Dual inhibitor kallikrein antibodies and uses thereof

Abstract

Aspects of the present application provide dual inhibitor anti-KLK5 / KLK7 antibodies and methods of using them to promote barrier function and reduce inflammation and to treat conditions such as Neethton Syndrome, eosinophilic esophagitis, and atopic dermatitis.

Description

Related Applications

[0001] This application is made pursuant to 35 U.S.C. This application claims priority under 35 U.S.C. 119(e) to U.S. Provisional Application U.S.S.N. 63 / 489,414, filed March 9, 2023, and U.S. Provisional Application U.S.S.N. 63 / 614,102, filed December 22, 2023, the entire contents of each of which are incorporated herein by reference.

[0002] Reference to Electronic Sequence Listing The contents of the electronic sequence listing (A104770001WO00-SEQ-LJG.xml; size: 36,826 bytes; and date created: March 8, 2024) is incorporated herein by reference in its entirety BACKGROUND

[0003] Kallikrein (KLK) regulates desquamation and innate immunity to maintain skin homeostasis and wound healing. In healthy skin, the outermost layer of the epidermis is shed periodically by a KLK-driven proteolytic cascade that leads to degradation of corneodesmosomes and desquamation. KLK5 is thought to be the primary activator of this proteolytic cascade. Autocatalytic KLK5 enzymatically converts proKLK7 and proKLK14 to active forms and stimulates a positive feedback loop that leads to more production of proKLK5 through KLK14. These KLK enzymes are controlled by endogenous serine protease inhibitors, such as lymphoepithelial Kazal-type-related inhibitor. KLK (including KLK5 and KLK7) dysregulation is associated with skin disorders, inflammatory diseases, and cancer. For example, overactive kallikrein 5 and 7 cause genetic and spontaneous epidermal barrier disorders (e.g., Netherton syndrome, eosinophilic esophagitis, atopic dermatitis). SUMMARY

[0004] Certain aspects of the present disclosure relate to the recognition that a loss of balance between endogenous KLK proteases and associated protease inhibitors can lead to barrier dysfunction and induce inflammation (see, e.g., Figure 1This can lead to inflammatory disorders such as Netherton's syndrome, eosinophilic esophagitis, and atopic dermatitis. In some embodiments, methods and related compositions are provided for inhibiting KLK5 and KLK7 with the aim of improving barrier function and reducing inflammation, thereby improving disease severity. In particular, several aspects of this disclosure provide dual inhibitory antibodies targeting KLK5 and KLK7 (referred to as anti-KLK5 / KLK7 antibodies) that have high binding affinity and specificity for both KLK5 and KLK7 through a common, unique antigen-specific binding site. Therefore, in some embodiments, this disclosure provides methods for treating disorders associated with KLK5 and KLK7 dysregulation, antibodies for use in the methods, and related antibody compositions, such as Netherton's syndrome, atopic dermatitis (with and without filaggrin mutations), eosinophilic esophagitis, nodular prurigo, chronic pruritus of unknown cause (CPUO), asthma (e.g., KLK5-associated asthma), and ichthyosis vulgaris.

[0005] In some aspects, this disclosure provides a dual inhibitor antibody that specifically binds to KLK5 and KLK7, the dual inhibitor antibody comprising HC CDR1, HCCDR2 and HC CDR3 having a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 7, and LC CDR1, LCCDR2 and LC CDR3 having a light chain variable domain having the amino acid sequence of SEQ ID NO: 8.

[0006] In some aspects, this disclosure provides a dual inhibitor antibody that specifically binds to KLK5 and KLK7, the dual inhibitor antibody comprising HC CDR1, HCCDR2 and HC CDR3 having a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 13, and LC CDR1, LCCDR2 and LC CDR3 having a light chain variable domain having the amino acid sequence of SEQ ID NO: 14.

[0007] In some aspects, this disclosure provides a dual inhibitor antibody that specifically binds to KLK5 and KLK7, the dual inhibitor antibody comprising HC CDR1, HCCDR2 and HC CDR3 having a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 17, and LC CDR1, LCCDR2 and LC CDR3 having a light chain variable domain having the amino acid sequence of SEQ ID NO: 14.

[0008] In some aspects, the present disclosure provides a dual inhibitor antibody that specifically binds to KLK5 and KLK7, the dual inhibitor antibody comprising a HC CDR1, a HC CDR2, and a HC CDR3 of a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 21, and a LC CDR1, a LC CDR2, and a LC CDR3 of a light chain variable domain having the amino acid sequence of SEQ ID NO: 14.

[0009] In some embodiments, the dual inhibitor antibody comprises a HC CDR1 having the amino acid sequence of SEQ ID NO: 1; a HC CDR2 having the amino acid sequence of SEQ ID NO: 2; a HC CDR3 having the amino acid sequence of SEQ ID NO: 3; a LC CDR1 having the amino acid sequence of SEQ ID NO: 4; a LC CDR2 having the amino acid sequence of SEQ ID NO: 5; and a LC CDR3 having the amino acid sequence of SEQ ID NO: 6.

[0010] In some embodiments, the dual inhibitor antibody comprises a HC CDR1 having the amino acid sequence of SEQ ID NO: 9; a HC CDR2 having the amino acid sequence of SEQ ID NO: 10; a HC CDR3 having the amino acid sequence of SEQ ID NO: 11; a LC CDR1 having the amino acid sequence of SEQ ID NO: 4; a LC CDR2 having the amino acid sequence of SEQ ID NO: 5; and a LC CDR3 having the amino acid sequence of SEQ ID NO: 12.

[0011] In some embodiments, the dual inhibitor antibody comprises a HC CDR1 having the amino acid sequence of SEQ ID NO: 9; a HC CDR2 having the amino acid sequence of SEQ ID NO: 15; a HC CDR3 having the amino acid sequence of SEQ ID NO: 16; a LC CDR1 having the amino acid sequence of SEQ ID NO: 4; a LC CDR2 having the amino acid sequence of SEQ ID NO: 5; and a LC CDR3 having the amino acid sequence of SEQ ID NO: 12.

[0012] In some embodiments, the dual inhibitor antibody comprises a HC CDR1 having the amino acid sequence of SEQ ID NO: 18; a HC CDR2 having the amino acid sequence of SEQ ID NO: 19; a HC CDR3 having the amino acid sequence of SEQ ID NO: 20; a LC CDR1 having the amino acid sequence of SEQ ID NO: 4; a LC CDR2 having the amino acid sequence of SEQ ID NO: 5; and a LC CDR3 having the amino acid sequence of SEQ ID NO: 12.

[0013] In some embodiments, the dual inhibitor antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 7; and a VL comprising the amino acid sequence of SEQ ID NO: 8.

[0014] In some embodiments, the dual inhibitor antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 13; and a VL comprising the amino acid sequence of SEQ ID NO: 14.

[0015] In some embodiments, the dual inhibitor antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 17; and a VL comprising the amino acid sequence of SEQ ID NO: 14.

[0016] In some embodiments, the dual inhibitor antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 21; and a VL comprising the amino acid sequence of SEQ ID NO: 14.

[0017] In some aspects, the disclosure provides a dual inhibitor antibody comprising a HC CDR1, a HC CDR2, a HC CDR3, a LC CDR1, a LC CDR2, and / or a LC CDR3 of any one of the dual inhibitor antibodies listed in Table la and Table lb.

[0018] In some aspects, the disclosure provides a dual inhibitor antibody comprising a VH and / or a VL of any one of the dual inhibitor antibodies listed in Table la and Table lb.

[0019] In some embodiments, the dual inhibitor antibody binds to the active site of KLK5 and the active site of KLK7.

[0020] In some embodiments, the dual inhibitor antibody competes with SPINK 5 and / or matriptase for binding to the active site of KLK5 and the active site of KLK7.

[0021] In some embodiments, the dual inhibitor antibody binds to active forms of KLK5 and KLK7, but not inactive forms of KLK5 or KLK7. In some embodiments, the antibody specifically binds to active forms of KLK5 and KLK7, but does not specifically bind to inactive forms of KLK5 or KLK7. In some embodiments, the antibody detectably binds to active forms of KLK5 and KLK7, but does not detectably bind to inactive forms of KLK5 or KLK7 under the same or comparable conditions.

[0022] In some embodiments, the dual inhibitor antibody inhibits protease activity of KLK5 and KLK7.

[0023] In some embodiments, the antibody is not cleaved in the heavy chain by KLK5 or KLK7 when bound to KLK5 or KLK7.

[0024] In some embodiments, the anti-KLK5 / KLK7 antibody is not a bispecific antigen binding molecule, wherein KLK5 binding is conferred by one binding site within the antibody and KLK7 binding is conferred by a different binding site.

[0025] In some embodiments, the anti-KLK5 / KLK7 antibody is a multispecific antigen binding molecule further comprising an antigen binding domain that binds to an antigen other than KLK5 or KLK7.

[0026] In some aspects, the present disclosure provides a composition comprising a dual inhibitor antibody described herein and an acceptable carrier.

[0027] In some aspects, the present disclosure provides a nucleic acid encoding a dual inhibitor antibody described herein.

[0028] In some aspects, the present disclosure provides a method of treating a skin barrier defect, the method comprising administering to a subject an effective amount of a dual inhibitor antibody described herein or a composition thereof. In some embodiments, the skin barrier defect is associated with Netherton Syndrome, atopic dermatitis, eosinophilic esophagitis, prurigo nodularis, chronic pruritus of unknown origin (CPUO), dry skin, asthma (particularly KLK5), ichthyosis vulgaris, or pruritus or chronic itch.

[0029] In some aspects, the present disclosure provides a dual inhibitor antibody against KLK5 and KLK7 (i.e., an anti-KLK5 / KLK7 antibody) or a composition thereof for use in a method of treating a skin barrier defect. In some embodiments, the anti-KLK5 / KLK7 antibody or a composition thereof is for use in a method of treating a skin barrier defect associated with Netherton syndrome, atopic dermatitis, eosinophilic esophagitis, prurigo nodularis, chronic prurigo of unknown origin (CPUO), dry skin, asthma (particularly KLK5), ichthyosis vulgaris, or pruritus or chronic prurigo.

[0030] The foregoing and other aspects, implementations, operations, functions, features, and embodiments of the present teachings can be more fully understood from the following description, taken in conjunction with the accompanying drawings. BRIEF DESCRIPTION OF DRAWINGS

[0031] The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate certain embodiments and together with the written description serve to provide a non-limiting example of the compositions and methods disclosed herein.

[0032] Figure 1 is a graphical illustration showing that aberrant protease activation (e.g., aberrant KLK5, KLK7, and KLK14) activation leads to skin barrier defect-associated diseases.

[0033] Figures 2A-2B shows the relative response curves of KLK5 / 7-Dual-Ab4 and comparative antibody #1 in binding to the active form of human KLK5 (huKLK5) or the pro-form of huKLK5 ( Figure 2A ) or the active form or pro-form of huKLK7 ( Figure 2B ). Comparative antibody #1 shows binding to both forms, while KLK5 / 7-Dual-Ab4 specifically binds to the active huKLK5.

[0034] Figures 3A-3B is SDS-PAGE results showing antibody control anti-KLK5-Ab1, KLK5 / 7-Dual-Ab1, KLK5 / 7-Dual-Ab2, KLK5 / 7-Dual-Ab3, KLK5 / 7-Dual-Ab4, and KLK5 / 7-Dual-Ab5 after incubation with KLK5 ( Figure 3A ) or KLK7 ( Figure 3B ) alone. Control anti-KLK5-Ab1 is a positive control for KLK5 cleavage activity (but it will not be cleaved by KLK7), as evidenced by two bands of approximately 38 kDa and 12 kDa weights. Neither KLK5 nor KLK7 cleaves the other antibodies.

[0035] Figure 4 Bispecific antibodies KLK5 / 7-Dual-Ab2, KLK5 / 7-Dual-Ab3 and KLK5 / 7-Dual-Ab4 were shown to have no inhibitory activity against other KLK family members and related proteases relative to isotype control. The tested antibodies do not specifically inhibit non-KLK5 / 7 family members or related proteases as the relative activity is not higher than the isotype control.

[0036] Figures 5A-5B The relative response of antibodies KLK5 / 7-Dual-Ab2 and KLK5 / 7-Dual-Ab4 binding to huKLK5 ( Figure 5A ) or huKLK7 ( Figure 5B ) in the presence of the serine protease inhibitor PMSF, leupeptin or SPINK5 is shown.

[0037] Figure 6 shows the competitive binding of anti-KLK5 / 7 antibodies versus SPINK5 to KLK5 (top) and KLK7 (bottom), SPINK5 binds to the active site of KLK5 and KLK7. In the left panel, antibody KLK5 / 7-Dual-Ab4 is bound to the chip and KLK5 or KLK7 is added, resulting in an increase in the binding curve. The addition of a second anti-KLK5 / 7 antibody or SPINK5 (indicated by brackets and "mAb #2") does not increase binding. In the right panel, SPINK5 is bound to the chip and KLK5 or KLK7 is added, resulting in an increase in the binding curve. The addition of an anti-KLK5 / 7 antibody (indicated by brackets and "mAb #2") does not increase binding as SPINK5 is already bound to the active site of KLK5 or KLK7.

[0038] Figures 7A-7B The effect of treatment with KLK5 / 7-Dual-Ab4 at 30 mg / kg ( Figure 7A ) or 3 mg / kg ( Figure 7B ) on stratum corneum thickness in the MC903 atopic dermatitis mouse model is shown. Treatment with KLK5 / 7-Dual-Ab4 results in a significant decrease in thickness.

[0039] Figures 8A-8I is a graph representing the therapeutic efficacy of anti-KLK5 / 7 antibodies on disease manifestations in the Nc / Nga atopic dermatitis mouse model as measured by clinical score ( Figure 8A ), histological score ( Figure 8D and Figure 8G ), stratum corneum thickness (ear thickness) ( Figure 8B and Figure 8H ), pruritus ( Figure 8C and Figure 8I ), epidermal area ( Figure 8E) and IgE antibody production ( Figure 8F ) was measured.

[0040] Figures 9A-9E The results of administering anti-KLK5 / 7 antibodies to a squamous tail mouse model are shown, such as by epidermal area ( Figure 9A Incomplete keratosis (a type of keratosis) Figure 9B ), spongy edema (a histological landmark of eczema epidermis) Figure 9C ) and IL-4 in the ear ( Figure 9D ) and TNFα ( Figure 9E The generation of ) is measured.

[0041] Figures 10A-10F This demonstrates the use of KLK5 / 7-Dual-Ab4 in disease-induced human epidermal equivalent air-liquid interface cultures (MC903 model). Figure 10C Comparative antibody #1 ( Figure 10D ), Comparative antibody #3 ( Figure 10E After treatment, compared with no MC903 ( Figure 10A ) and MC903+ control IgG ( Figure 10B (Comparison) Representative histological images and overview measurements of hyperkeratosis. Figure 10F A quantitative overview of stratum corneum thickness under each condition is shown.

[0042] Figure 11 This is the crystal structure of the KLK5 / 7-Dual-Ab1 antibody Fab that binds to the active site of the StoA variant of the human KLK7 antigen. The heavy chain is indicated in light gray, the Fab light chain in dark gray, and the antigen in black.

[0043] Figure 12 The heavy chain CDR3 ring residues of KLK5 / 7-Dual-Ab1 (dark gray) are shown, occupying the binding pocket of the active site of the human KLK7 antigen (light gray). The catalytic triplet residues of the antigen are shown in a rod shape, as are the tryptophan residues at the bottom of the S4 binding pocket. Detailed Implementation

[0044] This disclosure is based, at least in part, on the development of dual inhibitor antibodies and variants thereof targeting KLK5 and KLK7. These dual inhibitor antibodies target KLK5 and KLK7 through a common, unique antigen-specific binding site. Such dual inhibitor antibodies exhibit high binding affinity and specificity for KLK5 and KLK7 (anti-KLK5 / KLK7 antibodies). Methods for using anti-KLK5 / KLK7 antibodies and variants thereof in research, diagnostic / detection, and therapeutic applications are also provided, as well as anti-KLK5 / KLK7 antibodies for use in such methods.

[0045] The foregoing and other aspects, implementations, operations, functions, features, and embodiments of the present teachings can be more fully understood from the following description, taken in conjunction with the accompanying drawings.

[0046] I. DEFINITIONS Administering / administration: As used herein, the term "administering" or "administration" means providing an antibody or composition thereof to a subject in a physiologically and / or pharmacologically acceptable manner (e.g., to treat a condition in the subject).

[0047] Affinity matured antibody: The term "affinity matured antibody" is used herein to refer to an antibody with one or more alterations in one or more CDRs that result in improvement of the affinity (e.g., KD, kd, or ka) of the antibody for the target antigen compared to a parent antibody that does not have the alteration(s). In some embodiments, an exemplary affinity matured antibody can have nanomolar or even picomolar affinities for the target antigen. A variety of procedures for affinity maturation of antibodies are known, including screening of combinatorial antibody libraries using methods of biopanning. For example, Marks et al., BioTechnology, 10: 779-783 (1992) describes affinity maturation by VHand VLdomain shuffling. Random mutagenesis of CDR and / or framework residues is described by Barbas et al., Proc. Nat. Acad. Sci. USA, 91 : 3809-3813 (1994); Schier et al., Gene, 169: 147-155 (1995); Yelton et al., J. Immunol., 155: 1994-2004 (1995); Jackson et al., J. Immunol., 154(7): 3310-3319 (1995); and Hawkins et al., J. Mol. Biol., 226: 889-896 (1992). U.S. Patent No. 6,914,128 Bl describes selective mutagenesis at selective mutagenesis positions and at contact or hypermutation positions using amino acid residues that enhance activity.

[0048] Antibody: As used herein, the term "antibody" refers to a polypeptide comprising at least one immunoglobulin variable domain comprising at least one unique antigen-specific binding site; or a portion of an immunoglobulin variable domain comprising at least one unique antigen-specific binding site (e.g., a paratope or portion thereof). In some embodiments, an antibody comprises at least one unique antigen-specific binding site that specifically binds to an active site of an enzyme. In some embodiments, an antibody is a full-length antibody. In some embodiments, an antibody is a chimeric antibody. In some embodiments, an antibody is a humanized antibody. In some embodiments, however, an antibody is a Fab fragment, a F(ab')2 fragment, a Fv fragment, or a scFv fragment. In some embodiments, an antibody is a multispecific antibody (e.g., a bispecific antibody). In some embodiments, an antibody is a nanobody derived from a camelid antibody or a nanobody derived from a shark antibody. In some embodiments, an antibody is a diabody. In some embodiments, an antibody comprises a framework having a human germline sequence. In another embodiment, an antibody comprises a heavy chain constant domain selected from the group consisting of: an IgG, IgGl, IgG2, IgG2A, IgG2B, IgG2C, IgG3, IgG4, IgAl, IgA2, IgD, IgM, and IgE constant domain. In some embodiments, an antibody comprises a heavy (H) chain variable region (abbreviated herein as VH) and / or a light (L) chain variable region (abbreviated herein as VL). In some embodiments, an antibody comprises a constant domain, e.g., an Fc region. An immunoglobulin constant domain refers to a heavy chain or light chain constant domain. Human IgG heavy and light chain constant domain amino acid sequences and their functional variants are known. With respect to heavy chains, in some embodiments, a heavy chain of an antibody described herein can be an alpha, delta, epsilon, gamma, or mu heavy chain. In some embodiments, a heavy chain of an antibody described herein can comprise a human alpha, delta, epsilon, gamma, or mu heavy chain. In one particular embodiment, an antibody described herein comprises human gamma 1 CH1, CH2, and / or CH3 domains. In some embodiments, a VH H The amino acid sequence of the V HThe domains comprise an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 98%, or at least 99% identical to any of the variable chain constant regions provided herein. In some embodiments, the antibodies are modified, for example, by glycosylation, phosphorylation, sumoylation, and / or methylation. In some embodiments, the antibodies are glycosylated antibodies conjugated to one or more sugar or carbohydrate molecules. In some embodiments, the one or more sugar or carbohydrate molecules are conjugated to the antibody by N-glycosylation, O-glycosylation, C-glycosylation, glypiation (GPI anchor attachment), and / or phosphoglycosylation. In some embodiments, the one or more sugar or carbohydrate molecules are monosaccharides, disaccharides, oligosaccharides, or glycans. In some embodiments, the one or more sugar or carbohydrate molecules are branched oligosaccharides or branched glycans. In some embodiments, the one or more sugar or carbohydrate molecules comprise mannose units, glucose units, N-acetylglucosamine units, or phospholipid units. In some embodiments, the antibodies are constructs comprising polypeptides comprising one or more antigen binding fragments of the present disclosure linked to a linker polypeptide or an immunoglobulin constant domain. Linker polypeptides comprise two or more amino acid residues joined by a peptide bond and are used to link one or more antigen binding moieties. Examples of linker polypeptides have been reported (see, e.g., Holliger, P. et al. (1993) Proc. Natl. Acad. Sci. USA 90:6444-6448; Poljak, R. J. et al. (1994) Structure 2:1121-1123). Furthermore, the antibodies can be part of larger immunoadhesion molecules formed

[0049] Approximately: As used herein, the term “approximately” or “about,” when used in reference to one or more values of interest, denotes a range of values that fall within 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or less in either direction (greater than or less than) of the stated reference value unless otherwise stated or otherwise clear from the context (except where such numbers would exceed 100% of a possible value).

[0050] Bispecific antibody: As used herein, the term “bispecific antibody” refers to an antibody comprising two distinct antigen-specific binding sites or two linked (covalent or non-covalent) antibodies that together comprise two distinct antigen-specific binding sites. Non-limiting examples of bispecific antibody formats or architectures are provided in Labrijn, AF et al., Bispecific antibodies: a mechanistic review of the pipeline , Nature Reviews Drug Discovery vol. 18, pages 585-608 (2019), and Brinkmann U and Kontermann EE, The making of bispecific antibodies , MAbs. February / March 2017; 9(2): 182-212, the entire contents of each of which are incorporated herein by reference in their entireties.

[0051] CDR: As used herein, the term "CDR" refers to a complementarity determining region within an antibody variable sequence. A typical antibody molecule comprises a heavy chain variable region (VH) and a light chain variable region (VL), which generally are involved in antigen binding. The VHand VLregions can be further subdivided into regions of hypervariability, also called "complementarity determining regions" ("CDR"), interspersed with regions that are more conserved, called "framework regions" ("FR"). Each VHand VLis typically composed of three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The extent of the framework region and CDRs can be precisely defined using methods known in the art, e.g., by the Kabat definition, the IMGT definition, the Chothia definition, the AbM definition, and / or the contact definition, all of which are well known in the art.See, e.g., Kabat, E. A. et al. (1991) Sequences of Proteins of Immunological Interest, 5th Ed., U.S. Department of Health and Human Services, NIH Publication No. 91-3242; IMGT®, the international ImMunoGeneTics information system® http: / / www.imgt.org, Lefranc, M.-P. et al., Nucleic Acids Res., 27:209-212 (1999); Ruiz, M. et al., Nucleic Acids Res., 28:219-221 (2000); Lefranc, M.-P., Nucleic Acids Res., 29:207-209 (2001); Lefranc, M.-P., Nucleic Acids Res., 31:307-310 (2003); Lefranc, M.-P. et al., In Silico Biol., 5, 0006 (2004) [[Epub]], 5:45-60 (2005); Lefranc, M.-P. et al., Nucleic Acids Res., 33:D593-597 (2005); Lefranc, M.-P. et al., Nucleic Acids Res., 37:D1006-1012 (2009); Lefranc, M.-P. et al., Nucleic Acids Res., 43:D413-422 (2015); Chothia et al., (1989) Nature 342:877; Chothia, C. et al. (1987) J. Mol. Biol. 196:901-917, Al-lazikani et al. (1997) J. Molec. Biol. 273:927-948; and Almagro, J. Mol. Recognit. 17:132-143 (2004). See also hgmp.mrc.ac.uk and bioinf.org.uk / abs. As used herein, CDR can refer to CDRs defined by any method known in the art. Two antibodies have identical CDRs meaning that the two antibodies have the same amino acid sequence for the CDRs, as determined by the same method (e.g., IMGT definition).

[0052] In certain embodiments, there are three CDRs in each variable region of the heavy and light chains, which are designated CDR1, CDR2, and CDR3 for each variable region. As used herein, the term "CDR set" refers to a set of three CDRs that occur in a single variable region capable of binding an antigen. The exact boundaries of these CDRs are defined differently by different systems. The system described by Kabat (Kabat et al., Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, Md. (1987) and (1991)) not only provides an unambiguous residue numbering system applicable to any variable region of an antibody, but also provides precise residue boundaries defining the three CDRs. These CDRs can be referred to as Kabat CDRs. Sub-portions of CDRs can be designated as LI, L2, and L3 or HI, H2, and H3, where "L" and "H" designate light chain regions and heavy chain regions, respectively. These regions can be referred to as Chothia CDRs, which have boundaries that overlap with Kabat CDRs. Other boundaries defining CDRs that overlap with Kabat CDRs are described by Padlan (FASEB J. 9: 133-139 (1995)) and MacCallum (J Mol Biol 262(5): 732-45 (1996)). Other CDR boundary definitions can not strictly follow one of the above systems, but will still overlap with Kabat CDRs, although they can be shortened or lengthened according to predictions or experimental findings that particular residues or groups of residues or even entire CDRs do not significantly impact antigen binding. Methods used herein can utilize CDRs defined according to any of these systems, although preferred embodiments use Kabat or Chothia defined CDRs.

[0053] CDR-grafted antibody: As used herein, the term "CDR-grafted antibody" refers to an antibody that comprises heavy and light chain variable region sequences from one species, but in which the sequence of one or more CDR regions of the VH and / or VL has been replaced with CDR sequences of another species, e.g., an antibody with murine heavy and light chain variable regions in which one or more murine CDRs (e.g., CDR3) have been replaced with human CDR sequences.

[0054] Chimeric antibody: As used herein, the term "chimeric antibody" refers to an antibody that comprises heavy and light chain variable region sequences from one species and constant region sequences from another species, e.g., an antibody with murine heavy and light chain variable regions linked to human constant regions.

[0055] Complementary: As used herein, the term “complementary” refers to the ability of two nucleotides or two sets of nucleotides to pair precisely with one another. In particular, complementary is a term that characterizes the degree of hydrogen bond pairing between two nucleotides or two sets of nucleotides that binds them. For example, if a base at a position of an oligonucleotide is capable of hydrogen bonding with a base at a corresponding position of a target nucleic acid (e.g., mRNA), then the bases are considered to be complementary to one another at that position. Base pairing can include canonical Watson-Crick base pairing and non-Watson-Crick base pairing (e.g., wobble base pairing and Hoogsteen base pairing). For example, in some embodiments, for complementary base pairing, an adenosine-type base (A) is complementary to a thymidine-type base (T) or a uracil-type base (U), a cytosine-type base (C) is complementary to a guanosine-type base (G), and a universal base such as 3-nitropyrrole or 5-nitroindole can hybridize to and be considered complementary to any A, C, U, or T. Inosine (I) is also considered a universal base in the art and is considered complementary to any A, C, U, or T.

