Klk5 / 7 + il-13 targeting antibodies and uses thereof

Abstract

Aspects of the present application provide bispecific antibodies and methods for using the same to promote barrier function and reduce inflammation and to treat conditions such as Netherton syndrome, eosinophilic esophagitis, and atopic dermatitis. In some embodiments, the bispecific antibodies comprise at least one antigen-specific binding site that specifically binds to KLK5 and KLK7 and at least one antigen-specific binding site that specifically binds to IL-13 or a receptor thereof.

Description

Related applications

[0001] This application is based on 35 USC 119 (e) claims priority to U.S. Provisional Application No. 63 / 578,959, filed August 25, 2023; U.S. Provisional Application No. 63 / 549,274, filed February 2, 2024; and U.S. Provisional Application No. 63 / 671,694, filed July 15, 2024, the entire contents of which are incorporated herein by reference.

[0002] Reference to the electronic sequence list The contents of the electronic serial number (A140770004WO00-SEQ-LJG.xml; size: 241,765 bytes; and creation date: August 22, 2024) are incorporated herein by reference in their entirety. Background Technology

[0003] Skin contains tissue structures and cells (such as epithelial cells and immune cells), which together provide functional and physical barriers, playing a vital role in preventing allergens from entering the body and responding to pathogens. Kallikrein (KLK) regulates desquamation and innate immunity to support skin homeostasis and wound healing. In healthy skin, the outermost layer of the epidermis is regularly shed via a KLK-driven proteolytic cascade, leading to the degradation of keratinized desmosomes and desquamation. KLK5 is considered the major activator of this proteolytic cascade. Autoactivated KLK5 enzymatically converts proKLK7 and proKLK14 to their active forms and stimulates a positive feedback loop that, through KLK14, leads to the production of more proKLK5. These KLK enzymes are inhibited by endogenous serine protease inhibitors, such as lymphoepithelial Kazal-associated inhibitors. Dysregulation of KLK (including KLK5 and KLK7) is associated with skin conditions, inflammatory diseases, and cancer. For example, overactive kallikrein 5 and 7 can cause hereditary and spontaneous epidermal barrier disorders (e.g., Netherton syndrome, eosinophilic esophagitis, atopic dermatitis). Summary of the Invention

[0004] Certain aspects of this disclosure relate to the imbalance between endogenous KLK protease and related protease inhibitors leading to barrier dysfunction and inducing inflammation (e.g. Figure 1 This understanding (as shown) can lead to inflammatory disorders such as Natherton's syndrome, eosinophilic esophagitis, and atopic dermatitis. Furthermore, increased activity of T helper 2 (Th2) cytokines (e.g., IL-13) can further drive inflammation and lead to barrier dysfunction and related disorders (such as...). Figure 2(As shown). In some embodiments, the compositions and methods provided herein can be used to inhibit KLK5 / KLK7 and IL-13 signaling to improve barrier function and reduce inflammation, which alleviates the severity of the disease.

[0005] Furthermore, aspects of this disclosure provide multispecific antibodies comprising at least one antigen-specific binding site specifically binding to KLK5 and KLK7, and at least one antigen-specific binding site specifically binding to IL-13 or its receptor. In some embodiments, such multispecific antibodies are configured such that they comprise at least one arm containing an active site binding to KLK5 and KLK7, and at least one other arm binding to IL-13 or the IL-13 receptor (IL-13R). In some embodiments, such multispecific antibodies are advantageous because they inhibit both KLK5 / 7 activity and IL-13 activity in a single molecule. In some embodiments, regarding the KLK5 / 7 binding site within the multispecific antibody (e.g., a bispecific antibody), such antibodies specifically bind to the active form of the enzyme rather than the precursor form. In some embodiments, methods and related compositions, as well as antibodies used in the methods, are provided that can be used to inhibit KLK5 and KLK7 and IL-13 activity to improve barrier function and reduce inflammation, which alleviates the severity of disease. In particular, aspects of this disclosure provide bispecific antibodies, one arm of which contains an antigen-specific binding site for a dual inhibitor antibody targeting KLK5 and KLK7 (referred to as an anti-KLK5 / KLK7 antibody) having high binding affinity and specificity to the active sites of both KLK5 and KLK7, and the other arm specifically binds to IL-13 or its receptor (referred to as an anti-KLK5 / KLK7 + IL-13 targeting bispecific antibody) and inhibits its activity. Therefore, in some embodiments, this disclosure provides methods and related antibody compositions for treating disorders associated with KLK5 and KLK7 dysregulation and / or abnormal Th2 cytokine activity, such as Natherton'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.

[0006] In some embodiments, this disclosure provides a bispecific antibody comprising at least one antigen-specific binding site of a dual inhibitor antibody targeting KLK5 and KLK7 and at least one antigen-specific binding site that specifically binds to and inhibits IL-13 (referred to as an anti-KLK5 / KLK7 + IL-13 targeted bispecific antibody). Therefore, in some embodiments, this disclosure provides methods and related antibody compositions for treating disorders associated with KLK5 and KLK7 dysregulation, wherein Th2 cytokines (such as IL-13) also play a role in the pathogenesis, such as Natherton'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.

[0007] In some aspects, this disclosure provides a bispecific antibody comprising an antigen-specific binding site that specifically binds to KLK5 and KLK7, and an antigen-specific binding site that specifically binds to IL-13 or its receptor. In some embodiments, the antigen-specific binding site that specifically binds to KLK5 and KLK7 binds to the active sites of KLK5 and KLK7 and inhibits enzyme activity. In some embodiments, the antigen-specific binding site that specifically binds to KLK5 and KLK7 comprises any of the antibodies listed in Tables 1a and 1b, namely HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and / or LC CDR3.

[0008] In some embodiments, the antigen-specific binding sites that specifically bind to KLK5 and KLK7 comprise: HC CDR1, HC CDR2, and HC CDR3 of the heavy chain variable domain, having the amino acid sequence of SEQ ID NO: 7; and LC CDR1, LC CDR2, and LC CDR3 of the light chain variable domain, having the amino acid sequence of SEQ ID NO: 8. In some embodiments, the antigen-specific binding sites that specifically bind to KLK5 and KLK7 comprise: HC CDR1, HC CDR2, and HC CDR3 of the heavy chain variable domain, having the amino acid sequence of SEQ ID NO: 13; and LC CDR1, LC CDR2, and LC CDR3 of the light chain variable domain, having the amino acid sequence of SEQ ID NO: 14. In some embodiments, the antigen-specific binding sites that specifically bind to KLK5 and KLK7 comprise: HC CDR1, HC CDR2, and HC CDR3 of the heavy chain variable domain, having the amino acid sequence of SEQ ID NO: 17; and LC CDR1, LC CDR2, and LC CDR3 of the light chain variable domain, having the amino acid sequence of SEQ ID NO: 14. In some embodiments, the antigen-specific binding sites that specifically bind to KLK5 and KLK7 comprise: HC CDR1, HC CDR2, and HC CDR3 of the heavy chain variable domain, having the amino acid sequence of SEQ ID NO: 21; and LC CDR1, LC CDR2, and LC CDR3 of the light chain variable domain, having the amino acid sequence of SEQ ID NO: 14.

[0009] In some embodiments, the antigen-specific binding sites that specifically bind to KLK5 and KLK7 include: HCCDR1 having the amino acid sequence of SEQ ID NO: 1; HCCDR2 having the amino acid sequence of SEQ ID NO: 2; HCCDR3 having the amino acid sequence of SEQ ID NO: 3; LCCDR1 having the amino acid sequence of SEQ ID NO: 4; LCCDR2 having the amino acid sequence of SEQ ID NO: 5; and LCCDR3 having the amino acid sequence of SEQ ID NO: 6. In some embodiments, the antigen-specific binding sites that specifically bind to KLK5 and KLK7 include: 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. In some embodiments, the antigen-specific binding sites that specifically bind to KLK5 and KLK7 include: 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. In some embodiments, the antigen-specific binding sites that specifically bind to KLK5 and KLK7 include: 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.

[0010] In some embodiments, the antigen-specific binding sites that specifically bind to KLK5 and KLK7 have VH and / or VL of any of the antibodies listed in Table 1a.

[0011] In some embodiments, the antigen-specific binding site that specifically binds to KLK5 and KLK7 comprises: VH, which comprises the amino acid sequence of SEQ ID NO: 7; and VL, which comprises the amino acid sequence of SEQ ID NO: 8. In some embodiments, the antigen-specific binding site that specifically binds to KLK5 and KLK7 comprises: VH, which comprises the amino acid sequence of SEQ ID NO: 13; and VL, which comprises the amino acid sequence of SEQ ID NO: 14. In some embodiments, the antigen-specific binding site that specifically binds to KLK5 and KLK7 comprises: VH, which comprises the amino acid sequence of SEQ ID NO: 17; and VL, which comprises the amino acid sequence of SEQ ID NO: 14. In some embodiments, the antigen-specific binding site that specifically binds to KLK5 and KLK7 comprises: VH, which comprises the amino acid sequence of SEQ ID NO: 21; and VL, which comprises the amino acid sequence of SEQ ID NO: 14.

[0012] In some embodiments, the antigen-specific binding site that specifically binds to IL-13 or its receptor includes any of the antibodies listed in Table 2, namely HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2 and / or LC CDR3.

[0013] In some embodiments, the antigen-specific binding site that specifically binds to IL-13 or IL-13R comprises: heavy chain variable domains HCCDR1, HCCDR2, and HCCDR3 having the amino acid sequence of SEQ ID NO: 53; and light chain variable domains LCCDR1, LCCDR2, and LCCDR3 having the amino acid sequence of SEQ ID NO: 54. In some embodiments, the antigen-specific binding site that specifically binds to IL-13 or IL-13R comprises: heavy chain variable domains HCCDR1, HCCDR2, and HCCDR3 having the amino acid sequence of SEQ ID NO: 32; and light chain variable domains LCCDR1, LCCDR2, and LCCDR3 having the amino acid sequence of SEQ ID NO: 33. In some embodiments, the antigen-specific binding site that specifically binds to IL-13 or IL-13R comprises: heavy chain variable domains HC CDR1, HC CDR2, and HC CDR3 having the amino acid sequence of SEQ ID NO: 74; and light chain variable domains LC CDR1, LC CDR2, and LC CDR3 having the amino acid sequence of SEQ ID NO: 75. In some embodiments, the antigen-specific binding site that specifically binds to IL-13 or IL-13R comprises: heavy chain variable domains HC CDR1, HC CDR2, and HC CDR3 having the amino acid sequence of SEQ ID NO: 94; and light chain variable domains LC CDR1, LC CDR2, and LC CDR3 having the amino acid sequence of SEQ ID NO: 95. In some embodiments, the antigen-specific binding site that specifically binds to IL-13 or IL-13R comprises: HC CDR1, HC CDR2, and HC CDR3 of the heavy chain variable domain having the amino acid sequence of SEQ ID NO: 114; and LC CDR1, LC CDR2, and LC CDR3 of the light chain variable domain having the amino acid sequence of SEQ ID NO: 115. In some embodiments, the antigen-specific binding site that specifically binds to IL-13 or IL-13R comprises: HC CDR1, HC CDR2, and HC CDR3 of the heavy chain variable domain having the amino acid sequence of SEQ ID NO: 134; and LC CDR1, LC CDR2, and LC CDR3 of the light chain variable domain having the amino acid sequence of SEQ ID NO: 135.In some embodiments, the antigen-specific binding site that specifically binds to IL-13 or IL-13R comprises: HC CDR1, HC CDR2, and HC CDR3 of the heavy chain variable domain having the amino acid sequence of SEQ ID NO: 657; and LC CDR1, LC CDR2, and LC CDR3 of the light chain variable domain having the amino acid sequence of SEQ ID NO: 658.

