. A composition for use in a method for treating hereditary angioedema (HAE) attacks or reducing the incidence of HAE attacks, comprising an antibody that binds to human plasma kallikrein (pKal) and comprises a heavy chain complementarity determining region (CDR) shown as SEQ ID NOs. 5-7 and a light chain CDR shown as SEQ ID NOs. 8-10, The aforementioned method, This includes administering the antibody to a human subject requiring it during a first treatment period; During the first treatment period, the antibody is administered to the human subjects in multiple doses, approximately 300 mg every two weeks; The aforementioned human subjects are either infected with HAE, suspected of being infected, or at risk of being infected. (i) Being female; (ii) Being between 40 and 65 years old; (iii) Having experienced one to two, two to three, or more than three HAE attacks in the four weeks prior to the first dose of the first treatment period; and / or (iv) Having received treatment with C1-INH prior to the first treatment period; composition.

2. A composition for use in a method for treating hereditary angioedema (HAE) attacks or reducing the incidence of HAE attacks, comprising an antibody that binds to human plasma kallikrein (pKal) and comprises a heavy chain complementarity determining region (CDR) shown as SEQ ID NOs. 5-7 and a light chain CDR shown as SEQ ID NOs. 8-10, The aforementioned method, This includes administering the antibody to a human subject requiring it during a first treatment period. The aforementioned human subject, (i) being in adolescence between the ages of 12 and 18; and (ii) The patient has experienced two to three or more HAE attacks in the four weeks prior to the first dose of the antibody, The antibody is administered to the human subjects at a dose of approximately 300 mg every two weeks. composition.

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

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

4.

5. The composition according to any one of claims 1 to 4, wherein the antibody comprises a heavy chain indicated by SEQ ID NO: 1 and a light chain indicated by SEQ ID NO:

2.

6. The composition according to any one of claims 1 to 5, wherein the antibody is formulated in a pharmaceutical composition comprising a pharmaceutically acceptable carrier.

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

8. The composition according to claim 7, wherein the sodium phosphate is concentrated at a concentration of about 30 mM, the citric acid is concentrated at a concentration of about 19 mM, the histidine is concentrated at a concentration of about 50 mM, the sodium chloride is concentrated at a concentration of about 90 mM, and the polysorbate 80 is concentrated at a concentration of about 0.01%.

9. The composition according to any one of claims 1 to 8, wherein the antibody is administered subcutaneously.

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

11. The composition according to 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 according to any one of claims 1 to 11, wherein the human subject has received one or more pre-HAE treatments prior to the first treatment period.

13. The composition according to claim 12, wherein the pre-HAE treatment comprises C1-INH, a plasma kallikrein inhibitor, a bradykinin receptor antagonist, an androgen, an antifibrinlytic agent, or a combination thereof.

14. The composition according to claim 13, wherein the pre-HAE treatment comprises C1-INH, ecalantide, icatibant, danazol, tranexamic acid, or a combination thereof.

15. The composition according to any one of claims 12 to 14, wherein the method includes a gradual reduction period for the one or more pre-HAE treatments.

16. The composition according to claim 15, wherein the gradual reduction period is approximately 2 to 4 weeks.

17. The composition according to any one of claims 12 to 16, wherein the one or more pre-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 according to any one of claims 1 to 11, wherein the human subject has not undergone prior HAE treatment.

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

20. The composition according to any one of claims 1 to 19, wherein the human subject has experienced 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.

21. The composition according to any one of claims 1 to 20, further comprising administering the antibody to the subject during a second treatment period following the first treatment period.

22. The composition according to claim 21, wherein the initial dose of the second treatment period is approximately two weeks after the last dose of the first treatment period.

23. The composition according to claim 21 or claim 22, wherein the second treatment period comprises one or more administrations of the antibody at approximately 300 mg.

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

25. The composition according to any one of claims 1 to 24, wherein the human subject has not received long-term prophylaxis to HAE or HAE treatment containing an angiotensin-converting enzyme (ACE) inhibitor, estrogen-containing drug therapy, or androgen before and / or during the first treatment period.

26. The composition according to any one of claims 21 to 24, wherein the human subject has not received long-term prophylaxis to HAE or HAE treatment containing an angiotensin-converting enzyme (ACE) inhibitor, estrogen-containing drug therapy, or androgen during the second treatment period.

27. The method described above is (a) After the first treatment period, administer the antibody to a human subject in need at a single dose of approximately 300 mg; (b) If the subject experiences an HAE seizure after (a), administer the antibody to the subject in one or more doses of approximately 300 mg; The composition according to any one of claims 1 and 3 to 26, further comprising:

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

29. The composition according to claim 28, wherein the initial dose of stage (b) is within one week after the HAE seizure.

30. The composition according to any one of claims 27 to 29, wherein the single dose of (a) and the initial dose of (b) are separated by at least 10 days.

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