[0056] Conservative amino acid substitution: As used herein, “conservative amino acid substitution” refers to an amino acid substitution that does not change the relative charge or size characteristics of the protein in which the amino acid substitution is made. Variants can be prepared according to methods known to those of ordinary skill in the art for altering polypeptide sequences, for example, as found in references that compile such methods: for example, Molecular Cloning: A Laboratory Manual, J. Sambrook et al., eds., 4th ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 2012, or Current Protocols in Molecular Biology, F.M. Ausubel et al., eds., John Wiley & Sons, Inc., New York. Conservative substitutions of amino acids include substitutions within the following groups: (a) M, I, L, V; (b) F, Y, W; (c) K, R, H; (d) A, G; (e) S, T; (f) Q, N; and (g) E, D.

[0057] Cross-reactivity: As used herein, the term "cross-reactivity" refers to the property of an agent to specifically bind with similar affinity or avidity to more than one similar type or class of antigen (e.g., antigens of multiple homologs, paralogs, or orthologs). For example, in some embodiments, an antibody that is cross-reactive with similar types or classes of human and non-human primate antigens (e.g., human KLK5 and non-human primate KLK5, human KLK7 and non-human primate KLK7) is capable of binding to the human and non-human primate antigens with similar affinity or avidity. In some embodiments, an antibody is cross-reactive with similar types or classes of human and rodent antigens. In some embodiments, an antibody is cross-reactive with similar types or classes of rodent and non-human primate antigens. In some embodiments, an antibody is cross-reactive with similar types or classes of human, non-human primate, and rodent antigens.

[0058] Dual inhibitor antibody: As used herein, the term "dual inhibitor antibody" refers to an antibody that targets at least two (e.g., two, three) different antigens and inhibits the activity of those antigens through one common unique antigen-specific binding site. In some embodiments, a dual inhibitor antibody targets at least two different proteins (e.g., expressed by two different genes (e.g., endogenous genes, e.g., homologs, paralogs)) and inhibits the activity of the at least two different proteins (e.g., enzymes, e.g., proteases) through one common unique antigen-specific binding site. In some embodiments, a dual inhibitor antibody targets at least two different proteases (e.g., expressed by two different endogenous genes, e.g., KLK5 and KLK7) and inhibits the activity of the at least two different proteases through one common unique antigen-specific binding site. In some embodiments, the common unique antigen-specific binding site binds to a similar (e.g., homologous) domain shared between or among the at least two different antigens. For example, in some embodiments, the common unique antigen-specific binding site binds to a similar (e.g., homologous) catalytic domain or substrate binding site shared between or among the at least two different enzymes (e.g., proteases). In some embodiments, the common unique antigen-specific binding site of a dual inhibitor antibody comprises the amino acids of one or more complementarity determining regions of the antibody. In some embodiments, the common unique antigen-specific binding site of a dual inhibitor antibody is within the heavy chain variable region and / or the light chain variable region of the antibody. In some embodiments, the common unique antigen-specific binding site of a dual inhibitor antibody comprises one or more complementarity determining regions of the heavy chain variable region and / or the light chain variable region of the antibody. In some embodiments, the common unique antigen-specific binding site of a dual inhibitor antibody comprises the HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and LC CDR3 of the heavy chain variable region and the light chain variable region of the antibody. In some embodiments, a dual inhibitor antibody specifically binds to two different proteins (e.g., KLK5 and KLK7) expressed by two different genes.

[0059] Effective amount: As used herein, "effective amount" means the amount of each active agent (e.g., an anti-KLK5 / KLK7 antibody) alone or in combination with one or more other active agents required to exert a desired effect (e.g., to produce a therapeutic effect in a subject). In some embodiments, the therapeutic effect is reduced KLK5 and / or KLK7 activity and / or ameliorated disease (e.g., Netherton Syndrome, eosinophilic esophagitis, and atopic dermatitis) or associated symptoms, e.g., improved barrier function.

[0060] Frame: As used herein, the term “frame” or “frame sequence” refers to the remaining sequence in the variable region after subtracting the CDR. Because the exact definition of a CDR sequence can be determined by different systems, the meaning of a frame sequence may be interpreted differently. The six CDRs (CDR-L1, CDR-L2, and CDR-L3 for the light chain, and CDR-H1, CDR-H2, and CDR-H3 for the heavy chain) further divide the frame region on both the light and heavy chains into four subregions (FR1, FR2, FR3, and FR4) on each chain, where CDR1 is located between FR1 and FR2, CDR2 between FR2 and FR3, and CDR3 between FR3 and FR4. Without specifying a particular subregion as FR1, FR2, FR3, or FR4, the frame region, as used by others, represents a combination of FRs within the variable region of a single naturally occurring immunoglobulin chain. As used herein, FR represents one of the four subregions, and FR represents two or more of the four subregions that constitute the frame region. Human heavy chain and light chain receptor sequences are known in the art. In one embodiment, the receptor sequences known in the art can be used in the antibodies disclosed herein.

[0061] Human Antibody: As used herein, the term "human antibody" is intended to include antibodies having variable and constant regions derived from human germline immunoglobulin sequences. Human antibodies of this disclosure may include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutations in vivo), such as in CDRs, and particularly CDR3. However, as used herein, the term "human antibody" is not intended to include antibodies in which a CDR sequence derived from the germline of another mammalian species (e.g., mouse) has been grafted onto a human frame sequence.

[0062] Humanized antibodies: As used herein, the term "humanized antibody" refers to antibodies that contain components derived from non-human species (e.g., ， The heavy and light chain variable region sequences of mice, but V... H and / or V L At least a portion of the sequence has been altered to be more "human-like," i.e., more similar to human germline variable sequences. One type of humanized antibody is a CDR transplantation antibody, in which a human CDR sequence is introduced into non-human VH and VL sequences to replace the corresponding non-human CDR sequences. In one embodiment, a humanized antibody is provided. Such antibodies can be produced by obtaining murine monoclonal antibodies using conventional hybridoma techniques, followed by humanization using in vitro genetic engineering, such as those disclosed in PCT Publication No. WO 2005 / 123126 A2 by Kasaian et al.

[0063] Humanized antibodies are human immunoglobulins (recipient antibody) in which residues from a complementary determining region (CDR) of the recipient are replaced by residues from a CDR of a non-human species (donor antibody) such as mouse, rat, or rabbit having the desired specificity, affinity, and capacity. In some embodiments, Fv framework region (FR) residues of the human immunoglobulin are replaced by corresponding non-human residues. Furthermore, humanized antibodies can comprise residues that are found neither in the recipient antibody nor in the imported CDR or framework sequences, but are included to further refine and optimize antibody performance. In general, the humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin and all or substantially all of the FR regions are those of a human immunoglobulin consensus sequence. The humanized antibody optimally also will include at least a portion of an immunoglobulin constant region or domain (Fc), typically that of a human immunoglobulin. The antibody can have an Fc region modified as described in WO 99 / 58572. Other forms of humanized antibodies have one or more CDRs (one, two, three, four, five, six) altered with respect to the original antibody, which are also referred to as one or more CDRs derived from one or more CDRs of the original antibody. Humanized antibodies can also involve affinity maturation.

[0064] In some embodiments, humanization is achieved by grafting CDRs (e.g., as shown in Table la or lb) into a human variable domain (e.g., IGKV1-NL1 01 and IGHV1-3 01 human variable domain). In some embodiments, the antibodies of the present disclosure are humanized variants comprising one or more amino acid substitutions (e.g., in the VH framework regions) compared to any of the VHs listed in Table la or lb, and / or comprising one or more amino acid substitutions (e.g., in the VL framework regions) compared to any of the VLs listed in Table la or lb.

[0065] Isolated antibody: As used herein, “isolated antibody” is intended to refer to an antibody that is substantially free of other antibodies having different antigenic specificities (e.g., an isolated dual inhibitor antibody that specifically binds to KLK5 and KLK7 is substantially free of antibodies that specifically bind antigens other than KLK5 and KLK7). However, in some embodiments, an isolated antibody can have cross-reactivity with other antigens. Furthermore, an isolated antibody can be substantially free of other cellular material and / or chemicals.

[0066] Kabat Numbering: As used herein, the terms “Kabat numbering,” “Kabat definition,” and “Kabat labeling” are used interchangeably. These terms, as accepted in the art, refer to a system for numbering amino acid residues that are more variable (i.e., hypervariable) than other amino acid residues in the heavy and light chain variable regions of an antibody or its antigen-binding moiety (Kabat et al. (1971) Ann. NY Acad, Sci. 190:382-391, and Kabat, EA et al. (1991) Sequences of Proteins of Immunological Interest, 5th edition, US Department of Health and Human Services, NIH Publication No. 91-3242). For heavy chain variable regions, the hypervariable region is located at amino acid positions 31 to 35 (for CDR1), 50 to 65 (for CDR2), and 95 to 102 (for CDR3). For light chain variable regions, the hypervariable region is located at amino acid positions 24 to 34 (for CDR1), 50 to 56, and 95 to 102 (for CDR3). (For CDR2) and amino acid positions in the range of 89 to 97 (for CDR3).

[0067] Multispecific antigen-binding molecule: As used herein, the term "multispecific antigen-binding molecule" refers to a molecule containing two or more antigen-specific binding sites. In some embodiments, the multispecific antigen-binding molecule is a multispecific antibody (e.g., a bispecific antibody).

[0068] Multispecific antibody: As used herein, the term "multispecific antibody" refers to an antibody containing at least two unique antigen-specific binding sites or at least two linked (covalent or non-covalent) antibodies that together contain at least two unique antigen-specific binding sites. In some embodiments, multispecific antibodies are bispecific antibodies. The following literature provides non-limiting examples of multispecific antibody forms or structures: Sawant MS et al., Toward Drug-Like Multispecific Antibodies by Design , Int J Mol Sci. 2020 Oct 12;21(20):7496; Klein C et al, The use of CrossMAb technology for the generation of bi- and multispecific antibodies , Mabs 2016 Aug–Sep;8(6):1010–20; and Brinkmann U and Kontermann EE, The making of bispecific antibodies , Mabs. Feb / Mar 2017;9(2):182-212, the entire contents of each of which are incorporated herein by reference.

[0069] Recombinant Antibody: As used herein, the term “recombinant antibody” is intended to include all antibodies prepared, expressed, generated, or isolated in a recombinant manner, such as antibodies expressed using a recombinant expression vector transfected into host cells (described in more detail in this disclosure), including, for example, antibodies isolated from recombinant, combined human antibody libraries (Hoogenboom HR, (1997) TIB Tech. 15:62-70; Azzazy H. and Highsmith WE, (2002) Clin. Biochem. 35:425-445; Gavilondo JV and Larrick JW (2002) BioTechniques 29:128-145; Hoogenboom H. and Chames P. (2000) Immunology Today 21:371-378), and antibodies isolated from transgenic animals (e.g., mice) carrying human immunoglobulin genes (see, for example, Taylor, LD et al. (1992) Nucl. Acids Res.). 20:6287-6295; Kellermann SA. and Green LL (2002) Current Opinion in Biotechnology 13:593-597; Little M. et al. (2000) Immunology Today 21:364-370) or antibodies prepared, expressed, generated, or isolated by any other means involving splicing human immunoglobulin gene sequences into other DNA sequences. In some embodiments, recombinant human antibodies are provided herein. In some embodiments, such recombinant human antibodies have variable and constant regions derived from human germline immunoglobulin sequences. However, in some embodiments, such recombinant human antibodies have undergone in vitro mutagenesis (or, when using transgenic animals carrying human Ig sequences, in vivo somatic mutagenesis), and therefore, the V of the recombinant antibody H District and V L The amino acid sequence of the region is as follows, although it originates from human reproductive system V. H and V L The sequence is associated with, but may not be naturally present in, the human antibody germline library in vivo. One embodiment of this disclosure provides fully human antibodies, such as fully human antibodies capable of binding to human KLK5 or KLK7, which can be generated using appropriate techniques, such as, but not limited to, using human Ig phage libraries, such as those disclosed in Jermutus et al., PCT Publication No. WO 2005 / 007699 A2.

[0070] Selectivity: As used herein, the term "selectivity" or "selectively" refers to the ability of a molecule to produce an effect (e.g., inhibition, antagonism, agonism, etc.) on its target molecule as compared to a reference molecule. For example, a molecule that selectively inhibits its target molecule means that such molecule is capable of inhibiting its target molecule to a degree that distinguishes from a reference molecule in an inhibition assay or other inhibition context. For example, for an inhibitor, the term "selectively inhibits" refers to the ability of the inhibitor to inhibit its target molecule in an inhibition assay to a degree that distinguishes from a reference molecule that is substantially uninhibited, e.g., to a degree that allows for selective inhibition of the target molecule, as described herein. Once the reaction is terminated, the signal produced by the inhibition of the target molecule can be measured. The half-maximal inhibitor concentration for the target molecule and the reference molecule can be calculated.

[0071] Specific binding: As used herein, the term "specific binding" refers to the ability of a molecule to bind to a binding partner with a degree of affinity or avidity that allows for the use of the molecule to distinguish the binding partner from an appropriate control in a binding assay or other binding context. For an antibody, the term "specific binding" refers to the ability of the antibody to bind to a particular antigen with a degree of affinity or avidity that allows for the use of the antibody to distinguish the particular antigen from other antigens, as described herein. In some embodiments, an antibody specifically binds to a target if the Kd of the antibody for the target is at least about 10 D is at least about 10 -4 M, 10 -5 M, 10 -6 M, 10 -7 M, 10 -8 M, 10 -9 M, 10 -10 M, 10 -11 M, 10 -12 M, 10 -13 M or lower. In some embodiments, an antibody specifically binds to KLK5 or KLK7.

[0072] Subject: As used herein, the term "subject" refers to a mammal. In some embodiments, a subject is a non-human primate or a rodent. In some embodiments, a subject is a human. In some embodiments, a subject is a patient, e.g., a human patient having or suspected of having a disease.

[0073] Treatment: As used herein, the term“treating” or“treatment” refers to the application or administration of a composition including one or more active agents (e.g., an anti-KLK5 / KLK7 antibody) to a subject with the intent to cure, heal, alleviate, relieve, alter, remedy, improve, or influence the condition, symptoms of the disease, or the predisposition toward developing the disease. Alleviating the target disease / disorder includes delaying or preventing the development or progression of the disease, or lessening the severity of the disease. It is understood that reference to treatment also refers to antibodies, including dual inhibitor antibodies, used in such methods.

[0074] II. Dual Inhibitor Antibodies Targeting KLK5 and KLK7 (a) Anti-KLK5 / KLK7 antibodies In some embodiments, a dual inhibitor antibody targeting KLK5 and KLK7 (referred to as an anti-KLK5 / KLK7 antibody) is an antibody that is specific for both kallikrein-5 (KLK5) and KLK7 through one common specific antigen binding site. In some aspects, provided herein are antibodies that bind to KLK5 (e.g., human KLK5 or mouse KLK5) and KLK7 (e.g., human KLK7 or mouse KLK7) with high specificity and affinity through one common antigen binding site. In some embodiments, the anti-KLK5 / KLK7 antibodies described herein specifically bind to an epitope in KLK5 that is exposed or becomes exposed to the antibody and to an epitope in KLK7 that is exposed or becomes exposed to the antibody. In some embodiments, the anti-KLK5 / KLK7 antibodies provided herein specifically bind to KLK5 from a human, a non-human primate, a mouse, a rat, and the like. In some embodiments, the anti-KLK5 antibodies provided herein specifically bind to human KLK5. In some embodiments, the anti-KLK5 antibodies provided herein specifically bind to mouse KLK5.

[0075] In some embodiments, the anti-KLK5 / KLK7 antibody is not a bispecific antibody or bispecific antigen binding molecule, wherein KLK5 binding is conferred by one binding site within the antibody and KLK7 binding is conferred by another binding site within the antibody.

[0076] In some embodiments, the anti-KLK5 / KLK7 antibodies described herein can be characterized by reference to certain functional properties. In some embodiments, the anti-KLK5 / KLK7 antibodies described herein specifically bind to KLK5 and KLK7. In some embodiments, the anti-KLK5 / KLK7 antibodies specifically bind to active forms of KLK5 and KLK7. In some embodiments, the anti-KLK5 / KLK7 antibodies do not bind to inactive forms (pro-forms) of KLK5 and KLK7. In some embodiments, the antibodies specifically bind to active forms of KLK5 and KLK7, but do not specifically bind to inactive forms of KLK5 or KLK7. In some embodiments, the antibodies detectably bind to active forms of KLK5 and KLK7, but do not detectably bind to inactive forms of KLK5 or KLK7 under the same or comparable conditions. In some embodiments, the anti-KLK5 / KLK7 antibodies inhibit KLK5 and KLK7 protease activity. In some embodiments, the anti-KLK5 / KLK7 antibodies are not cleaved by KLK5 or KLK7 when bound to KLK5 or KLK7. In some embodiments, the anti-KLK5 / KLK7 antibodies compete with SPINK5 and / or leupeptin for binding to the active site of KLK5 and KLK7. In some embodiments, the anti-KLK5 / KLK7 antibodies reduce hyperkeratosis and desquamation. In some embodiments, the anti-KLK5 / KLK7 antibodies reduce stratum corneum thickness. In some embodiments, the anti-KLK5 / KLK7 antibodies reduce inflammation and epidermal effects. Kallikrein-5, also known as stratum corneum chymotryptic enzyme (SCTE), is a serine protease expressed in the epidermis, encoded by the KLK5 gene. The KLK5 gene is one of fifteen kallikrein subfamily members located on a gene cluster. Its expression is upregulated by estrogen and progesterone. KLK5 is expressed in the stratum granulosum and stratum corneum. In some embodiments, KLK5 modulates epidermal desquamation. In some embodiments, KLK5 modulates epidermal desquamation with another member of the kallikrein family of proteases (e.g., KLK7 and / or KLK14). In some embodiments, KLK5 degrades proteins that form the epidermis (e.g., stratum corneum, stratum lucidum, stratum granulosum, stratum spinosum, or stratum basale). In some embodiments, KLK5 degrades proteins that form the stratum corneum and / or stratum granulosum (e.g., corneodesmosin (CDSN), desmoglein 1 (DSG1), and desmocollin 1 (DSC1), etc.). In the epidermis (e.g., stratum granulosum and stratum corneum), KLK5 is expressed in an inactive form (sometimes referred to as a pro-form, proKLK5), and can auto-activate itself. When activated, KLK5 can convert both proKLK7 and proKLK14 to active forms through proteolytic cleavage.Next, active KLK14 is able to activate newly produced proKLK5, thereby forming a positive feedback loop (see, e.g., Nauroy et al., Kallikreins: Essential epidermal messengers for regulation of the skin microenvironment during homeostasis, repair and disease, Matrix Biol Plus. 2019;6-7:100019). KLK7 and KLK14 also degrade proteins that form the stratum corneum and / or the granular layer (e.g., corneodesmosin (CDSN), desmoglein 1 (DSG1), and desmocollin 1 (DSC1), etc.). The structural proteins (e.g., CDSN, DSG1, DSC1) are adhesion proteins of the extracellular portion of the corneodesmosomes, which are junctional structures that mediate the cohesion of keratinocytes. Degradation of these proteins at the surface of the epidermis causes desquamation, which can result in skin barrier defects (e.g., corneodesmosome disorganization, decreased permeability barrier, allergy and inflammation, etc.). KLK5 and KLK7 are involved in this process (see, e.g., Caubet et al., Degradation of Corneodesmosome Proteins by Two Serine Proteases of the Kallikrein Family, SCTE / KLK5 / hK5 and SCCE / KLK7 / hK7, Journal of Investigative Dermatology, Vol. 122, No. 5, May 2004, pp. 1235-1244). Inhibition of KLK5 and / or KLK7 promotes improvement of skin barrier integrity and reduction of inflammation (e.g., Chavarria-Smith et al., Dual antibody inhibition of KLK5 and KLK7 for Netherton syndrome and atopic dermatitis, SCIENCE TRANSLATIONAL MEDICINE , December 14, 2022, Vol. 14, No. 675).

[0077] Kallikrein-7 is a serine protease encoded by the KLK7 gene in humans. KLK7 is characterized as stratum corneum chymotrypsin (SCCE). [It is the seventh member of the human kallikrein family of 15 homologous serine proteases located on chromosome 19. KLK7 is secreted in an inactive zymogen form (e.g., in the granular layer of the epidermis), requiring proteolytic cleavage to be activated. In some embodiments, KLK5 or matriptase activates KLK7. Once active, KLK7 is able to cleave proteins that form the stratum corneum and / or the granular layer (e.g., corneodesmosin (CDSN), desmoglein 1 (DSG1), and desmocollin 1 (DSC1), etc.) (see, e.g., Caubet et al. (May 2004). Degradation of corneodesmosome proteins by two serine proteases of the kallikrein family, SCTE / KLK5 / hK5 and SCCE / KLK7 / hK7. The Journal of Investigative Dermatology . 122 (5): 1235-1244). These proteins constitute the extracellular component of the corneodesmosomes, which are intercellular cohesive structures that link the intermediate filaments of adjacent cells in the stratum corneum. In some embodiments, proteolysis of the corneodesmosomes leads to epidermal desquamation (i.e., the shedding of corneocytes from the outer layer of the epidermis). In some embodiments, the combined action of KLK5 and KLK7 suggests that the KLK skin cascade is responsible for coordinating desquamation. KLK7 is a chymotrypsin-like serine protease that cleaves proteins at tyrosine, phenylalanine, or leucine residues. In some embodiments, KLK7 dysregulation is associated with several skin disorders, including atopic dermatitis, psoriasis, and Netherton syndrome. These diseases are characterized by excessively dry, scaly, and inflamed skin due to the disruption of skin homeostasis and proper barrier function.

[0078] In some embodiments, the anti-KLK5 / KLK7 antibodies described herein specifically bind to an epitope on human KLK5. An exemplary amino acid sequence of human KLK5 is set forth in NCBI Accession Nos. NP_001070959.1, NP_001070960.1, or NP_036559.1, and UniProt Accession Nos. Q8IU55, Q6S9W8, M0QXX2, Q9P0G3, A0A2I2MP48, or A0A2I2MP49, the complete sequences of which are incorporated herein by reference.

[0079] In some embodiments, the anti-KLK5 / KLK7 antibodies described herein specifically bind to an epitope on mouse KLK5 by the same antigen binding site that binds to KLK5 (e.g., human KLK5 or mouse KLK5). Exemplary amino acid sequences of mouse KLK5 are set forth in NCBI Accession Nos. NP_081082.1, XP_006541213.1, XP_006541214.1, XP_006541215.1, XP_036009294.1, or XP_036009295.1, and UniProt Accession Nos. P15945 or Q9D140, the entire sequences of which are incorporated herein by reference.

[0080] In some embodiments, the anti-KLK5 / KLK7 antibodies described herein specifically bind to an epitope on human KLK7 by the same antigen binding site that binds to KLK5 (e.g., human KLK5 or mouse KLK5). Exemplary amino acid sequences of human KLK7 are set forth in NCBI Accession Nos. NP_001193982.1, NP_001230055.1, NP_005037.1, NP_644806.1, and UniProt Accession Nos. M0QYU8, Q6DTY1, X2J289, X2J4X7, A0A024R4H6, P49862, A0A2H4GDB2, and A0A2H4GDB6, the entire sequences of which are incorporated herein by reference.

[0081] In some embodiments, the anti-KLK5 / KLK7 antibodies described herein specifically bind to an epitope on mouse KLK7 by the same antigen binding site that binds to KLK5 (e.g., human KLK5 or mouse KLK5). Exemplary amino acid sequences of mouse KLK7 are set forth in NCBI Accession No. NP_036002.1 and UniProt Accession No. Q91VE3, the entire sequences of which are incorporated herein by reference.

[0082] In some embodiments, the anti-KLK5 / KLK7 antibodies described herein specifically bind to an epitope on KLK5 (e.g., catalytic domain / pocket of human KLK5 or mouse KLK5) and an epitope on KLK7 (e.g., catalytic domain / pocket of human KLK7 or mouse KLK7). In some embodiments, the anti-KLK5 / KLK7 antibodies described herein prevent KLK5 (e.g., human or mouse KLK5) and KLK7 (e.g., human or mouse KLK7) from cleaving their substrates. In some embodiments, the anti-KLK5 antibodies described herein bind to a fragment of KLK5 (e.g., human or mouse KLK5) and a fragment of KLK7 (e.g., human or mouse KLK7). The fragment of KLK5 and / or KLK7 (e.g., human or mouse) can be between about 5 and about 425 amino acids, between about 10 and about 400 amino acids, between about 50 and about 350 amino acids, between about 100 and about 300 amino acids, between about 150 and about 250 amino acids, between about 200 and about 300 amino acids, between about 75 and about 150 amino acids, between about 25 and about 100 amino acids, between about 10 and about 30 amino acids in length. Without wishing to be bound by any particular theory, and in some embodiments, the heavy chain (HC) complementarity determining region 3 (CDR3) of any of the anti-KLK5 / KLK7 antibodies described herein inhibits KLK5 (e.g., human or mouse KLK5) and KLK7 (e.g., human or mouse KLK7) by binding to the catalytic domain / pocket of KLK5.