[0014] In some embodiments, the antigen-specific binding site that specifically binds to IL-13 or IL-13R comprises: HCCDR1 having the amino acid sequence of SEQ ID NO: 55; HCCDR2 having the amino acid sequence of SEQ ID NO: 56; HCCDR3 having the amino acid sequence of SEQ ID NO: 57; LCCCDR1 having the amino acid sequence of SEQ ID NO: 58; LCCCDR2 having the amino acid sequence of SEQ ID NO: 59; and LCCCDR3 having the amino acid sequence of SEQ ID NO: 60. In some embodiments, the antigen-specific binding site that specifically binds to IL-13 or IL-13R comprises: HC CDR1 having the amino acid sequence of SEQ ID NO: 61; HC CDR2 having the amino acid sequence of SEQ ID NO: 62; HC CDR3 having the amino acid sequence of SEQ ID NO: 63; LC CDR1 having the amino acid sequence of SEQ ID NO: 64; LC CDR2 having the amino acid sequence of SEQ ID NO: 65; and LC CDR3 having the amino acid sequence of SEQ ID NO: 66. In some embodiments, the antigen-specific binding site that specifically binds to IL-13 or IL-13R comprises: HC CDR1 having the amino acid sequence of SEQ ID NO: 67; HC CDR2 having the amino acid sequence of SEQ ID NO: 68; HC CDR3 having the amino acid sequence of SEQ ID NO: 69; LC CDR1 having the amino acid sequence of SEQ ID NO: 70; LC CDR2 having the amino acid sequence of SEQ ID NO: 71; and LC CDR3 having the amino acid sequence of SEQ ID NO: 72. In some embodiments, the antigen-specific binding site that specifically binds to IL-13 or IL-13R comprises: HC CDR1 having the amino acid sequence of SEQ ID NO: 34; HC CDR2 having the amino acid sequence of SEQ ID NO: 35; HC CDR3 having the amino acid sequence of SEQ ID NO: 36; LC CDR1 having the amino acid sequence of SEQ ID NO: 37; LC CDR2 having the amino acid sequence of SEQ ID NO: 38; and LC CDR3 having the amino acid sequence of SEQ ID NO: 39.In some embodiments, the antigen-specific binding site that specifically binds to IL-13 or IL-13R comprises: HC CDR1 having the amino acid sequence of SEQ ID NO: 40; HC CDR2 having the amino acid sequence of SEQ ID NO: 41; HC CDR3 having the amino acid sequence of SEQ ID NO: 42; LC CDR1 having the amino acid sequence of SEQ ID NO: 43; LC CDR2 having the amino acid sequence of SEQ ID NO: 44; and LC CDR3 having the amino acid sequence of SEQ ID NO: 45. In some embodiments, the antigen-specific binding site that specifically binds to IL-13 or IL-13R comprises: HC CDR1 having the amino acid sequence of SEQ ID NO: 46; HC CDR2 having the amino acid sequence of SEQ ID NO: 47; HC CDR3 having the amino acid sequence of SEQ ID NO: 48; LC CDR1 having the amino acid sequence of SEQ ID NO: 49; LC CDR2 having the amino acid sequence of SEQ ID NO: 50; and LC CDR3 having the amino acid sequence of SEQ ID NO: 51. In some embodiments, the antigen-specific binding site that specifically binds to IL-13 or IL-13R comprises: HC CDR1 having the amino acid sequence of SEQ ID NO: 76; HC CDR2 having the amino acid sequence of SEQ ID NO: 77; HC CDR3 having the amino acid sequence of SEQ ID NO: 78; LC CDR1 having the amino acid sequence of SEQ ID NO: 79; LC CDR2 having the amino acid sequence of SEQ ID NO: 80; and LCCDR3 having the amino acid sequence of SEQ ID NO: 81. In some embodiments, the antigen-specific binding site that specifically binds to IL-13 or IL-13R comprises: HC CDR1 having the amino acid sequence of SEQ ID NO: 82; HC CDR2 having the amino acid sequence of SEQ ID NO: 83; HC CDR3 having the amino acid sequence of SEQ ID NO: 84; LC CDR1 having the amino acid sequence of SEQ ID NO: 85; LC CDR2 having the amino acid sequence of SEQ ID NO: 86; and LC CDR3 having the amino acid sequence of SEQ ID NO: 87.In some embodiments, the antigen-specific binding site that specifically binds to IL-13 or IL-13R comprises: HC CDR1 having the amino acid sequence of SEQ ID NO: 88; HC CDR2 having the amino acid sequence of SEQ ID NO: 89; HC CDR3 having the amino acid sequence of SEQ ID NO: 90; LCCDR1 having the amino acid sequence of SEQ ID NO: 91; LCCDR2 having the amino acid sequence of SEQ ID NO: 92; and LCCDR3 having the amino acid sequence of SEQ ID NO: 93. In some embodiments, the antigen-specific binding site that specifically binds to IL-13 or IL-13R comprises: HC CDR1 having the amino acid sequence of SEQ ID NO: 96; HCCDR2 having the amino acid sequence of SEQ ID NO: 97; HC CDR3 having the amino acid sequence of SEQ ID NO: 98; LC CDR1 having the amino acid sequence of SEQ ID NO: 99; LC CDR2 having the amino acid sequence of SEQ ID NO: 100; and LC CDR3 having the amino acid sequence of SEQ ID NO: 101. In some embodiments, the antigen-specific binding site that specifically binds to IL-13 or IL-13R comprises: HC CDR1 having the amino acid sequence of SEQ ID NO: 102; HC CDR2 having the amino acid sequence of SEQ ID NO: 103; HC CDR3 having the amino acid sequence of SEQ ID NO: 104; LC CDR1 having the amino acid sequence of SEQ ID NO: 105; LC CDR2 having the amino acid sequence of SEQ ID NO: 106; and LC CDR3 having the amino acid sequence of SEQ ID NO: 107. In some embodiments, the antigen-specific binding site that specifically binds to IL-13 or IL-13R comprises: HC CDR1 having the amino acid sequence of SEQ ID NO: 108; HC CDR2 having the amino acid sequence of SEQ ID NO: 109; HC CDR3 having the amino acid sequence of SEQ ID NO: 110; LC CDR1 having the amino acid sequence of SEQ ID NO: 111; LC CDR2 having the amino acid sequence of SEQ ID NO: 112; and LC CDR3 having the amino acid sequence of SEQ ID NO: 113.In some embodiments, the antigen-specific binding site that specifically binds to IL-13 or IL-13R comprises: HC CDR1 having the amino acid sequence of SEQ ID NO: 116; HC CDR2 having the amino acid sequence of SEQ ID NO: 117; HC CDR3 having the amino acid sequence of SEQ ID NO: 118; LC CDR1 having the amino acid sequence of SEQ ID NO: 119; LCCDR2 having the amino acid sequence of SEQ ID NO: 120; and LC CDR3 having the amino acid sequence of SEQ ID NO: 121. In some embodiments, the antigen-specific binding site that specifically binds to IL-13 or IL-13R comprises: HCCDR1 having the amino acid sequence of SEQ ID NO: 122; HCCDR2 having the amino acid sequence of SEQ ID NO: 123; HCCDR3 having the amino acid sequence of SEQ ID NO: 124; LCCCDR1 having the amino acid sequence of SEQ ID NO: 125; LCCCDR2 having the amino acid sequence of SEQ ID NO: 126; and LCCCDR3 having the amino acid sequence of SEQ ID NO: 127. In some embodiments, the antigen-specific binding site that specifically binds to IL-13 or IL-13R comprises: HC CDR1 having the amino acid sequence of SEQ ID NO: 128; HC CDR2 having the amino acid sequence of SEQ ID NO: 129; HC CDR3 having the amino acid sequence of SEQ ID NO: 130; LC CDR1 having the amino acid sequence of SEQ ID NO: 131; LC CDR2 having the amino acid sequence of SEQ ID NO: 132; and LC CDR3 having the amino acid sequence of SEQ ID NO: 133. In some embodiments, the antigen-specific binding site that specifically binds to IL-13 or IL-13R comprises: HC CDR1 having the amino acid sequence of SEQ ID NO: 136; HC CDR2 having the amino acid sequence of SEQ ID NO: 137; HC CDR3 having the amino acid sequence of SEQ ID NO: 138; LC CDR1 having the amino acid sequence of SEQ ID NO: 139; LC CDR2 having the amino acid sequence of SEQ ID NO: 140; and LCCDR3 having the amino acid sequence of SEQ ID NO: 141.In some embodiments, the antigen-specific binding site that specifically binds to IL-13 or IL-13R comprises: HC CDR1 having the amino acid sequence of SEQ ID NO: 142; HC CDR2 having the amino acid sequence of SEQ ID NO: 143; HC CDR3 having the amino acid sequence of SEQ ID NO: 144; LCCDR1 having the amino acid sequence of SEQ ID NO: 145; LCCDR2 having the amino acid sequence of SEQ ID NO: 146; and LCCDR3 having the amino acid sequence of SEQ ID NO: 147. In some embodiments, the antigen-specific binding site that specifically binds to IL-13 or IL-13R comprises: HC CDR1 having the amino acid sequence of SEQ ID NO: 148; HC CDR2 having the amino acid sequence of SEQ ID NO: 149; HC CDR3 having the amino acid sequence of SEQ ID NO: 150; LC CDR1 having the amino acid sequence of SEQ ID NO: 151; LC CDR2 having the amino acid sequence of SEQ ID NO: 152; and LC CDR3 having the amino acid sequence of SEQ ID NO: 153. In some embodiments, the antigen-specific binding site that specifically binds to IL-13 or IL-13R comprises: HC CDR1 having the amino acid sequence of SEQ ID NO: 655; HC CDR2 having the amino acid sequence of SEQ ID NO: 41; HC CDR3 having the amino acid sequence of SEQ ID NO: 42; LC CDR1 having the amino acid sequence of SEQ ID NO: 43; LC CDR2 having the amino acid sequence of SEQ ID NO: 44; and LC CDR3 having the amino acid sequence of SEQ ID NO: 45. In some embodiments, the antigen-specific binding site that specifically binds to IL-13 or IL-13R comprises: HC CDR1 having the amino acid sequence of SEQ ID NO: 656; HC CDR2 having the amino acid sequence of SEQ ID NO: 47; HC CDR3 having the amino acid sequence of SEQ ID NO: 48; LC CDR1 having the amino acid sequence of SEQ ID NO: 49; LC CDR2 having the amino acid sequence of SEQ ID NO: 50; and LC CDR3 having the amino acid sequence of SEQ ID NO: 51.

[0015] In some embodiments, the antigen-specific binding site that specifically binds to IL-13 or IL-13R comprises: VH, which comprises the amino acid sequence of SEQ ID NO: 53; and VL, which comprises the amino acid sequence of SEQ ID NO: 54. In some embodiments, the antigen-specific binding site that specifically binds to IL-13 or IL-13R comprises: VH, which comprises the amino acid sequence of SEQ ID NO: 32; and VL, which comprises the amino acid sequence of SEQ ID NO: 33. In some embodiments, the antigen-specific binding site that specifically binds to IL-13 or IL-13R comprises: VH, which comprises the amino acid sequence of SEQ ID NO: 74; and VL, which comprises the amino acid sequence of SEQ ID NO: 75. In some embodiments, the antigen-specific binding site that specifically binds to IL-13 or IL-13R comprises: VH, which comprises the amino acid sequence of SEQ ID NO: 94; and VL, which comprises the amino acid sequence of SEQ ID NO: 95. In some embodiments, the antigen-specific binding site that specifically binds to IL-13 or IL-13R comprises: VH, which comprises the amino acid sequence of SEQ ID NO: 114; and VL, which comprises the amino acid sequence of SEQ ID NO: 115. In some embodiments, the antigen-specific binding site that specifically binds to IL-13 or IL-13R comprises: VH, which comprises the amino acid sequence of SEQ ID NO: 134; and VL, which comprises the amino acid sequence of SEQ ID NO: 135. In some embodiments, the antigen-specific binding site that specifically binds to IL-13 or IL-13R comprises: VH, which comprises the amino acid sequence of SEQ ID NO: 657; and VL, which comprises the amino acid sequence of SEQ ID NO: 658.

[0016] In some embodiments, the antigen-specific binding site that specifically binds to IL-13 or its receptor has a VH and / or VL of any of the antibodies in Table 2. In some embodiments, the binding affinity of the bispecific antibody to KLK5 and KLK7 is not more than 20% different from that of an anti-KLK5 / KLK7 antibody containing the same CDR and / or VH / VL as the bispecific antibody. In some embodiments, the binding affinity of the bispecific antibody to IL-13 signaling is not more than 20% different from that of an anti-IL-13 antibody containing the same CDR and / or VH / VL as the bispecific antibody. In some embodiments, the bispecific antibody retains at least 80% of the inhibitory activity against KLK5 and KLK7 compared to an anti-KLK5 / KLK7 antibody containing the same CDR and / or VH / VL as the bispecific antibody. In some embodiments, the bispecific antibody retains at least 80% of the inhibitory activity against IL-13 signaling relative to an anti-IL-13 antibody containing the same CDR and / or VH / VL as the bispecific antibody.

[0017] In some embodiments, this disclosure provides a composition comprising the bispecific antibody provided herein and an acceptable vector.

[0018] In some embodiments, this disclosure provides a method comprising administering to a subject the bispecific antibody or composition provided herein. In some embodiments, the subject has a skin barrier defect.

[0019] In some embodiments, this disclosure provides a method for treating skin barrier defects, the method comprising administering to a subject an effective amount of the bispecific antibody or composition provided herein. In some embodiments, the skin barrier defect is associated with Natherton syndrome, atopic dermatitis, eosinophilic esophagitis, nodular prurigo, chronic pruritus of unknown cause (CPUO), dry skin, asthma (particularly KLK5), ichthyosis vulgaris, or itching or chronic pruritus.

[0020] In some embodiments, the subject has atopic dermatitis. In some embodiments, the administration of the antibodies or compositions provided herein reduces ear thickness by more than 30% compared to the administration of anti-KLK5 / KLK7 antibodies containing the same CDR and / or VH / VL as the bispecific antibody. In some embodiments, the administration reduces ear thickness by more than 30% compared to the administration of anti-IL-13 antibodies containing the same CDR and / or VH / VL as the bispecific antibody. In some embodiments, the administration reduces skin erythema / bleeding by more than 30% compared to the administration of anti-KLK5 / KLK7 antibodies containing the same CDR and / or VH / VL as the bispecific antibody. In some embodiments, the administration reduces skin erythema / bleeding by more than 30% compared to the administration of anti-IL-13 antibodies containing the same CDR and / or VH / VL as the bispecific antibody. In some embodiments, this application reduces skin erythema / bleeding by more than 30% compared to subjects receiving anti-KLK5 / KLK7 antibodies containing the same CDR and / or VH / VL as the bispecific antibody. In some embodiments, this application reduces skin peeling / erosion by more than 30% compared to subjects receiving anti-IL-13 antibodies containing the same CDR and / or VH / VL as the bispecific antibody. In some embodiments, this application reduces skin desquamation / dryness by more than 30% compared to subjects receiving anti-IL-13 antibodies containing the same CDR and / or VH / VL as the bispecific antibody. In some embodiments, this application reduces skin edema by more than 30% compared to subjects receiving anti-KLK5 / KLK7 antibodies containing the same CDR and / or VH / VL as the bispecific antibody. In some implementations, this administration reduces skin edema by more than 30% compared to subjects receiving anti-IL-13 antibodies containing the same CDR and / or VH / VL as the bispecific antibody.

[0021] The foregoing and other aspects, embodiments, operations, functions, features, and implementations of the present invention can be more fully understood by taking into account the accompanying drawings in the following description. Attached Figure Description

[0022] 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 non-limiting examples of certain aspects of the compositions and methods disclosed herein.

[0023] Figure 1This is a graph showing the activation of abnormal proteases (e.g., abnormal KLK5, KLK7, and KLK14) that lead to diseases related to skin barrier defects.

[0024] Figure 2 This is a graph showing that in disorders associated with epidermal barrier dysfunction, elevated KLK5 / KLK7 activity leads to Th2 cell activation (which further increases KLK5 / KLK7 activity).

[0025] Figures 3A to 3B This figure shows that in the Nc / Nga model of atopic dermatitis, inhibiting KLK5 / KLK7 and Th2 signaling has a superior effect compared to anti-IL-4R antibodies. Figure 3A The results showed that mice treated with KLK5 / KLK7-Dual-Ab4 had a further reduction in ear thickness compared to mice treated with anti-IL-4R antibody. Figure 3B The results showed that mice treated with KLK5 / KLK7-Dual-Ab4 had lower clinical skin scores compared to mice treated with anti-IL-4R antibodies.

[0026] Figures 4A to 4I The binding affinity relates to an exemplary anti-KLK5 / KLK7 + IL-13 bispecific antibody (referred to as K13-0004). Figures 4A to 4C It shows the relationship with KLK5 ( Figure 4A ), KLK7 ( Figure 4B ) and IL-13 ( Figure 4C ) combined K D value. Figures 4D to 4H Showing KLK5 ( Figure 4D ;and Figure 4E Comparison of exemplary anti-KLK5 / KLK7 biantibodies), KLK7 ( Figure 4F ;and Figure 4G Comparison of exemplary anti-KLK5 / KLK7 biantibodies), IL-13 ( Figure 4H ) and extracellular matrix proteins (ECM) periosteal proteins ( Figure 4I The inhibition of ).

[0027] Figures 5A to 5D It involves eliminating KLK5 and KLK7-mediated primary human keratinocyte barrier disruption. Figure 5A This demonstrates the use of different concentrations of anti-KLK5 / KLK7+IL-13 bispecific antibody ( Figure 5A ) or different concentrations of anti-KLK5 / KLK7 dual antibody ( Figure 5B Transepithelial resistance (TEER) after treatment. Figures 5C to 5D This demonstrates the use of different concentrations of anti-KLK5 / KLK7+ IL-13 bispecific antibody ( Figure 5C) or different concentrations of anti-KLK5 / KLK7 dual antibody ( Figure 5D Migration of FITC-dextran dyes after treatment. Detailed Implementation

[0028] This disclosure is based, at least in part, on the development of a bispecific antibody comprising one arm and another arm, wherein one arm contains an antigen-binding site for a dual inhibitor antibody or variant thereof targeting KLK5 and KLK7, and the other arm contains an antigen-binding site for an anti-IL-13 or anti-IL-13R antibody (referred to as an anti-KLK5 / KLK7+IL-13 bispecific antibody). 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). In some embodiments, a bispecific antibody is provided wherein one arm contains an antigen-specific binding site for a dual inhibitor antibody targeting KLK5 and KLK7, and wherein the other arm specifically binds to IL-13 and inhibits its activity. It also provides methods for using anti-KLK5 / KLK7+IL-13 bispecific antibodies and their variants in research, diagnostic / detection and therapeutic applications, as well as anti-KLK5 / KLK7+IL-13 bispecific antibodies for use in such methods.

[0029] The foregoing and other aspects, embodiments, operations, functions, features, and implementations of the present invention can be more fully understood by taking into account the accompanying drawings in the following description.

[0030] I. Definition Administration: As used herein, the term “administering / administration” means to provide an antibody or a combination thereof to a subject in a physiologically and / or pharmacologically useful manner (e.g., to treat a subject’s condition).