[0083] In some embodiments, the anti-KLK5 / KLK7 antibodies described herein inhibit KLK5 protease activity, KLK7 protease activity, or KLK5 and KLK7 protease activity. In some embodiments, the anti-KLK5 / KLK7 antibodies inhibit KLK5 (e.g., human KLK5 or mouse KLK5) cleavage of BOC-Val-Pro-Arg-AMC with an IC50 of less than 30 nM, less than 25 nM, less than 20 nM, less than 15 nM, less than 10 nM, less than 5 nM, less than 3 nM, less than 2.5 nM, less than 2 nM, or less than 1.5 nM, less than 1 nM, less than 0.5 nM, less than 0.3 nM, less than 0.25 nM, less than 0.2 nM, or less than 0.1 nM. In some embodiments, the anti-KLK5 / KLK7 antibodies inhibit KLK5 (e.g., human KLK5 or mouse KLK5) cleavage of BOC-Val-Pro-Arg-AMC with an IC50 of between 0.1 nM and 30 nM, 0.1 nM and 20 nM, 0.1 nM and 10 nM, 0.1 nM and 5 nM, 0.1 nM and 2.5 nM, 0.1 nM and 2 nM, 0.1 nM and 1 nM, 0.1 nM and 0.5 nM, 0.1 nM and 0.25 nM, 0.1 nM and 50 nM, 0.1 nM and 40 nM, 0.1 nM and 30 nM, 0.1 nM and 20 nM, 0.1 nM and 10 nM, 0.1 nM and 5 nM, 0.1 nM and 2.5 nM, 0.1 nM and 2 nM, 0.1 nM and 1 nM, 0.1 nM and 0.9 nM, 0.1 nM and 0.8 nM, 0.1 nM and 0.7 nM, 0.1 nM and 0.6 nM, 0.1 nM and 0.5 nM, 0.1 nM and 0.4 nM, 0.1 nM and 0.3 nM, 0.1 nM and 0.25 nM, 0.1 nM and 0.2 nM, 0.1 nM and 0.15 nM, 0.15 nM and 0.2 nM, 0.15 nM and 0.25 nM, 0.15 nM and 0.3 nM, 0.15 nM and 0.4 nM, 0.15 nM and 0.5 nM, 0.15 nM and 1 nM, 0.2 nM and 30 nM, 0.2 nM and 20 nM, 0.2 nM and 10 nM, 0.2 nM and 5 nM, 0.2 nM and 2.5 nM, 0.2 nM and 2 nM, 0.2 nM and 1 nM, 0.2 nM and 0.5 nM, 0.2 nM and 0.2 nM, 0.2 nM and 50 nM, 0.2 nM and 40 nM, 0.2 nM and 30 nM, 0.2 nM and 20 nM, 0.2 nM and 10 nM, 0.2 nM to 5 nM, 0.2 nM to 2.5 nM, 0.2 nM to 2 nM, 0.2 nM to 1 nM, 0.2 nM to 0.9 nM, 0.2 nM to 0.8 nM, 0.2 nM to 0.7 nM, 0.2 nM to 0.6 nM, 0.2 nM to 0.5 nM, 0.2 nM to 0.4 nM, 0.2 nM to 0.3 nM, 0.2 nM to 0.25 nM, 1 nM to 30 nM, 1 nM to 20 nM, 1 nM to 10 nM, 1 nM to 5 nM, 1 nM to 2.5 nM, 1 nM to 2 nM, 1 nM to 3 nM, 1 nM to 5.5 nM, 1.5 nM to 2 nM, 1.5 nM to 3 nM, 1.5 nM to 5.5 nM, 2 nM to 5 nM, 2 nM to 4 nM, 2 nM to 5.5 nM, 3 nM to 5.5 nM, 4 nM to 5.5 nM, 3 nM to 30 nM, 3 nM to 20 nM, 3 nM to 10 nM, 3 nM to 5 nM, 3 nM to 2.5 nM, 3 nM to 4 nM, 3 nM to 5.5 nM, 5 nM to 30 nM, 5 nM to 20 nM, 5 nM to 10 nM, 5 nM to 9 nM, 5 nM to 8 nM, 5 nM to 7 nM, 5 nM to 6 nM, 5 nM to 5.5 nM, 10 nM to 30 nM, 10 nM to 25 nM, 10 nM to 20 nM, 10 nM to 18 nM, 10 nM to 15 nM, 10 nM to 12 nM, 12 nM to 20 nM, 12 nM to 25 nM, 12 nM to 16 nM, 12 nM to 18 nM, 12 nM to 20 nM, 12 nM to 24 nM, 12 nM to 28 nM, 12 nM to 30 nM, 15 nM to 30 nM, 15 nM to 25 nM, 15 nM to 20 nM, 15 nM to 18 nM, 18 nM to 30 nM, 18 nM to 25 nM, 18 nM to 20 nM, 20 nM to 30 nM, 20 nM to 25 nM, 20 nM to 22 nM, 20 nM to 24 nM, 20 nM to 26 nM, 20 nM to 28 nM, 22 nM to 30 nM, 22 nM to 25 nM, 22 nM to 28 nM, 24 nM to 30 nM, 24 nM to 25 nM, 24 nM to 26 nM, or 24 nM to 28 nM. In some embodiments, the anti-KLK5 / KLK7 antibody inhibits cleavage of KHLF-AMC by KLK7 with an IC50 of less than 6 nM, less than 5 nM, less than 4 nM, less than 3 nM, less than 2.5 nM, less than 2 nM, or less than 1.5 nM, less than 1 nM, less than 0.5 nM, less than 0.4 nM, less than 0.3 nM, less than 0.2 nM, less than 0.16 nM, less than 0.1 nM, or less than 0.05 nM. In some embodiments, the anti-KLK5 / KLK7 antibody inhibits cleavage of KHLF-AMC by KLK7 with an IC50 of 0.1 nM to 30 nM, 0.1 nM to 20 nM, 0.1 nM to 10 nM, 0.1 nM to 5 nM, 0.1 nM to 2.5 nM, 0.1 nM to 2 nM, 0.1 nM to 1 nM, 0.1 nM to 0.5 nM, 0.1 nM to 0.25 nM, 0.1 nM to 50 nM, 0.1 nM to 40 nM, 0.1 nM to 30 nM, 0.1 nM to 20 nM, 0.1 nM to 10 nM, 0.1 nM to 5 nM, 0.1 nM to 2.5 nM, 0.1 nM to 2 nM, 0.1 nM to 1 nM, 0.1 nM to 0.9 nM, 0.1 nM to 0.8 nM, 0.1 nM to 0.7 nM, 0.1 nM to 0.6 nM, 0.1 nM to 0.5 nM, 0.1 nM to 0.4 nM, 0.1 nM to 0.3 nM, 0.1 nM to 0.25 nM, 0.1 nM to 0.2 nM, 0.1 nM to 0.15 nM, 0.15 nM to 0.2 nM, 0.15 nM to 0.25 nM, 0.15 nM to 0.3 nM, 0.15 nM to 0.4 nM, 0.15 nM to 0.5 nM, 0.15 nM to 1 nM, 0.2 nM to 30 nM, 0.2 nM to 20 nM, 0.2 nM to 10 nM, 0.2 nM to 5 nM, 0.2 nM to 2.5 nM, 0.2 nM to 2 nM, 0.2 nM to 1 nM, 0.2 nM to 0.5 nM, 0.2 nM to 0.2 nM, 0.2 nM to 50 nM, 0.2 nM to 40 nM, 0.2 nM to 30 nM, 0.2 nM to 20 nM, 0.2 nM to 10 nM, 0.2 nM to 5 nM, 0.2 nM to 2.5 nM, 0.2 nM to 2 nM, 0.2 nM to 1 nM, 0.2 nM to 0.9 nM, 0.2 nM to 0.8 nM, 0.2 nM to 0.7 nM, 0.2 nM to 0.6 nM, 0.2 nM to 0.5 nM, 0.2 nM to 0.4 nM, 0.2 nM to 0.3 nM, 0.2 nM to 0.25 nM, 1 nM to 30 nM, 1 nM to 20 nM, 1 nM to 10 nM, 1 nM to 5 nM, 1 nM to 2.5 nM, 1 nM to 2 nM, 1 nM to 1 nM, 1 nM to 0.9 nM, 1 nM to 0.8 nM, 1 nM to 0.7 nM, 1 nM to 0.6 nM, 1 nM to 0.5 nM, 1 nM to 0.4 nM, 1 nM to 0.3 nM, 1 nM to 0.25 nM, 1 nM to 0.2 nM, 1 nM to 0.15 nM, 1 nM to 0.1 nM, 1 nM to 0.05 nM, 2 nM to 30 nM, 2 nM to 20 nM, 2 nM to 10 nM, 2 nM to 5 nM, 2 nM to 2.5 nM, 2 nM to 1 nM, 2 nM to 0.9 nM, 2 nM to 0.8 nM, 2 nM to 0.7 nM, 2 nM to 0.6 nM, 2 nM to 0.5 nM, 2 nM to 0.4 nM, 2 nM to 0.3 nM, 2 nM to 0.25 nM, 2 nM to 0.2 nM, 2 nM to 0.15 nM, 2 nM to 0.1 nM, 2 nM to 0.05 nM, 5 nM to 30 nM, 5 nM to 20 nM, 5 nM to 10 nM, 5 nM to 2.5 nM, 5 nM to 2 nM, 5 nM to 1 nM, 5 nM to 0.9 nM, 5 nM to 0.8 nM, 5 nM to 0.7 nM, 5 nM to 0.6 nM, 5 nM to 0.5 nM, 5 nM to 0.4 nM, 5 nM to 0.3 nM, 5 nM to 0.25 nM, 5 nM to 0.2 nM, 5 nM to 0.15 nM, 5 nM to 0.1 nM, 5 nM to 0.05 nM, 10 nM to 30 nM, 10 nM to 20 nM, 10 nM to 10 nM, 10 nM to 5 nM, 10 nM to 2.5 nM, 10 nM to 2 nM, 10 nM to 1 nM, 10 nM to 0.9 nM, 10 nM to 0.8 nM, 10 nM to 0.7 nM, 10 nM to 0.6 nM, 10 nM to 0.5 nM, 10 nM to 0.4 nM, 10 nM to 0.3 nM, 10 nM to 0.25 nM, 10 nM to 0.2 nM, 10 nM to 0.15 nM, 10 nM to 0.1 nM, 10 nM to 0.05 nM, 20 nM to 30 nM, 20 nM to 20 nM, 20 nM to 10 nM, 20 nM to 5 nM, 20 nM to 2.5 nM, 20 nM to 2 nM, 20 nM to 1 nM, 20 nM to 0.9 nM, 20 nM to 0.8 nM, 20 nM to 0.7 nM, 20 nM to 0.6 nM, 20 nM to 0.5 nM, 20 nM to 0.4 nM, 20 nM to 0.3 nM, 20 nM to 0.25 nM, 20 nM to 0.2 nM, 20 nM to 0.15 nM, 20 nM to 0.1 nM, 20 nM to 0.05 nM, 30 nM to 30 nM, 30 nM to 20 nM, 30 nM to 10 nM, 30 nM to 5 nM, 30 nM to 2.5 nM, 30 nM to 2 nM, 30 nM to 1 nM, 30 nM to 0.9 nM, 30 nM to 0.8 nM, 305 nM, 1 nM to 2 nM, 1 nM to 3 nM, 1 nM to 5.5 nM, 1.5 nM to 2 nM, 1.5 nM to 3 nM, 1.5 nM to 5.5 nM, 2 nM to 5 nM, 2 nM to 4 nM, 2 nM to 5.5 nM, 3 nM to 5.5 nM, 4 nM to 5.5 nM, 3 nM to 30 nM, 3 nM to 20 nM, 3 nM to 10 nM, 3 nM to 5 nM, 3 nM to 2.5 nM, 3 nM to 4 nM, 3 nM to 5.5 nM, 5 nM to 30 nM, 5 nM to 20 nM, 5 nM to 10 nM, 5 nM to 9 nM, 5 nM to 8 nM, 5 nM to 7 nM, 5 nM to 6 nM, 5 nM to 5.5 nM, 10 nM to 30 nM, 10 nM to 25 nM, 10 nM to 20 nM, 10 nM to 18 nM, 10 nM to 15 nM, 10 nM to 12 nM, 12 nM to 20 nM, 12 nM to 25 nM, 12 nM to 16 nM, 12 nM to 18 nM, 12 nM to 20 nM, 12 nM to 24 nM, 12 nM to 28 nM, 12 nM to 30 nM, 15 nM to 30 nM, 15 nM to 25 nM, 15 nM to 20 nM, 15 nM to 18 nM, 18 nM to 30 nM, 18 nM to 25 nM, 18 nM to 20 nM, 20 nM to 30 nM, 20 nM to 25 nM, 20 nM to 22 nM, 20 nM to 24 nM, 20 nM to 26 nM, 20 nM to 28 nM, 22 nM to 30 nM, 22 nM to 25 nM, 22 nM to 28 nM, 24 nM to 30 nM, 24 nM to 25 nM, 24 nM to 26 nM, or 24 nM to 28 nM.

[0084] In some embodiments, the anti-KLK5 / KLK7 antibodies described herein specifically bind to active forms of KLK5, KLK7, or KLK5 and KLK7. In some embodiments, the anti-KLK5 / KLK7 antibodies described herein do not bind to inactive forms of KLK5, KLK7, or KLK5 and KLK7. In some embodiments, the anti-KLK5 / KLK7 antibodies described herein specifically bind to active forms of KLK5, active forms of KLK7, or active forms of KLK5 and KLK7, but do not specifically bind to inactive forms of KLK5, inactive forms of KLK7, or inactive forms of KLK5 and KLK7. In some embodiments, the anti-KLK5 / KLK7 antibodies described herein detectably bind to active forms of KLK5, active forms of KLK7, or active forms of KLK5 and KLK7, but do not detectably bind to inactive forms of KLK5, inactive forms of KLK7, or inactive forms of KLK5 and KLK7 under the same or comparable conditions. In some embodiments, the anti-KLK5 / KLK7 antibodies described herein specifically bind to active sites of KLK5, KLK7, or KLK5 and KLK7. The active site of KLK5 and / or KLK7 is the site at which a KLK5 and / or KLK7 substrate molecule binds and is cleaved. The active site can also be referred to as the catalytic domain or catalytic triad. In some embodiments, the active site of KLK5 or KLK7 (i.e., the catalytic domain or catalytic triad) is composed of the amino acids Ser195, His57, and Asp102 of KLK5 or KLK7 (see, e.g., Goettig et al., Natural and synthetic inhibitors of kallikrein-related peptidases (KLKs), Biochimica et Biophysica Acta 1824 (2012) 1998-2015). Biochimie . 2010 Nov; 92(11): 1546-1567).

[0085] In some embodiments, the antibodies described herein are optimized versions (e.g., affinity matured) of the parent antibodies. In some embodiments, the antibodies described herein bind to KLK5, KLK7, or KLK5 and KLK7 with a binding affinity (e.g., as Kd) of less than about 10 -4 M, less than 10 -5 M, less than 10 -6 M, less than 10 -7 M, less than 10 -8 M, less than 10 -9 M, less than 10 -10 M, less than 10 -11 M, less than 10 -12 M, less than 10 -13 M, or lower (e.g., as K DKLK5 (e.g., human or mouse KLK5) and KLK7 (e.g., human or mouse KLK7). In some embodiments, the antibodies described herein bind KLK5 (e.g., human or mouse KLK5) and KLK7 (e.g., human or mouse KLK7) with a binding affinity (e.g., as K -10 M between 1 x 10 -9 M and 1 x 10 -10 M between 1 x 10 -9 M and 1 x 10 -10 M between 1 x 10 -9 M and 1 x 10 -11 M between 1 x 10 -10 M and 1 x 10 -11 M between 1 x 10 -10 M and 1 x 10 -13 M between 1 x 10 - 12 M between 1 x 10 D KLK5 (e.g., human or mouse KLK5) and KLK7 (e.g., human or mouse KLK7). For example, the antibodies of the present disclosure can bind to KLK5 protein (e.g., human or mouse KLK5) and KLK7 (e.g., human or mouse KLK7) with an affinity between 1 pM and 500 nM, for example, between 50 pM and 100 nM, between 500 pM and 50 nM, between 1 pM and 100 pM, between 10 pM and 100 pM, between 50 pM and 100 pM, between 100 pM and 500 pM, between 500 pM and 1 nM, between 1 nM and 5 nM, between 1 nM and 10 nM, between 5 nM and 25 nM, between 10 nM and 50 nM, between 50 nM and 100 nM, between 100 nM and 500 nM. The present disclosure also includes antibodies that compete with any of the antibodies described herein for binding to KLK5 protein (e.g., human or mouse KLK5) and KLK7 (e.g., human or mouse KLK7) and have an affinity of 100 nM or less (e.g., 80 nM or less, 50 nM or less, 20 nM or less, 10 nM or less, 1 nM or less, 500 pM or less, 50 pM or less, or 5 pM or less). The affinity and binding kinetics of an antibody can be tested using any suitable method, including but not limited to biosensor technology (e.g., OCTET or BIACORE). In some embodiments, the antibodies described herein bind KLK5 (e.g., human or mouse KLK5) and KLK7 (e.g., human or mouse KLK7) with a K D KLK5 and KLK7.

[0086] Binding affinity (or binding specificity) can be determined by a variety of methods, including equilibrium dialysis, equilibrium binding, gel filtration, ELISA, surface plasmon resonance (SPR), fluorescence activated cell sorting (FACS), or spectroscopy (e.g., using a fluorescence assay). Exemplary conditions for assessing binding affinity are in HBS-P buffer (10 mM HEPES pH 7.4, 150 mM NaCl, 0.005% (v / v) surfactant P20) and PBS buffer (10 mM PO4-3, 137 mM NaCl, and 2.7 mM KCl). These techniques can be used to measure the concentration of bound protein as a function of the concentration of target protein. The concentration of bound protein ([bound]) is generally related to the concentration of free target protein ([free]) by the following equation: [bound] = [free] / (Kd + [free]) However, it is not always necessary to determine K A as it is sometimes sufficient to obtain a quantitative measure of affinity (e.g., determined using a method such as ELISA or FACS analysis) that is proportional to K A determination of a higher affinity, e.g., 2-fold higher, to obtain a qualitative measure of affinity, or to obtain an inference of affinity, e.g., by activity in a functional assay (e.g., an in vitro or in vivo assay).

[0087] Table 1a and Table 1b provide exemplary anti-KLK5 / KLK7 antibody sequences (e.g., heavy chain (HC) and light chain (LC) sequences, heavy chain variable domain (VH) and light chain variable domain (VL), CDR sequences.

[0088] Table 1a. Examples of anti-KLK5 / KLK7 antibodies In some embodiments, certain amino acid positions in the antibodies described herein (e.g., amino acids in the VH / VL regions and / or CDR regions) are substitutable, and substitution results in an antibody having substantially similar binding and biological activity (e.g., substantially similar binding affinity, binding specificity, protease activity inhibitory activity, anti-inflammatory activity, or a combination thereof) to the reference antibody. To identify substitutable positions of an antibody, the amino acid sequence of the antibody is compared to the sequences of other antibodies belonging to the same group as the antibody. If the identity of the amino acid differs at any particular position between different related antibodies of a group, then that position is a substitutable position of the antibody. In other words, a substitutable position is a position where the identity of the amino acid differs between related antibodies. Positions containing constant amino acids are not substitutable positions.

[0089] In some embodiments, the above-described methods can be employed to provide consensus antibody sequences. In such consensus sequences, non-substitutable positions are indicated by the amino acid present at that position, and substitutable positions are indicated as "X".

[0090] Depending on how the antibody is to be used, X can be a) any amino acid, b) any amino acid present at that position in any of the related antibodies in the set or a conservatively substituted variant thereof, or c) any amino acid present at that position in any of the related antibodies in the set. Any antibody having a sequence encompassed by a consensus sequence should bind to the same antigen as any of the related antibodies.

[0091] In some embodiments, the above-described methods can be used in methods of designing and making variants of a parent antibody that at least maintain (e.g., maintain or increase) the antigen binding activity of the parent antibody. Because antibodies have been produced and tested that contain substitutions at substitutable positions, the substitutions can be made with the knowledge that those substitutions at those positions should not significantly decrease the binding activity of the antibody. Generally, an antibody variant of a parent antibody has an antigen binding affinity that is at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100% (e.g., at least 150%, at least 200%, at least 500%, at least 1000%, and typically up to at least 10,000%) of the binding affinity of the parent antibody for a particular antigen.

[0092] In some embodiments, substitutable positions of a parent antibody can be substituted with a) any of the 20 naturally occurring amino acids to produce a random substitution, b) an amino acid having similar biochemical properties as the amino acid already present at the substitutable position to produce a conservative substitution, c) an amino acid present at the same position in a related antibody to produce a directed substitution, or d) an amino acid present at the same position in a similar human antibody to produce a humanized substitution. The substitutions can be made at any portion of the antibody variable region, including any framework region or CDR. In certain embodiments, a single substitutable amino acid can be substituted. However, in other embodiments, multiple substitutable amino acids (e.g., up to about 5 or 10 or more) can be substituted. In particular embodiments, the type of substitution that can be made at each substitutable position can be indicated by the type of amino acid present at that position in the related antibodies. For example, if a certain position in a set of related antibodies is present with unrelated amino acids (e.g., Ala, Gly, Cys, Glu, and Thr), then any amino acid can be substituted at that position without significantly decreasing the binding activity of the antibody. Exemplary amino acid substitutions for the anti-KLK5 / KLK7 antibodies described herein are shown in Table lb: Table lb. Exemplary anti-KLK5 / KLK7 antibody amino acid substitutions

[0093] In some embodiments, an antibody of the present disclosure comprises a HC CDR1 comprising the amino acid sequence of GSISSXiDYYWX2(SEQ ID NO: 28), wherein Xi is S, D, or L, and X2is G or V; a HC CDR2 comprising the amino acid sequence of SIX3YX4X5X6TYYX7PSLKS (SEQ ID NO: 29), wherein X3is Y or D, X4is S, F, or Y, X5is G or A, X6is S or D, or X7is N or S; a HC CDR3 comprising the amino acid sequence of ARGRPLGYGAX8HX9YYGMDV (SEQ ID NO: 30), wherein X8is R or K, or X9is Y or D; a LC CDR1 comprising the amino acid sequence of SEQ ID NO: 4; a LC CDR2 comprising the amino acid sequence of SEQ ID NO: 5; and / or a LC CDR3 comprising the amino acid sequence of QQSPX 10 FPPLT (SEQ ID NO: 31), wherein X 10 is P or Y.

[0094] In some embodiments, an antibody of the present disclosure comprises one or more HC CDR (e.g., HC CDR1, HC CDR2, or HC CDR3) amino acid sequences from any anti-KLK5 / KLK7 antibody selected from Table la and Table lb. In some embodiments, an antibody of the present disclosure comprises a HC CDR3 amino acid sequence from any anti-KLK5 / KLK7 antibody selected from Table la and Table lb. In some embodiments, an antibody of the present disclosure comprises a HC CDR1, HC CDR2, and HC CDR3 as provided for any antibody selected from Table la and Table lb. In some embodiments, an antibody of the present disclosure comprises a LC CDR3 amino acid sequence from any anti-KLK5 / KLK7 antibody selected from Table la and Table lb. In some embodiments, an antibody of the present disclosure comprises one or more LC CDR (e.g., LC CDR1, LC CDR2, or LC CDR3) amino acid sequences from any anti-KLK5 / KLK7 antibody selected from Table la and Table lb. In some embodiments, an antibody of the present disclosure comprises a LC CDR1, LC CDR2, and LC CDR3 as provided for any anti-KLK5 antibody selected from Table la and Table lb.

[0095] In some embodiments, the antibodies of the present disclosure comprise HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and LC CDR3 as provided for any anti-KLK5 / KLK7 antibody selected from Table la and Table lb. In some embodiments, antibody heavy and / or light chain CDR3 domains can play a particularly important role in the binding specificity / affinity of an antibody to an antigen. Accordingly, the antibodies of the present disclosure can include at least the heavy and / or light chain CDR3 of any anti-KLK5 / KLK7 antibody selected from Table la and Table lb.

[0096] Variants of any of the exemplary anti-KLK5 / KLK7 antibodies as disclosed herein are also within the scope of the present disclosure. Variants can contain one or more amino acid residue variations in the VH and / or VL or in one or more HC CDRs and / or one or more LC CDRs relative to a reference antibody, while retaining substantially similar binding and biological activities (e.g., substantially similar binding affinity, binding specificity, protease activity inhibitory activity, anti-inflammatory activity, or a combination thereof) to the reference antibody.

[0097] In some embodiments, an antibody of the present disclosure has one or more CDR (e.g., HC CDR or LC CDR) sequences substantially similar to any one of the HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and / or LC CDR3 sequences from an anti-KLK5 / KLK7 antibody selected from Table la and Table lb. In some embodiments, the position of one or more CDRs along the VH (e.g., HC CDR1, HC CDR2, or HC CDR3) and / or VL (e.g., LC CDR1, LC CDR2, or LC CDR3) region of an antibody described herein can vary by one, two, three, four, five, or six amino acid positions, so long as specific binding to KLK5 (e.g., human or mouse KLK5) and KLK7 (e.g., human or mouse KLK7) is maintained (e.g., substantially maintains at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% of the binding of the original antibody from which it is derived). For example, in some embodiments, the position of a CDR defining any antibody described herein can be varied by shifting the N-terminal and / or C-terminal boundary of the CDR by one, two, three, four, five, or six amino acids relative to the CDR position of any antibody described herein, so long as specific binding to KLK5 (e.g., human or mouse KLK5) and KLK7 (e.g., human or mouse KLK7) is maintained (e.g., substantially maintains at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% of the binding of the original antibody from which it is derived). In another embodiment, the length of one or more CDRs along the VH (e.g., HC CDR1, HC CDR2, or HC CDR3) and / or VL (e.g., LC CDR1, LC CDR2, or LC CDR3) region of an antibody described herein can vary (e.g., be shortened or lengthened) by one, two, three, four, five, or more amino acids, so long as immunospecific binding to KLK5 (e.g., human or mouse KLK5) and KLK7 (e.g., human or mouse KLK7) is maintained (e.g., substantially maintains at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% of the binding of the original antibody from which it is derived).

[0098] Accordingly, in some embodiments, the HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and / or LC CDR3 described herein can be one, two, three, four, five or more amino acids shorter than one or more CDRs described herein (e.g., from any anti-KLK5 / KLK7 antibody selected from Tables la and lb) so long as specific binding to KLK5 (e.g., human or mouse KLK5) and KLK7 (e.g., human or mouse KLK7) is maintained (e.g., binding is substantially maintained, e.g., at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% relative to the binding of the original antibody from which it is derived). In some embodiments, the HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and / or LC CDR3 described herein can be one, two, three, four, five or more amino acids longer than one or more CDRs described herein (e.g., from any anti-KLK5 / KLK7 antibody selected from Tables la and lb) so long as specific binding to KLK5 (e.g., human or mouse KLK5) and KLK7 (e.g., human or mouse KLK7) is maintained (e.g., binding is substantially maintained, e.g., at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% relative to the binding of the original antibody from which it is derived). In some embodiments, the amino portion of the HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and / or LC CDR3 described herein can be one, two, three, four, five or more amino acids longer than one or more CDRs described herein (e.g., from any anti-KLK5 / KLK7 antibody selected from Tables la and lb) so long as specific binding to KLK5 (e.g., human or mouse KLK5) and KLK7 (e.g., human or mouse KLK7) is maintained (e.g., binding is substantially maintained, e.g., at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% relative to the binding of the original antibody from which it is derived).In some embodiments, the carboxyl portion of the HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and / or LC CDR3 described herein can be extended by one, two, three, four, five, or more amino acids compared to one or more CDRs described herein (e.g., from a CDR selected from any of the anti-KLK5 / KLK7 antibodies of Table la and Table lb) so long as specific binding to KLK5 (e.g., human or mouse KLK5) and KLK7 (e.g., human or mouse KLK7) is maintained (e.g., binding is substantially maintained, e.g., at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% relative to the binding of the original antibody from which it is derived). In some embodiments, the amino portion of the HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and / or LC CDR3 described herein can be shortened by one, two, three, four, five, or more amino acids compared to one or more CDRs described herein (e.g., from a CDR selected from any of the anti-KLK5 / KLK7 antibodies of Table la and Table lb) so long as specific binding to KLK5 (e.g., human or mouse KLK5) and KLK7 (e.g., human or mouse KLK7) is maintained (e.g., binding is substantially maintained, e.g., at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% relative to the binding of the original antibody from which it is derived). In some embodiments, the carboxyl portion of the HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and / or LC CDR3 described herein can be shortened by one, two, three, four, five, or more amino acids compared to one or more CDRs described herein (e.g., from a CDR selected from any of the anti-KLK5 / KLK7 antibodies of Table la and Table lb) so long as specific binding to KLK5 (e.g., human or mouse KLK5) and KLK7 (e.g., human or mouse KLK7) is maintained (e.g., binding is substantially maintained, e.g., at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% relative to the binding of the original antibody from which it is derived). Whether specific binding to KLK5 (e.g., human or mouse KLK5) and KLK7 (e.g., human or mouse KLK7) is maintained can be determined using any method, e.g., using binding assays and conditions described in the art.