[0031] Affinity-maturating antibody: The term "affinity-maturating antibody" is used herein to refer to an antibody having one or more alterations in one or more CDRs, which, compared to a parent antibody without said alterations, result in an improvement in the antibody's affinity (e.g., KD, kd, or ka) for the target antigen. In some embodiments, exemplary affinity-maturating antibodies may have nanomolar or even picomolar affinity for the target antigen. Various procedures are available for generating affinity-maturating antibodies, including screening combinatorial antibody libraries already prepared using biodisplay. For example, Marks et al., BioTechnology, 10: 779-783 (1992) describe affinity maturation achieved through VH and VL domain shuffling. The following literature describes random mutagenesis of CDR and / or framework residues: 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). US Patent No. 6,914,128 B1 describes selective mutagenesis using amino acid residues that enhance activity at selective mutagenesis sites and at contact or hypermutation sites.

[0032] Antibody: As used herein, the term "antibody" refers to a polypeptide comprising at least one immunoglobulin variable domain containing at least one unique antigen-specific binding site; or a portion of an immunoglobulin variable domain (e.g., a complementary site or a portion thereof) containing at least one unique antigen-specific binding site. In some embodiments, the antibody comprises at least one unique antigen-specific binding site that specifically binds to the active site of an enzyme. In some embodiments, the antibody is an IL-13-targeting antibody comprising at least one antigen-specific binding site that specifically binds to IL-13 or its receptor. In some embodiments, the antibody is a full-length antibody. In some embodiments, the antibody is a chimeric antibody. In some embodiments, the antibody is a humanized antibody. However, in some embodiments, the antibody is a Fab fragment, an F(ab')2 fragment, an Fv fragment, or a scFv fragment. In some embodiments, the antibody is a multispecific antibody (e.g., a bispecific antibody). In some embodiments, the antibody is a nanobody derived from a camel antibody or a shark antibody. In some embodiments, the antibody is a bifunctional antibody. In some embodiments, the antibody comprises a framework having a human germline sequence. In another embodiment, the antibody comprises a heavy chain constant domain selected from the group consisting of IgG, IgG1, IgG2, IgG2A, IgG2B, IgG2C, IgG3, IgG4, IgA1, IgA2, IgD, IgM, and IgE constant domains. In some embodiments, the antibody comprises a heavy (H) chain variable region or domain (abbreviated herein as VH) and / or a light (L) chain variable region or domain (abbreviated herein as VL). In some embodiments, the antibody comprises a constant domain. An immunoglobulin constant domain refers to a heavy chain or light chain constant domain. The amino acid sequences of the human IgG heavy chain and light chain constant domains and their functional variations are known. In some embodiments, the immunoglobulin heavy chain constant domain comprises an Fc region. Regarding the heavy chain, in some embodiments, the heavy chain of the antibody described herein may be an α (alpha), Δ (delta), ε (epsilon), γ (gamma), or µ (mu) heavy chain. In some embodiments, the heavy chain of the antibody described herein may comprise a human α (alpha), Δ (delta), ε (epsilon), γ (gamma), or μ (mu) heavy chain. In some embodiments, the antibody described herein comprises human γ1 CH1, CH2, and / or CH3 domains. In some embodiments, the amino acid sequence of the antibody comprises the amino acid sequence of the human γ (gamma) heavy chain constant region, such as any sequence known in the art. Non-limiting examples of human constant region sequences have been described in the art, for example, see U.S. Patent No. 5,693,780 and Kabat EA et al., (1991), ibid.In some embodiments, the antibody comprises a heavy chain containing an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 98%, or at least 99% identity with any of the variable chain constant regions provided herein. In some embodiments, the antibody comprises a light chain containing an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 98%, or at least 99% identity with any of the light chain constant regions provided herein. In some embodiments, the antibody is modified, for example by glycosylation, phosphorylation, SUMOylation, and / or methylation. In some embodiments, the antibody is a glycosylated antibody 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 via N-glycosylation, O-glycosylation, C-glycosylation, glypiation (GPI anchoring), and / or phosphoglycosylation. In some embodiments, the one or more sugar or carbohydrate molecules are monosaccharides, disaccharides, oligosaccharides, or polysaccharides. In some embodiments, the one or more sugar or carbohydrate molecules are branched-chain oligosaccharides or branched-chain polysaccharides. In some embodiments, the one or more sugar or carbohydrate molecules include mannose units, glucose units, N-acetylglucosamine units, or phospholipid units. In some embodiments, the antibody is a construct comprising a polypeptide containing one or more antigen-binding fragments of this disclosure linked to a linker polypeptide or an immunoglobulin constant domain. The linker polypeptide comprises two or more amino acid residues linked by peptide bonds and is used to link one or more antigen-binding moieties. Examples of linker polypeptides have been reported (see, for example, Holliger, P. et al. (1993) Proc. Natl. Acad. Sci. USA 90:6444-6448; Poljak, RJ et al. (1994) Structure 2:1121-1123). Furthermore, the antibody may be a portion of a larger immunoadhesion molecule formed by covalent or non-covalent association of the antibody or antibody moiety with one or more other proteins or peptides. Examples of such immunoadhesion molecules include the preparation of tetrameric scFv molecules using the streptavidin core region (Kipriyanov, SM et al. (1995) Human Antibodies and Hybridomas 6:93-101) and the preparation of divalent and biotinylated scFv molecules using cysteine ​​residues, biomarker peptides, and C-terminal multihistidine tags (Kipriyanov, SM et al. (1994) Mol. Immunol. 31:1047-1058).

[0033] Approximately: As used herein, the terms “approximately” or “about” when applied to one or more values ​​of interest refer to a value similar to a specified reference value. In some embodiments, unless otherwise stated or clearly apparent from the context (unless such a figure would exceed 100% of a possible value), the terms “approximately” or “about” refer to a range of values ​​within 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less of the stated reference value in either direction (greater or less).

[0034] Bispecific antibody: As used herein, the term "bispecific antibody" refers to an antibody containing two distinct antigen-specific binding sites or two linked (covalent or non-covalent) antibodies that together contain two distinct antigen-specific binding sites. Non-limiting examples of bispecific antibody forms or structures are provided by Labrijn, AF, et al. Bispecific antibodies: a mechanistic review of the pipeline Nature Reviews Drug Discovery, Vol. 18, pp. 585-608 (2019) and Brinkmann U and Kontermann EE, The making of bispecific antibodies In MAbs. Feb / Mar 2017;9(2):182-212, the entire contents of each of them are incorporated herein by reference.

[0035] CDR: As used herein, the term "CDR" refers to the complementarity-determining region within the variable sequence of an antibody. A typical antibody molecule contains a heavy chain variable region or heavy chain variable domain (VH) and a light chain variable region or light chain variable domain (VL), which are typically involved in antigen binding. The VH and VL regions can be further subdivided into hypervariable regions (also known as "complementarity-determining regions" ("CDRs")) interspersed with more conserved regions (called "frame regions" ("FRs")). Each VH and VL typically consists of three CDRs and four FRs, arranged in the following order from the amino terminus to the carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The extent of the frame regions and CDRs can be precisely identified using methods known in the art, such as the Kabat definition, IMGT definition, Chothia definition, AbM definition, and / or contact definition, all of which are well-known in the art.See, for example, Kabat, E.A. et al. (1991), Sequences of Proteins of Immunological Interest, 5th edition, 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 a CDR defined by any method known in the art. Two antibodies having the same CDR means that the two antibodies have the same amino acid sequence of the CDR, as determined by the same method (e.g., IMGT definition).

[0036] In some implementations, each variable region of the heavy and light chains contains three CDRs, designated CDR1, CDR2, and CDR3 for each variable region. As used herein, the term "CDR set" refers to a group of three CDRs present in a single variable region capable of binding the antigen. The exact boundaries of these CDRs are defined differently depending on the system. 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 a definitive residue numbering system applicable to any variable region of an antibody, but also provides precise residue boundaries defining three CDRs. These CDRs can be referred to as Kabat CDRs. Sub-regions of a CDR can be designated as L1, L2, and L3 or H1, H2, and H3, where “L” and “H” designate the light chain region and the heavy chain region, respectively. These regions can be referred to as Chothia CDRs, whose boundaries overlap with Kabat CDRs. Padlan (FASEB J.9:133-139 (1995)) and MacCallum (J Mol Biol 262(5):732-45 (1996)) describe other boundaries defining CDRs that overlap with Kabat CDRs. Other CDR boundary definitions may not strictly follow one of the systems described above, but will still overlap with Kabat CDRs. CDRs overlap, however, they may shorten or lengthen based on predictions or experimental findings that specific residues or groups of residues, or even the entire CDR, do not significantly affect antigen binding. The methods used herein can utilize CDRs defined according to any of these systems, although preferred embodiments use CDRs defined by Kabat or Chothia.

[0037] CDR transplantation antibody: As used herein, the term “CDR transplantation antibody” refers to an antibody that contains heavy and light chain variable region sequences from one species, but in which one or more of the CDR regions of VH and / or VL are replaced by CDR sequences from another species, such as antibodies with murine heavy and light chain variable regions in which one or more of the murine CDR (e.g., CDR3) have been replaced by human CDR sequences.

[0038] 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, such as antibodies with murine heavy and light chain variable regions linked to human constant regions.

[0039] Complementarity: As used herein, the term “complementarity” refers to the ability of two nucleotides or groups of nucleotides to pair precisely. Specifically, complementarity is a term characterizing the degree to which hydrogen bonds bind two nucleotides or groups of nucleotides. For example, if a base at a position on an oligonucleotide can form a hydrogen bond with a base at a corresponding position on a target nucleic acid (e.g., mRNA), the two bases are considered complementary at that position. Base pairing can include canonical Watson-Crick base pairing and non-Watson-Crick base pairing (e.g., wobbly base pairing and Hoogsteen base pairing). For example, in some embodiments, for complementary base pairing, adenosine-type bases (A) are complementary to thymidine-type bases (T) or uracil-type bases (U), cytosine-type bases (C) are complementary to guanosine-type bases (G), and universal bases such as 3-nitropyrrole or 5-nitroindole can hybridize with any A, C, U, or T and are 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.

[0040] Conservative amino acid substitution: As used herein, “conservative amino acid substitution” refers to an amino acid substitution that does not alter the relative charge or size characteristics of the protein to which the substitution is made. Variants can be prepared according to methods known to those skilled in the art for altering polypeptide sequences, such as those found in the following compilations of references: e.g., *Molecular Cloning: A Laboratory Manual*, J. Sambrook et al., 4th ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 2012; or *Current Protocols in Molecular Biology*, FM Ausubel et al., John Wiley & Sons, Inc., New York. Conservative substitution of amino acids includes substitutions between amino acids 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.

[0041] Cross-reactivity: As used herein, the term "cross-reactivity" refers to the ability of an agent to specifically bind with similar affinity or co-affinity to up to one similar type or class of antigens (e.g., antigens of multiple homologs, paralogs, or orthologs). For example, in some embodiments, antibodies that are 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) are able to bind to human and non-human primate antigens with similar affinity or co-affinity. In some embodiments, antibodies are cross-reactive with similar types or classes of human and rodent antigens. In some embodiments, antibodies are cross-reactive with similar types or classes of rodent and non-human primate antigens. In some embodiments, antibodies are cross-reactive with similar types or classes of human, non-human primate, and rodent antigens.

[0042] 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 through a common, unique antigen-specific binding site and inhibits the activity of those antigens. In some embodiments, a dual-inhibitor antibody targets at least two different proteins (e.g., expressed by two different genes (e.g., endogenous genes, such as homologs, parahomologs)) through a common, unique antigen-specific binding site and inhibits the activity of at least two different proteins (e.g., enzymes, such as proteases). 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)) through a common, unique antigen-specific binding site and inhibits the activity of at least two different proteases. In some embodiments, the common, unique antigen-specific binding site binds to a similar (e.g., homologous) domain shared between or therein of 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 therein of at least two different enzymes (e.g., proteases). In some embodiments, the common unique antigen-specific binding site of the dual inhibitor antibody comprises amino acids from one or more complementarity-determining regions of the antibody. In some embodiments, the common unique antigen-specific binding site of the dual inhibitor antibody is located within the heavy chain variable region and / or light chain variable region of the antibody. In some embodiments, the common unique antigen-specific binding site of the dual inhibitor antibody comprises one or more complementarity-determining regions from the heavy chain variable region and / or light chain variable region of the antibody. In some embodiments, the common unique antigen-specific binding site of the dual inhibitor antibody comprises HC CDR1, HC CDR2, HC CDR3, LC CDR1, LCCDR2, and LC CDR3 from the heavy chain variable region and light chain variable region of the antibody. In some embodiments, the dual inhibitor antibody specifically binds to two different proteins expressed by two different genes (e.g., KLK5 and KLK7).

[0043] Effective amount: As used herein, "effective amount" refers to the amount of each active agent (e.g., anti-KLK5 / KLK7 + IL-13 bispecific antibody) required, alone or in combination with one or more other active agents, to impart the desired effect (e.g., a therapeutic effect on a subject). In some embodiments, the therapeutic "effect" is a reduction in KLK5 and / or KLK7 activity and / or a reduction in IL-13 activity and / or an alleviation of disease (e.g., Natherton's syndrome, eosinophilic esophagitis, and atopic dermatitis) or related symptoms, such as improved barrier function.

[0044] 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 FRs represent 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, receptor sequences known in the art can be used in the antibodies disclosed herein.

[0045] 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.

[0046] Humanized antibodies: As used herein, the term "humanized antibody" refers to antibodies that contain variable region sequences of heavy and light chains derived from a non-human species (e.g., mouse), but in which 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.

[0047] Humanized antibodies are human immunoglobulins (recipient antibodies) in which residues of the complementarity-determining region (CDR) from the recipient are replaced by residues of the CDR from a non-human species (donor antibody) (such as mouse, rat, or rabbit) having the desired specificity, affinity, and capability. In some embodiments, Fv frame region (FR) residues of the human immunoglobulin are replaced by corresponding non-human residues. Furthermore, humanized antibodies may include residues not found in either the recipient antibody or the input CDR or frame sequence, but such residues are included for further refinement and optimization of antibody performance. Generally, humanized antibodies will contain at least one and typically substantially all of two variable domains, wherein all or substantially all of the CDR regions correspond to those of non-human immunoglobulins, and all or substantially all of the FR regions are those of the human immunoglobulin common sequence. Optionally, humanized antibodies will also contain at least a portion of the immunoglobulin constant region or domain (Fc), typically at least a portion of the constant region or domain (Fc) of the human immunoglobulin. Antibodies may 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, or six) that are altered relative to the original antibody, referred to as one or more CDRs derived from one or more CDRs of the original antibody. Humanized antibodies may also involve affinity maturation.

[0048] In some implementations, CDRs (e.g., as shown in Table 1a or Table 1b) are transplanted into human variable structural domains (e.g., IGKV1-NL1). 01 and IGHV1-3 Humanization is achieved in the variable domain (01). In some embodiments, the antibody of this disclosure is a humanized variant containing one or more amino acid substitutions (e.g., in the VH frame region) compared to any of the VH listed in Table 1a or Table 1b, and / or containing one or more amino acid substitutions (e.g., in the VL frame region) compared to any of the VL listed in Table 1a or Table 1b.

[0049] Isolated antibody: As used herein, "isolated antibody" refers to an antibody that is substantially free of other antibodies with different antigen specificities. However, in some embodiments, isolated antibodies may be cross-reactive to other antigens. Furthermore, isolated antibodies may be substantially free of other cellular material and / or chemicals.

[0050] Kabat Numbering: As used herein, the terms “Kabat numbering,” “Kabat definition,” and “Kabat labeling” are used interchangeably. These terms, as recognized 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 ed., US Department of Health and Human Services, NIH Publication No. 91-3242). For the heavy chain variable region, the hypervariable region ranges from amino acid positions 31 to 35 of CDR1, amino acid positions 50 to 65 of CDR2, and amino acid positions 95 to 102 of CDR3. For the light chain variable regions, the range of the hypervariable regions is between amino acid positions 24 to 34 of CDR1, amino acid positions 50 to 56 of CDR2, and amino acid positions 89 to 97 of CDR3.

[0051] 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).

[0052] 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, a multispecific antibody is a bispecific antibody. Non-limiting examples of the form or structure of multispecific antibodies are provided by 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 August to September 2016;8(6):1010-20; and Brinkmann U and Kontermann EE, The making of bispecific antibodies In MAbs. Feb / Mar 2017;9(2):182-212, the entire contents of each of them are incorporated herein by reference.

[0053] Recombinant Antibody: As used herein, the term “recombinant antibody” is intended to include all antibodies prepared, expressed, generated, or isolated by recombinant means, such as antibodies expressed using recombinant expression vectors transfected into host cells (described in more detail in this disclosure), including, for example, antibodies isolated from recombinant 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 animals (e.g., mice) transgenic to target 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, produced, 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 are subjected to in vitro mutagenesis (or, when using animals transgenic to target human Ig sequences, in vivo somatic cell mutagenesis), and thus the amino acid sequences of the VH and VL regions of the recombinant antibody are sequences that, although derived from and associated with human germline VH and VL sequences, may not be naturally present in the in vivo human antibody germline library. One embodiment of this disclosure provides a fully human antibody (e.g., a fully human antibody 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. WO2005 / 007699 A2.

[0054] Selectivity: As used herein, the term "selective" or "selectively" refers to the ability of a molecule to influence its target molecule (e.g., inhibit, antagonize, agonize, etc.) compared to a reference molecule. For example, a molecule that selectively inhibits its target molecule means that it is able to inhibit its target molecule to a degree distinct from that of a reference molecule in an inhibition assay or other inhibition context. For example, with respect to inhibitors, the term "selectively inhibiting" refers to the ability of an inhibitor to inhibit its target molecule to a degree different from that of a reference molecule that is substantially uninhibited in an inhibition assay, such as reaching a degree that allows for selective inhibition of the target molecule, as described herein. Once the reaction is terminated, the signal generated by inhibiting the target molecule can be measured. The half-maximum inhibitor concentrations of the target molecule and the reference molecule can be calculated.

[0055] Specific binding: As used herein, the term "specific binding" refers to the ability of a molecule to bind to a conjugate with a degree of affinity or cohesion such that the molecule can be used in a binding assay or other binding context to distinguish the conjugate from an appropriate control. With respect to antibodies, the term "specific binding" refers to the ability of an antibody to bind to a specific antigen with a degree of affinity or cohesion relative to the binding of an antibody to one or more appropriate reference antigens, such that the antibody can be used to distinguish the specific antigen from other antigens, as described herein. In some embodiments, if the antibody binds to the K of the target... D It 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 If M is lower than a certain value, the antibody specifically binds to the target. In some embodiments, the antibody specifically binds to KLK5 or KLK7. In some embodiments, the antibody specifically binds to a Th2 cytokine (e.g., IL-13). In some embodiments, the bispecific antibody provided herein specifically binds to KLK5 and KLK7 on one arm and specifically binds to a Th2 cytokine or its receptor (e.g., IL-13 or IL-13R) on the other arm.

[0056] Subject: As used herein, the term "subject" refers to a mammal. In some embodiments, the subject is a non-human primate or rodent. In some embodiments, the subject is a human. In some embodiments, the subject is a patient, such as a person who has or is suspected of having a disease.

[0057] Th2-targeting antibody: As used herein, the term "Th2-targeting antibody" refers to an antibody that binds to Th2 cytokines or their receptors. In some embodiments, Th2-targeting antibodies inhibit Th2 cytokine-mediated signaling. In some embodiments, Th2-targeting antibodies inhibit Th2 cytokine-mediated inflammation.

[0058] Treatment: As used herein, the term "treatment" refers to the application or administration of a composition comprising one or more active agents (e.g., an anti-KLK5 / KLK7 + IL-13 bispecific antibody) to a subject suffering from a target disease or condition, symptoms of the disease / condition, or susceptibility to the disease / condition, with the aim of curing, healing, alleviating, mitigating, altering, remedying, reducing, improving, or influencing the disease, its symptoms, or susceptibility. Alleviating a target disease / condition includes delaying or preventing the development or progression of the disease, or reducing its severity. It should be understood that references to treatment may also refer to antibodies, including KLK5 / KLK7 + IL-13 bispecific antibodies used in such methods.

[0059] II. Antibodies (a) Dual inhibitor antibody against KLK5 and KLK7 In some embodiments, dual inhibitor antibodies targeting KLK5 and KLK7 (referred to as anti-KLK5 / KLK7 antibodies) are antibodies that are specific to both kallikrein-5 (KLK5) and KLK7 through a common specific antigen-binding site. Dual inhibitor KLK5 / KLK7 antibodies are described in PCT / US2024 / 019231, the entire contents of which are incorporated herein by reference. In some aspects, this document provides 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 a common antigen-binding site. In some embodiments, the anti-KLK5 / KLK7 antibodies described herein specifically bind to both a KLK5 epitope exposed to or becoming exposed to the antibody and a KLK7 epitope exposed to or becoming exposed to the antibody. In some embodiments, the anti-KLK5 / KLK7 antibody described herein binds to the active sites of KLK5 and KLK7. In some embodiments, the anti-KLK5 / KLK7 antibody provided herein specifically binds to KLK5 and KLK7 derived from humans, non-human primates, mice, rats, etc. In some embodiments, the anti-KLK5 / KLK7 antibody provided herein specifically binds to human KLK5. In some embodiments, the anti-KLK5 / KLK7 antibody provided herein specifically binds to mouse KLK5. In some embodiments, the anti-KLK5 / KLK7 antibody provided herein specifically binds to human KLK7. In some embodiments, the anti-KLK5 / KLK7 antibody provided herein specifically binds to mouse KLK7. In some embodiments, a multispecific antibody is provided herein comprising an arm having an antigen-specific binding site of a dual inhibitor antibody targeting KLK5 and KLK7 and an arm having an antigen-specific binding site specifically binding to IL-13 or its receptor. For example, in some embodiments, this document provides a bispecific antibody comprising an arm having an antigen-specific binding site for a dual inhibitor antibody targeting KLK5 and KLK, and an arm having an antigen-specific binding site specifically binding to IL-13 or IL-13R.

[0060] In some implementations, the anti-KLK5 / KLK7 antibody is not a bispecific antibody or a 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.

[0061] 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 the active forms of KLK5 and KLK7. In some embodiments, the anti-KLK5 / KLK7 antibodies do not bind to inactive forms (precursor forms) of KLK5 and KLK7. In some embodiments, the antibody specifically binds to the active forms of KLK5 and KLK7, but does not specifically bind to the inactive forms of KLK5 or KLK7. In some embodiments, the antibody detectably binds to the active forms of KLK5 and KLK7, but undetectably binds to the inactive forms of KLK5 or KLK7 under the same or equivalent conditions. In some embodiments, the anti-KLK5 / KLK7 antibody inhibits the activity of KLK5 and KLK7 proteases. In some embodiments, when the anti-KLK5 / KLK7 antibody binds to KLK5 or KLK7, it is not cleaved by KLK5 or KLK7. In some embodiments, the anti-KLK5 / KLK7 antibody competes with SPINK5 and / or leucine for binding to the active sites of KLK5 and KLK7. In some embodiments, the anti-KLK5 / KLK7 antibody reduces hyperkeratosis and desquamation. In some embodiments, the anti-KLK5 / KLK7 antibody reduces stratum corneum thickness. In some embodiments, the anti-KLK5 / KLK7 antibody reduces inflammation and epidermal effects.

[0062] Kallikrein-5 (also known as stratum corneum trypsin (SCTE)) is a serine protease expressed in the epidermis, encoded by the KLK5 gene. The KLK5 gene is one of fifteen members of the kallikrein subfamily located on a chromosome cluster. Its expression is upregulated by estrogen and progesterone. KLK5 is expressed in the granular layer and stratum corneum. In some embodiments, KLK5 regulates epidermal desquamation. In some embodiments, KLK5 regulates epidermal desquamation in conjunction 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., corneal lockstrand protein (CDSN), desmosome core glycoprotein 1 (DSG1), and desmosome glialin 1 (DSC1), etc.). In the epidermis (e.g., the granular layer and stratum corneum), KLK5 is expressed in an inactive form (sometimes referred to as the proform / pro-form) proKLK5 and is self-activated. When activated, KLK5 can be proteolytically cleaved, converting both proKLK7 and proKLK14 into their active forms. The active KLK14 can then activate the newly generated proKLK5, creating a positive feedback loop (see, for example, Nauroy et al., Kallikreins: Essentialepidermal 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 granular layer (e.g., corneal chain-locked protein (CDSN), desmosome core glycoprotein 1 (DSG1), and desmosome glialin 1 (DSC1), etc.). Structural proteins (such as CDSN, DSG1, and DSC1) are adhesion proteins of the extracellular portion of keratinocytes, which are connective structures that mediate keratinocyte adhesion. Degradation of these proteins at the epidermal surface leads to desquamation, which can result in skin barrier defects (e.g., stratum corneum shedding, decreased barrier permeability, allergies, and inflammation).KLK5 and KLK7 are involved in this process (see, for example, 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 improved skin barrier integrity and reduced 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, Volume 14, Issue 675).

[0063] 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, which includes fifteen homologous serine proteases located on chromosome 19.] KLK7 is secreted as an inactive zymogen (e.g., in the granular layer of the epidermis), thus requiring activation for proteolytic cleavage. In some embodiments, KLK7 is activated by KLK5 or a matriptase. Once active, KLK7 is able to cleave proteins that form the stratum corneum and / or granular layer (e.g., corneal lock-chain protein (CDSN), desmosome core glycoprotein 1 (DSG1), and desmosome glialin 1 (DSC1), etc.) (see, for example, Caubet et al. (May 2004). Degradation of corneodesmosome proteins by two serine proteases of thekallikrein family, SCTE / KLK5 / hK5 and SCCE / KLK7 / hK7. The Journal of Investigative Dermatology. 122 (5): 1235-1244). These proteins constitute the extracellular component of keratinocytes, which are intercellular adhesion structures consisting of intermediate filaments connecting adjacent cells in the stratum corneum. In some embodiments, proteolysis of keratinocytes leads to epidermal desquamation (i.e., the shedding of keratinocytes 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, dysregulation of KLK7 is associated with several skin conditions, including atopic dermatitis, psoriasis, and Natherton's syndrome. These conditions are characterized by excessive dryness, scaling, and inflammation of the skin due to disruption of skin homeostasis and proper barrier function.

[0064] In some embodiments, the anti-KLK5 / KLK7 antibodies described herein specifically bind to epitopes on human KLK5. Exemplary amino acid sequences of human KLK5 are shown in the following: NCBI accession numbers NP_001070959.1, NP_001070960.1, or NP_036559.1, and UniProt accession numbers Q8IU55, Q6S9W8, M0QXX2, Q9P0G3, A0A2I2MP48, or A0A2I2MP49, the entire sequences of which are incorporated herein by reference.

[0065] In some embodiments, the anti-KLK5 / KLK7 antibody described herein specifically binds to an epitope on mouse KLK5. Exemplary amino acid sequences of mouse KLK5 are shown in the following: NCBI accession numbers NP_081082.1, XP_006541213.1, XP_006541214.1, XP_006541215.1, XP_036009294.1 or XP_036009295.1, and UniProt accession number P15945 or Q9D140, the entire sequences of which are incorporated herein by reference.

[0066] In some embodiments, the anti-KLK5 / KLK7 antibodies described herein specifically bind to epitopes on human KLK7 via the same antigen-binding site that binds to KLK5 (e.g., human KLK5 or mouse KLK5). Exemplary amino acid sequences of human KLK7 are shown in the following: NCBI accession numbers NP_001193982.1, NP_001230055.1, NP_005037.1, NP_644806.1, and UniProt accession numbers M0QYU8, Q6DTY1, X2J289, X2J4X7, A0A024R4H6, P49862, A0A2H4GDB2, and A0A2H4GDB6, the entire sequences of which are incorporated herein by reference.

[0067] In some embodiments, the anti-KLK5 / KLK7 antibodies described herein specifically bind to epitopes on mouse KLK7 via the same antigen-binding site that binds to KLK5 (e.g., human KLK5 or mouse KLK5). Exemplary amino acid sequences of mouse KLK7 are shown in the following: NCBI Registry No. NP_036002.1 and UniProt Registry No. Q91VE3, the entire sequences of which are incorporated herein by reference.

[0068] In some embodiments, the anti-KLK5 / KLK7 antibody described herein specifically binds to epitopes on KLK5 (e.g., the active site of an enzyme, also referred to as the catalytic domain / pocket of human KLK5 or mouse KLK5) and epitopes on KLK7 (e.g., the catalytic domain / pocket of human KLK7 or mouse KLK7). In some embodiments, the anti-KLK5 / KLK7 antibody described herein prevents 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 antibody described herein binds to fragments of KLK5 (e.g., human or mouse KLK5) and KLK7 (e.g., human or mouse KLK7). The length of the KLK5 and / or KLK7 (e.g., human or mouse) fragments can be between about 5 and about 425 amino acids, about 10 and about 400 amino acids, about 50 and about 350 amino acids, about 100 and about 300 amino acids, about 150 and about 250 amino acids, about 200 and about 300 amino acids, about 75 and about 150 amino acids, about 25 and about 100 amino acids, or about 10 and about 30 amino acids. Without wishing to be bound by any particular theory, in some embodiments, the heavy chain (HC) complementarity-determining region 3 (CDR3) of either 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.

[0069] In some embodiments, the anti-KLK5 / KLK7 antibody described herein inhibits KLK5 protease activity, KLK7 protease activity, or both KLK5 and KLK7 protease activities. In some embodiments, the anti-KLK5 / KLK7 antibody inhibits 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 implementations, the anti-KLK5 / KLK7 antibody is present in concentrations 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, and 0.1 nM to 0.7 nM. 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 0.5nM, 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 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 IC50 values ​​in the ranges of 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 inhibit the cleavage of KLK5 (e.g., human KLK5 or mouse KLK5) by BOC-Val-Pro-Arg-AMC. In some implementations, the anti-KLK5 / KLK7 antibody is present in amounts 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, or less than 0.IC50 of 1 nM or less than 0.05 nM inhibits KLK7 cutting of KHLF-AMC. In some embodiments, the anti-KLK5 / KLK7 antibody inhibits the cleavage of KLK7 by KHLF-AMC, with an IC50 ranging from 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, and 0.1 nM to 0.9 nM. 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 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 Within the ranges of 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.

[0070] In some embodiments, the anti-KLK5 / KLK7 antibody described herein specifically binds to the active forms of KLK5, KLK7, or KLK5 and KLK7. In some embodiments, the anti-KLK5 / KLK7 antibody described herein does not bind to the inactive forms of KLK5, KLK7, or KLK5 and KLK7. In some embodiments, the anti-KLK5 / KLK7 antibody described herein specifically binds to the active sites of KLK5, KLK7, or KLK5 and KLK7. In some embodiments, the anti-KLK5 / KLK7 antibody described herein specifically binds to the active forms of KLK5, KLK7, or KLK5 and KLK7, but does not specifically bind to the inactive forms of KLK5, KLK7, or KLK5 and KLK7. In some embodiments, the anti-KLK5 / KLK7 antibody described herein detectably binds to the active form of KLK5, the active form of KLK7, or both active forms of KLK5 and active forms of KLK7, but undetectably binds to the inactive form of KLK5, the inactive form of KLK7, or both active forms of KLK5 and inactive forms of KLK7 under the same or equivalent conditions. The active site of KLK5 and / or KLK7 is the site at which the KLK5 and / or KLK7 substrate molecule binds for cleavage. The active site may also be referred to as a catalytic domain or catalytic triad. In some embodiments, the active site of KLK5 or KLK7 (i.e., the catalytic domain or catalytic triad) consists of the amino acids Ser195, His57, and Asp102 of KLK5 or KLK7 (see, for example, Goettig et al., Natural and synthetic inhibitors of kallikrein-related peptidases (KLKs)). Biochimie . November 2010; 92(11): 1546–1567).