[0099] In some embodiments, an antibody of the present disclosure has one or more CDR (e.g., HC CDR or LC CDR) sequences substantially similar to any of the anti-KLK5 / KLK7 antibodies selected from Table la and Table lb. For example, an antibody described herein can include one or more CDR sequences from any of the anti-KLK5 / KLK7 antibodies selected from Table la and Table lb that contain up to 5, 4, 3, 2, or 1 amino acid residue variations from the corresponding CDR region in any of the CDRs provided herein (e.g., a CDR from any of the anti-KLK5 / KLK7 antibodies selected from Table la and Table lb), so long as specific binding to KLK5 (e.g., human or mouse KLK5) and KLK7 (e.g., human or mouse KLK7) is maintained (e.g., substantially maintained, e.g., at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% relative to the binding of the original antibody from which it was derived). In some embodiments, any amino acid variations in any of the CDRs provided herein can be conservative variations. Conservative variations can be introduced into a CDR at positions where residues are less likely to be involved in interactions with KLK5 (e.g., human or mouse KLK5) and / or KLK7 (e.g., human or mouse KLK7), e.g., as determined based on crystal structures. Some aspects of the present disclosure provide antibodies comprising one or more heavy chain variable (VH) and / or light chain variable (VL) domains provided herein. In some embodiments, any of the VH domains provided herein include one or more HC CDR sequences (e.g., HC CDR1, HC CDR2, and HC CDR3) provided herein, e.g., any of the HC CDR sequences provided in any of the anti-KLK5 / KLK7 antibodies selected from Table la and Table lb. In some embodiments, any of the VL domains provided herein include one or more LC CDR sequences (e.g., LC CDR1, LC CDR2, and LC CDR3) provided herein, e.g., any of the LC CDR sequences provided in any of the anti-KLK5 / KLK7 antibodies selected from Table la and Table lb.

[0100] In some embodiments, an antibody of the present disclosure includes any of the antibodies comprising a heavy chain variable domain and / or a light chain variable domain of any of the anti-KLK5 / KLK7 antibodies selected from Table la and Table lb, and variants thereof. In some embodiments, an antibody of the present disclosure includes any of the antibodies comprising a heavy chain variable and a light chain variable pair of any of the anti-KLK5 / KLK7 antibodies selected from Table la and Table lb.

[0101] Aspects of the disclosure provide antibodies having heavy chain variable (VH) and / or light chain variable (VL) domain amino acid sequences that are homologous to any of those described herein. In some embodiments, an antibody comprises a heavy chain variable sequence or a light chain variable sequence that is at least 75% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to the heavy chain variable sequence and / or any light chain variable sequence of any anti-KLK5 / KLK7 antibody selected from Table la and Table lb. In some embodiments, the homologous heavy chain variable and / or light chain variable amino acid sequences are not changed within any of the CDR sequences provided herein. For example, in some embodiments, the degree of sequence variation (e.g., 75%, 80%, 85%, 90%, 95%, 98%, or 99%) can occur within the heavy chain variable and / or light chain variable sequence, excluding any of the CDR sequences provided herein. In some embodiments, an antibody provided herein comprises a heavy chain variable sequence and a light chain variable sequence comprising a framework sequence that is at least 75%, 80%, 85%, 90%, 95%, 98%, or 99% identical to the framework sequence of any anti-KLK5 / KLK7 antibody selected from Table la and Table lb.

[0102] In some embodiments, an antibody of the disclosure is a humanized antibody (e.g., a humanized variant containing one or more CDRs of Table la and Table lb). In some embodiments, an antibody of the disclosure comprises a HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and LC CDR3 identical to the HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and LC CDR3 shown in Table la and Table lb, and comprises a humanized heavy chain variable region and / or a humanized light chain variable region.

[0103] In some embodiments, an antibody of the disclosure is a humanized antibody comprising a VH containing no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variations) compared to the VH of any anti-KLK5 / KLK7 antibody listed in Table la and Table lb. Alternatively or additionally, an antibody of the disclosure is a humanized antibody comprising a VL containing no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variations) compared to the VL of any anti-KLK5 / KLK7 antibody listed in Table la and Table lb.

[0104] In some embodiments, an anti-KLK5 / KLK7 antibody of the present disclosure comprises an HC CDR1, an HC CDR2, and an HC CDR3 of a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 7. Alternatively or additionally, an anti-KLK5 / KLK7 antibody of the present disclosure comprises an LC CDR1, an LC CDR2, and an LC CDR3 of a light chain variable domain having the amino acid sequence of SEQ ID NO: 8.

[0105] In some embodiments, an anti-KLK5 / KLK7 antibody of the present disclosure comprises an HC CDR1 having the amino acid sequence of SEQ ID NO: 1; an HC CDR2 having the amino acid sequence of SEQ ID NO: 2; an HC CDR3 having the amino acid sequence of SEQ ID NO: 3; an LC CDR1 having the amino acid sequence of SEQ ID NO: 4; an LC CDR2 having the amino acid sequence of SEQ ID NO: 5; and an LC CDR3 having the amino acid sequence of SEQ ID NO: 6.

[0106] In some embodiments, an anti-KLK5 / KLK7 antibody of the present disclosure comprises an HC CDR1, an HC CDR2, and an HC CDR3 that collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variations) compared to an HC CDR1 having the amino acid sequence of SEQ ID NO: 1, an HC CDR2 having the amino acid sequence of SEQ ID NO: 2, and an HC CDR3 having the amino acid sequence of SEQ ID NO: 3. Alternatively or additionally, an anti-KLK5 / KLK7 antibody of the present disclosure comprises an LC CDR1, an LC CDR2, and an LC CDR3 that collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variations) compared to an LC CDR1 having the amino acid sequence of SEQ ID NO: 4, an LC CDR2 having the amino acid sequence of SEQ ID NO: 5, and an LC CDR3 having the amino acid sequence of SEQ ID NO: 6.

[0107] In some embodiments, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a HC CDR1, a HC CDR2, and a HC CDR3 that are collectively at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to a HC CDR1 having the amino acid sequence of SEQ ID NO: 1, a HC CDR2 having the amino acid sequence of SEQ ID NO: 2, and a HC CDR3 having the amino acid sequence of SEQ ID NO: 3. Alternatively or additionally, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a LC CDR1, a LC CDR2, and a LC CDR3 that are collectively at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to a LC CDR1 having the amino acid sequence of SEQ ID NO: 4, a LC CDR2 having the amino acid sequence of SEQ ID NO: 5, and a LC CDR3 having the amino acid sequence of SEQ ID NO: 6.

[0108] In some embodiments, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a HC CDR1 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variations) compared to a HC CDR1 having the amino acid sequence of SEQ ID NO: 1; a HC CDR2 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variations) compared to a HC CDR2 having the amino acid sequence of SEQ ID NO: 2; and / or a HC CDR3 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variations) compared to a HC CDR3 having the amino acid sequence of SEQ ID NO: 3. Alternatively or additionally, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a LC CDR1 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variations) compared to a LC CDR1 having the amino acid sequence of SEQ ID NO: 4; a LC CDR2 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variations) compared to a LC CDR2 having the amino acid sequence of SEQ ID NO: 5; and / or a LC CDR3 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variations) compared to a LC CDR3 having the amino acid sequence of SEQ ID NO: 6.

[0109] In some embodiments, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a VH comprising the amino acid sequence of SEQ ID NO: 7. Alternatively or additionally, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a VL comprising the amino acid sequence of SEQ ID NO: 8.

[0110] In some embodiments, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a VH that contains no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variations) compared to the VH as set forth in SEQ ID NO: 7. Alternatively or additionally, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a VL that contains no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variations) compared to the VL as set forth in SEQ ID NO: 8.

[0111] In some embodiments, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a VH that is at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to the VH as set forth in SEQ ID NO: 7. Alternatively or additionally, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a VL that is at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to the VL as set forth in SEQ ID NO: 8.

[0112] In some embodiments, an anti-KLK5 / KLK7 antibody of the present disclosure comprises an HC CDR1, an HC CDR2, and an HC CDR3 of a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 13. Alternatively or additionally, an anti-KLK5 / KLK7 antibody of the present disclosure comprises an LC CDR1, an LC CDR2, and an LC CDR3 of a light chain variable domain having the amino acid sequence of SEQ ID NO: 14.

[0113] In some embodiments, an anti-KLK5 / KLK7 antibody of the present disclosure comprises an HC CDR1 having the amino acid sequence of SEQ ID NO: 9; an HC CDR2 having the amino acid sequence of SEQ ID NO: 10; an HC CDR3 having the amino acid sequence of SEQ ID NO: 11; an LC CDR1 having the amino acid sequence of SEQ ID NO: 4; an LC CDR2 having the amino acid sequence of SEQ ID NO: 5; and an LC CDR3 having the amino acid sequence of SEQ ID NO: 12.

[0114] In some embodiments, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a HC CDR1, a HC CDR2, and a HC CDR3 that collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variations) compared to a HC CDR1 having the amino acid sequence of SEQ ID NO: 9, a HC CDR2 having the amino acid sequence of SEQ ID NO: 10, and a HC CDR3 having the amino acid sequence of SEQ ID NO: 11. As used anywhere in the present disclosure, “collectively” means that the total number of amino acid variations in all three heavy chain CDRs is within the defined range. Alternatively or additionally, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a LC CDR1, a LC CDR2, and a LC CDR3 that collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variations) compared to a LC CDR1 having the amino acid sequence of SEQ ID NO: 4, a LC CDR2 having the amino acid sequence of SEQ ID NO: 5, and a LC CDR3 having the amino acid sequence of SEQ ID NO: 12.

[0115] In some embodiments, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a HC CDR1, a HC CDR2, and a HC CDR3 that collectively are at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to a HC CDR1 having the amino acid sequence of SEQ ID NO: 9, a HC CDR2 having the amino acid sequence of SEQ ID NO: 10, and a HC CDR3 having the amino acid sequence of SEQ ID NO: 11. Alternatively or additionally, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a LC CDR1, a LC CDR2, and a LC CDR3 that collectively are at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to a LC CDR1 having the amino acid sequence of SEQ ID NO: 4, a LC CDR2 having the amino acid sequence of SEQ ID NO: 5, and a LC CDR3 having the amino acid sequence of SEQ ID NO: 12.

[0116] In some embodiments, an anti-KLK5 / KLK7 antibody of the present disclosure comprises: a HC CDR1 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variations) compared to a HC CDR1 having the amino acid sequence of SEQ ID NO: 9; a HC CDR2 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variations) compared to a HC CDR2 having the amino acid sequence of SEQ ID NO: 10; and / or a HC CDR3 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variations) compared to a HC CDR3 having the amino acid sequence of SEQ ID NO: 11. Alternatively or additionally, an anti-KLK5 / KLK7 antibody of the present disclosure comprises: a LC CDR1 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variations) compared to a LC CDR1 having the amino acid sequence of SEQ ID NO: 4; a LC CDR2 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variations) compared to a LC CDR2 having the amino acid sequence of SEQ ID NO: 5; and / or a LC CDR3 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variations) compared to a LC CDR3 having the amino acid sequence of SEQ ID NO: 12.

[0117] In some embodiments, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a VH comprising the amino acid sequence of SEQ ID NO: 13. Alternatively or additionally, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a VL comprising the amino acid sequence of SEQ ID NO: 14.

[0118] In some embodiments, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a VH containing no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variations) compared to a VH as set forth in SEQ ID NO: 13. Alternatively or additionally, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a VL containing no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variations) compared to a VL as set forth in SEQ ID NO: 14.

[0119] In some embodiments, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a VH comprising an amino acid sequence that is at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to the VH as set forth in SEQ ID NO: 13. Alternatively or additionally, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a VL comprising an amino acid sequence that is at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to the VL as set forth in SEQ ID NO: 14.

[0120] In some embodiments, an anti-KLK5 / KLK7 antibody of the present disclosure comprises an HC CDR1, an HC CDR2, and an HC CDR3 of a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 17. Alternatively or additionally, an anti-KLK5 / KLK7 antibody of the present disclosure comprises an LC CDR1, an LC CDR2, and an LC CDR3 of a light chain variable domain having the amino acid sequence of SEQ ID NO: 14.

[0121] In some embodiments, an anti-KLK5 / KLK7 antibody of the present disclosure comprises an HC CDR1 having the amino acid sequence of SEQ ID NO: 9; an HC CDR2 having the amino acid sequence of SEQ ID NO: 15; an HC CDR3 having the amino acid sequence of SEQ ID NO: 16; an LC CDR1 having the amino acid sequence of SEQ ID NO: 4; an LC CDR2 having the amino acid sequence of SEQ ID NO: 5; and an LC CDR3 having the amino acid sequence of SEQ ID NO: 12.

[0122] In some embodiments, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a HC CDR1, a HC CDR2, and a HC CDR3 that collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variations) compared to a HC CDR1 having the amino acid sequence of SEQ ID NO: 9, a HC CDR2 having the amino acid sequence of SEQ ID NO: 15, and a HC CDR3 having the amino acid sequence of SEQ ID NO: 16. As used anywhere in the present disclosure, “collectively” means that the total number of amino acid variations in all three heavy chain CDRs is within the defined range. Alternatively or additionally, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a LC CDR1, a LC CDR2, and a LC CDR3 that collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variations) compared to a LC CDR1 having the amino acid sequence of SEQ ID NO: 4, a LC CDR2 having the amino acid sequence of SEQ ID NO: 5, and a LC CDR3 having the amino acid sequence of SEQ ID NO: 12.

[0123] In some embodiments, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a HC CDR1, a HC CDR2, and a HC CDR3 that collectively are at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to a HC CDR1 having the amino acid sequence of SEQ ID NO: 9, a HC CDR2 having the amino acid sequence of SEQ ID NO: 15, and a HC CDR3 having the amino acid sequence of SEQ ID NO: 16. Alternatively or additionally, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a LC CDR1, a LC CDR2, and a LC CDR3 that collectively are at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to a LC CDR1 having the amino acid sequence of SEQ ID NO: 4, a LC CDR2 having the amino acid sequence of SEQ ID NO: 5, and a LC CDR3 having the amino acid sequence of SEQ ID NO: 12.

[0124] In some embodiments, an anti-KLK5 / KLK7 antibody of the present disclosure comprises: a HC CDR1 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variations) compared to a HC CDR1 having the amino acid sequence of SEQ ID NO: 9; a HC CDR2 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variations) compared to a HC CDR2 having the amino acid sequence of SEQ ID NO: 15; and / or a HC CDR3 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variations) compared to a HC CDR3 having the amino acid sequence of SEQ ID NO: 16. Alternatively or additionally, an anti-KLK5 / KLK7 antibody of the present disclosure comprises: a LC CDR1 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variations) compared to a LC CDR1 having the amino acid sequence of SEQ ID NO: 4; a LC CDR2 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variations) compared to a LC CDR2 having the amino acid sequence of SEQ ID NO: 5; and / or a LC CDR3 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variations) compared to a LC CDR3 having the amino acid sequence of SEQ ID NO: 12.

[0125] In some embodiments, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a VH comprising the amino acid sequence of SEQ ID NO: 17. Alternatively or additionally, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a VL comprising the amino acid sequence of SEQ ID NO: 14.

[0126] In some embodiments, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a VH containing no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variations) compared to a VH as set forth in SEQ ID NO: 17. Alternatively or additionally, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a VL containing no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variations) compared to a VL as set forth in SEQ ID NO: 14.

[0127] In some embodiments, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a VH comprising an amino acid sequence that is at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to the VH as set forth in SEQ ID NO: 17. Alternatively or additionally, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a VL comprising an amino acid sequence that is at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to the VL as set forth in SEQ ID NO: 14.

[0128] In some embodiments, an anti-KLK5 / KLK7 antibody of the present disclosure comprises an HC CDR1, an HC CDR2, and an HC CDR3 of a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 21. Alternatively or additionally, an anti-KLK5 / KLK7 antibody of the present disclosure comprises an LC CDR1, an LC CDR2, and an LC CDR3 of a light chain variable domain having the amino acid sequence of SEQ ID NO: 14.

[0129] In some embodiments, an anti-KLK5 / KLK7 antibody of the present disclosure comprises an HC CDR1 having the amino acid sequence of SEQ ID NO: 18; an HC CDR2 having the amino acid sequence of SEQ ID NO: 19; an HC CDR3 having the amino acid sequence of SEQ ID NO: 20; an LC CDR1 having the amino acid sequence of SEQ ID NO: 4; an LC CDR2 having the amino acid sequence of SEQ ID NO: 5; and an LC CDR3 having the amino acid sequence of SEQ ID NO: 12.

[0130] In some embodiments, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a HC CDR1, a HC CDR2, and a HC CDR3 that collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variations) compared to a HC CDR1 having the amino acid sequence of SEQ ID NO: 18, a HC CDR2 having the amino acid sequence of SEQ ID NO: 19, and a HC CDR3 having the amino acid sequence of SEQ ID NO: 20. As used anywhere in the present disclosure, “collectively” means that the total number of amino acid variations in all three heavy chain CDRs is within the defined range. Alternatively or additionally, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a LC CDR1, a LC CDR2, and a LC CDR3 that collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variations) compared to a LC CDR1 having the amino acid sequence of SEQ ID NO: 4, a LC CDR2 having the amino acid sequence of SEQ ID NO: 5, and a LC CDR3 having the amino acid sequence of SEQ ID NO: 12.

[0131] In some embodiments, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a HC CDR1, a HC CDR2, and a HC CDR3 that collectively are at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to a HC CDR1 having the amino acid sequence of SEQ ID NO: 18, a HC CDR2 having the amino acid sequence of SEQ ID NO: 19, and a HC CDR3 having the amino acid sequence of SEQ ID NO: 20. Alternatively or additionally, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a LC CDR1, a LC CDR2, and a LC CDR3 that collectively are at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to a LC CDR1 having the amino acid sequence of SEQ ID NO: 4, a LC CDR2 having the amino acid sequence of SEQ ID NO: 5, and a LC CDR3 having the amino acid sequence of SEQ ID NO: 12.

[0132] In some embodiments, an anti-KLK5 / KLK7 antibody of the present disclosure comprises: a HC CDR1 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variations) compared to a HC CDR1 having the amino acid sequence of SEQ ID NO: 18; a HC CDR2 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variations) compared to a HC CDR2 having the amino acid sequence of SEQ ID NO: 19; and / or a HC CDR3 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variations) compared to a HC CDR3 having the amino acid sequence of SEQ ID NO: 20. Alternatively or additionally, an anti-KLK5 / KLK7 antibody of the present disclosure comprises: a LC CDR1 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variations) compared to a LC CDR1 having the amino acid sequence of SEQ ID NO: 4; a LC CDR2 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variations) compared to a LC CDR2 having the amino acid sequence of SEQ ID NO: 5; and / or a LC CDR3 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variations) compared to a LC CDR3 having the amino acid sequence of SEQ ID NO: 12.

[0133] In some embodiments, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a VH comprising the amino acid sequence of SEQ ID NO: 21. Alternatively or additionally, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a VL comprising the amino acid sequence of SEQ ID NO: 14.

[0134] In some embodiments, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a VH that contains no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variations) compared to a VH as set forth in SEQ ID NO: 21. Alternatively or additionally, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a VL that contains no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variations) compared to a VL as set forth in SEQ ID NO: 14.

[0135] In some embodiments, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a VH comprising an amino acid sequence that is at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to a VH as set forth in SEQ ID NO: 21. Alternatively or additionally, an anti-KLK5 / KLK7 antibody of the present disclosure comprises a VL comprising an amino acid sequence that is at least 80% (e.g., 80%, 85%, 90%, 95%, 98%, or 99%) identical to a VL as set forth in SEQ ID NO: 14.

[0136] The antibodies described herein can be in any antibody format, including but not limited to, intact (i.e., full-length) antibodies, antigen-binding fragments thereof (e.g., Fab, F(ab'), F(ab')2, Fv), single-chain antibodies, bispecific antibodies, or nanobodies. In some embodiments, an anti-KLK5 / KLK7 antibody described herein is a scFv. In some embodiments, an anti-KLK5 / KLK7 antibody described herein is a scFv-Fab (e.g., a scFv fused to a portion of a constant region).

[0137] In some embodiments, an anti-KLK5 / KLK7 antibody of the present disclosure is a chimeric antibody, which can include heavy and light chain constant regions from a human antibody. Chimeric antibodies refer to antibodies that have variable regions or portions of variable regions from a first species and constant regions from a second species. Typically, in these chimeric antibodies, the variable regions of both the light and heavy chains mimic the variable regions of antibodies derived from one mammalian species (e.g., a non-human mammal, such as a mouse, rabbit, and rat), while the constant portions are homologous to sequences in antibodies derived from another mammal (e.g., human). In some embodiments, amino acid modifications can be made in the variable and / or constant regions.

[0138] In some embodiments, an antibody of the present disclosure comprises a VL domain and / or a VH domain selected from any of the anti-KLK5 / KLK7 antibodies of Table la and Table lb, and comprises a constant region comprising an amino acid sequence of a constant region of an IgG, IgE, IgM, IgD, IgA, or IgY immunoglobulin molecule, an immunoglobulin molecule of any class (e.g., IgGl, IgG2, IgG3, IgG4, IgAl, and IgA2), or any subclass (e.g., IgG2a and IgG2b). Non-limiting examples of human constant regions have been described in the art, see, e.g., Kabat E A et al., (1991) supra.

[0139] In some embodiments, the light chain of any of the anti-KLK5 / KLK7 antibodies described herein can further comprise a light chain constant region (CL), which can be any CL known in the art. In some embodiments, the CL is a kappa light chain. In other embodiments, the CL is a lambda light chain. In some embodiments, the CL is a kappa light chain.

[0140] Other antibody heavy and light chain constant regions are well known in the art, such as those provided in the IMGT database (www.imgt.org) or www.vbase2.org / vbstat.php., both of which are incorporated herein by reference.

[0141] In some embodiments, conservative mutations can be introduced into antibody sequences (e.g., CDR or framework sequences) at positions less likely to be involved in interactions with a target antigen (e.g., human or mouse KLK5 and / or human or mouse KLK7), e.g., as determined based on crystal structures. In some embodiments, one, two, or more mutations (e.g., amino acid substitutions) are introduced into the Fc region of an anti-KLK5 / KLK7 antibody described herein (e.g., in the CH2 domain (residues 231-340 of human IgGl) and / or the CH3 domain (residues 341-447 of human IgGl) and / or the hinge region, wherein numbering is according to the Kabat numbering system (e.g., the EU index in Kabat)) to alter one or more functional properties of the antibody, such as serum half-life, complement fixation, Fc receptor binding, and / or antigen-dependent cellular cytotoxicity.

[0142] In some embodiments, one, two, or more mutations (e.g., amino acid substitutions) are introduced into the hinge region of the Fc region (CH1 domain) such that the number of cysteine residues in the hinge region is altered (e.g., increased or decreased), as described in, e.g., U.S. Patent No. 5,677,425. The number of cysteine residues in the hinge region of the CH1 domain can be altered, e.g., to facilitate assembly of light and heavy chains, or to alter (e.g., increase or decrease) the stability of the antibody, or to facilitate linker conjugation.

[0143] In some embodiments, one, two, or more mutations (e.g., amino acid substitutions) are introduced into the Fc region of an antibody described herein (e.g., in the CH2 domain (residues 231-340 of human IgGl) and / or the CH3 domain (residues 341-447 of human IgGl) and / or the hinge region, where numbering is according to the Kabat numbering system (e.g., the EU index in Kabat)) to increase or decrease the affinity of the antibody for an Fc receptor on the surface of an effector cell (e.g., an activating Fc receptor). Mutations in the Fc region of an antibody that decrease or increase the affinity of the antibody for an Fc receptor, and techniques for introducing such mutations into an Fc receptor or fragment thereof, are known to those of skill in the art. Examples of mutations made in the Fc receptor of an antibody that can alter the affinity of the antibody for an Fc receptor are described in, e.g., Smith P, et al. (2012) PNAS 109: 6181-6186; U.S. Patent No. 6,737,056; and International Publication Nos. WO 02 / 060919, WO 98 / 23289, and WO 97 / 34631, which are incorporated herein by reference.

[0144] In some embodiments, one, two, or more amino acid mutations (i.e., substitutions, insertions, or deletions) are introduced into an IgG constant domain or FcRn binding fragment thereof (preferably an Fc or hinge-Fc domain fragment) to alter (e.g., decrease or increase) the half-life of the antibody in vivo. For examples of mutations that alter (e.g., decrease or increase) the half-life of an antibody in vivo, see, e.g., International Publication Nos. WO 02 / 060919, WO 98 / 23289, and WO 97 / 34631; and U.S. Patent Nos. 5,869,046, 6,121,022, 6,277,375, and 6,165,745.

[0145] In some embodiments, one, two or more amino acid mutations (i.e., substitutions, insertions or deletions) are introduced into an IgG constant domain or FcRn binding fragment thereof (preferably an Fc or hinge-Fc domain fragment) to reduce the half-life of an anti-KLK5 / KLK7 antibody in vivo. In some embodiments, one, two or more amino acid mutations (i.e., substitutions, insertions or deletions) are introduced into an IgG constant domain or FcRn binding fragment thereof (preferably an Fc or hinge-Fc domain fragment) to increase the half-life of an antibody in vivo. In some embodiments, the antibody can have one or more amino acid mutations (e.g., substitutions) in the second constant (CH2) domain (residues 231-340 of human IgGl) and / or the third constant (CH3) domain (residues 341-447 of human IgGl), wherein numbering is according to the EU index in Kabat (Kabat E A et al., (1991) supra). In some embodiments, the constant region of IgGl of an antibody described herein comprises a substitution of methionine (M) to tyrosine (Y) at position 252, a substitution of serine (S) to threonine (T) at position 254, and a substitution of threonine (T) to glutamic acid (E) at position 256, numbered according to the EU index in Kabat. See U.S. Patent No. 7,658,921, which is incorporated herein by reference. This type of mutant IgG (termed a “YTE mutant”) has been shown to exhibit a four-fold increase in half-life compared to the wild-type form of the same antibody (see Dall’Acqua W F et al., (2006) J Biol Chem 281: 23514-24). In some embodiments, the antibody comprises an IgG constant domain comprising one, two, three or more amino acid substitutions of amino acid residues at positions 251-257, 285-290, 308-314, 385-389, and 428-436, numbered according to the EU index in Kabat.

[0146] In some embodiments, the antibody comprises an Fc region that has been engineered for the purpose of half-life extension, for example by introduction of M428L and / or N434A substitutions. The following reference provides non-limiting examples of such Fc variants that affect circulating half-life: Saunders KO, Conceptual Approaches to Modulating Antibody Effector Functions and Circulation Half-Life, Front Immunol. 2019; 10: 1296, the contents of which are incorporated herein by reference.

[0147] In some embodiments, one, two, or more amino acid substitutions are introduced into the IgG constant domain Fc region to alter the effector function of an anti-KLK5 / KLK7 antibody, for example by introducing Leu234Ala and Leu235Ala mutations (commonly referred to as LALA mutations). The effector ligand whose affinity is altered can be, for example, an Fc receptor or the Cl component of complement. Such an approach is described in greater detail in U.S. Patent Nos. 5,624,821 and 5,648,260. In some embodiments, deletion or inactivation (by point mutation or otherwise) of a constant region domain can reduce Fc receptor binding of the circulating antibody, thereby increasing tumor localization. See, e.g., U.S. Patent Nos. 5,585,097 and 8,591,886 for a description of mutations that result in deletion or inactivation of a constant domain and thereby increase tumor localization. In some embodiments, one or more amino acid substitutions can be introduced into the Fc region of an antibody described herein to remove potential glycosylation sites on the Fc region, which can reduce Fc receptor binding (see, e.g., Shields R L et al., (2001) J Biol Chem 276: 6591-604).

[0148] In some embodiments, one or more amino acids in the constant region of an anti-KLK5 / KLK7 antibody described herein can be replaced with a different amino acid residue such that the antibody has altered Clq binding and / or reduced or abolished complement dependent cytotoxicity (CDC). Such an approach is described in greater detail in U.S. Patent No. 6,194,551 (Idusogie et al.). In some embodiments, one or more amino acid residues in the N-terminal region of the CH2 domain of an antibody described herein are altered, thereby altering the ability of the antibody to fix complement. Such an approach is described further in International Publication No. WO 94 / 29351. In some embodiments, the Fc region of an antibody described herein is modified to increase the ability of the antibody to mediate antibody dependent cellular cytotoxicity (ADCC) and / or to increase the affinity of the antibody for the Fc gamma receptor. Such an approach is described further in International Publication No. WO 00 / 42072.

[0149] In some embodiments, the antibody comprises an Fc variant comprising amino acid substitutions L234A, L235E, and P329G, wherein numbering is according to the EU index. In some embodiments, the antibody comprising the Fc variant exhibits reduced affinity for one or more or each of FcyRJ, FcyRIIA, FcyRIIIA, and Clq compared to an antibody comprising a wild-type human Fc region. Examples of such Fc variants are provided in International Patent Application Publication No. WO 2021 / 055669, entitled FC VARIANTS WITH REDUCED EFFECTOR FUNCTION, published March 25, 2021; and U.S. Patent Application Publication No. US 2021-0087271, entitled FC VARIANTS WITH REDUCED EFFECTOR FUNCTION, published March 25, 2021, the contents of which are incorporated herein by reference.

[0150] In some embodiments, the heavy and / or light chain variable domain sequences of the antibodies provided herein can be used to generate, e.g., CDR-grafted, chimeric, humanized, or composite human antibodies or antigen-binding fragments, as described elsewhere herein. As will be appreciated by one of ordinary skill in the art, any variants, CDR-grafted, chimeric, humanized, or composite antibodies derived from any of the antibodies provided herein can be used in the compositions and methods described herein, and will maintain the ability to specifically bind KLK5 and KLK7, such that the variants, CDR-grafted, chimeric, humanized, or composite antibodies have at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or more KLK5 and KLK7 binding relative to the original antibody from which they are derived.

[0151] In some embodiments, the antibodies provided herein comprise mutations that confer desirable properties to the antibodies. For example, to avoid potential complications due to Fab arm exchange that is known to occur in native IgG4 mAbs, the antibodies provided herein can comprise a stabilizing ‘Adair’ mutation (Angal S. et al., “A single amino acid substitution abolishes the heterogeneity of chimeric mouse / human (IgG4) antibody,” Mol Immunol 30, 105-108; 1993), in which serine 228 (EU; residue 241 Kabat numbering) is converted to proline, resulting in an IgG1-like hinge sequence. Thus, any of the antibodies can include a stabilizing ‘Adair’ mutation.

[0152] In some embodiments, the antibody is modified, e.g., by glycosylation, phosphorylation, SUMOylation, and / or methylation. In some embodiments, the antibody is a glycosylated antibody, which is conjugated to one or more sugar or carbohydrate molecules. In some embodiments, the one or more sugar or carbohydrate molecules is conjugated to the antibody by N-glycosylation, O-glycosylation, C-glycosylation, glycosylphosphatidylinositolization (GPI anchor attachment), and / or phosphosugylation. In some embodiments, the one or more sugar or carbohydrate molecules is a monosaccharide, disaccharide, oligosaccharide, or glycan. In some embodiments, the one or more sugar or carbohydrate molecules is a branched oligosaccharide or branched glycan. In some embodiments, the one or more sugar or carbohydrate molecules comprises a mannose unit, a glucose unit, an N-acetylglucosamine unit, an N-acetylgalactosamine unit, a galactose unit, a fucose unit, or a phospholipid unit. In some embodiments, there are about 1-10, about 1-5, about 5-10, about 1-4, about 1-3, or about 2 sugar molecules. In some embodiments, the glycosylated antibody is fully or partially glycosylated. In some embodiments, the antibody is glycosylated by a chemical reaction or enzymatically. In some embodiments, the antibody is glycosylated in vitro or in a cell, which can optionally lack an enzyme in the N-glycosylation or O-glycosylation pathway, e.g., a glycosyltransferase. In some embodiments, the antibody is functionalized with a sugar or carbohydrate molecule, as described in International Patent Application Publication WO2014065661, published May 1, 2014, entitled “Modified antibody, antibody-conjugate and process for the preparation thereof.”

[0153] In some embodiments, any of the anti-KLK5 / KLK7 antibodies described herein can comprise a signal peptide (e.g., an N-terminal signal peptide) in the heavy chain and / or light chain sequence. In some embodiments, the anti-KLK5 / KLK7 antibodies described herein comprise any of the VH and VL sequences described herein, any of the IgG heavy and light chain sequences, or any of the F(ab’) heavy and light chain sequences, and further comprise a signal peptide (e.g., an N-terminal signal peptide).

[0154] (b) Multispecific antibodies In some embodiments, the antibodies provided herein are multispecific antibodies (e.g., bispecific antibodies). For example, in some embodiments, one or more anti-KLK5 / KLK7 antibodies can be combined with one or more different anti-KLK5 / KLK7 antibodies to produce a multispecific or bispecific anti-KLK5 / KLK7 antibody. For example, one or more anti-KLK5 / KLK7 antibodies as described herein (Table la and Table lb) can be combined with one or more different anti-KLK5 / KLK7 antibodies as described herein (Table la and Table lb) to produce a multispecific antibody.

[0155] In some embodiments, one or more anti-KLK5 / KLK7 antibodies can be combined with any other appropriate therapeutic antibody to produce a multispecific or bispecific anti-KLK5 / KLK7 / additional target antibody. For example, an anti-KLK5 / KLK7 antibody as described herein (Table la and Table lb) can be combined with any appropriate antibody to produce a bispecific antibody. Such additional therapeutic antibodies include, but are not limited to: anti-IL4R antibodies (e.g., dupilumab), anti-IL-13 antibodies, TNF inhibitors (e.g., anti-TNF antibodies), IL-12 / 23 antibodies, IL-17 antibodies, adalimumab, infliximab, golimumab, ustekinumab, secukinumab, ixekizumab, budelumab, abatacept, tildrakizumab-asmn, risankizumab-rzaa, and guselkumab.

[0156] In some embodiments, one or more anti-KLK5 / KLK7 antibodies can be combined with any other appropriate anti-KLK7 antibody to produce a multispecific or bispecific anti-KLK5 / KLK7 antibody. For example, an anti-KLK5 / KLK7 antibody as described herein (Tables la and lb) can be combined with any other appropriate anti-KLK7 antibody to produce a bispecific antibody. The following references provide non-limiting examples of appropriate anti-KLK7 antibodies: U.S. Patent Application Publication No. 2021-0130492, entitled “ANTI-KLK7 ANTIBODIES, ANTI-KLK5 ANTIBODIES, MULTISPECIFIC ANTI-KLK5 / KLK7 ANTIBODIES, AND METHODS OF USE,” published May 6, 2021; International Patent Application Publication No. WO2021226695, entitled RECOMBINANT HUMAN ANTIBODIES FOR INHIBITING HUMAN TISSUE KALLIKREIN 7 (KLK7) AND USE IN DISEASES RELATED TO THE PROCESS OF SKIN DESQUAMATION,” published November 18, 2021; and International Patent Application Publication No. WO2005075667, entitled “DIAGNOSTICS AND THERAPEUTICS FOR DISEASES ASSOCIATED WITH KALLIKREIN 7 (KLK7),” published August 18, 2005, the contents of which are incorporated herein by reference.

[0157] In some embodiments, one or more anti-KLK5 / KLK7 antibodies can be combined with any appropriate anti-KLK5 antibody to generate a multi-specific or bispecific anti-KLK5 / KLK7 antibody. For example, a KLK5 / KLK7 antibody as described herein (Table la and Table lb) can be combined with any appropriate anti-KLK5 antibody. The following references provide non-limiting examples of anti-KLK5 antibodies: U.S. Patent No. 11,292,828, entitled “KLK5 INHIBITORY PEPTIDE,” issued April 5, 2022; U.S. Patent Application Publication No. 2022-0306725, entitled “KLK5 INHIBITORY PEPTIDE,” published September 29, 2022; U.S. Patent Application Publication No. 2019-0078160, entitled “USE OF KLK5 ANTAGONISTS FOR TREATMENT OF A DISEASE,” published March 14, 2019; International Patent Application Publication No. WO2021156171, entitled “ANTIBODIES AGAINST KLK5,” published August 12, 2021; International Patent Application Publication No. WO2021156170, entitled “ANTIBODIES AGAINST KLK5,” published August 12, 2021; U.S. Patent Application Publication No. 2021-0301032, entitled “ANTI-KLK5 ANTIBODIES AND METHODS OF USE,” published September 30, 2021; and U.S. Patent Application Publication No. 2021-0130492, entitled “ANTI-KLK7 ANTIBODIES, ANTI-KLK5 ANTIBODIES, MULTISPECIFIC ANTI-KLK5 / KLK7 ANTIBODIES, AND METHODS OF USE,” published May 6, 2021, the contents of which are incorporated herein by reference.

[0158] In some embodiments, the multispecific antibody comprises three, four, five, six, seven, eight or more unique antigen-specific binding sites. In some embodiments, each unique antigen-specific binding site of the multispecific antibody targets a different antigen. In some embodiments, each unique antigen-specific binding site of the multispecific antibody targets a different region of the same antigen. In some embodiments, the multispecific antibody comprises unique antigen-specific binding sites that target different antigens and / or unique antigen-specific binding sites that target different regions of the same antigen. In some embodiments, the multispecific antibody comprises at least one antigen-specific binding site that targets a first antigen and at least one antigen-specific binding site that targets a second antigen. In some embodiments, the multispecific antibody comprises two or more antigen-specific binding sites that target different regions of a first antigen and / or two or more antigen-specific binding sites that target different regions of a second antigen.

[0159] In some embodiments, the multispecific antibody targets two antigens and contains one antigen-specific binding site for each antigen (1 + 1). In some embodiments, the multispecific antibody targets two antigens and contains two antigen-specific binding sites for each antigen (2 + 2). In some embodiments, the multispecific antibody targets two antigens and contains one antigen-specific binding site for one antigen and two antigen-specific binding sites for the other antigen (1 + 2). In some embodiments, the multispecific antibody targets two antigens and contains two antigen-specific binding sites for one antigen and three antigen-specific binding sites for the other antigen (2 + 3). In some embodiments, the multispecific antibody targets two antigens and contains three antigen-specific binding sites for one antigen and three antigen-specific binding sites for the other antigen (3 + 3).

[0160] In some embodiments, the multispecific antibody lacks Fc-mediated effector functions, e.g., antibody-dependent cell-mediated cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), complement fixation, and FcRn-mediated recycling. However, in some embodiments, the multispecific antibody comprises one or more Fc regions that support Fc-mediated effector functions, e.g., antibody-dependent cell-mediated cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), complement fixation, and FcRn-mediated recycling.

[0161] In some embodiments, an antibody provided herein is a bispecific antibody. In some embodiments, a bispecific antibody comprises at least two different Fv regions. In some embodiments, a bispecific antibody comprises two different heavy chains and two different light chains. In some embodiments, a bispecific antibody comprises one or more IgG molecules. In some embodiments, a bispecific antibody comprises one or more IgG molecules containing an additional antigen-specific binding site, e.g., an IgG molecule comprising an additional or modified Ig-like structure.

[0162] In some embodiments, a bispecific antibody comprises two single chain variable fragments (scFv) connected by a linker. In some embodiments, a bispecific antibody comprises two single domain antibodies, e.g., VH or VL domains, VHH, VNAR, or nanobodies, connected by a linker, e.g., a flexible glycine-rich linker, e.g., a (G4S)3 linker. In some embodiments, a bispecific antibody is in the form of a diabody, e.g., as described in P Holliger, T Prospero, and G Winter, “Diabodies”: small bivalent and bispecific antibody fragments, Proc Natl Acad Sci U S A. 1993 Jul 15; 90(14): 6444-6448, the entire contents of which are incorporated herein by reference in their entirety. In some embodiments, a bispecific antibody is a Fab fusion protein, e.g., a Fab-Fab fusion protein, a Fab-scFv fusion protein, or a Fab-Fv fusion protein. In some embodiments, a bispecific antibody comprises one antigen binding site, e.g., a scFv, which is modified to contain a second and distinct antigen-specific binding site as an integral part of the antibody, e.g., a scFv.

[0163] In some embodiments, the bispecific antibody is in a fragment-based format, symmetric format, or asymmetric format. In some embodiments, the bispecific antibody in a fragment-based format does not comprise an Fc region. In some embodiments, the bispecific antibody is in a tandem VHH, tandem scFv, DART, diabody, F(ab)2, scFv-Fab, tandem VHH, (scFv)2-Fab, or tandem diabody format. In some embodiments, the bispecific antibody is in an asymmetric format selected from the group consisting of rat-mouse hybrid IgG, hetero H HL exchange and / or assembly IgG, hetero H forced HL IgG, cH IgG, hetero HCrossMab, scFv-Fab IgG, DART-Fc, LP-DART, CODV-Fab-TL, HLE-BiTE, and F(ab)3CrossMab formats. In some embodiments, the bispecific antibody is in a symmetric format selected from the group consisting of IgG-(scFv)2, Bs4Ab, DVD-Ig, tetravalent DART-Fc, (scFV)4-Fc, CODV-Ig, two-in-one, mAb2, F(ab)4 CrossMab, and tandem VHH-Fc formats.

[0164] In some embodiments, the bispecific antibody is engineered to facilitate formation by knobs-into-holes technology, for example, to facilitate heterodimerization. In some embodiments, knobs-into-holes technology can be used to generate bispecific IgG molecules, trivalent Ig-like antibodies, bispecific Fc and CH3 fusion proteins, and other formats as discussed in Ridgway JB et al., ‘Knobs-into-holes’ engineering of antibody CH3 domains for heavy chain heterodimerization. Protein Eng 1996; 9:617-21; Atwell S et al., Stable heterodimers from remodeling the domain interface of a homodimer using a phage display library, J Mol Biol 1997; 270:26-35; and Merchant AM et al., An efficient route to human bispecific IgG, Nat Biotechnol 1998; 16:677-681, the entire contents of each of which are incorporated herein by reference in their entirety.

[0165] In some embodiments, the bispecific antibody lacks Fc-mediated effector functions, e.g., antibody-dependent cell-mediated cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), complement fixation, and / or FcRn-mediated recycling. However, in some embodiments, the bispecific antibody comprises one or more Fc regions that support Fc-mediated effector functions, e.g., antibody-dependent cell-mediated cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), complement fixation, and FcRn-mediated recycling.

[0166] III. Preparation of Anti-KLK5 / KLK7 Antibodies Antibodies described herein can be prepared by any method known in the art. See, e.g., Harlow and Lane, (1998) Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, New York.

[0167] In some embodiments, antibodies specific for a target antigen (e.g., KLK5 and / or KLK7) can be prepared by conventional hybridoma technology. Full-length target antigen or fragments thereof, optionally coupled to a carrier protein (e.g., KLH), can be used to immunize a host animal to generate antibodies that bind to the antigen. The route and schedule of immunization of the host animal is generally consistent with established and conventional techniques for antibody stimulation and production, as further described herein. General techniques for the production of mouse, humanized, and human antibodies are known in the art and are described herein. It is contemplated that any mammalian subject, including humans, or antibody-producing cells therefrom, can be manipulated for use as a basis for the production of mammalian, including human, hybridoma cell lines. Typically, the host animal is inoculated with an amount of immunogen, including as described herein, intraperitoneally, intramuscularly, orally, subcutaneously, intraplantarly, and / or intradermally.

[0168] If desired, an antibody of interest (monoclonal or polyclonal) (e.g., produced by a hybridoma) can be sequenced, and then the polynucleotide sequence can be cloned into a vector for expression or propagation. The sequence encoding the antibody of interest can be maintained in the vector in a host cell, and then the host cell can be expanded and frozen for future use. In an alternative, the polynucleotide sequence can be used for genetic manipulation to "humanize" the antibody or to improve affinity (affinity maturation) or other characteristics of the antibody. For example, if the antibody is to be used in human clinical trials and treatments, the constant region can be engineered to more closely resemble a human constant region to avoid an immune response. Genetic manipulation of the antibody sequence can be required to obtain greater affinity and higher efficacy for the target antigen. It will be apparent to those skilled in the art that one or more polynucleotide changes can be made to the antibody and still maintain its binding specificity for the target antigen.

[0169] In other embodiments, fully human antibodies can be obtained by using commercially available mice that have been engineered to express specific human immunoglobulins. Transgenic animals designed to produce more desirable (e.g., fully human antibodies) or more robust immune responses can also be used to generate humanized or human antibodies. Examples of such technologies are XenomouseRTMfrom Amgen, Inc. (Fremont, CA), and HuMAb-MouseRTMand TC MouseTMfrom Medarex, Inc. (Princeton, NJ), or H2L2 mice from Harbour Antibodies BV (Holland). In another alternative, antibodies can be made recombinantly by phage display or yeast technologies. See, e.g., U.S. Patent Nos. 5,565,332, 5,580,717, 5,733,743, and 6,265,150; and Winter et al. (1994) Annu. Rev. Immunol. 12:433-455. Alternatively, phage display technology (McCafferty et al. (1990) Nature 348:552-553) can be used to produce human antibodies and antibody fragments in vitro, from immunoglobulin variable (V) domain gene repertoires from unimmunized donors.

[0170] Antigen-binding fragments of intact antibodies (full length antibodies) can be produced by conventional methods. For example, F(ab')2 fragments can be generated by pepsin digestion of the antibody molecule, and Fab fragments can be generated by reducing the disulfide bridges of F(ab')2 fragments. Genetically engineered antibodies, such as humanized antibodies, chimeric antibodies, single-chain antibodies, and bispecific antibodies, can be produced by, for example, conventional recombinant techniques. In one embodiment, DNA encoding a monoclonal antibody specific to a target antigen can be readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that are specific to the heavy and light chain of the monoclonal antibody). The hybridoma cells serve as a preferred source of such DNA. Once isolated, the DNA can be placed into one or more expression vectors, which are then transfected into host cells such as E. coli cells, simian COS cells, Chinese hamster ovary (CHO) cells, human HEK293 cells, or myeloma cells that do not otherwise produce immunoglobulins, to obtain the synthesis of monoclonal antibodies in the recombinant host cells. See, e.g., PCT Publ. No. WO 87 / 04462. The DNA can then be modified, for example, by substituting the coding sequence for human heavy and light chain constant domains in place of the analogous murine sequences (Morrison et al. (1984) Proc. Nat. Acad. Sci. 81 :6851), or by covalently joining all or a portion of the coding sequence for a polypeptide to that for an immunoglobulin to create a chimeric antibody. In this manner, it is possible to produce a genetically engineered antibody having the binding specificity of the target antigen, such as a "chimeric" or "hybrid" antibody.

[0171] A single chain antibody can be prepared via recombinant techniques by linking a nucleotide sequence encoding the variable region of the heavy chain to a nucleotide sequence encoding the variable region of the light chain. Preferably, a flexible linker is incorporated between the two variable regions.

[0172] Antibodies obtained following methods known in the art and described herein can be characterized using methods well known in the art. For example, one method is to identify the epitope to which the antibody binds, or "epitope mapping." A number of methods are known in the art for mapping and characterizing the location of epitopes on proteins, including solving the crystal structure of the antibody-antigen complex, competition assays, gene segment expression assays, and synthetic peptide-based assays, as described, for example, in Chapter 11 of Harlow and Lane, Using Antibodies, a Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1999. In one embodiment, epitope mapping can be accomplished using H / D-Ex (hydrogen deuterium exchange) binding proteolysis and mass spectrometry. In another embodiment, epitope mapping can be used to determine the sequence to which the antibody binds. Epitopes can be linear epitopes, contained in a single amino acid segment, or conformational epitopes, formed by three-dimensional interactions of amino acids, which can not necessarily be contained in a single segment (primary structure linear sequence). Peptides of varying lengths (e.g., at least 4-6 amino acids long) can be isolated or synthesized (e.g., recombinantly) and used in binding assays with the antibody. In another embodiment, the epitope to which the antibody binds can be determined in a systematic screen by using overlapping peptides derived from the sequence of the target antigen and determining binding of the antibody. According to gene segment expression assays, the open reading frame encoding the target antigen is fragmented, either randomly or by specific genetic construction, and the expressed antigen fragments are assayed for reactivity with the antibody to be tested. For example, gene segments can be generated by PCR, and then transcribed and translated into protein in vitro in the presence of radioactive amino acids. Binding of the antibody to the radiolabeled antigen fragments is then determined by immunoprecipitation and gel electrophoresis. Large libraries of random peptide sequences displayed on the surface of bacteriophage particles (phage libraries) can also be used to identify certain epitopes. Alternatively, a defined library of overlapping peptide segments can be tested for binding to the test antibody in a simple binding assay. In another embodiment, mutagenesis of the antigen binding domain, domain swap experiments, and alanine scanning mutagenesis can be performed to identify residues required, sufficient, and / or necessary for epitope binding. Alternatively, competition assays can be performed using other antibodies known to bind to the same antigen to determine whether the antibody binds to the same epitope as the other antibodies. Competition assays are well known to those of skill in the art.

[0173] In some embodiments, the antibodies described herein are produced by recombinant techniques, as exemplified below. Nucleic acids encoding the heavy and light chains of an antibody as described herein can be cloned into an expression vector, each nucleotide sequence operably linked to a suitable promoter. In one embodiment, each nucleotide sequence encoding a heavy and light chain is operably linked to a unique promoter. Alternatively, the nucleotide sequences encoding the heavy and light chains can be operably linked to a single promoter, such that both the heavy and light chains are expressed from the same promoter. If desired, an internal ribosome entry site (IRES) can be inserted between the heavy and light chain encoding sequences.

[0174] In some embodiments, the nucleotide sequences encoding the two chains of an antibody are cloned into two vectors, which can be introduced into the same or different cells. When the two chains are expressed in different cells, each can be isolated from the host cells expressing them, and the isolated heavy and light chains can be mixed and incubated under suitable conditions to allow the antibody to form.

[0175] Generally, the nucleic acid sequences encoding one or all chains of an antibody can be cloned into a suitable expression vector and operably linked to a suitable promoter using methods known in the art. For example, the nucleotide sequences and vectors are contacted with restriction enzymes under suitable conditions to produce complementary ends on each molecule, which can pair with each other and be joined together by a ligase. Alternatively, synthetic nucleic acid linkers can be attached to the ends of the genes. These synthetic linkers contain nucleic acid sequences that correspond to specific restriction sites in the vector. The choice of expression vector / promoter will depend on the type of host cell used to produce the antibody.

[0176] A variety of promoters can be used to express the antibodies described herein, including but not limited to the cytomegalovirus (CMV) intermediate early promoter, viral LTRs (e.g., Rous sarcoma virus LTR, HIV-LTR, HTLV-1 LTR), simian virus 40 (SV40) early promoter, E. coli lac UV promoter, and herpes simplex tk virus promoter.

[0177] Adjustable promoters can also be used. Such adjustable promoters include those that use the lac repressor from E. coli as a transcriptional regulator to modulate transcription of a mammalian cell promoter carrying the lac operator [[Brown, M. et al., Cell, 49:603-612 (1987)], those that use the tetracycline repressor (tetR) [[Gossen, M. and Bujard, H., Proc. Natl. Acad. Sci. USA 89:5547-555115 (1992); Yao, F. et al., Human Gene Therapy, 9:1939-1950 (1998); Shockelt, P. et al., Proc. Natl. Acad. Sci. USA, 92:6522-6526 (1995)]]. Other systems include FK506 dimer, VP16 or p65 using astradiol, RU486, dienestrol or rapamycin. Inducible systems are available from Invitrogen, Clontech and Ariad, among others.

[0178] An inducible promoter can be used. In one embodiment, the lac repressor from E. coli can act as a transcriptional regulator to modulate transcription from a mammalian cell promoter carrying the lac operator [[M. Brown et al., Cell, 49:603-612 (1987)]; Gossen and Bujard (1992); [[M. Gossen et al., Natl. Acad. Sci. USA, 89:5547-5551 (1992)]], the tetracycline repressor (tetR) combined with a transcriptional activator (VP 16) to produce a tetR-mammalian cell transcriptional activator fusion protein tTa (tetR-VP 16), combined with tetO carrying a minimal promoter derived from the human cytomegalovirus (hCMV) promoter to produce a tetR-tet operator system to control gene expression in mammalian cells. In one embodiment, a tetracycline-inducible switch is used. When the tetracycline operator is positioned correctly downstream of the TATA element of the CMV IE promoter, the tetracycline repressor (tetR) alone, rather than the tetR-mammalian cell transcription factor fusion derivative, can act as an effective transregulator to modulate gene expression in mammalian cells (Yao et al., Human Gene Therapy). One particular advantage of this tetracycline-inducible switch is that it does not require the use of tetracycline repressor-mammalian cell transactivator or repressor fusion proteins to achieve its regulatable effect, which in some cases can be toxic to cells (Gossen 5 et al., Natl. Acad. Sci. USA, 89:5547-5551 (1992); Shockett et al., Proc. Natl. Acad. Sci. USA, 92:6522-6526 (1995)).

[0179] Additionally, the vector can contain some or all of, for example, selectable marker genes, such as a neomycin gene for selection of stable or transient transfectants in mammalian cells; an enhancer / promoter sequence from the immediate early gene of human CMV for high level transcription; transcription termination and RNA processing signals from SV40 for mRNA stability; SV40 polyomavirus origin of replication and ColEl for proper episomal replication; internal ribosome binding sites (IRES), universal multiple cloning sites; and T7 and SP6 RNA promoters for in vitro transcription of sense and antisense RNAs. Suitable vectors, as well as methods for producing vectors containing transgenes, are well known and available in the art. Examples of polyadenylation signals that can be used in practicing the methods described herein include, but are not limited to, the human collagen I polyadenylation signal, the human collagen II polyadenylation signal, and the SV40 polyadenylation signal.

[0180] One or more vectors (e.g., expression vectors) comprising nucleic acids encoding any of the described antibodies (e.g., the nucleic acid coding sequences listed in Table 3) can be introduced into a suitable host cell to produce the antibody. Non-limiting examples of host cells include Chinese hamster ovary (CHO) cells, dhfr- CHO cells, human embryonic kidney (HEK)-293 cells, African green monkey kidney (VERO) cells, non- secreting empty (NS0) cells, human embryonic retina (PER.C6) cells, Sp2 / 0 cells, baby hamster kidney (BHK) cells, Madin-Darby canine kidney (MDCK) cells, Madin-Darby bovine kidney (MDBK) cells, and monkey kidney CV1 line transformed by SV40 (COS) cells. In some embodiments, the host cell expressing an antibody described herein is a CHO cell. The host cell can be cultured under suitable conditions to express the antibody or any polypeptide chain thereof. Such an antibody or polypeptide chain thereof can be recovered from the cultured cells (e.g., cells or culture supernatant) by conventional methods (e.g., affinity purification). If necessary, the polypeptide chains of the antibody can be incubated under suitable conditions for a suitable period of time to produce the antibody. In some embodiments, the host cell comprises a nucleic acid encoding a heavy chain of an antibody described herein. In some embodiments, the host cell comprises a nucleic acid encoding a light chain of an antibody described herein. In some embodiments, the host cell comprises a nucleic acid encoding a heavy chain and a nucleic acid encoding a light chain.