[0071] In some embodiments, the antibodies described herein are optimized forms of parental antibodies (e.g., affinity-matured). In some embodiments, the antibodies described herein have a concentration of at least about 10... -4 M, at least about 10 -5 M, at least about 10 -6 M, at least about 10 -7 M, at least about 10 -8 M, at least about 10 -9 M, at least about 10 -10 M, at least about 10 -11 M, at least about 10 -12 M, at least about 10 -13 M or smaller binding affinity (e.g., as by K)D The antibody (as indicated) specifically binds to KLK5 (e.g., human or mouse KLK5) and KLK7 (e.g., human or mouse KLK7). In some embodiments, the antibody described herein is in 1×10 -10 M and 5×10 -9 Between M, 1×10 -10 M and 1×10 -9 Between M, 5×10 -10 With 1×10 -9 Between M, 5×10 -11 With 1×10 -10 M, 1×10 -11 With 5×10 - 10 Between M or 5×10 -13 With 1×10 -12 The binding affinity between M (e.g., as by K) D The antibodies of this disclosure can specifically bind to KLK5 (e.g., human or mouse KLK5) and KLK7 (e.g., human or mouse KLK7), as indicated. For example, the antibodies of this disclosure can bind to KLK5 proteins (e.g., human or mouse KLK5) and KLK7 (e.g., human or mouse KLK7) with an affinity between 1 pM and 500 nM, such as between 50 pM and 100 nM, 500 pM and 50 nM, 1 pM and 100 pM, 10 pM and 100 pM, 50 pM and 100 pM, 100 pM and 500 nM, 500 pM and 1 nM, 1 nM and 5 nM, 1 nM and 10 nM, 5 nM and 25 nM, 10 nM and 50 nM, 50 nM and 100 nM, and 100 nM and 500 nM. This disclosure also includes antibodies that compete with any of the antibodies described herein for binding to the 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 the antibodies can be tested using any suitable method, including but not limited to biosensor technologies (e.g., OCTET or BIACORE). In some embodiments, the antibodies described herein are expressed in sub-nanomolar K... D Combined with KLK5 and KLK7.

[0072] 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 spectroscopic methods (e.g., using fluorescence assays). 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 the bound protein as a function of the target protein concentration. The concentration of the bound protein ([[bound]]) is generally correlated with the concentration of the free target protein ([[free]]), as shown in the following formula: [[Binding]] = [[Free]] / (Kd + [[Free]]) However, it is not always necessary to determine K precisely. A Because sometimes you get K A A quantitative measurement of affinity proportional to the amount of affinity (e.g., determined using methods such as ELISA or FACS analysis) is sufficient and can therefore be used for comparison, such as determining whether a higher affinity is, for example, 2 times higher, to obtain a qualitative measurement of affinity, or to obtain an inference of affinity, such as by activity in a functional assay (e.g., in vitro or in vivo assay).

[0073] Tables 1a and 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) sequences, and CDR sequences).

[0074] Table 1a. Examples of anti-KLK5 / KLK7 antibodies

[0075] In some embodiments, certain amino acid positions in the antibody described herein (e.g., amino acids in the VH / VL region and / or CDR region) are substituted, and this substitution results in the antibody having substantially similar binding and biological activities to a reference antibody (e.g., substantially similar binding affinity, binding specificity, protease activity inhibitory activity, anti-inflammatory activity, or combinations thereof). To identify substituted positions of an antibody, the amino acid sequence of that antibody is compared with the sequences of other antibodies belonging to the same group as that antibody. If the identity of that amino acid changes at any particular position among different related antibodies in a group, that position is a substituted position of that antibody. In other words, a substituted position is a position where the identity of the amino acid differs between related antibodies. Positions containing constant amino acids are non-substituted positions.

[0076] In some embodiments, the above method can be used to provide a shared antibody sequence. In such a shared sequence, non-substitutable positions are indicated by the amino acids present at those positions, and substitutable positions are indicated by "X".

[0077] Depending on how the antibody is used, X can be a) any amino acid, b) any amino acid present at that position in any of the relevant antibodies in the group, or a variant thereof with a conserved substitution, or c) any amino acid present at that position in any of the relevant antibodies in the group. Any antibody having a sequence covered by a common denominator should bind to the same antigen as any of the relevant antibodies.

[0078] In some embodiments, the above-described methods can be used in the design and preparation of variants of the parent antibody that at least maintain (e.g., maintain or increase) the antigen-binding activity of the parent antibody. Because antibodies containing substitutions at substituted sites have already been produced and tested, substitutions at those sites can be performed knowing that they will not significantly reduce the binding activity of the antibody. Typically, the antigen-binding affinity of the antibody variant of the parent antibody 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% of the binding affinity of the parent antibody to a specific antigen (e.g., at least 150%, at least 200%, at least 500%, at least 1000%, typically up to at least 10,000%).

[0079] In some embodiments, the substitutable position of the parent antibody can be substituted by: a) any of 20 naturally occurring amino acids to produce a random substitution; b) an amino acid having similar biochemical properties to an amino acid already present at the substitutable position to produce a conserved substitution; c) an amino acid present at the same position as in the associated 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. Substitution can occur at any portion of the antibody variable region (including any frame region or CDR). In some 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 a particular embodiment, the type of substitution that can occur at each substitutable position can be indicated by the type of amino acid present at that position in the associated antibody. For example, if an unrelated amino acid (e.g., Ala, Gly, Cys, Glu, and Thr) is present at a position in a group of associated antibodies, then any amino acid can be substituted at that position without significantly reducing the antibody binding activity. Exemplary amino acid substitutions for the anti-KLK5 / KLK7 antibodies described herein are shown in Table 1b: Table 1b. Exemplary amino acid substitutions in anti-KLK5 / KLK7 antibodies

[0080] In some embodiments, the antibodies of this disclosure comprise HC CDR1, which comprises the amino acid sequence GSISSX1DYYWX2 (SEQ ID NO: 28), wherein X1 is S, D, or L, and X2 is G or V; HC CDR2, which comprises the amino acid sequence SIX3YX4X5X6TYYX7PSLKS (SEQ ID NO: 29), wherein X3 is Y or D, X4 is S, F, or Y, X5 is G or A, X6 is S or D, or X7 is N or S; HC CDR3, which comprises the amino acid sequence ARGRPLGYGAX8HX9YYGMDV (SEQ ID NO: 30), wherein X8 is R or K, or X9 is Y or D; LC CDR1, which comprises the amino acid sequence SEQ ID NO: 4; LC CDR2, which comprises the amino acid sequence SEQ ID NO: 5; and / or LC CDR3, which comprises QQSPX 10 The amino acid sequence of FPPLT (SEQ ID NO: 31), where X 10 It is either P or Y.

[0081] In some embodiments, the antibodies of this disclosure comprise one or more of the HC CDR (e.g., HC CDR1, HC CDR2, or HC CDR3) amino acid sequences selected from any of the anti-KLK5 / KLK7 antibodies selected from Tables 1a and 1b. In some embodiments, the antibodies of this disclosure comprise the HC CDR3 amino acid sequence selected from any of the anti-KLK5 / KLK7 antibodies selected from Tables 1a and 1b. In some embodiments, the antibodies of this disclosure comprise the HC CDR1, HC CDR2, and HC CDR3 as provided for any of the antibodies selected from Tables 1a and 1b. In some embodiments, the antibodies of this disclosure comprise the LC CDR3 amino acid sequence selected from any of the anti-KLK5 / KLK7 antibodies selected from Tables 1a and 1b. In some embodiments, the antibodies of this disclosure comprise one or more of the LC CDR (e.g., LC CDR1, LC CDR2, or LC CDR3) amino acid sequences selected from any of the anti-KLK5 / KLK7 antibodies selected from Tables 1a and 1b. In some embodiments, the antibodies disclosed herein comprise LC CDR1, LC CDR2, and LC CDR3 as provided for any of the anti-KLK5 antibodies selected from Tables 1a and 1b.

[0082] In some embodiments, the antibodies of this disclosure comprise HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and LC CDR3 as provided for any of the anti-KLK5 / KLK7 antibodies selected from Tables 1a and 1b. In some embodiments, the CDR3 domains of the antibody heavy chain and / or light chain may play a particularly important role in the antibody's binding specificity / affinity to the antigen. Therefore, the antibodies of this disclosure may at least comprise the heavy chain and / or light chain CDR3 of any of the anti-KLK5 / KLK7 antibodies selected from Tables 1a and 1b.

[0083] Variants of any of the exemplary anti-KLK5 / KLK7 antibodies disclosed herein are also within the scope of this disclosure. Relative to the reference antibody, the variant may contain one or more amino acid residue variations in VH and / or VL, or in one or more of the HC CDR and / or one or more of the LC CDR, while retaining substantially similar binding and biological activities to the reference antibody (e.g., substantially similar binding affinity, binding specificity, protease activity, inhibitory activity, anti-inflammatory activity, or combinations thereof).

[0084] In some embodiments, the antibody of this disclosure has one or more CDR (e.g., HC CDR or LC CDR) sequences that are substantially similar to any of the HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2 and / or LC CDR3 sequences selected from the anti-KLK5 / KLK7 antibodies of Tables 1a and 1b. In some embodiments, the positions 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) regions of the antibody described herein may vary by one, two, three, four, five or six amino acid positions, as 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 maintaining, for example, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% binding to the original antibody from which it is derived). For example, in some embodiments, the position of the 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 of the antibodies described herein, as 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 maintaining at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% of the binding to 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, LCCDR2, or LC CDR3) regions of the antibody described herein may vary (e.g., shorten or lengthen) by one, two, three, four, five, or more amino acids, as long as immune-specific binding to KLK5 (e.g., human or mouse KLK5) and KLK7 (e.g., human or mouse KLK7) is maintained (e.g., substantially maintaining at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% of the binding to the original antibody from which it is derived).

[0085] Therefore, in some embodiments, the HC CDR1, HC CDR2, HC CDR3, LC CDR1, LCCDR2 and / or LC CDR3 described herein may be one, two, three, four, five or more amino acids shorter than one or more of the CDRs described herein (e.g., CDRs from any of the anti-KLK5 / KLK7 antibodies selected from Tables 1a and 1b), as 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 maintaining binding relative to the original antibody from which it is derived, for example, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%). In some embodiments, the HC CDR1, HC CDR2, HC CDR3, LCCDR1, LC CDR2 and / or LC CDR3 described herein may be one, two, three, four, five or more amino acids longer than one or more of the CDRs described herein (e.g., CDRs from any of the anti-KLK5 / KLK7 antibodies selected from Tables 1a and 1b), as 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 maintaining binding relative to the original antibody from which it is derived, for example, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%). In some embodiments, the amino moiety of the HC CDR1, HC CDR2, HCCDR3, LC CDR1, LC CDR2 and / or LC CDR3 described herein may be extended by one, two, three, four, five or more amino acids compared to one or more of the CDRs described herein (e.g., CDRs from any of the anti-KLK5 / KLK7 antibodies selected from Tables 1a and 1b), as 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 maintaining binding relative to the original antibody from which it is derived, for example, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%).In some embodiments, the carboxyl moiety of the HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2 and / or LC CDR3 described herein may be extended by one, two, three, four, five or more amino acids compared to one or more of the CDRs described herein (e.g., CDRs from any of the anti-KLK5 / KLK7 antibodies selected from Tables 1a and 1b), as 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 maintaining binding relative to the original antibody from which it is derived, for example, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%). In some embodiments, the amino moiety of the HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2 and / or LC CDR3 described herein may be shortened by one, two, three, four, five or more amino acids compared to one or more of the CDRs described herein (e.g., CDRs from any of the anti-KLK5 / KLK7 antibodies selected from Tables 1a and 1b), as 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 maintaining binding relative to the original antibody from which it is derived, for example, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%). In some embodiments, the carboxyl moiety of the HC CDR1, HC CDR2, HC CDR3, LCCDR1, LC CDR2, and / or LC CDR3 described herein may be shortened by one, two, three, four, five, or more amino acids compared to one or more of the CDRs described herein (e.g., CDRs from any of the anti-KLK5 / KLK7 antibodies selected from Tables 1a and 1b), as 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 maintaining binding relative to the original antibody from which it is derived, for example, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%). Any method may be used to determine whether specific binding to KLK5 (e.g., human or mouse KLK5) and KLK7 (e.g., human or mouse KLK7) is maintained, for example, using binding assays and conditions described in the art.

[0086] In some embodiments, the antibodies of this disclosure have one or more CDR (e.g., HC CDR or LC CDR) sequences that are substantially similar to any of the anti-KLK5 / KLK7 antibodies selected from Tables 1a and 1b. For example, the antibodies described herein may include one or more CDR sequences from any of the anti-KLK5 / KLK7 antibodies selected from Tables 1a and 1b, which contain up to 5, 4, 3, 2, or 1 amino acid residue variations compared to the corresponding CDR region in any of the CDRs provided herein (e.g., CDRs from any of the anti-KLK5 / KLK7 antibodies selected from Tables 1a and 1b), provided that specific binding to KLK5 (e.g., human or mouse KLK5) and KLK7 (e.g., human or mouse KLK7) is maintained (e.g., substantially maintaining binding relative to the original antibody from which it is derived, for example, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%). In some embodiments, any amino acid variation in any of the CDRs provided herein may be a conserved variation. A conserved variation may be introduced into a CDR at a location where residues are unlikely to interact with KLK5 (e.g., human or mouse KLK5) and / or KLK7 (e.g., human or mouse KLK7), for example, as determined based on the crystal structure. Some aspects of this disclosure provide antibodies comprising one or more of the heavy chain variable (VH) and / or light chain variable (VL) domains provided herein. In some embodiments, any of the VH domains provided herein includes one or more of the HC CDR sequences (e.g., HC CDR1, HC CDR2, and HC CDR3) provided herein, for example, any of the HC CDR sequences provided in any of the anti-KLK5 / KLK7 sequences selected from Tables 1a and 1b. In some embodiments, any of the VL domains provided herein includes one or more of the LC CDR sequences (e.g., LC CDR1, LC CDR2, and LC CDR3) provided herein, for example, any of the HC CDR sequences provided in any of the anti-KLK5 / KLK7 antibodies selected from Tables 1a and 1b.

[0087] In some embodiments, the antibodies of this disclosure include any antibody comprising a heavy chain variable domain and / or a light chain variable domain of any of the anti-KLK5 / KLK7 antibodies selected from Tables 1a and 1b and their variants. In some embodiments, the antibodies of this disclosure include any antibody comprising a heavy chain variable pair and a light chain variable pair of any of the anti-KLK5 / KLK7 antibodies selected from Tables 1a and 1b.

[0088] This disclosure provides antibodies having heavy chain variable (VH) and / or light chain variable (VL) domain amino acid sequences homologous to any of the amino acid sequences described herein. In some embodiments, the antibody comprises a heavy chain variable sequence or light chain variable sequence having at least 75% (e.g., at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity to any of the anti-KLK5 / KLK7 antibodies selected from Tables 1a and 1b. In some embodiments, the homologous heavy chain variable amino acid sequence and / or light chain variable amino acid sequence remains unchanged within any of the CDR sequences provided herein. For example, in some embodiments, the degree of sequence variation (e.g., at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) may appear in heavy chain variable sequences and / or light chain variable sequences other than any of the CDR sequences provided herein. In some embodiments, the antibodies provided herein comprise heavy chain variable sequences and light chain variable sequences containing a frame sequence having at least 75% (e.g., at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with any anti-KLK5 / KLK7 antibody selected from Tables 1a and 1b.