[0181] In some embodiments, the method for making an antibody described herein involves a recombinant expression vector that encodes both the heavy and light chains of an antibody described herein, as otherwise described herein. The recombinant expression vector can be introduced into a suitable host cell (e.g., a dhfr- CHO cell) by conventional means (e.g., calcium phosphate-mediated transfection). Positive transformant host cells can be selected and cultured under suitable conditions that allow for expression of the two polypeptide chains that form the antibody, which can be recovered from the cells or culture medium. If necessary, the two chains recovered from the host cells can be incubated under suitable conditions that allow for formation of the antibody.

[0182] In one embodiment, two recombinant expression vectors are provided, one encoding the heavy chain of the antibody and the other encoding the light chain of the antibody. Both recombinant expression vectors can be introduced into a suitable host cell (e.g., a dhfr- CHO cell) by conventional means (e.g., calcium phosphate-mediated transfection).

[0183] Alternatively, each expression vector can be introduced into a suitable host cell. Positive transformants can be selected and cultured under suitable conditions that allow for expression of the polypeptide chains of the antibody. When both expression vectors are introduced into the same host cell, the antibody produced therein can be recovered from the host cell or the culture medium. If necessary, the polypeptide chains can be recovered from the host cell or the culture medium and then incubated under suitable conditions that allow for formation of the antibody. When the two expression vectors are introduced into different host cells, each of which can be recovered from the respective host cell or the respective culture medium. Then, the two polypeptide chains can be incubated under suitable conditions to form the antibody.

[0184] Standard molecular biology techniques are used to prepare the recombinant expression vectors, transfect the host cells, select for transformants, culture the host cells, and recover the antibody from the culture medium. For example, some antibodies can be isolated by affinity chromatography using a protein A or protein G coupled matrix.

[0185] Any nucleic acid encoding a heavy chain, a light chain, or both, of an antibody as described herein (e.g., as provided in Table 3), vectors containing them (e.g., expression vectors), and host cells comprising the vectors are within the scope of the present disclosure.

[0186] Table 3: Nucleic acid sequences encoding the VH / VL of the anti-KLK5 / KLK7 antibodies listed in Table la In some embodiments, the present disclosure provides an isolated nucleic acid comprising a sequence that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to any one of SEQ ID NOs: 22, 24, 26, or 27. In some embodiments, the present disclosure provides an isolated nucleic acid comprising a sequence that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to any one of SEQ ID NOs: 23, 25, 32, or 33.

[0187] In some embodiments, the present disclosure provides an expression vector encoding an anti-KLK5 / KLK7 antibody described herein. In some embodiments, the expression vector comprises an isolated nucleic acid that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to any one of SEQ ID NOs: 22, 24, 26, or 27. In some embodiments, the expression vector comprises an isolated nucleic acid that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to any one of SEQ ID NOs: 23, 25, 32, or 33.

[0188] In some embodiments, an anti-KLK5 / KLK7 antibody described herein is produced by expressing in a recombinant cell: (i) an isolated nucleic acid that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to any one of SEQ ID NOs: 22, 24, 26, or 27, and / or (ii) an isolated nucleic acid that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to any one of SEQ ID NOs: 23, 25, 32, or 33.

[0189] In some embodiments, an anti-KLK5 / KLK7 antibody described herein is produced by expression of an expression vector in a recombinant cell, the expression vector comprising: (i) an isolated nucleic acid that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to any one of SEQ ID NOs: 22, 24, 26, and 27, and / or (ii) an isolated nucleic acid that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to any one of SEQ ID NOs: 23, 25, 32, or 33.

[0190] In some embodiments, the present disclosure provides an isolated nucleic acid comprising a sequence that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO: 22, and / or an isolated nucleic acid that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO: 23.

[0191] In some embodiments, the present disclosure provides an isolated nucleic acid comprising a sequence that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO: 24, and / or an isolated nucleic acid that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO: 25.

[0192] In some embodiments, the present disclosure provides an isolated nucleic acid comprising a sequence that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO: 26, and / or an isolated nucleic acid that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO: 32.

[0193] In some embodiments, the present disclosure provides an isolated nucleic acid comprising a sequence that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO: 27, and / or an isolated nucleic acid that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO: 33.

[0194] In some embodiments, the present disclosure provides an isolated nucleic acid comprising a sequence that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO: 22, and / or an isolated nucleic acid that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO: 23.

[0195] In some embodiments, the present disclosure provides an expression vector comprising an isolated nucleic acid comprising a sequence that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO: 24, and / or an isolated nucleic acid that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO: 25.

[0196] In some embodiments, the present disclosure provides an expression vector comprising an isolated nucleic acid comprising a sequence that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO: 26, and / or an isolated nucleic acid that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO: 32.

[0197] In some embodiments, the present disclosure provides an expression vector comprising an isolated nucleic acid comprising a sequence that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO: 27, and / or an isolated nucleic acid that is at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to SEQ ID NO: 33.

[0198] In some embodiments, the antibodies described herein are used to modulate the activity or function of at least one gene, protein, and / or nucleic acid. In some embodiments, the molecular payload is responsible for modulating the gene, protein, and / or nucleic acid. The molecular payload can be a small molecule, a protein, a nucleic acid, an oligonucleotide, or any molecular entity capable of modulating the activity or function of a gene, protein, and / or nucleic acid in a cell.

[0199] In some embodiments, multispecific antibodies comprise direct fusions or linkages of different antigen-specific binding sites. In some embodiments, multispecific antibodies comprise immunoglobulin-derived heterodimerization domains to generate multispecific antibodies. In some embodiments, multispecific antibodies can be formed by co-expression of different heavy chains and two different light chains. In some embodiments, multispecific antibodies can be formed by co-expression of different heavy chains and one common light chain. In some embodiments, multispecific antibodies comprise engineered CH1 domains (first constant Ig domain of the heavy chain) that promote correct heavy chain-light chain pairing, e.g., in such co-expression systems, as disclosed in International Patent Application Publication No. WO2021067404, “CH1 DOMAIN VARIANTS ENGINEERED FOR PREFERENTIAL LIGHT CHAIN PAIRING AND MULTISPECIFIC ANTIBODIES COMPRISING THE SAME”, published April 8, 2021, the contents of which are incorporated by reference herein.

[0200] In some embodiments, multispecific antibodies comprise variant CH1 domains that pair (e.g., preferentially pair) with particular variant CL domains. For example, in some embodiments, multispecific antibodies comprise heavy chains comprising variant CH1 domains that preferentially pair with variant CLK or CLl domains. Non-limiting examples of such multispecific antibody configurations are provided in International Patent Application Publication No. WO2022150787, “VARIANT CH1 DOMAINS AND VARIANT CL DOMAINS ENGINEERED FOR PREFERENTIAL CHAIN PAIRING AND MULTI-SPECIFIC ANTIBODIES COMPRISING THE SAME”, published July 14, 2022, the contents of which are incorporated by reference herein in their entirety.

[0201] In some embodiments, the multispecific antibody comprises variant CH3 domains that preferentially form CH3-CH3 heterodimers rather than CH3-CH3 homodimers. Incorporation of such variant CH3 domains facilitates, for example, heterodimerization of different antibodies, thereby forming a multispecific antibody. The following references provide non-limiting examples of such multispecific antibody configurations: International Patent Application Publication No. WO2022150785, “VARIANT CH3 DOMAINS ENGINEERED FOR PREFERENTIAL CH3 HETERODIMERIZATION, MULTI-SPECIFIC ANTIBODIES COMPRISING THE SAME, AND METHODS OF MAKING THEREOF”; published July 14, 2022, the contents of which are incorporated herein by reference in their entirety.

[0202] In some embodiments, the bispecific antibody comprises a direct fusion or linkage of different antigen-specific binding sites. In some embodiments, the bispecific antibody comprises an immunoglobulin-derived heterodimerization domain to generate the bispecific antibody. For example, in some embodiments, the bispecific antibody can be formed by co-expression of two different heavy chains and two different light chains. In some embodiments, the bispecific antibody can be formed by co-expression of two different heavy chains and one common light chain. In some embodiments, fusion of two antibody-producing cell lines allows the heavy and light chain combinations of two different antibodies, such that the resulting bispecific antibody comprises the heavy and light chains of the first antibody and the heavy and light chains of the second antibody. In some embodiments, the heavy and light chain constant regions are of the same isotype. In some embodiments, the heavy and light chain constant regions are of different isotypes.

[0203] In some embodiments, the bispecific antibody comprises variant heavy and / or light chains that force or facilitate the correct assembly of two heavy chains with a cognate heavy and light chain, or purification of the correctly assembled bispecific antibody (see, e.g., Figure 3 and Figure 4and Table 1). In some embodiments, bispecific antibodies are formed using knobs-into-holes technology, e.g., to facilitate heterodimerization. In some embodiments, knobs-into-holes technology can be used to generate bispecific IgG molecules, trivalent Ig-like antibodies, bispecific Fc and CH3 fusion proteins and other formats as discussed in Ridgway JB et al., ‘Knobs-into-holes’ engineering of antibody CH3 domains for heavy chain heterodimerization. Protein Eng 1996; 9:617-21; Atwell S et al., Stable heterodimers from remodeling the domain interface of a homodimer using a phage display library, J Mol Biol 1997; 270:26-35; and Merchant AM et al., An efficient route to human bispecific IgG, Nat Biotechnol 1998; 16:677-681, the entire contents of each of which are incorporated herein by reference in their entirety.

[0204] In some embodiments, bispecific antibodies comprise engineered CH1 domains (the first constant Ig domain of the heavy chain) that promote correct heavy chain-light chain pairing, e.g., in such co-expression systems as disclosed in International Patent Application Publication No. WO2021067404, “CH1 DOMAIN VARIANTS ENGINEERED FOR PREFERENTIAL LIGHT CHAIN PAIRING AND MULTISPECIFIC ANTIBODIES COMPRISING THE SAME”, published April 8, 2021, the contents of which are incorporated herein by reference.

[0205] In some embodiments, the bispecific antibody comprises a variant CH1 domain paired (e.g., preferentially paired) with a particular variant CL domain. For example, in some embodiments, the bispecific antibody comprises a heavy chain comprising a variant CH1 domain that preferentially pairs with a variant CLK or CLl domain. The following reference provides non-limiting examples of such bispecific antibody configurations: International Patent Application Publication No. WO2022150787, “VARIANT CH1 DOMAINS AND VARIANT CL DOMAINS ENGINEERED FOR PREFERENTIAL CHAIN PAIRING AND MULTI-SPECIFIC ANTIBODIES COMPRISING THE SAME,” published July 14, 2022, the contents of which are incorporated herein by reference in their entirety.

[0206] In some embodiments, the bispecific antibody comprises a variant CH3 domain that preferentially forms a CH3-CH3 heterodimer rather than a CH3-CH3 homodimer. Incorporation of such variant CH3 domains facilitates, for example, heterodimerization of different antibodies, thereby forming a bispecific antibody. The following reference provides non-limiting examples of such bispecific antibody configurations: International Patent Application Publication No. WO2022150785, “VARIANT CH3 DOMAINS ENGINEERED FOR PREFERENTIAL CH3 HETERODIMERIZATION, MULTI-SPECIFIC ANTIBODIES COMPRISING THE SAME, AND METHODS OF MAKING THEREOF”; 2022 July 14, the contents of which are incorporated herein by reference in their entirety.

[0207] In some embodiments, non-immunoglobulin heterodimerization modules can be used to combine different antigen-specific binding sites in a non-covalent or covalent manner to form bispecific antibodies. For example, in some embodiments, bispecific antibodies are formed by the dock-and-lock method (DNL) which uses a heterodimeric assembly of the regulatory subunit of cAMP-dependent protein kinase (PKA) and the anchoring domain (AD) of A-kinase anchoring proteins (AKAPs). In some embodiments, non-immunoglobulin heterodimerization modules can be used to combine different antigen-specific binding sites to form bispecific antibodies, such as the barnase-barstar system, the adapter / docking tag module based on a mutant RNase I fragment, and the SNARE module based on the interaction of three synaptic fusion proteins, small synaptic vesicle proteins, and SNAP25.

[0208] IV. Pharmaceutical Compositions The antibodies as described herein, as well as the encoding nucleic acids or sets of nucleic acids, vectors comprising them, or host cells comprising the vectors, can be mixed with a pharmaceutically acceptable carrier (excipient) to form a pharmaceutical composition for treating a disease of interest. By "acceptable" it is meant that the carrier must be compatible with the active ingredient of the composition (and preferably capable of stabilizing the active ingredient) and not deleterious to the subject to be treated. Pharmaceutically acceptable excipients (carriers), including buffers, are well known in the art. See, e.g., Remington: The Science and Practice of Pharmacy 20th Ed. (2000) Lippincott Williams and Wilkins, K. E. Hoover, Ed.

[0209] The anti-KLK5 / KLK7 antibody-containing pharmaceutical compositions disclosed herein can also include a suitable buffer. A buffer is a weak acid or base used to maintain the pH of a solution near a chosen value upon the addition of another acid or base. In some embodiments, the buffers disclosed herein can be buffers that are able to maintain a physiological pH even as the concentration of carbon dioxide (produced by cellular respiration) changes. Exemplary buffers include, but are not limited to, HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid) buffer, Dulbecco's Phosphate Buffered Saline (DPBS) buffer, or Phosphate Buffered Saline (PBS) buffer. Such buffers can include disodium hydrogen phosphate and sodium chloride, or potassium dihydrogen phosphate and potassium chloride.

[0210] The pharmaceutical compositions described herein comprise one or more suitable salts. A salt is an ionic compound that can be formed by the neutralization reaction of an acid and a base. (Skoog, D.A; West, D.M.; Holler, J.F.; Crouch, S.R. (2004). “Chapters 14-16”. Fundamentals of Analytical Chemistry (8th ed.)). A salt is composed of an associated number of cations (positively charged ions) and anions (negative ions) such that the product is electrically neutral (no net charge).

[0211] In some embodiments, the pharmaceutical compositions can comprise a pharmaceutically acceptable carrier, excipient, or stabilizer in lyophilized formulation or aqueous solution. (Remington: The Science and Practice of Pharmacy 20th Ed. (2000) Lippincott Williams and Wilkins, K. E. Hoover, Ed.). In some embodiments, the pharmaceutical compositions can be formulated for intravenous injection. In some embodiments, the pharmaceutical compositions can be formulated for subcutaneous injection.

[0212] Pharmaceutical compositions to be used for in vivo administration must be sterile. This is readily accomplished by, for example, filtration through sterile filtration membranes. Therapeutic antibody compositions are generally placed into a container having a sterile access port, such as an intravenous or subcutaneous solution bag or vial having a stopper pierceable by a hypodermic injection needle.

[0213] V. Methods of Use In certain aspects, the present disclosure provides methods and related compositions for treating conditions associated with KLK5- and KLK7-related disorders, including, for example, Netherton Syndrome, atopic dermatitis (with and without filaggrin mutations), eosinophilic esophagitis, prurigo nodularis, chronic prurigo of unknown origin (CPUO), dry skin, asthma (e.g., KLK5-associated asthma in particular), ichthyosis vulgaris, and skin pruritus.

[0214] Aspects of the present disclosure relate to methods and compositions (e.g., anti-KLK5 / KLK7 dual inhibitor antibodies) that can be used to promote proper barrier function (e.g., epidermal barrier function). Hyperactive kallikrein 5 / 7 causes genetic and spontaneous disruption of epidermal barrier function and is associated with related disorders such as Netherton Syndrome, Eosinophilic Esophagitis, Atopic Dermatitis. Accordingly, in some embodiments, the methods provided herein include administering to a subject an effective amount of one or more of the anti-KLK5 / KLK7 antibodies provided herein to restore epithelial barrier in a subject in need thereof. In other aspects, methods are provided for addressing one or more aspects of altered barrier function. For example, in some embodiments, the methods provided herein include administering to a subject an effective amount of one or more of the anti-KLK5 / KLK7 antibodies provided herein to reduce dermal infiltration in a subject in need thereof. In some embodiments, the methods provided herein include administering to a subject an effective amount of one or more of the anti-KLK5 / KLK7 antibodies provided herein to reduce epithelial inflammation in a subject in need thereof. In some embodiments, the methods provided herein include administering to a subject an effective amount of one or more of the anti-KLK5 / KLK7 antibodies provided herein to reduce epithelial permeability in a subject in need thereof. In some embodiments, the methods provided herein include administering to a subject an effective amount of the anti-KLK5 / KLK7 antibodies provided herein to reduce parakeratosis in a subject in need thereof. In some embodiments, the methods provided herein include administering to a subject an effective amount of one or more of the anti-KLK5 / KLK7 antibodies provided herein to reduce skin inflammatory cytokines in a subject in need thereof. In some embodiments, the methods provided herein include administering to a subject an effective amount of one or more of the anti-KLK5 / KLK7 antibodies provided herein to reduce transepidermal water loss in a subject in need thereof.

[0215] Other aspects of the present disclosure relate to methods and compositions (e.g., anti-KLK5 / KLK7 antibodies) useful for treating atopic dermatitis. Atopic dermatitis (AD), also known as eczema, is a common chronic pruritic inflammatory skin disease. In some embodiments, atopic dermatitis begins in the infant or toddler stage (e.g., at 2 years of age or about 2 years of age) of a subject. Accordingly, in some embodiments, the methods provided herein are useful for treating a subject having atopic dermatitis who is 2 years of age or older. Atopic dermatitis can be associated with elevated total serum IgE concentrations. Accordingly, in some embodiments, the methods provided herein are useful for treating a subject having atopic dermatitis and elevated levels of total serum IgE concentrations (e.g., as compared to normal IgE levels in a subject who does not have atopic dermatitis or a related condition). In some embodiments, atopic dermatitis is associated with chronic recurrent forms of skin inflammation, epidermal barrier dysfunction (e.g., leading to dry skin), and / or IgE-mediated sensitivity to allergens (e.g., food and environmental allergens). Accordingly, in some embodiments, provided herein are methods of treating a subject having atopic dermatitis, the methods comprising administering to the subject an effective amount of an anti-KLK5 / KLK7 antibody. Further, in some embodiments, the subject to be treated exhibits chronic recurrent forms of skin inflammation, epidermal barrier dysfunction, and / or IgE-mediated sensitivity to allergens.

[0216] Other aspects of the present disclosure relate to methods and compositions (e.g., anti-KLK5 / KLK7 antibodies) useful for treating a subject having Netherton Syndrome. In some embodiments, provided herein are methods of treating a subject having Netherton Syndrome, the methods comprising administering to the subject an effective amount of one or more anti-KLK5 / KLK7 antibodies. Netherton Syndrome is a rare and severe autosomal recessive skin disorder. In some embodiments, Netherton Syndrome is associated with congenital erythroderma, specific hair shaft abnormalities, and / or atopic manifestations (e.g., with high IgE levels) associated with the subject not having Netherton Syndrome or a related condition. In some embodiments, a subject having Netherton Syndrome exhibits atopic manifestations, including eczematous skin rash, atopic dermatitis, pruritus, hay fever, angioedema, urticaria, high levels of IgE in serum, and / or high eosinophilia. In some embodiments, Netherton Syndrome is caused by mutations in the serine protease inhibitor Kazal-type 5 (SPINK5) gene, which encodes the proteinase inhibitor lymphoepithelial Kazal-type-related inhibitor. In some embodiments, the lack of this proteinase inhibitor leads to secondary corneocyte detachment due to epidermal protease hyperactivity. Accordingly, in some embodiments, the methods provided herein comprise administering to the subject an effective amount of one or more of the anti-KLK5 / KLK7 antibodies provided herein to ameliorate one or more aspects or symptoms associated with Netherton Syndrome (e.g., atopic manifestations, such as skin rash scaling, corneocyte detachment).

[0217] Other aspects of the present disclosure relate to methods and compositions (e.g., anti-KLK5 / KLK7 antibodies) useful for treating eosinophilic esophagitis. In some embodiments, symptoms of eosinophilic esophagitis include feeding difficulties, failure to thrive, vomiting, upper abdominal or chest pain, dysphagia, and food impaction. In some embodiments, a subject having eosinophilic esophagitis is a young male with relatively high atopic disease predisposition. In some embodiments, a subject having eosinophilic esophagitis is diagnosed by endoscopy and / or biopsy results of eosinophils isolated from the esophagus. In some embodiments, eosinophilic esophagitis is defined histologically by the presence of proliferative changes, including in some embodiments, thickening of the basal epithelial layer and / or papillary elongation, a minimum of 24 eosinophils per high power field in the distal esophagus, and / or an absence of eosinophilia in any other intestinal segment assessed. In some embodiments, a subject having eosinophilic esophagitis exhibits low levels or a lack of certain serine protease inhibitors belonging to the lymphoepithelial Kazal-type inhibitor protein family (e.g., SPINK7), e.g., in an esophageal biopsy. In some embodiments, eosinophilic esophagitis is distinguished from reflux esophagitis by the degree of mucosal eosinophilia and lack of response to acid suppression. In some embodiments, provided herein are methods of treating a subject having eosinophilic esophagitis, the methods comprising administering to the subject an effective amount of one or more anti-KLK5 / KLK7 antibodies.

[0218] Other aspects of the present disclosure relate to methods and compositions (e.g., anti-KLK5 / KLK7 antibodies) useful for treating nodular prurigo. Nodular prurigo is a chronic inflammatory skin disease in which intense itching, a symmetrically distributed skin rash is most commonly seen on the arms, legs, upper back, and / or abdomen. In some embodiments, nodular prurigo occurs spontaneously. However, in some embodiments, nodular prurigo is associated with other skin diseases or medical conditions, such as cancer, diabetes, chronic kidney disease, or AIDS. In some embodiments, altered functioning of the immune system and nerves in the skin is thought to contribute to the intensification of the sensation of itch (pruritus) that leads to frequent scratching; from this, such frequent scratching and picking at the skin leads to further thickening and formation of lesions. Thus, in some embodiments, provided herein are methods of treating a subject having nodular prurigo, the methods comprising administering to the subject an effective amount of one or more anti-KLK5 / KLK7 antibodies.

[0219] Other aspects of the present disclosure relate to methods and compositions (e.g., anti-KLK5 / KLK7 antibodies) useful for treating chronic pruritus. In some embodiments, chronic pruritus is associated with itch that persists for more than 6 weeks (e.g., up to 3 months, up to 6 months, up to 1 year, or more). In some embodiments, the occurrence of chronic pruritus is associated with an underlying unrelated disease, including chronic kidney disease, hepatobiliary disease, and neuropathic conditions such as acrodynia and allodynic back pain. In some embodiments, when the underlying cause of the pruritus cannot be determined, a diagnosis of chronic pruritus of unknown origin (CPUO) is made. In some embodiments, chronic pruritus is associated with severe itch and prominent scratch marks. In some embodiments, provided herein are methods of treating a subject having chronic pruritus, including CPUO, comprising administering to the subject an effective amount of one or more anti-KLK5 / KLK7 antibodies.

[0220] Other aspects of the present disclosure relate to methods and compositions (e.g., anti-KLK5 / KLK7 antibodies) useful for treating ichthyosis vulgaris. In some embodiments, ichthyosis vulgaris is caused by a heterozygous mutation in the filaggrin gene. In some embodiments, subjects having homozygous or compound heterozygous mutations in this gene have a more severe phenotype. In some embodiments, a histological feature of ichthyosis vulgaris is the absence or reduction of lamellar granules in the epidermis and mild hyperkeratosis. Lamellar granules contain a histidine-rich protein, which is the precursor form of filaggrin, a keratin filament aggregation protein (pre-filaggrin). In some embodiments, pre-filaggrin and filaggrin are reduced or absent in subjects having ichthyosis vulgaris. In some embodiments, ichthyosis vulgaris includes accentuated palm lines, periorificial keratosis, and fine scaling most apparent on the lower abdomen, arms, and legs. In some embodiments, a subject can exhibit pronounced desquamation. In some embodiments, a subject can exhibit accentuated palm lines, periorificial keratosis, and in some cases, fine scaling. Accordingly, in some embodiments, provided herein are methods of treating a subject having ichthyosis vulgaris or one or more symptoms or phenotypic features thereof, comprising administering to the subject an effective amount of one or more anti-KLK5 / KLK7 antibodies.

[0221] Other aspects of this disclosure relate to methods and compositions (e.g., anti-KLK5 / KLK7 antibodies) that can be used to treat psoriasis. Psoriasis (or psoriasis vulgaris) is a chronic inflammatory skin disease. In some embodiments, psoriasis is characterized by red, scaly patches of skin that can be found on the scalp, elbows, and / or knees of a subject. In some embodiments, psoriasis is associated with severe arthritis in a subject. In some embodiments, psoriasis-related lesions are caused by abnormal keratinocyte proliferation and inflammatory cell infiltration into the dermis and epidermis. In some embodiments, the subject experiences a psoriasis flare-up between the ages of 15 and 30. In some embodiments, psoriatic lesions are characterized by induration, scaling, and / or erythema, which may be accompanied by histological evidence of inflammation, abnormal keratinocyte proliferation / terminal differentiation, and / or dermal angiogenesis. In some embodiments, psoriatic inflammatory infiltration may be evident at the dermal-epidermal junction, containing activated T cells and antigen-presenting cells (APCs). In some embodiments, the presence of activated T cells and APCs in such infiltration precedes the occurrence of epidermal hyperproliferation. In some embodiments, increased levels of inflammatory cytokines can be detected in the psoriatic epidermis, which can lead to enhanced T cell activation and excessive proliferation and accelerated differentiation of keratinocytes. In some embodiments, this document provides a method for treating a subject with psoriasis, the method comprising administering to the subject an effective amount of one or more anti-KLK5 / KLK7 antibodies. In some embodiments, the subject receives treatment prior to the occurrence of epidermal hyperproliferation. However, in some embodiments, the subject receives treatment after the occurrence of epidermal hyperproliferation and accelerated differentiation of keratinocytes.

[0222] Other aspects of this disclosure relate to methods and compositions (e.g., anti-KLK5 / KLK7 antibodies) that can be used to treat rosacea. Rosacea is an inflammatory disease characterized by erythema, papules, pustules, and / or telangiectasia. In some embodiments, a subject with rosacea expresses abnormally high levels of cathelicidin in their facial skin. In some embodiments, the proteolytically processed form of the cathelicidin peptide found in rosacea differs from that present in normal subjects. In some embodiments, a method of treating a subject with rosacea is provided herein, the method comprising administering to the subject an effective amount of one or more anti-KLK5 / KLK7 antibodies.

[0223] Other aspects of the present disclosure relate to methods of treating a subject having asthma provided herein, the methods comprising administering to the subject an effective amount of one or more antibodies disclosed herein (e.g., an anti-KLK5 / KLK7 antibody). In some embodiments, provided herein are methods of treating a subject having asthma, the methods comprising administering an effective amount of one or more anti-KLK5 / KLK7 antibodies. In some embodiments, the subject has asthma (e.g., a subject to be treated with a KLK5-targeting antibody provided herein). In some embodiments, the asthma is selected from the group consisting of: allergic asthma, aspirin-sensitive / aggravated asthma, smoking-induced asthma, asthma that is uncontrolled by corticosteroids or other chronic asthma control medications, atopic asthma, bronchial obstruction-related asthma, pathogenesis-related asthma, chronic asthma, corticosteroid-naive asthma, corticosteroid- refractory asthma, corticosteroid-resistant asthma, high-eosinophil-level asthma, eosinophilic asthma, low-eosinophil-level asthma, exercise-induced asthma, mild asthma, moderate-to-severe asthma, Netherton syndrome asthma, newly diagnosed and / or untreated asthma, non-allergic asthma, non-Th2-driven asthma, periostin-high-level asthma, periostin-low-level asthma, Th2-low-level asthma, type 2 (T2)-driven asthma, and type 2 low-inflammation asthma. In some embodiments, the subject has atopic asthma or allergic asthma. In some embodiments, the subject has aspirin-sensitive or aspirin-aggravated asthma. In some embodiments, the subject has asthma associated with non-steroidal anti-inflammatory drugs (NSAIDs). Thus, in some embodiments, the subject has asthma triggered by aspirin or similar NSAIDs (e.g., a recent intake of aspirin or similar NSAIDs). In some embodiments, the subject has bronchospasm, which can or can not be characterized as asthma. For example, in some embodiments, the subject has exercise-induced bronchospasm.

[0224] In some embodiments, the subject has eosinophilic asthma. In some embodiments, the subject has eosinophilic inflammation positive (EIP) asthma. In some embodiments, the subject has eosinophilic inflammation negative (EIN) asthma. In some embodiments, the subject has eosinophilic high level asthma (e.g., any of at least about 150, 200, 250, 300, 350, or 400 eosinophil counts per ml of blood). In some embodiments, the subject has eosinophilic low level asthma (e.g., less than about 150 eosinophil counts per pL of whole blood or less than about 100 eosinophil counts per pL of blood). These and other examples of asthma-related conditions that can be treated using the compositions (e.g., anti-KLK5 / KLK7 antibodies) provided herein are disclosed in WO2015 / 061441, METHODS OF DIAGNOSING AND TREATING EOSINOPHILIC DISORDERS, published April 30, 2015, the relevant content of which is incorporated herein by reference.

[0225] In some embodiments, the subject has exercise-induced asthma, intermittent or exercise-induced mild asthma, mild or corticosteroid naive asthma, moderate to severe asthma, Netherton syndrome asthma, newly diagnosed asthma, untreated asthma, or severe asthma. In some embodiments, the subject has asthma that was previously uncontrolled or unmanaged without or without the use (e.g., long-term use) of inhaled topical or systemic corticosteroids to control or manage symptoms (e.g., symptoms such as cough, wheeze, shortness of breath / dyspnea, and chest pain) In some embodiments, the subject has periostin low level asthma (e.g., has a periostin level of less than about 20 ng / mL of serum). In some embodiments, the subject has periostin high level asthma (e.g., has a periostin level of at least about any of 20 ng / mL, 25 ng / mL, or 50 ng / mL of serum). In some embodiments, the subject has non-allergic asthma (e.g., which can or can not be triggered by infection or relatively cold dry air, such as infection with respiratory viruses (e.g., influenza virus, coronavirus, parainfluenza virus, rhinovirus, human metapneumovirus, and respiratory syncytial virus) or inhaled irritants (air pollutants, smoke, combustion particles (e.g., diesel particles), volatile chemicals, indoor or outdoor gases). In some embodiments, the subject has asthma due to acute or chronic primary or second-hand smoke exposure (cigarettes, cigars, pipes, or other combustion products) or due to inhalation or vaping (nicotine or other similar substances). In some embodiments, the subject has persistent chronic severe asthma with possible life-threatening acute exacerbation events (flare-ups or bursts).

[0226] In some embodiments, the subject has a T helper lymphocyte type 2 (Th2) or type 2 (Th2) high asthma condition. In some embodiments, the subject has a Th2-induced asthma. For example, in some embodiments, Th2 cells and / or their secreted effector molecules mediate the immune response to allergens and are triggered by exposure to a specific allergen that causes allergic asthma in the subject. In some embodiments, the subject has activated Th2 cell-mediated asthma, which can be caused in part by the secretion of interleukins (e.g., IL-4, IL-5, and IL-13). Thus, in some embodiments, KLK5 antibodies (or other antibodies) can be used alone or in combination with one or more antibodies targeting cytokines (e.g., IL-13, IL-17, IL-5, and IL-4) and allergen-related targets (e.g., IgE) in a multispecific antibody format. Examples of such antibodies for treating asthma include, but are not limited to, omalizumab (XOLAIR®) (targets soluble IgE); reslizumab (targets IL-13); mepolizumab (targets IL-5); and quilizumab (targets membrane-bound IgE).

[0227] Additionally, in other embodiments, the antibodies as described herein are used to treat, but are not limited to, inflammatory disorders, infectious diseases, allergic diseases, and autoimmune disorders. In some embodiments, the inflammatory disorder is selected from, but not limited to, rosacea, nodular prurigo, Crohn’s disease, ankylosing spondylitis, ulcerative colitis, hidradenitis suppurativa, and uveitis or one or more barrier function-related symptoms thereof. In some embodiments, the allergic disease is selected from, but not limited to, eczema, atopic dermatitis, asthma, sinusitis, and eosinophilic esophagitis. In some embodiments, the autoimmune disease is selected from, but not limited to, rheumatoid arthritis, psoriatic arthritis, juvenile idiopathic arthritis, Behcet’s disease, and plaque psoriasis or one or more barrier function-related symptoms thereof.

[0228] It will be apparent to those skilled in the art, based on the teachings provided herein, to determine an amount of an antibody (e.g., an anti-KLK5 / KLK7 antibody) that achieves a therapeutic effect. As recognized by those skilled in the art, the effective amount varies depending on the particular condition being treated, the severity of the condition, individual patient parameters including age, physical condition, size, gender and weight, the duration of the treatment, the nature of concurrent therapy (if any), the specific route of administration and like factors within the knowledge and expertise of the health practitioner. The specific dosing regimen (i.e., dosages, scheduling, and repetition) employed in the methods described herein will depend on the particular subject and that subject’s medical history, as discussed herein.

[0229] Empirical considerations will generally help determine the dosage, e.g., the time to reach maximum effect, the half-life, and / or the time to exceed a particular concentration. For example, antibodies that are compatible with the human immune system, such as humanized or fully human antibodies, can be used to prolong the half-life of the antibody and prevent the antibody from being attacked by the host immune system. Other reasons for dosage adjustment include differences in pharmacokinetics or pharmacodynamics responses driven by gender, age, individual response, polymorphisms on the antibody target, and / or receptors involved in antibody clearance. The frequency of administration can be determined and adjusted over the course of therapy, and is generally, but not necessarily, based on the treatment and / or inhibition and / or amelioration and / or delay of the target disease / condition. Alternatively, a sustained continuous release formulation of the antibody can be appropriate. A variety of formulations and devices are known in the art for achieving sustained release.

[0230] The frequency of administration can vary depending on the claimed method. In some embodiments, the composition can be administered once. In some embodiments, the composition will be administered multiple times. In some embodiments, the frequency of administration is every week, every 2 weeks, every 3 weeks, every 4 weeks, every 5 weeks, every 6 weeks, every 7 weeks, every 8 weeks, every 9 weeks, or every 10 weeks; or every month, every 2 months, or every 3 months or longer. In some embodiments, the composition will be administered every day, every two weeks, every week, every two months, every month, or at any interval of time that provides suitable (e.g., maximal) efficacy while minimizing the safety risks to the subject. Generally, the efficacy of the therapy as well as the therapeutic and safety risks can be monitored throughout the course of therapy.

[0231] In some embodiments, the compositions (e.g., anti-KLK5 / KLK7 antibodies) provided herein can be administered to a subject at one or more time intervals over a set period of time. In some cases, the period of time during which the composition is administered to the subject at one or more time intervals can be separated by a period of time during which the composition is not administered to the subject. In some embodiments, the relative duration of each period of time can depend on the subject’s response to the therapy or the severity of the disease or both, and / or can be determined based on the judgment of the treating physician.

[0232] In some embodiments, the antibodies can be administered parenterally. For example, the parenterally administered compositions can be administered topically, transmucosally, by subcutaneous, intradermal, intravenous, intraperitoneal, intratumoral, intramuscular, intraarticular, intraarterial, or infusion techniques.

[0233] In some embodiments, the antibodies (e.g., anti-KLK5 / KLK7 antibodies) are administered intravenously. In some embodiments, the antibodies (e.g., anti-KLK5 / KLK7 antibodies) are administered subcutaneously or topically.

[0234] For intravenous injection, water soluble antibodies can be administered by drip infusion, whereby a pharmaceutical preparation containing the antibody and a physiologically acceptable excipient is infused. Physiologically acceptable excipients can include, for example, 5% dextrose, 0.9% normal saline, Ringer's solution, or other suitable excipients. Other injectable compositions can contain various carriers such as vegetable oils, dimethylacetamide, dimethylformamide, ethyl lactate, ethyl carbonate, isopropyl myristate, ethyl alcohol, and polyhydric alcohols (glycerol, propylene glycol, liquid polyethylene glycol, and the like). In some instances, a formulation (e.g., a sterile formulation of a suitable soluble salt form of the antibody) can be dissolved and administered in a pharmaceutical excipient (e.g., water for injection, 0.9% normal saline, or a 5% glucose solution).

[0235] In one embodiment, the antibody is administered by site-specific or targeted local delivery techniques. Examples of site-specific or targeted local delivery techniques include various implantable, transdermal, or transmucosal reservoir sources or local delivery systems of the antibody.

[0236] Anti-KLK5 / KLK7 antibodies as described in the present disclosure, and therapeutic methods involving them, can be used in combination with other types of therapies directed to the target diseases or conditions disclosed herein. In this context, the antibody compositions and therapeutic agents can be administered simultaneously or sequentially. Such therapies can be administered simultaneously or sequentially (in any order) with the treatment according to the present disclosure.

[0237] Accordingly, aspects of the present disclosure relate to methods and compositions (e.g., anti-KLK5 / KLK7 antibodies). In some embodiments, the antibodies as described herein can be administered as a combination therapy (concomitantly or sequentially, e.g., over time). In some embodiments, the combination therapy comprises administration of one or more antibodies as described herein (e.g., anti-KLK5 / KLK7 antibodies) and at least one additional therapeutic agent (e.g., one, two, three, four, five, six, or seven therapeutic agents). In some embodiments, the one or more antibodies as described herein and the at least one additional therapeutic agent (e.g., one, two, three, four, five, six, or seven therapeutic agents) are administered together. In some embodiments, the one or more antibodies as described herein and the at least one additional therapeutic agent (e.g., one, two, three, four, five, six, or seven therapeutic agents) are administered separately.

[0238] In some embodiments, the additional therapeutic agent is an anti-inflammatory agent. In some embodiments, the anti-inflammatory agent is selected from, but not limited to, low dose antibiotics, steroids, corticosteroids, tacrolimus, anti-IL4R antibodies (e.g., dupilumab), anti-IL-13 antibodies, TNF inhibitors (e.g., anti-TNF), IL-12 / 23 inhibitors, IL-17 inhibitors, and IL-4 receptor inhibitors, doxycycline, methotrexate, prednisone, cyclosporine, mycophenolate mofetil, dupilumab, pexildizumab, etanercept, adalimumab, infliximab, golimumab, ustekinumab, secukinumab, ixekizumab, brolucizumab, abatacept, rilzabrutinib-asmn, risankizumab-rzaa, and guselkumab. In some embodiments, the anti-inflammatory agent is administered orally. In some embodiments, the anti-inflammatory agent is administered topically. In some embodiments, the anti-inflammatory agent is administered by injection (e.g., intravenously, subcutaneously, or intramuscularly).

[0239] In some embodiments, the therapeutic combination comprises one or more antibodies (e.g., anti-KLK5 / KLK7 antibodies) as described herein, delivered with one or more additional antibodies or fragments thereof (e.g., one, two, three, four, five, six, or seven antibodies). In some embodiments, the therapeutic combination comprising one or more antibodies as described herein and one or more additional antibodies or fragments thereof (e.g., one, two, three, four, five, six, or seven antibodies) are delivered separately. In some embodiments, the therapeutic combination comprising one or more antibodies as described herein and one or more additional antibodies or fragments thereof (e.g., one, two, three, four, five, six, or seven antibodies) are delivered together.

[0240] In some embodiments, the therapeutic combination comprising one or more antibodies as described herein and one or more additional antibodies or fragments thereof (e.g., one, two, three, four, five, six, or seven antibodies) is a multispecific antibody combination. In some embodiments, the multispecific antibody combination comprises a KLK5 / KLK7 antigen binding site, and one or more additional unique antigen binding sites from one or more additional antibodies. In some embodiments, the multispecific antibody comprises a direct fusion or linkage of different antigen-specific binding sites. In some embodiments, the additional antibody or fragment thereof is selected from, but not limited to, an anti-IL4R antibody, an anti-IL-13 antibody, an anti-TNF antibody, an anti-IL-12 / 23 antibody, an anti-IL-17 antibody, doxycycline, dupilumab, pexildizumab, etanercept, adalimumab, infliximab, golimumab, ustekinumab, secukinumab, ixekizumab, brolucizumab, abatacept, rilzabrutinib-asmn, risankizumab-rzaa, and / or guselkumab.

[0241] In some embodiments, in the treatment of rosacea, in one non-limiting example, the additional therapeutic agent can be selected from a topically applied steroid; an orally applied methotrexate; an orally applied cyclosporine; and / or an applied TNF inhibitor. In some embodiments, in the treatment of atopic dermatitis, in one non-limiting example, the additional therapeutic agent can be selected from a subcutaneously applied dupilumab, and / or a topically applied steroid.

[0242] Any of the anti-KLK5 / KLK7 antibodies disclosed herein can also be used to detect the presence of KLK5 and / or KLK7 in vitro or in vivo. The results obtained from such detection methods can be used for diagnostic purposes (e.g., diagnosing a disease associated with KLK5 and / or KLK7) or scientific research purposes (e.g., identifying new KLK5 secreting cell types, studying the regulation of biologically active and / or secreted KLK5 and / or KLK7). For assay uses (e.g., diagnostic uses), the anti-KLK5 / KLK7 antibodies as described herein can be conjugated to a detectable label (e.g., an imaging agent, e.g., a contrast agent) for detecting the presence of KLK5 and / or KLK7 in vivo or in vitro.

[0243] As used herein, “conjugated” or “attached” means that two entities are associated, preferably with sufficient affinity so as to achieve a therapeutic / diagnostic benefit of the association between the two entities. The association between the two entities can be direct, or achieved through a linker (e.g., a polymeric linker). Conjugation or attachment can include covalent or non-covalent bonding as well as other forms of association, such as entrapment, e.g., entrapment of one entity on or in another entity, or entrapment of either or both entities on or in a third entity (e.g., a micelle).

[0244] In other embodiments, the anti-KLK5 / KLK7 antibodies as described herein can be attached to a detectable label, which is a compound capable of directly or indirectly releasing a detectable signal, such that the aptamer can be detected, measured, and / or identified in vitro or in vivo. Examples of such “detectable labels” include, but are not limited to, fluorescent labels, chemiluminescent labels, colorimetric labels, enzymatic labels, radioisotopes, and affinity labels (e.g., biotin). Such labels can be conjugated to the aptamer directly or indirectly by conventional methods.

[0245] The reporter agent can also be a dye (e.g., a fluorophore), which can be used to detect diseases mediated by KLK5 and / or KLK7 expressing cells, respectively, in a tissue sample.

[0246] For in vitro diagnostic assays, an anti-KLK5 / KLK7 antibody can be contacted with a sample suspected of containing KLK5 and / or KLK7, such as a KLK5-expressing cell or soluble KLK5 in a disease microenvironment. The antibody and sample can be incubated under suitable conditions for a suitable time to allow the antibody to bind to KLK5 antigen. Such interactions can then be detected by conventional methods, such as ELISA, histological staining, or FACS. For in vivo diagnostic assays, an appropriate amount of an anti-KLK5 / KLK7 antibody conjugated to a label (e.g., an imaging agent or a contrast agent) can be administered to a subject in need of examination. The presence of the labeled antibody can be detected based on a signal released from the label by conventional methods.

[0247] For scientific research assays, an anti-KLK5 / KLK7 antibody can be used to study the biological activity of KLK5 and / or KLK7, detect the presence of intracellular or extracellular KLK5 and / or KLK7, and or modulate the effects of KLK5. For example, an appropriate amount of an anti-KLK5 / KLK7 can be contacted with a sample suspected of producing KLK5 and / or KLK7 (e.g., a new cell type that has not been previously identified as a KLK5 and / or KLK7-producing cell). The cells are permeabilized prior to contacting the anti-KLK5 / KLK7 antibody. The antibody and sample can be incubated under suitable conditions for a suitable time to allow the antibody to bind to KLK5 antigen. Such interactions can then be detected by conventional methods, such as ELISA, histological staining, or FACS.

[0248] VI. Kits for therapeutic and diagnostic applications The present disclosure also provides kits for therapeutic or diagnostic applications as disclosed herein. Such kits can include one or more containers comprising an antibody, such as any of those described herein.

[0249] In some embodiments, the kits can include instructions for use in accordance with any of the methods described herein. The included instructions can include a description of administering the antibody to treat, delay onset, or ameliorate a target disease, such as those described herein. The kits can also include a description of selecting an individual suitable for treatment based on identifying whether the individual has a target disease. In other embodiments, the instructions include a description of administering the antibody to an individual at risk for a target disease.

[0250] Instructions relating to the use of the antibody generally include information as to dosage, dosing schedule, and route of administration for the intended treatment. The containers can be unit doses, bulk packages (e.g., multi-dose packages), or sub-unit doses. Instructions provided in the kits of the application are generally written instructions, but machine-readable instructions (e.g., instructions carried on magnetic or optical storage disks) are also acceptable.

[0251] The label or package insert indicates that the composition is used for treating, delaying the onset of, and / or ameliorating a disease or condition. Instructions for practicing any of the methods described herein can be provided.

[0252] The kits of the application are in suitable packaging. Suitable packaging includes, but is not limited to, vials, bottles, jars, flexible packaging (e.g., sealed Mylar or plastic bags), and the like.

[0253] Packaging for use in combination with a particular device, such as an infusion device, e.g., a mini-pump, is also contemplated. The kit can have a sterile access port (e.g., the container can be an intravenous solution bag, or a vial having a stopper pierceable by a hypodermic injection needle). The container can also have a sterile access port (e.g., the container can be an intravenous solution bag, or a vial having a stopper pierceable by a hypodermic injection needle). At least one active agent in the composition is an antibody, such as those described herein.

[0254] The kits can optionally provide additional components, such as buffers and explanatory information. Typically, the kit includes a container and a label or package insert on or associated with the container. In some embodiments, the application provides articles of manufacture which include the contents of the above-described kits.

[0255] Also provided herein are kits for detecting a target protein (e.g., KLK5 and / or KLK7) in a sample. Such kits can include any of the antibodies described herein. In some cases, the antibodies can be conjugated to a detectable label, such as those described herein. As used herein, “conjugated” or “attached” means that two entities are associated, preferably with sufficient affinity so as to achieve a therapeutic / diagnostic benefit of the association between the two entities. The association between the two entities can be direct, or achieved through a linker (e.g., a polymeric linker). Conjugation or attachment can include covalent or non-covalent bonding as well as other forms of association, such as entrapment, e.g., entrapment of one entity on or in another entity, or entrapment of either or both entities on or in a third entity (e.g., a micelle).

[0256] Alternatively or additionally, the kit can include a secondary antibody capable of binding to the antibodies described herein. The kit can also include instructions for using the antibodies to detect target proteins (e.g., KLK5 and / or KLK7).

[0257] EMBODIMENTS Example 1: Generation and selection of anti-KLK5 and anti-KLK7 antibodies (i) Affinity KLK5 and KLK7 are pathological mediators observed in the context of aberrant protease activation Figure 1 Antibody binding kinetics experiments were performed by Biacore to screen for anti-KLK5 / KLK7 antibodies with high affinity for the respective target for inhibiting KLK5 / 7 activity and subsequently ameliorating or eliminating symptoms associated with aberrant protease activation. All screening assays were performed at 25 °C. The running buffer used was 20 mM HEPES, 300 mM NaCl, 0.01% Tween-20, pH 7.5. Antibodies (1 ug / mL) were captured by flowing at 10 uL / min for 30 s on a Series S Protein A sensor chip (Cytiva) on Fc2-4. Kinetic measurements were performed in single-cycle kinetics mode using a series of 4-5 concentrations with the highest concentration being 25-100 nM and 4-fold serial dilutions. Contact times were typically 300 s and dissociation times ranged between 1800-3600 s. A flow rate of 30 uL / min was typically used. The sensor chip was regenerated with 10 mM Glycine, pH 1.5 with a contact time of 30 s, flowing at 50 ul / mL. The results show that the dual inhibitor antibodies described in Table la show binding specificity to both KLK5 and KLK7. Table 2 below provides an overview of the binding kinetics of the anti-KLK5 antibodies.

[0258] Table 2 Anti-KLK5 / KLK7 antibody affinity and potency

[0259] (ii) Protease inhibition assay To assess the ability of the antibodies described in Table la to inhibit KLK5 and / or KLK7 protease activity, respectively, 1.5 nM human or mouse KLK5 or 0.5 nM human or mouse KLK7 was prepared in assay buffer (0.1 M NaH2P04 pH 7.5 for KLK5; and 50 mM Tris, 150 mM NaCl, pH 7.5 for KLK7). Antibodies were added diluted in PBS to the test concentration. Substrate was then added (50 uM BOC-Val-Pro-Arg-AMC for KLK5; and 30 uM KHLF-AMC for KLK7). The fluorescence signal was then measured every min for 30 min at room temperature.

[0260] The results show that the anti-KLK5 / KLK7 antibodies are able to inhibit KLK5 and KLK7 protease activity, as described in Table 2.

[0261] Example 2. Anti-KLK5 / KLK7 antibodies specifically bind to the active form of human KLK5.

[0262] KLK5 / 7-Dual-Ab4 or Comparative Antibody #1 (a bispecific antibody with a first arm that binds KLK5 and a second arm that binds KLK7) was captured on a Series S Protein A sensor chip (Cytiva) at a concentration of 1 ug / ml. The active or precursor form of huKLK5 or huKLK7 was flowed through at 50 uL / min at the following concentrations (0.156 nM, 0.625 nM, 2.5 nM, 10 nM, or 40 nM) with a contact time of 5 min and a dissociation time of 1 hour. The sensor chip was regenerated with 10 mM glycine, pH 1.5, with a contact time of 30 seconds, flowing at 50 uL / min. KLK5 / 7-Dual-Ab4 was found to bind to active huKLK5 ( Figure 2A ) and active huKLK7 ( Figure 2B ), but not to the precursor form of either. Comparative Antibody #1 bound to the precursor and active forms of KLK5 ( Figure 2A ) and KLK7 ( Figure 2B ) with similar potency. These data indicate that KLK5 / 7-Dual-Ab4 binds to the active site of huKLK5 and huKLK7, as the proper structure of the active site is not present in the precursor form of the proteases. Comparative Antibody #1 binds to an allosteric site that is present in both the active and precursor forms, but does not bind to the active site of the enzymes.

[0263] Example 3. Anti-KLK5 / KLK7 antibodies do not undergo cleavage when binding to their target.

[0264] One issue to consider when developing antibodies that bind proteases is that proteolytic activity of the antibody's target can result in cleavage of the antibody after binding, thereby rendering the antibody inactive. To determine whether KLK5 or KLK7 can cleave anti-KLK5 / KLK7 antibodies, stoichiometric amounts of KLK and mAb were incubated at 37°C for 18 h. 40 pmol of KLK5 or KLK7 was used per reaction and mixed with 20 pmol of each test antibody to a final volume of 20 uL. After overnight incubation, each reaction was analyzed by reducing SDS-PAGE gel.

[0265] Cleavage of the antibody heavy chain (HC) at the complementarity determining region 3 (CDR3) produced 12 kDa and 38 kDa fragments. SDS-PAGE analysis showed that while a control anti-KLK5 antibody that binds to the active site of KLK5 and is cleaved by KLK5 (referred to as "control anti-KLK5 antibody" in this example) produced 12 kDa and 38 kDa fragments upon co-incubation with active KLK5, incubation of the anti-KLK5 / 7 antibodies with active KLK5 did not produce the fragments, indicating that they were not cleaved by KLK5 Figure 3A ). SDS-PAGE analysis of co-incubation of active KLK7 with the anti-KLK5 / 7 antibodies showed that neither of the anti-KLK5 / 7 antibodies were cleaved by KLK7 Figure 3B ).

[0266] Recent crystal structures of the control anti-KLK5-Ab1 Fab and KLK5 / 7-Dual-Ab1 Fab show that the HC CDR3 of these anti-KLK5 / 7 antibodies insert into the respective KLK active site Figure 11 ). The control anti-KLK5-Ab1 HC CDR3 inserts in the forward (N→C) direction, while the KLK5 / 7-Dual-Ab1 CDR3 inserts in the reverse (C→N) direction Figure 12 ). This reverse insertion can be the reason that the anti-KLK5 / 7 antibodies are not cleaved by their targets.

[0267] Example 4. Anti-KLK5 / KLK7 antibodies are specific for KLK5 and KLK7 and do not bind other KLK family members.

[0268] Members of the kallikrein family and the kallikrein-like family are quite similar at the gene and protein level. Therefore, developing antibodies that specifically bind to one or more selected members of the KLK family presents a challenge. To determine whether the anti-KLK5 / 7 antibodies bind to other KLK family members or related proteases, 10 ug / mL of the anti-KLK5 / 7 antibodies were tested for inhibition of plasma KLK, KLK1, KLK2, KLK4, KLK6, KLK12, KLK14, trypsin, chymotrypsin, and urokinase using substrate cleavage as readout.

[0269] Substrates for protease assays were as follows: plasma KLK, KLK1, KLK2 (Pro-Phe-Arg-AMC); KLK4, KLK12, KLK13, and KLK14 (Boc-VPR-AMC); KLK6 (Boc-QAR-AMC); trypsin (MCA-RPKPVG-NVAL(DNP)-NH2); chymotrypsin (Suc-AAPF-AMC); and urokinase (Z-Gly-Gly-Arg-AMC).

[0270] Enzymes were prepared in assay buffer at the following concentrations: plasma KLK (10 nM), KLK1 (10 nM), KLK2 (10 nM), KLK4 (5 nM), KLK6 (5 nM), KLK12 (2.5 nM), KLK14 (2.5 nM), trypsin (0.25 nM), chymotrypsin (17 nM), and urokinase (20 nM).