[0089] In some embodiments, the antibodies of this disclosure are humanized antibodies (e.g., humanized variants containing one or more CDRs of Tables 1a and 1b). In some embodiments, the antibodies of this disclosure comprise the same HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and LC CDR3 as shown in Tables 1a and 1b, and comprise humanized heavy chain variable regions and / or humanized light chain variable regions.

[0090] In some embodiments, the antibody of this 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 variation) compared to the VH of any anti-KLK5 / KLK7 antibody listed in Tables 1a and 1b. Alternatively, the antibody of this 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 variation) compared to the VL of any anti-KLK5 / KLK7 antibody listed in Tables 1a and 1b.

[0091] In some embodiments, the anti-KLK5 / KLK7 antibody of this disclosure comprises HC CDR1, HC CDR2, and HC CDR3 having a heavy chain variable domain of the amino acid sequence of SEQ ID NO: 7. Alternatively, the anti-KLK5 / KLK7 antibody of this disclosure comprises LC CDR1, LC CDR2, and LC CDR3 having a light chain variable domain of the amino acid sequence of SEQ ID NO: 8.

[0092] In some embodiments, the anti-KLK5 / KLK7 antibody of this disclosure comprises 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.

[0093] In some embodiments, the anti-KLK5 / KLK7 antibody of this disclosure comprises HC CDR1, HC CDR2, and HC CDR3, which together contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variation) compared to HC CDR1 having the amino acid sequence of SEQ ID NO: 1, HC CDR2 having the amino acid sequence of SEQ ID NO: 2, and HC CDR3 having the amino acid sequence of SEQ ID NO: 3. As used anywhere in this disclosure, “total” means that the total number of amino acid variations in all three heavy chain CDRs is within the defined range. Alternatively, the anti-KLK5 / KLK7 antibody of this disclosure comprises LC CDR1, LC CDR2, and LC CDR3, which together contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variation) compared to 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.

[0094] In some embodiments, the anti-KLK5 / KLK7 antibody of this disclosure comprises HC CDR1, HC CDR2, and HC CDR3, all of which have at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with HC CDR1 having the amino acid sequence of SEQ ID NO: 1, HC CDR2 having the amino acid sequence of SEQ ID NO: 2, and HC CDR3 having the amino acid sequence of SEQ ID NO: 3. Alternatively, the anti-KLK5 / KLK7 antibody of this disclosure comprises LC CDR1, LCCDR2, and LC CDR3, all of which have at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with 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.

[0095] In some embodiments, the anti-KLK5 / KLK7 antibody of this disclosure comprises: HC CDR1 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to HC CDR1 having the amino acid sequence of SEQ ID NO: 1; HC CDR2 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to HC CDR2 having the amino acid sequence of SEQ ID NO: 2; and / or HCCDR3 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to HC CDR3 having the amino acid sequence of SEQ ID NO: 3. Alternatively, the anti-KLK5 / KLK7 antibody of this disclosure comprises: an LC CDR1 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to an LC CDR1 having the amino acid sequence of SEQ ID NO: 4; an LC CDR2 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to an LC CDR2 having the amino acid sequence of SEQ ID NO: 5; and / or an LC CDR3 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to an LC CDR3 having the amino acid sequence of SEQ ID NO: 6.

[0096] In some embodiments, the anti-KLK5 / KLK7 antibody of this disclosure comprises VH containing the amino acid sequence of SEQ ID NO: 7. Alternatively, the anti-KLK5 / KLK7 antibody of this disclosure comprises VL containing the amino acid sequence of SEQ ID NO: 8.

[0097] In some embodiments, the anti-KLK5 / KLK7 antibody of this 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 variation) compared to the VH shown in SEQ ID NO: 7. Alternatively, the anti-KLK5 / KLK7 antibody of this 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 variation) compared to the VL shown in SEQ ID NO: 8.

[0098] In some embodiments, the anti-KLK5 / KLK7 antibody of this disclosure comprises VH, said VH comprising an amino acid sequence having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the VH shown in SEQ ID NO: 7. Alternatively, the anti-KLK5 / KLK7 antibody of this disclosure comprises VL, said VL comprising an amino acid sequence having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the VL shown in SEQ ID NO: 8.

[0099] In some embodiments, the anti-KLK5 / KLK7 antibody of this disclosure comprises heavy chain variable domains HC CDR1, HC CDR2, and HC CDR3, having the amino acid sequence of SEQ ID NO: 13. Alternatively, the anti-KLK5 / KLK7 antibody of this disclosure comprises light chain variable domains LC CDR1, LC CDR2, and LC CDR3, having the amino acid sequence of SEQ ID NO: 14.

[0100] In some embodiments, the anti-KLK5 / KLK7 antibody of this disclosure comprises 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.

[0101] In some embodiments, the anti-KLK5 / KLK7 antibody of this disclosure comprises HC CDR1, HC CDR2, and HC CDR3, which together contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variation) compared to HC CDR1 having the amino acid sequence of SEQ ID NO: 9, HC CDR2 having the amino acid sequence of SEQ ID NO: 10, and HC CDR3 having the amino acid sequence of SEQ ID NO: 11. As used anywhere in this disclosure, “total” means the total number of amino acid variations in all three heavy chain CDRs is within the specified range. Alternatively, the anti-KLK5 / KLK7 antibody of this disclosure comprises LC CDR1, LC CDR2, and LC CDR3, which together contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variation) compared to 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.

[0102] In some embodiments, the anti-KLK5 / KLK7 antibody of this disclosure comprises HC CDR1, HC CDR2, and HC CDR3, all of which have at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with HC CDR1 having the amino acid sequence of SEQ ID NO: 9, HC CDR2 having the amino acid sequence of SEQ ID NO: 10, and HC CDR3 having the amino acid sequence of SEQ ID NO: 11. Alternatively, the anti-KLK5 / KLK7 antibody of this disclosure comprises LC CDR1, LCCDR2, and LC CDR3, all of which have at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with 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.

[0103] In some embodiments, the anti-KLK5 / KLK7 antibody of this disclosure comprises: HC CDR1 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to HC CDR1 having the amino acid sequence of SEQ ID NO: 9; HC CDR2 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to HC CDR2 having the amino acid sequence of SEQ ID NO: 10; and / or HCCDR3 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to HC CDR3 having the amino acid sequence of SEQ ID NO: 11. Alternatively, the anti-KLK5 / KLK7 antibody of this disclosure comprises: an LC CDR1 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to an LC CDR1 having the amino acid sequence of SEQ ID NO: 4; an LC CDR2 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to an LC CDR2 having the amino acid sequence of SEQ ID NO: 5; and / or an LC CDR3 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to an LC CDR3 having the amino acid sequence of SEQ ID NO: 12.

[0104] In some embodiments, the anti-KLK5 / KLK7 antibody of this disclosure comprises VH containing the amino acid sequence of SEQ ID NO: 13. Alternatively, the anti-KLK5 / KLK7 antibody of this disclosure comprises VL containing the amino acid sequence of SEQ ID NO: 14.

[0105] In some embodiments, the anti-KLK5 / KLK7 antibody of this 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 variation) compared to the VH shown in SEQ ID NO: 13. Alternatively, the anti-KLK5 / KLK7 antibody of this 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 variation) compared to the VL shown in SEQ ID NO: 14.

[0106] In some embodiments, the anti-KLK5 / KLK7 antibody of this disclosure comprises VH, said VH comprising an amino acid sequence having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the VH shown in SEQ ID NO: 13. Alternatively, the anti-KLK5 / KLK7 antibody of this disclosure comprises VL, said VL comprising an amino acid sequence having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the VL shown in SEQ ID NO: 14.

[0107] In some embodiments, the anti-KLK5 / KLK7 antibody of this disclosure comprises heavy chain variable domains HC CDR1, HC CDR2, and HC CDR3, having the amino acid sequence of SEQ ID NO: 17. Alternatively, the anti-KLK5 / KLK7 antibody of this disclosure comprises light chain variable domains LC CDR1, LC CDR2, and LC CDR3, having the amino acid sequence of SEQ ID NO: 14.

[0108] In some embodiments, the anti-KLK5 / KLK7 antibody of this disclosure comprises 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.

[0109] In some embodiments, the anti-KLK5 / KLK7 antibody of this disclosure comprises HC CDR1, HC CDR2, and HC CDR3, which together contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variation) compared to HC CDR1 having the amino acid sequence of SEQ ID NO: 9, HC CDR2 having the amino acid sequence of SEQ ID NO: 15, and HC CDR3 having the amino acid sequence of SEQ ID NO: 16. As used anywhere in this disclosure, “total” means the total number of amino acid variations in all three heavy chain CDRs is within the specified range. Alternatively, the anti-KLK5 / KLK7 antibody of this disclosure comprises LC CDR1, LC CDR2, and LC CDR3, which together contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variation) compared to 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.

[0110] In some embodiments, the anti-KLK5 / KLK7 antibody of this disclosure comprises HC CDR1, HC CDR2, and HC CDR3, all of which have at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with HC CDR1 having the amino acid sequence of SEQ ID NO: 9, HC CDR2 having the amino acid sequence of SEQ ID NO: 15, and HC CDR3 having the amino acid sequence of SEQ ID NO: 16. Alternatively, the anti-KLK5 / KLK7 antibody of this disclosure comprises LC CDR1, LCCDR2, and LC CDR3, all of which have at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with 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.

[0111] In some embodiments, the anti-KLK5 / KLK7 antibody of this disclosure comprises: HC CDR1 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to HC CDR1 having the amino acid sequence of SEQ ID NO: 9; HC CDR2 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to HC CDR2 having the amino acid sequence of SEQ ID NO: 15; and / or HCCDR3 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to HC CDR3 having the amino acid sequence of SEQ ID NO: 16. Alternatively, the anti-KLK5 / KLK7 antibody of this disclosure comprises: an LC CDR1 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to an LC CDR1 having the amino acid sequence of SEQ ID NO: 4; an LC CDR2 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to an LC CDR2 having the amino acid sequence of SEQ ID NO: 5; and / or an LC CDR3 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to an LC CDR3 having the amino acid sequence of SEQ ID NO: 12.

[0112] In some embodiments, the anti-KLK5 / KLK7 antibody of this disclosure comprises VH containing the amino acid sequence of SEQ ID NO: 17. Alternatively, the anti-KLK5 / KLK7 antibody of this disclosure comprises VL containing the amino acid sequence of SEQ ID NO: 14.

[0113] In some embodiments, the anti-KLK5 / KLK7 antibody of this 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 variation) compared to the VH shown in SEQ ID NO: 13. Alternatively, the anti-KLK5 / KLK7 antibody of this 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 variation) compared to the VL shown in SEQ ID NO: 14.

[0114] In some embodiments, the anti-KLK5 / KLK7 antibody of this disclosure comprises a VH containing an amino acid sequence having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the VH shown in SEQ ID NO: 17. Alternatively, the anti-KLK5 / KLK7 antibody of this disclosure comprises a VL containing an amino acid sequence having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the VL shown in SEQ ID NO: 14.

[0115] In some embodiments, the anti-KLK5 / KLK7 antibody of this disclosure comprises heavy chain variable domains HC CDR1, HC CDR2, and HC CDR3, having the amino acid sequence of SEQ ID NO: 21. Alternatively, the anti-KLK5 / KLK7 antibody of this disclosure comprises light chain variable domains LC CDR1, LC CDR2, and LC CDR3, having the amino acid sequence of SEQ ID NO: 14.

[0116] In some embodiments, the anti-KLK5 / KLK7 antibody of this disclosure comprises 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.

[0117] In some embodiments, the anti-KLK5 / KLK7 antibody of this disclosure comprises HC CDR1, HC CDR2, and HC CDR3, which together contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variation) compared to HC CDR1 having the amino acid sequence of SEQ ID NO: 18, HC CDR2 having the amino acid sequence of SEQ ID NO: 19, and HC CDR3 having the amino acid sequence of SEQ ID NO: 20. As used anywhere in this disclosure, “total” means the total number of amino acid variations in all three heavy chain CDRs is within the specified range. Alternatively, the anti-KLK5 / KLK7 antibody of this disclosure comprises LC CDR1, LC CDR2, and LC CDR3, which together contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variation) compared to 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.

[0118] In some embodiments, the anti-KLK5 / KLK7 antibody of this disclosure comprises HC CDR1, HC CDR2, and HC CDR3, all of which have at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with HC CDR1 having the amino acid sequence of SEQ ID NO: 18, HC CDR2 having the amino acid sequence of SEQ ID NO: 19, and HC CDR3 having the amino acid sequence of SEQ ID NO: 20. Alternatively, the anti-KLK5 / KLK7 antibody of this disclosure comprises LC CDR1, LCCDR2, and LC CDR3, all of which have at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with 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.

[0119] In some embodiments, the anti-KLK5 / KLK7 antibody of this disclosure comprises: HC CDR1 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to HC CDR1 having the amino acid sequence of SEQ ID NO: 18; HC CDR2 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to HC CDR2 having the amino acid sequence of SEQ ID NO: 19; and / or HCCDR3 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to HC CDR3 having the amino acid sequence of SEQ ID NO: 20. Alternatively, the anti-KLK5 / KLK7 antibody of this disclosure comprises: an LC CDR1 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to an LC CDR1 having the amino acid sequence of SEQ ID NO: 4; an LC CDR2 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to an LC CDR2 having the amino acid sequence of SEQ ID NO: 5; and / or an LC CDR3 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to an LC CDR3 having the amino acid sequence of SEQ ID NO: 12.

[0120] In some embodiments, the anti-KLK5 / KLK7 antibody of this disclosure comprises VH containing the amino acid sequence of SEQ ID NO: 21. Alternatively, the anti-KLK5 / KLK7 antibody of this disclosure comprises VL containing the amino acid sequence of SEQ ID NO: 14.

[0121] In some embodiments, the anti-KLK5 / KLK7 antibody of this 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 variation) compared to the VH shown in SEQ ID NO: 21. Alternatively, the anti-KLK5 / KLK7 antibody of this 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 variation) compared to the VL shown in SEQ ID NO: 14.

[0122] In some embodiments, the anti-KLK5 / KLK7 antibody of this disclosure comprises VH, said VH comprising an amino acid sequence having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the VH shown in SEQ ID NO: 21. Alternatively, the anti-KLK5 / KLK7 antibody of this disclosure comprises VL, said VL comprising an amino acid sequence having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the VL shown in SEQ ID NO: 14.

[0123] The antibodies described herein can be in any antibody form, including but not limited to full-length antibodies, their antigen-binding fragments (such as Fab, F(ab'), F(ab')2, Fv), single-chain antibodies, bispecific antibodies, or nanobodies. In some embodiments, the anti-KLK5 / KLK7 antibody described herein is an scFv. In some embodiments, the anti-KLK5 / KLK7 antibody described herein is an scFv-Fab (e.g., an scFv fused to a portion of a constant region).

[0124] In some embodiments, the anti-KLK5 / KLK7 antibody of this disclosure is a chimeric antibody, which may include a heavy constant region and a light constant region derived from a human antibody. A chimeric antibody is an antibody having a variable region or a portion of a variable region from a first species and a constant region 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., non-human mammals such as mice, rabbits, and rats), while the constant portion is sequence homologous to an antibody derived from another mammal (such as a human). In some embodiments, amino acid modifications may be made in the variable and / or constant regions.