[0271] Assay buffers were as follows: KLK1 and 2 (50 mM Tris, 150 mM NaCl, 10 mM CaCl2, 0.05% (w / v) Brij-35, pH 7.5); KLK3 (50 mM Tris, 1 M NaCl, pH 8.0); KLK4 (50 mM Tris, 1 M NaCl, pH 8.0); KLK6 (50 mM Tris, 1 M sodium citrate, pH 7.5); KLK12 and 13 (100 mM Tris, 150 mM NaCl, 10 mM CaCl2, 0.05% (w / v) Brij-35, pH 7.5); KLK14 (50 mM Tris, 150 mM NaCl, 0.05% (w / v) Brij-35, pH 8.0), Trypsin (lx HBS + 1 mM CaCl2+ 0.05% Tween 20, pH 7.4); Chymotrypsin (25 mM Tris, 0.5 mM CaCl2, pH 8.0); Urokinase (50 mM Tris + 0.01% Tween 20 pH 8.5; and Plasma kallikrein KLKb1 (50 mM Tris, 250 mM NaCl, pH 7.5).

[0272] Antibodies were diluted in PBS to the test concentration before substrate was added. Fluorescence was then measured every minute for 30 minutes at room temperature.

[0273] The following KLK family members were screened for affinity by Biacore: hKLK3, hKLK8, hKLK9, hKLK10, hKLK11, hKLK12, hKLK13, and hKLK15. All screening assays were performed at 25 °C. The running buffer used was 20 mM HEPES, 300 mM NaCl, 0.01% Tween-20, pH 7.5. Antibodies (1 pg / mL) were captured on Fc2-4 of a Series S Protein A sensor chip (Cytiva) by flowing at 10 uL / min for 30 seconds. Kinetic measurements were made using 25 and 100 nM analyte in single-cycle kinetic mode. Contact time was 120 seconds and dissociation was 300 seconds. Flow rate was typically 30 uL / min. The Protein A sensor chip was regenerated with 10 mM glycine, pH 1.5, with a contact time of 30 seconds, flowing at 50 uL / min.

[0274] KLK6, KLK14 and KLK13 were followed up because they showed some binding / inhibition activity with the anti-KLK5 / 7 antibody. All IC50 curves were at least 100-fold higher than the activity against KLK5 or KLK7, demonstrating that the anti-KLK5 / 7 antibody has high specificity for KLK5 and KLK7, but no specificity for other KLK family members or related proteases. Figure 4 ).

[0275] Example 5. Anti-KLK5 / KLK7 antibody binds to the active site of KLK5 and KLK7.

[0276] The inhibitory capacity of an antibody is related to the ability of the antibody to compete with its target substrate for binding to the active site of its target. The binding activity of the anti-KLK5 / 7 antibody to KLK5 and KLK7 was tested in the presence of small molecule inhibitors known to bind to the active site of KLK5 and KLK7.

[0277] Binning was performed by immobilizing the dual specific KLK5 / 7 antibody (2 ug / mL) on a Protein A chip surface. The flow rate was 30 uL / min and the contact time was 30 seconds. huKLK5 ( Figure 5A ) or huKLK7 ( Figure 5B ) were then pre-incubated with inhibitors and then applied to the chip. 20 nM KLK5 or KLK7 were incubated with 0.35 mg / mL of the small molecule inhibitor PMSF (2 mM final concentration, made from a 100x stock), 1 mg / mL of the peptide inhibitor leupeptin (1 mM final concentration) or 25 ug / mL SPINK5. The flow rate was 30 uL / min, the contact time was 300 seconds and the dissociation time was 120 seconds.

[0278] When KLK5 or KLK7 were pre-incubated with various inhibitors that bind to the active site, almost no binding of the anti-KLK5 / 7 antibody to KLK5 ( Figure 5A ) or KLK7 ( Figure 5B ) was observed, indicating that the anti-KLK5 / 7 antibody binds to the active site of KLK5 and KLK7, as they are unable to bind when the active site has already been bound by another molecule.

[0279] Example 6. Anti-KLK5 / KLK7 antibody binds to the same epitope on KLK5 and KLK7 as SPINK5.

[0280] In this example, the binding of anti-KLK5 / 7 antibodies to the active sites of KLK5 and KLK7 was further analyzed. The binning was performed in a sandwich format. In the first experiment, anti-KLK5 / 7 antibody KLK5 / 7-Dual-Ab4 was covalently coupled to the CM5 chip surface, after which huKLK5 or huKLK7 was flowed over. Next, the second anti-KLK5 / 7 antibody was flowed over the chip surface at a concentration of 10 μg / mL (Figure 6, left panel). In the second experiment, recombinant human SPINK5 was covalently coupled to the CM5 chip surface, after which huKLK5 or huKLK7 was flowed over. Next, the anti-KLK5 / 7 antibody was flowed over the surface (Figure 6, right panel). The running buffer was 20 mM HEPES, 300 mM NaCl, 0.05% Tween-20, pH 7.5. Biacore was performed with a flow rate of 50 uL / min. The contact time for each antibody or protein was 60 seconds and the dissociation time was 60 seconds.

[0281] The data indicate that anti-KLK5 / 7 antibody KLK5 / 7-Dual-Ab4 competes with human SPINK5 for binding to huKLK5 and huKLK7. Since SPINK5 binds to the active sites of both KLK5 and KLK7, this further indicates that anti-KLK5 / 7 antibody KLK5 / 7-Dual-Ab4 also binds to the active sites of KLK5 and KLK7, and that SPINK5 and the inhibitory antibody cannot bind to KLK5 or KLK7 at the same time (Figure 6).

[0282] Example 7. Anti-KLK5 / 7 dual antibody treatment reduces barrier defects in MC903 atopic dermatitis mouse model.

[0283] On days 1-8, topical application of 40 μL MC903 (Sigma, calcipotriol hydrate, 2 ng / 10 μL in ethanol) in the shaved dorsal area of C57 / B6 mice was performed once daily to induce murine atopic dermatitis.

[0284] From day -6, mice were pre-treated with MC903 vehicle control, control IgG (non-specific antibody), KLK5 / 7-Dual-Ab4 or comparative antibody #2 (anti-mouse IL-4R antibody) at 30 mg / kg IP, 3 times per week. Alternatively, control IgG was administered at 30 mg / kg Q3D SC from day -6; KLK5 / 7-Dual-Ab4 was administered at 3 mg / kg Q3D, Q7D or Q14D SC from day -6; and comparative antibody #2 was injected at 30 mg / kg Q3D IP from day -6.

[0285] On day 9, a piece of dorsal skin was excised from each mouse and preserved in 10% neutral buffered formalin. The skin was trimmed, embedded in paraffin, sectioned, and stained with hematoxylin and eosin (H&E). The stratum corneum thickness was measured every 800 pm according to the grid set in Qupath, and then averaged for each animal Figures 7A-7B

[0286] Treatment with KLK5 / 7-Dual-Ab4 reduced hyperkeratosis and desquamation (as measured by stratum corneum thickness) to a greater extent than comparative antibody #2. The lowest effective subcutaneous dose was 3 mg / kg every 7 days.

[0287] Example 8. Treatment with anti-KLK5 / 7 dual antibodies reduces disease in the Nc / Nga atopic dermatitis mouse model.

[0288] Murine atopic dermatitis was induced in 8- to 10-week-old Nc / Nga mice, a strain with intrinsic barrier defects, by topical application of house dust mite (HDM) allergen. Animals received Biostir-AD ointment (120 mg / mouse) on both ears and dorsal skin areas, including the neck, containing HDM allergen derived from house dust mites (Dermatophagoides farina) at a total of 480 mg per mouse, twice a week for two weeks. Sodium dodecyl sulfate (SDS) treatment was applied 2 hours prior to the second treatment with Biostir-AD (day 4).

[0289] Mice were pre-treated with control IgG, KLK5 / 7-Dual-Ab4, or comparative antibody #2 at 30 mg / kg intraperitoneally three times a week for a total of 9 doses starting on day -6. Tacrolimus, a steroidal compound that treats atopic dermatitis but cannot be used long term due to severe withdrawal effects, was topically applied once a day for 21 days starting on day -6, and 1 hour after each Biostir-AD application.

[0290] To assess treatment efficacy, mice were scored for scratching ( Figure 8C ), macroscopic skin lesions, and ear thickness ( Figure 8B ​Evaluation was conducted. Mice were isolated and acclimatized for 30 minutes before the assessment to assess scratching. Scratching behavior was recorded visually: 60 minutes on day 0 and 30 minutes after drug administration on day 15. Ear thickness was measured using a Dyer micrometer. The severity of skin lesions was assessed using four parameters: erythema, hemorrhage, edema, epidermal abscess / erosion, and desquamation / dryness. Skin lesion parameters were assessed on the ears, neck, and back. The total clinical skin severity score was defined as the sum of individual scores (0: none; 1: mild; 2: moderate; 3: severe). Figure 8A ).

[0291] 1 cm was removed from each mouse 2 A piece of back skin was collected and preserved in 10% neutral buffered formalin. The skin was trimmed, embedded in paraffin, sectioned, and stained with H&E. The epidermal area was measured using QuPath software. Figure 8E Based on a grid in QuPath, keratin thickness was measured every 800 μm, and then averaged to generate values ​​for each animal. Samples were examined microscopically by a veterinary pathologist, who scored inflammation, necrosis, hyperplasia, and hyperkeratosis. Figure 8D The presence of HDM-specific IgE antibodies in serum was also analyzed. Figure 8F ).

[0292] Treatment with KLK5 / 7-Dual-Ab4 reduced ear thickness, clinical skin lesions, and pruritus to a similar degree to treatment with comparative antibody #2 or tacrolimus. Histological analysis showed that treatment with KLK5 / 7-Dual-Ab4 resulted in a greater reduction in hyperkeratosis and desquamation (as measured by stratum corneum thickness) compared to treatment with tacrolimus or comparative antibody #2, indicating that KLK5 / 7-Dual-Ab4 is a potent inhibitor of atopic dermatitis.

[0293] The above experiment was repeated using KLK5 / 7-Dual-Ab2, and it was found that it reduced itching compared to control IgG. Figure 8I ), stratum corneum thickness ( Figure 8H ) and histological score ( Figure 8G ), and is proportionate to the effect of competing antibody #2 ( Figure 8B ).

[0294] Example 9. Anti-KLK5 / 7 antibody treatment reduced disease in a mouse model of squamous tail atopic dermatitis.

[0295] Murine atopic dermatitis was induced in male scurfy mice (scurfy mice have intrinsic barrier defects due to loss of function of filaggrin, a filament-associated protein, and the transmembrane protein mattrin) by topical application of HDM allergen. Prior to HDM allergen administration, all mice were shaved on the nape of the neck. The no disease control group received petrolatum topically applied 3 times per week for 6 weeks on the shaved area of the nape of the neck and on both ears. All other treatment groups received Biostir cream topically applied 3 times per week for 6 weeks on the shaved area of the nape of the neck and on both ears. Each application of Biostir used approximately 100 mg of cream per mouse. For both the control and test groups, control IgG or KLK5 / 7-Dual-Ab4 was injected intraperitoneally at 30 mg / kg 3 times per week starting on day 0.

[0296] A 1 cm 2 piece of dorsal skin was excised from each mouse and preserved in 10% neutral buffered formalin. The skin was trimmed, embedded in paraffin, sectioned, and stained with H&E. Epidermal area Figure 9A ) was measured by Qupath software analysis. Samples were microscopically examined by a veterinary pathologist and scored for parakeratosis Figure 9B ) and spongiotic edema Figure 9C ). Cytokine expression was measured from lysed ear tissue.

[0297] Histological analysis showed that treatment of atopic dermatitis with KLK5 / 7-Dual-Ab4 reduced epidermal thickness, parakeratosis, and spongiotic edema. Cytokine analysis showed a reduction in the allergic-related cytokine interleukin (IL)-4 Figure 9D ) and the inflammatory cytokine tumor necrosis factor (TNFa) Figure 9E ). These data indicate that KLK5 / 7 inhibition can reduce inflammation and epidermal effects in a mouse model of atopic dermatitis.

[0298] Example 10. Treatment with anti-KLK5 / 7 antibody reduced hyperkeratosis in a human epidermis equivalent air-liquid interface culture induced for disease.

[0299] EpiDerm FT cultures with fibroblast-seeded dermis layer and primary human keratinocytes overlaid on top, differentiated to form epidermis in transwell, were incubated overnight in assay buffer. On day 0, 80 mM Mc903 treatment was applied on top of the transwell to induce damage and hyperkeratosis. Cultures were placed in presence of control IgG Figure 10B ; no MC903 control, Figure 10A ), KLK5 / 7-Dual-Ab4 Figure 10CComparative antibody #1 ( Figure 10D Or compare antibody #3 (an anti-KLK5 monospecific antibody that does not bind to the active site of KLK5) Figure 10E In the culture medium, 10 μg / mL was added to the top and bottom of the wells, for a total of 20 μg / mL. Antibody treatment began on day 0 and lasted for 5 days. Each condition was tested in 6 separate cell culture inserts. Histogel was added to the top of the cell inserts, and the inserts were fixed in formalin and embedded in paraffin for sectioning and H&E staining. Structural thickness was measured at 100 μM intervals using Qupath image analysis software.

[0300] stratum corneum thickness ( Figure 10F The results indicated that MC903 treatment induced hyperkeratosis in human EpiDermFT cultures. Treatment with KLK5 / 7-Dual-Ab4 or comparative antibody #1 (bispecific) reduced this hyperkeratosis because they inhibited both KLK5 and KLK7. Treatment with comparative antibody #3 (which inhibits only KLK5) showed some effect on hyperkeratosis, but to a lesser extent. The data suggest that inhibition of both KLK5 and KLK7 is necessary to completely salvage epidermal dysfunction.

[0301] Example 11. Crystal structure of KLK5 / 7-Dual-Ab1.

[0302] The crystallization conditions for the complex between KLK5 / 7-Dual-Ab1 and human KLK7 (StoA variant) were 25% (w / v) polyethylene glycol, 0.1 M PCTP buffer (pH 8), 20°C, and a protein-to-storage ratio of 1:1. The structure was fully refined using autoBUSTER and validated using MolProbity. KLK5 / 7-Dual-Ab1 and its derivatives are characterized by a longer CDR3 ring than the average heavy chain (indicated by the arrow), which occupies the active site of the antigen. Figure 11 The inhibition of KLK enzyme catalytic activity was achieved through competitive inhibition. The endogenous KLK7 substrate occupied the S4-S1 active site from the N-terminus to the C-terminus. In contrast, KLK5 / 7-Dual-Ab1 Fab occupied the active site from the C-terminus to the N-terminus (indicated by the arrow). Figure 12 This allows Fab to be protected from protein hydrolysis and degradation through reverse occupation. Tyrosine residues occupy the critical S1 binding pocket and are anchored by the polar interaction between the hydroxyl group of the Tyr residue and the side chain of the Asn residue found on the back of the pocket.

[0303] Other implementation plans All features disclosed in this specification may be combined in any combination. Where the disclosure of features in this specification refers to those as "comprising" one or more stated features, it will be understood that the subject matter includes at least one of those features unless the context clearly indicates otherwise. Furthermore, structure disclosed in this specification may be combined in any combination, and any implication of structure implies the possibility of inversion (i.e., structure can be combined in any order).

[0304] From the foregoing description, one skilled in the art can easily ascertain the essential characteristics of this application, and without departing from the spirit and scope thereof, can make various changes and modifications of the application to adapt it to various uses and conditions. Thus, other embodiments are also within the scope of the following claims: Equivalents In the claims, unless otherwise indicated the use of an "or" as limiting means a non-exclusive "or", nor should such a determination be taken with reference to any statement of elements or limitations recited using the term "comprising". Likewise, as used herein, the article "a" is intended to include "one or more" unless otherwise indicated. As used herein, the article "the" is intended to include "the one or more" or "at least one" unless otherwise indicated. In addition, to recite an element or a limitation as "means" for performing a specified function, or "step" for performing a specified function, if any is implied by context, means that the function is performed "resulting from" and "attributable to" the means or step, whether or not there is a specific listing of

[0305] Further, the application encompasses all variations, combinations, and permutations of one or more of the limitations, elements, clauses, and descriptive terms recited in one or more of the listed claims. For example, any claim that is dependent on another claim can be modified to include one or more limitations, elements, clauses, and descriptive terms from any other claim that is dependent on the same base claim. Where the deposition of features is presented in a list, e.g., in a Markush Group, each individual feature can be replaced by alternative features that serve the same, equivalent, or similar purpose, and the application is intended to cover all such possible variations. It is therefore intended that the application be considered as encompassing all such possible variations, including any variation that would be apparent to one of ordinary skill in the art in light of the disclosure herein. It is intended that the scope of the application embraces all such alterations and modifications of the technology already described and falling within the ambit of the following claims. Original phrasing Specifically listed.

[0306] As used herein in the specification and claims, the phrase “and / or” should be understood to mean “either or both” of the entities so conjoined, i.e., one or the other but not both operating simultaneously, unless otherwise indicated by context. As used herein in the specification and claims, the phrase “and / or,” alongside listings of items, e.g., in a list of items prefaced by “comprising” or “including,” indicates that at least one, and optionally more than one, of the listed items can be present in the composition, process, method, or machine. If only one such item is present, those other items can optionally be present. If more than one such item is present, those other items can also be present, whether or not paired with the one present. The term “comprising” is used herein to mean that the compositions and methods include the recited elements, but not excluding others. “Consisting essentially of’ when used herein in reference to a composition, process, method, or machine, refers to compositions, processes, methods, or machines that do not exclude an additional element of matter unless the additional element of matter materially affects the basic and novel characteristics of the compositions, processes, methods, or machines. “Consisting of’ refers to compositions, processes, methods, or machines that exclude additional elements of matter.

[0307] As used herein in the specification and claims, “or” should be understood to have the same meaning as “and / or” as defined above. For example, when separating items in a list, “or” or “and / or” shall be interpreted as being inclusive, i.e., the inclusion of at least one, but also including more than one, of a number or list of elements, and, optionally, additional unlisted items. Only terms clearly

[0308] As used herein in the specification and claims, the phrase "at least one," in reference to a list of one or more elements, should be understood to mean at least one element selected from any one, or more of the elements in the list of elements, but not necessarily including at least one of each and every element specifically listed within the list of elements and not excluding any combinations of elements in the list of elements. This definition also allows that the

[0309] It should also be understood that, regardless of whether the use of the term "at least one" in the description and / or claims of the present disclosure is explicitly recited, the present disclosure should be understood to be in the nature of a generic disclosure of a generic embodiment followed by a disclosure of a specific embodiment (or specific embodiments) followed by a disclosure of a further specific embodiment (or further specific embodiments). Thus, the use of the term "at least one" in the description and / or claims should, independently, be understood to refer to the specific embodiment or specific embodiments before the use of the term "at least one" and the use of the term "at least one" after the use of the term "at least one" should, independently, be understood to refer to the further specific embodiment or further specific embodiments.

[0310] In the claims, as well as in the specification above, all transitional phrases such as "comprising," "including," "carrying," "having," "containing," "involving," "holding," "composed of," and the like are to be understood to be open-ended, i.e., to mean including but not limited to. Only the transitional phrases "consisting of" and "consisting essentially of" shall be closed or semi-closed transitional phrases, respectively, as set forth in the United States Patent Office Manual of Patent Examining Procedures, Section 2111.03. It should be understood that embodiments described in the specification using open-ended transitional phrases (e.g., "comprising") are also contemplated as embodiments "consisting of" and "consisting essentially of" the features recited using the open-ended transitional phrases. For example, if the application describes a "composition comprising A and B," then the application also contemplates alternative embodiments "a composition consisting of A and B" and "a composition consisting essentially of A and B."

[0311] When reciting ranges of values, the endpoints are included in the range. Further, in the various embodiments of the application, the values stated as ranges can assume any specific value or sub-range within the stated range, unless otherwise explicitly stated in the context, to the tenth of the lower limit of the range, unless otherwise explicitly stated in the context, and any combination of ranges or specific values thereof unless otherwise explicitly stated in the context.

[0312] This application relates to a number of issued patents, published patent applications, journal articles, and other publications, all of which are incorporated herein by reference. If any incorporated reference conflicts with the present specification, the present specification shall control. In addition, any specific embodiment of the application within the prior art can be explicitly excluded from any one or more of the claims. Because such embodiments are deemed to be known to those of ordinary skill in the art, even if not expressly set forth in the present description, they can be excluded even if not expressly disclaimed. Any particular embodiment of the application can be excluded from any claim for any reason, whether or not related to the existence of prior art.

[0313] Those of ordinary skill in the art will recognize or be able to ascertain using no more than routine experimentation many equivalents of the specific embodiments described herein. The scope of the embodiments of the application described herein is not intended to be limited to the above description but is only indicated by the appended claims.

[0314] The recitation of a list of chemical groups in any definition of a variable herein includes the definition of that variable as any single group or combination of listed groups. The recitation of one embodiment of a variable herein includes that embodiment as any single embodiment or in combination with any other embodiment or portion thereof. The recitation of one embodiment herein includes that embodiment as any single embodiment or in combination with any other embodiment or portion thereof.

Claims

1. A dual inhibitor antibody that specifically binds to KLK5 and KLK7, said dual inhibitor antibody comprising: (a) HC CDR1, HC CDR2 and HCCDR3 having the heavy chain variable domain of the amino acid sequence of SEQ ID NO: 7, and LC CDR1, LC CDR2 and LCCDR3 having the light chain variable domain of the amino acid sequence of SEQ ID NO: 8; (b) HC CDR1, HC CDR2 and HCCDR3 having the amino acid sequence of SEQ ID NO: 13, and LC CDR1, LC CDR2 and LCCDR3 having the amino acid sequence of SEQ ID NO: 14; (c) HC CDR1, HC CDR2, and HCCDR3 having the heavy chain variable domain of the amino acid sequence of SEQ ID NO: 17, and LC CDR1, LC CDR2, and LCCDR3 having the light chain variable domain of the amino acid sequence of SEQ ID NO: 14; or (d) HC CDR1, HC CDR2 and HCCDR3 having the amino acid sequence of SEQ ID NO: 21, and LC CDR1, LC CDR2 and LCCDR3 having the amino acid sequence of SEQ ID NO:

14.

2. The dual inhibitor antibody of claim 1, wherein the dual inhibitor antibody comprises: (a) HC CDR1 having the amino acid sequence of SEQ ID NO: 1; HC CDR2 having the amino acid sequence of SEQ ID NO: 2; HC CDR3 having the amino acid sequence of SEQ ID NO: 3; LC CDR1 having the amino acid sequence of SEQ ID NO: 4; LC CDR2 having the amino acid sequence of SEQ ID NO: 5; and LC CDR3 having the amino acid sequence of SEQ ID NO: 6; (b) HC CDR1 having the amino acid sequence of SEQ ID NO: 9; HC CDR2 having the amino acid sequence of SEQ ID NO: 10; HC CDR3 having the amino acid sequence of SEQ ID NO: 11; LC CDR1 having the amino acid sequence of SEQ ID NO: 4; LC CDR2 having the amino acid sequence of SEQ ID NO: 5; and LC CDR3 having the amino acid sequence of SEQ ID NO: 12; (c) HC CDR1 having the amino acid sequence of SEQ ID NO: 9; HC CDR2 having the amino acid sequence of SEQ ID NO: 15; HC CDR3 having the amino acid sequence of SEQ ID NO: 16; LC CDR1 having the amino acid sequence of SEQ ID NO: 4; LC CDR2 having the amino acid sequence of SEQ ID NO: 5; and LC CDR3 having the amino acid sequence of SEQ ID NO: 12; or (d) HC CDR1 having the amino acid sequence of SEQ ID NO: 18; HC CDR2 having the amino acid sequence of SEQ ID NO: 19; HC CDR3 having the amino acid sequence of SEQ ID NO: 20; LC CDR1 having the amino acid sequence of SEQ ID NO: 4; LC CDR2 having the amino acid sequence of SEQ ID NO: 5; and LC CDR3 having the amino acid sequence of SEQ ID NO:

12.

3. The dual inhibitor antibody of claim 1 or 2, wherein the antibody comprises: (a) VH, which contains the amino acid sequence of SEQ ID NO: 7; and VL, which contains the amino acid sequence of SEQ ID NO: 8; (b) VH, which contains the amino acid sequence of SEQ ID NO: 13; and VL, which contains the amino acid sequence of SEQ ID NO: 14; (c) VH, comprising the amino acid sequence of SEQ ID NO: 17; and VL, comprising the amino acid sequence of SEQ ID NO: 14; or (d) VH, which contains the amino acid sequence of SEQ ID NO: 21; and VL, which contains the amino acid sequence of SEQ ID NO:

14.

4. A dual inhibitor antibody comprising HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2 and / or LC CDR3 of any dual inhibitor antibody listed in Tables 1a and 1b.

5. A dual inhibitor antibody comprising the VH and / or VL of any dual inhibitor antibody listed in Tables 1a and 1b.

6. The dual inhibitor antibody according to any one of claims 1-5, wherein the dual inhibitor antibody binds to the active site of KLK5 and the active site of KLK7.

7. The dual inhibitor antibody according to any one of claims 1-6, wherein the dual inhibitor antibody competitively binds to the KLK5 active site and the KLK7 active site with SPINK 5 and / or leucopeptide.

8. The dual inhibitor antibody according to any one of claims 1-7, wherein the dual inhibitor antibody binds to the active form of KLK5 and the active form of KLK7, but not to the inactive form of KLK5 or the inactive form of KLK7.

9. The dual inhibitor antibody according to any one of claims 1-8, wherein the dual inhibitor antibody inhibits the protease activity of KLK5 and KLK7.

10. The dual inhibitor antibody according to any one of claims 1-9, wherein the antibody is not cleaved by KLK5 or KLK7 in the heavy chain when it binds to KLK5 or KLK7.

11. The dual inhibitor antibody according to any one of claims 1-10, wherein the antibody is not a bispecific antigen-binding molecule, wherein KLK5 binding is conferred by one binding site within the antibody, and KLK7 binding is conferred by a different binding site.

12. The dual inhibitor antibody according to any one of claims 1-11, wherein the antibody is a multispecific antigen-binding molecule, and the antigen-binding molecule further comprises an antigen-binding domain that binds to antigens other than KLK5 or KLK7.

13. A composition comprising a dual inhibitor antibody as described in any one of claims 1-12 and an acceptable vector.

14. A nucleic acid encoding a dual inhibitor antibody as described in any one of claims 1-12.

15. A method for treating skin barrier defects, the method comprising administering to a subject an effective amount of a dual inhibitor antibody as described in any one of claims 1-2, or a composition as described in claim 13.

16. The method of claim 15, wherein the skin barrier defect is associated with Netherton syndrome, atopic dermatitis, eosinophilic esophagitis, nodular prurigo, chronic pruritus of unknown cause (CPUO), dry skin, asthma (especially KLK5), ichthyosis vulgaris, or pruritus or chronic pruritus.

17. The dual inhibitor antibody according to any one of claims 1-12, or the composition according to claim 13, in a method for treating skin barrier defects.

18. The dual inhibitor antibody for use according to claim 17, wherein the skin barrier defect is associated with Netherton syndrome, atopic dermatitis, eosinophilic esophagitis, nodular prurigo, chronic pruritus of unknown cause (CPUO), dry skin, asthma (especially KLK5), ichthyosis vulgaris, or pruritus or chronic pruritus.

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