[0125] In some embodiments, the antibodies of this disclosure comprise a VL domain and / or a VH domain selected from any of the anti-KLK5 / KLK7 antibodies listed in Tables 1a and 1b, and include a constant region comprising the amino acid sequence of the constant region of an immunoglobulin molecule of IgG, IgE, IgM, IgD, IgA, or IgY, any class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2) or any subclass (e.g., IgG2a and IgG2b). Non-limiting examples of human constant regions have been described in the art, see, for example, Kabat EA et al., (1991), ibid.

[0126] In some embodiments, the light chain of either of the anti-KLK5 / KLK7 antibodies described herein may further include a light chain constant region (CL), which may be any CL known in the art. In some instances, the CL is a κ light chain. In other instances, the CL is a λ light chain. In some embodiments, the CL is a κ light chain.

[0127] Other antibody heavy chain constant regions and light chain constant regions are well known in the art, for example, those provided in the IMGT database (www.imgt.org) or www.vbase2.org / vbstat.php, both of which are incorporated herein by reference.

[0128] In some embodiments, conserved mutations may be introduced into antibody sequences (e.g., CDRs or framework sequences) at sites where residues are unlikely to interact with the target antigen (e.g., human or mouse KLK5 and / or human or mouse KLK7), for example, as determined by crystal structure. In some embodiments, one, two, or more mutations (e.g., amino acid substitutions) are introduced into the Fc region of the anti-KLK5 / KLK7 antibody described herein (e.g., in the CH2 domain (residues 231-340 of human IgG1) and / or the CH3 domain (residues 341-447 of human IgG1) and / or the hinge region, which are numbered according to the Kabat numbering system (e.g., Kabat's EU index)) to alter one or more functional properties of the antibody, such as serum half-life, complement binding, Fc receptor binding, and / or antigen-dependent cytotoxicity.

[0129] 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, for example, in U.S. Patent No. 5,677,425. The number of cysteine ​​residues in the hinge region of the CH1 domain can be altered to, for example, promote the assembly of light and heavy chains, or alter (e.g., increase or decrease) antibody stability, or promote linker conjugation.

[0130] In some embodiments, one, two, or more mutations (e.g., amino acid substitutions) are introduced into the Fc region of the antibody described herein (e.g., in the CH2 domain (residues 231-340 of human IgG1) and / or the CH3 domain (residues 341-447 of human IgG1) and / or the hinge region, where numbering is performed according to the Kabat numbering system (e.g., Kabat's EU index)) to increase or decrease the antibody's affinity for Fc receptors (e.g., activated Fc receptors) on the surface of effector cells. Mutations in the Fc region of antibodies that decrease or increase the antibody's affinity for Fc receptors, and techniques for introducing such mutations into Fc receptors or fragments thereof, are known to those skilled in the art. Examples of mutations in the Fc receptor of antibodies that can alter the affinity of antibodies for the Fc receptor are described, for example, in Smith P et al., (2012) PNAS 109: 6181-6186, U.S. Patent No. 6,737,056, and International Patent Publications Nos. WO 02 / 060919, WO 98 / 23289, and WO 97 / 34631, which are incorporated herein by reference.

[0131] In some embodiments, one, two, or more amino acid mutations (i.e., substitution, insertion, or deletion) are introduced into the constant domain of IgG or its FcRn binding fragment (preferably an Fc or hinge-Fc domain fragment) to alter (e.g., reduce or increase) the half-life of the antibody in vivo. Examples of mutations that alter (e.g., reduce or increase) the half-life of the antibody in vivo can be found, for example, International Publications WO 02 / 060919, WO 98 / 23289, and WO 97 / 34631, and U.S. Patents 5,869,046, 6,121,022, 6,277,375, and 6,165,745.

[0132] In some embodiments, one, two, or more amino acid mutations (i.e., substitution, insertion, or deletion) are introduced into the constant domain of IgG or its FcRn binding fragment (preferably Fc or hinge-Fc domain fragment) to reduce the half-life of the anti-KLK5 / KLK7 antibody in vivo. In some embodiments, one, two, or more amino acid mutations (i.e., substitution, insertion, or deletion) are introduced into the constant domain of IgG or its FcRn binding fragment (preferably Fc or hinge-Fc domain fragment) to increase the half-life of the antibody in vivo. In some embodiments, the antibody may have one or more amino acid mutations (e.g., substitution) in the second constant (CH2) domain (residues 231-340 of human IgG1) and / or the third constant (CH3) domain (residues 341-447 of human IgG1), which are numbered according to Kabat's EU index (Kabat EA et al., (1991), ibid.). In some embodiments, the constant region of the antibody IgG1 described herein comprises a methionine (M) to tyrosine (Y) substitution at position 252, a serine (S) to threonine (T) substitution at position 254, and a threonine (T) to glutamic acid (E) substitution at position 256, numbered according to the Kabat EU index. See U.S. Patent No. 7,658,921, which is incorporated herein by reference. This type of mutant IgG, referred to as the “YTE mutant,” has been shown to exhibit a fourfold increase in half-life compared to the wild-type of the same antibody (see Dall'Acqua WF 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 at positions 251-257, 285-290, 308-314, 385-389, and 428-436, numbered according to the Kabat EU index.

[0133] In some implementations, for example by introducing M428L and / or N434A substitutions, the antibody comprises an Fc region that has been engineered for the purpose of extending the half-life. Non-limiting examples of Fc variants affecting the half-life in circulation are provided in 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.

[0134] In some embodiments, for example, by introducing Leu234Ala and Leu235Ala mutations (commonly referred to as LALA mutations), one, two, or more amino acid substitutions are introduced into the Fc region of the IgG constant domain to alter the effector function of anti-KLK5 / KLK7 antibodies. The effector ligand whose affinity is altered can be, for example, an Fc receptor or a C1 component of complement. This approach is described in more detail in U.S. Patent Nos. 5,624,821 and 5,648,260. In some embodiments, the deletion or inactivation of the constant domain (through point mutations or other means) can reduce Fc receptor binding of circulating antibodies, thereby increasing tumor localization. For a description of mutations that delete or inactivate the constant domain and thus increase tumor localization, see, for example, U.S. Patent Nos. 5,585,097 and 8,591,886. In some implementations, one or more amino acid substitutions may be introduced into the Fc region of the antibody described herein to remove potential glycosylation sites on the Fc region, which can reduce Fc receptor binding (see, for example, Shields RL et al., (2001) J Biol Chem 276: 6591-604).

[0135] In some embodiments, one or more amino groups in the constant region of the anti-KLK5 / KLK7 antibody described herein may be replaced with different amino acid residues, such that the antibody exhibits altered Clq binding and / or reduced or eliminated complement-dependent cytotoxicity (CDC). This method is described in more 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 the antibody described herein are altered, thereby changing the antibody's ability to fix complement. This method is further described in International Publication No. WO 94 / 29351. In some embodiments, the Fc region of the antibody described herein is modified to increase the antibody's ability to mediate antibody-dependent cytotoxicity (ADCC) and / or increase the antibody's affinity for the Fcγ receptor. This method is further described in International Publication No. WO 00 / 42072.

[0136] In some embodiments, the antibody comprises an Fc variant containing amino acid substitutions for L234A, L235E, and P329G, which are numbered according to the EU index. In some embodiments, antibodies comprising Fc variants exhibit reduced affinity for one or more, or each, of FcyRJ, FcyRIIA, FcyRIIIA, and Clq compared to antibodies comprising the 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 REDUCEDEFFECTOR 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.

[0137] In some embodiments, the heavy chain domain sequence and / or light chain variable domain sequence of the antibodies provided herein can be used to generate, for example, CDR-grafted, chimeric, humanized, or complexed human antibodies or antigen-binding fragments, as described elsewhere herein. As will be understood by those skilled in the art, any variant, CDR-grafted, chimeric, humanized, or complexed antibody 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 to KLK5 and KLK7 such that the variant, CDR-grafted, chimeric, humanized, or complexed antibody has at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or more binding to KLK5 and KLK7 relative to the original antibody from which it is derived.

[0138] In some embodiments, the antibodies provided herein contain mutations that confer the desired properties to the antibody. For example, to avoid potential complications due to Fab-arm exchange (which is known to occur in natural IgG4 mAb), the antibodies provided herein may contain a stable 'Adair' mutation (ngal 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 number; residue 241 Kabat number) is converted to proline, producing an IgG1-like hinge sequence. Therefore, any of the antibodies may contain a stable 'Adair' mutation.

[0139] In some embodiments, the antibody is modified, for example, by glycosylation, phosphorylation, SUMOylation, and / or methylation. In some embodiments, the antibody is a glycosylated antibody 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 via N-glycosylation, O-glycosylation, C-glycosylation, glypiation (GPI anchoring), and / or phosphoglycosylation. In some embodiments, the one or more sugar or carbohydrate molecules are monosaccharides, disaccharides, oligosaccharides, or polysaccharides. In some embodiments, the one or more sugar or carbohydrate molecules are branched oligosaccharides or branched polysaccharides. In some embodiments, the one or more sugar or carbohydrate molecules include mannose units, glucose units, N-acetylglucosamine units, N-acetylglucosamine units, galactose units, fucose units, or phospholipid units. In some embodiments, about 1-10, about 1-5, about 5-10, about 1-4, about 1-3, or about 2 sugar molecules are present. In some embodiments, the glycosylated antibody is fully or partially glycosylated. In some embodiments, the antibody is glycosylated by a chemical reaction or enzymatic process. In some embodiments, the antibody is glycosylated in vitro or intracellularly, and the cells may optionally lack enzymes in the N-glycosylation or O-glycosylation pathway, such as glycosyltransferases. In some embodiments, the antibody is functionalized with sugar or carbohydrate molecules, as described in the following literature: International Patent Application Publication WO2014065661, published May 1, 2014, entitled "Modified antibody, antibody-conjugate and process for the preparation thereof".

[0140] In some embodiments, any of the anti-KLK5 / KLK7 antibodies described herein may include 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 chain and light chain sequences, or any of the F(ab') heavy chain and light chain sequences, and further comprise a signal peptide (e.g., an N-terminal signal peptide).

[0141] (b) IL-13 / IL-13R targeting antibody Disorders associated with barrier dysfunction are characterized by barrier disruption and T helper cell 2 (Th2)-driven inflammation, such as inflammation mediated by Th2 cytokines and their receptors, including IL-13, IL-13 receptor (IL-13R), IL-4, IL-4R, IL-5, IL-5R, IL-6, IL-6R, IL-9, IL-9R, IL-31, IL-31R, IL-17E (IL-25), and IL-17ER (IL-25R). In some cases, unregulated KLK activity in the epidermis leads to and / or is associated with Th2 cell activation, resulting in the secretion of Th2 cytokines. Elevated Th2 cytokine levels may further lead to KLK overactivation. Figure 2 Therapeutic antibodies have been developed to inhibit Th2 cytokine-mediated inflammation by targeting Th2 cytokines and / or their respective receptors (called Th2-targeting antibodies, such as anti-IL-13 antibodies or anti-IL-13R antibodies).

[0142] In some embodiments, the antibodies provided herein comprise at least one antigen-specific binding site that specifically binds to IL-13 or its receptor. Antibodies targeting Th2 cytokines or their receptors include, but are not limited to, anti-IL-13 antibodies and anti-IL-13R antibodies. Any suitable IL-13-targeting antibody may be used in the methods and compositions, including, but not limited to, those described in Table 2.

[0143] Table 2: Examples of IL-13-targeting antibodies

[0144] In some embodiments, the IL-13-targeting antibody contains an antigen-specific binding site for IL-13 or its receptor. In some embodiments, the IL-13-targeting antibody contains an antigen-specific binding site for IL-13 or IL-13R derived from any known anti-IL-13 or its receptor, such as the anti-IL-13 / anti-IL-13R antibodies described below: WO2005062972, published on July 14, 2005, entitled “Treatment of cancer with novel anti-il 13 monoclonal antibodies”; WO 2005062967, published on July 14, 2005, entitled “Novel anti-il 13 antibodies and uses thereof”; WO 2008140455, published on May 15, 2007, entitled “Treatment of radiation and chemotherapy-induced fibrosis using novel anti-il 13 monoclonal antibody”. Antibodies; or WO2023245187, published on December 21, 2023, entitled "Antibodies that bind interleukin 13 and methods of use"; US6703488, published on March 9, 2004, entitled "Antibody / Receptor Targeting Moiety For Enhanced Delivery Of Armed Ligand"; US2005058645, published on March 17, 2005, entitled "Monoclonal Antibody Against Interleukin-13 Receptor Alpha 1 (IL-13Ralpha1)"; US2005154192, published on July 14, 2005, entitled "Anti-IL13Receptor Alpha1 Neutralizing" "Antibody"; US2006099652, published on May 11, 2006, and titled "IL 13 Receptor Alpha 2 Antibody And Methods Of Use"; US2006140948, published on June 29, 2006.And its name is "Fully Human Monoclonal Antibodies To IL-13"; US2006263356, which was published on November 23, 2006, and its name is "Antibodies Against IL-13 Receptor Alpha1 And Uses Thereof"; US2007048785, which was published on March 1, 2007, and its name is "Anti-IL-13 Antibodies And Complexes"; US2007128192, which was published on June 7, 2007, and its name is "Human Antibody Molecules For IL-13"; WO07080174, which was published on July 19, 2007, and its name is "Immunoglobulins"; US2007258979, which was published on November 8, 2007, and its name is "Chimeric And Humanised Monoclonal". Antibodies Against Interleukin-13; US2008008648, published January 10, 2008, titled "Treatment of Cancer With ANovel Anti-IL13 Monoclonal Antibody"; US2008044420, published February 21, 2008, titled "Anti-IL-13 Antibodies, Compositions, Methods and Uses"; US2008166343, published July 10, 2008, titled "High Affinity Antibody Antagonists Of Interleukin-13 Receptor Alpha 1"; US2008171014, published July 17, 2008, titled "Interleukin-13 Binding "Proteins"; US2008248048, published on October 9, 2008, and titled "Interleukin-13 Antibody Composition"; US2008267959, published on October 30, 2008, and titled "Anti-IL13 Human Antibodies"; US2009060906, published on March 5, 2009.And its name is "Anti-IL-13 Antibody Formulations And Uses Thereof"; US2009148905, published on June 11, 2009, and its name is "Antigen-Binding Constructs"; US2009285799, published on November 19, 2009, and its name is "Antibody Antagonists Of Interleukin-13 Receptor Alpha1"; WO09138413, published on November 19, 2009, and its name is "Single Domain Antibodies That Bind IL-13"; US2010226923, published on September 9, 2010, and its name is "Antibodies That Bind IL-4 And / Or IL-13 And Their Uses of Anti-IL-13: US2010260773, published on October 14, 2010, entitled "Antibody Molecules Having Binding Specificity For Human IL-13"; US2011052597, published on March 3, 2011, entitled "Anti-IL-13R Alpha1 Antibodies And Their Uses Thereof"; US2011206687, published on August 25, 2011, entitled "Isolation And Purification Of Anti-IL-13 Antibodies Using Protein A Affinity Chromatography"; US2012045438, published on February 23, 2012, entitled "Engineered Anti-IL-13 Antibodies, Compositions, Methods and..." Uses; US2012093830, published on April 19, 2012, entitled "SingleDomain Antibodies That Bind IL-13"; WO13103783, published on July 11, 2013, entitled "Murine IL-13 Antibodies"; US2014341913, published on November 20, 2014.The following are related patents: US2014348855, published on November 27, 2014, titled "Methods and Compositions For Treating Asthma Using Anti-IL-13 Antibodies"; US2015225479, published on August 13, 2015, titled "Anti-IL-4 / Anti-IL-13 Bispecific Antibody / Polyglutamate Formulations"; US2015266962, published on September 24, 2015, titled "Anti-IL-13 Receptor Alpha 2 Antibodies And Antibody-Drug Conjugates"; and US2016075777, published on March 17, 2016, titled "Anti-IL-4 / Anti-IL-13 Bispecific Antibody". Formulations; US2016319012, published November 3, 2016, entitled "Methods And Compositions Comprising Purified Recombinant Polypeptides"; US2017145089, published May 25, 2017, entitled "Anti-IL4-IL 13 Bispecific Antibodies"; WO17191627, published November 9, 2017, entitled "Antibodies To Interleukin 13 Receptor Alpha-1 (IL-13R Alpha 1) And Uses Thereof"; US2017334985, published November 23, 2017, entitled "Anti-IL-13 / IL-17 Bispecific Antibodies And Uses Thereof; WO19096219, published on May 23, 2019, and titled "Humanized Anti-IL-13 Antibody And Preparation Method And Use Thereof"; US2019247303, published on August 15, 2019.The names of these are as follows: US2019309059, published on October 10, 2019, titled "Inhalable Powder Composition Comprising IL-13 Antibody"; US2020165347, published on May 28, 2020, titled "Method Of Treatment Using IL-13R Antibody"; US2021277131, published on September 9, 2021, titled "Treatment Employing Anti-IL-13R Antibody Or Binding Fragment Thereof"; and US2021380674, published on December 9, 2021, titled "Methods Of Treatment Of Diseases In Which IL-13 Activity Is Detrimental Using Anti-IL-13". Antibodies; US2022033508, published on February 3, 2022, titled "Anti-IL13R Alpha2 Antibodies"; WO22174808, published on August 25, 2022, titled "Antibody Against IL-13Ra2 And Use Thereof"; US2022305104, published on September 29, 2022, titled "Methods Of Treating Atopic Dermatitis"; US2023002484, published on January 5, 2023, titled "Treatment Of Atopic Dermatitis Employing Anti-IL-13Ra1 Antibody Or Binding Fragment Thereof"; WO23287590, published on January 19, 2023, titled "IL-13 Antibodies For The Treatment WO23019260, published on February 16, 2023, is titled "IL-13 Antibodies For The Treatment of Atopic Dermatitis"; WO23023497.The following are listed: US2023084464, published on February 23, 2023, titled "Anti-IL-13 Antibody Formulation"; US2023087378, published on March 23, 2023, titled "Multi-Specific Antibody With Binding Specificity For Human IL-13 and IL-17"; WO23075700, published on May 4, 2023, titled "Anti-IL-13R Antibody Formulation"; and WO23075702, published on May 4, 2023, titled "Anti-IL-13R Antibody". Formulation; US2023183364, published on June 15, 2023, entitled "Anti-IL13R-Alpha2 Antibodies, Antigen-Binding Fragments And Uses Thereof"; WO23163659, published on August 31, 2023, entitled "Glycosylated Form Of Anti-IL13R Antibody"; US2023357381, published on November 9, 2023, entitled "Multispecific Antibodies Targeting IL-13 And IL-18"; WO23215769, published on November 9, 2023, entitled "IL-13 Antibodies For The Treatment Of Atopic Dermatitis; WO23245187, published on December 21, 2023, entitled "Antibodies That Bind Interleukin 13 And Methods Of Use"; WO24043837, published on February 29, 2024, entitled "High Concentration Anti-IL13R Antibody Formulation"; US2024117030, published on April 11, 2024.The descriptions of anti-IL-13 antibodies in each of these publications are incorporated herein by reference. US2024158521, published on May 16, 2024, is titled “Multispecific Antibodies And Uses Thereof”; US2024158521, published on May 16, 2024, is titled “Dna-Encoded Bispecific Antibodies Targeting IL13Ra2 And Methods Of Use In Cancer Therapeutics”; WO08140455, published on November 20, 2008, is titled “Treatment Of Radiation And Chemo-Therapy Induced Fibrosis Using Novel Anti-IL 13 Monoclonal Antibodies”; and WO24099310, published on May 16, 2024, is titled “Anti-IL-13 Long-Acting Nanobody Sequence And Use Thereof”. In some embodiments, the IL-13 targeting antibody comprises the HC CDR1, HC CDR2, and HC CDR3 of the VH of any of the anti-IL-13 antibodies or anti-IL-13R antibodies listed in Table 2, and the LC CDR1, LC CDR2, and LC CDR3 of the VL of any of the anti-IL-13 antibodies or anti-IL-13R antibodies listed in Table 2. In some embodiments, the IL-13 targeting antibody comprises the VH of any of the anti-IL-13 antibodies or anti-anti-IL-13R antibodies listed in Table 2, and the VL of any of the anti-IL-13 antibodies or anti-anti-IL-13R antibodies listed in Table 2. In some embodiments, the IL-13 targeting antibody comprises the HC CDR1, HC CDR2, and HC CDR3 of anluzumab, and the LC CDR1, LC CDR2, and LC CDR3 of anluzumab. In some embodiments, the IL-13 targeting antibody comprises the VH and VL of anluzumab. In some embodiments, the IL-13-targeting antibody comprises HC CDR1, HC CDR2, HC CDR3 of leprizumab and LC CDR1, LC CDR2, and LC CDR3 of leprizumab. In some embodiments, the IL-13-targeting antibody comprises VH and VL of leprizumab. In some embodiments, the IL-13-targeting antibody comprises HCCDR1, HC CDR2, HC CDR3 of trorocalumab and LC CDR1, LC CDR2, and LC CDR3 of trorocalumab. In some embodiments,The IL-13 targeting antibody comprises the VH and VL of trorocalumab. In some embodiments, the IL-13 targeting antibody comprises HC CDR1, HC CDR2, HC CDR3 of ixazolizumab and LC CDR1, LC CDR2, and LC CDR3 of ixazolizumab. In some embodiments, the IL-13 targeting antibody comprises the VH and VL of ixazolizumab. In some embodiments, the IL-13 targeting antibody comprises HC CDR1, HC CDR2, HC CDR3 of sendazolizumab and LC CDR1, LC CDR2, and LC CDR3 of sendazolizumab. In some embodiments, the IL-13 targeting antibody comprises the VH and VL of sendazolizumab. In some embodiments, the IL-13 targeting antibody comprises HC CDR1, HCCDR2, HC CDR3 of detkumab and LC CDR1, LC CDR2, and LC CDR3 of detkumab. In some embodiments, the IL-13 targeting antibody comprises the VH and VL of detcluzumab. In some embodiments, the IL-13 targeting antibody comprises HC CDR1, HC CDR2, HC CDR3 of APG777 and LC CDR1, LC CDR2, and LC CDR3 of APG777. In some embodiments, the IL-13 targeting antibody comprises the VH and VL of APG777.

[0145] (c) 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 may be combined with one or more different anti-KLK5 / KLK7 antibodies to produce multispecific or bispecific anti-KLK5 / KLK7 antibodies. For example, one or more anti-KLK5 / KLK7 antibodies described herein (Tables 1a and 1b) may be combined with one or more different anti-KLK5 / KLK7 antibodies described herein (Tables 1a and 1b) to produce multispecific antibodies.

[0146] In some implementations, one or more anti-KLK5 / KLK7 antibodies can be combined with any other suitable therapeutic antibody to produce a multispecific or bispecific anti-KLK5 / KLK7 / additional target antibody. For example, the anti-KLK5 / KLK7 antibodies described herein (Tables 1a and 1b) can be combined with any suitable antibody (e.g., an IL-13-targeting antibody as described in Table 2) to produce a bispecific antibody.

[0147] In some aspects, this disclosure provides a bispecific antibody comprising at least one antigen-specific binding site that specifically binds to KLK5 and KLK7 and at least one antigen-specific binding site that specifically binds to IL-13 or its receptor.

[0148] Table 6 below illustrates the possible arm configurations of the anti-KLK5 / KLK7 + IL-13 targeting bispecific antibody. In some embodiments, the anti-KLK5 / KLK7 + IL-13 targeting bispecific antibody includes an arm comprising a CDR and / or VH and / or VL, the sequence of which is derived from or corresponds to, or is identical to, the CDR and / or VH and / or VL sequences of the antibodies listed in the left column of Table 6, and a second arm comprising a CDR and / or VH / VL, the sequence of which is derived from or corresponds to, or is identical to, the CDR and / or VH and / or VL sequences of the anti-IL-13 antibody or another suitable antibody disclosed herein, listed in the corresponding row of the right column of Table 6. The CDRs and VH / VL sequences of the various antibodies are listed in Tables 1a and 2.

[0149] Table 6: Arm configuration of anti-KLK5 / KLK7 + Th2 bispecific antibody

[0150] In some embodiments, the anti-KLK5 / KLK7 + Th2 targeting bispecific antibody includes an arm comprising a CDR and / or VH / VL pair, the sequence of which is derived from or corresponds to, or is identical to, the CDR and / or VH / VL sequences of the anti-KLK5 / KLK7 antibodies listed in Table 1a, and a second arm comprising a CDR and / or VH / VL, the sequence of which is derived from or corresponds to, or is identical to, the CDR and / or VH / VL sequences of the anti-IL-13 or anti-IL-13R antibodies listed in Table 2, Table 6, or elsewhere herein. In some embodiments, the anti-KLK5 / KLK7 + Th2 bispecific antibody may include an arm comprising an antigen-specific binding site targeting KLK5 and KLK7, and a second arm comprising an antigen-specific binding site not targeting IL-13 or IL-13R.

[0151] In some embodiments, the bispecific antibody described herein comprises at least one antigen-specific binding site that specifically binds to KLK5 and KLK7 (e.g., KLK5 / KLK7-Dual-Ab4) and at least one antigen-specific binding site that specifically binds to IL-13 or IL-13R (anti-KLK5 / KLK7 + IL-13 bispecific antibody). Such antigen-specific binding sites may be derived from or correspond to either or the same as any of the anti-KLK5 / KLK7 antibody and IL-13 / IL-13R targeting antibody disclosed herein.

[0152] Interleukin (IL)-13 is a cytokine involved in the pathogenesis of skin barrier diseases, such as atopic dermatitis (AD). In some embodiments, IL-13 is a driver of type 2 T helper cell inflammation. In some embodiments, IL-13 is overexpressed in the lesioned skin of a subject with a skin barrier disease. In some embodiments, upon release in the peripheral skin, IL-13 activates its receptors, recruits inflammatory cells, and / or alters the skin microbiota. In some embodiments, IL-13 also reduces the expression of epidermal barrier proteins and activates sensory nerves mediating itch transmission signals. In some embodiments, blocking IL-13 signaling in combination with inhibition of KLK5 / KLK7 is effective in treating any of the diseases described herein.

[0153] In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody described herein comprises at least one antigen-specific binding site that specifically binds to the active sites of KLK5 and KLK7. In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody described herein comprises at least one antigen-specific binding site that specifically binds to KLK5 and KLK7, which is a binding site of a dual inhibitor antibody that specifically binds to KLK5 and KLK7 (e.g., any one of KLK5 / KLK7-Dual-Ab1, KLK5 / KLK7-Dual-Ab2, KLK5 / KLK7-Dual-Ab3, KLK5 / KLK7-Dual-Ab1). In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody comprises an arm containing at least one antigen-specific binding site specifically binding to both KLK5 and KLK7, said antigen-specific binding site comprising HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and / or LC CDR3 of any of the antibodies listed in Tables 1a and 1b (e.g., any of KLK5 / KLK7-Dual-Ab1, KLK5 / KLK7-Dual-Ab2, KLK5 / KLK7-Dual-Ab3, KLK5 / KLK7-Dual-Ab4), and an arm containing at least one antigen-specific binding site specifically binding to IL-13 or IL-13R, said antigen-specific binding site comprising HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and / or LC CDR3 of any of the anti-IL-13 or anti-IL-13R antibodies listed in Table 2. CDR2 and / or LC CDR3.In some embodiments, the anti-KLK5 / KLK7-IL-13 bispecific antibody comprises: an arm containing at least one antigen-specific binding site that specifically binds to both KLK5 and KLK7, said antigen-specific binding site comprising a VH having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with any of the antibodies listed in Tables 1a and 1b, and / or a VL having at least 80% identity with any of the antibodies listed in Tables 1a and 1b. (e.g., a VL with at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity); and an arm comprising at least one antigen-specific binding site that specifically binds to IL-13 or IL-13R, said antigen-specific binding site comprising a VH with at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with any of the anti-IL-13 or anti-IL-13R antibodies listed in Table 2, and / or a VL with at least 80% identity with any of the anti-IL-13 or anti-IL-13R antibodies listed in Table 2. (For example, VL of at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity.

[0154] In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody comprising at least one antigen-specific binding site specifically binding to KLK5 and KLK7 comprises: HC CDR1, HC CDR2, and HCCDR3 of the heavy chain variable domain having the amino acid sequence of SEQ ID NO: 7; and LC CDR1, LC CDR2, and LCCDR3 of the light chain variable domain having the amino acid sequence of SEQ ID NO: 8. In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody comprising at least one antigen-specific binding site specifically binding to KLK5 and KLK7 comprises: HC CDR1, HC CDR2, and HC CDR3 of the heavy chain variable domain having the amino acid sequence of SEQ ID NO: 13; and LC CDR1, LC CDR2, and LC CDR3 of the light chain variable domain having the amino acid sequence of SEQ ID NO: 14. In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody comprising at least one antigen-specific binding site specifically binding to KLK5 and KLK7 comprises: HC CDR1, HC CDR2, and HC CDR3 of the heavy chain variable domain having the amino acid sequence of SEQ ID NO: 17; and LC CDR1, LC CDR2, and LC CDR3 of the light chain variable domain having the amino acid sequence of SEQ ID NO: 14. In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody comprising at least one antigen-specific binding site specifically binding to KLK5 and KLK7 comprises: HC CDR1, HC CDR2, and HC CDR3 of the heavy chain variable domain having the amino acid sequence of SEQ ID NO: 21; and LC CDR1, LC CDR2, and LC CDR3 of the light chain variable domain having the amino acid sequence of SEQ ID NO: 14.

[0155] In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody comprising at least one antigen-specific binding site specifically binding to KLK5 and KLK7 comprises: 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. In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody comprising at least one antigen-specific binding site specifically binding to KLK5 and KLK7 comprises: 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. In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody comprising at least one antigen-specific binding site specifically binding to KLK5 and KLK7 comprises: 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.In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody comprising at least one antigen-specific binding site specifically binding to KLK5 and KLK7 comprises: 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.

[0156] In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody comprising at least one antigen-specific binding site specifically binding to KLK5 and KLK7 comprises: VH, which comprises the amino acid sequence of SEQ ID NO: 7; and VL, which comprises the amino acid sequence of SEQ ID NO: 8. In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody comprising at least one antigen-specific binding site specifically binding to KLK5 and KLK7 comprises: VH, which comprises the amino acid sequence of SEQ ID NO: 13; and VL, which comprises the amino acid sequence of SEQ ID NO: 14. In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody comprising at least one antigen-specific binding site specifically binding to KLK5 and KLK7 comprises: VH, which comprises the amino acid sequence of SEQ ID NO: 17; and LC CDR1, LC CDR2, and VL, which comprise the amino acid sequence of SEQ ID NO: 14. In some embodiments, an anti-KLK5 / KLK7+IL-13 bispecific antibody comprising at least one antigen-specific binding site that specifically binds to KLK5 and KLK7 comprises: VH, which comprises the amino acid sequence of SEQ ID NO: 21; and VL, which comprises the amino acid sequence of SEQ ID NO: 14.

[0157] In some embodiments, the anti-KLK5 / KLK7+IL-13 bispecific antibody comprises at least one antigen-specific binding site that specifically binds to IL-13. In some embodiments, the anti-KLK5 / KLK7+IL-13 bispecific antibody comprises an antigen-specific binding site for IL-13 derived from any known anti-IL-13 or its receptor, such as the anti-IL-13 / anti-IL-13R antibodies described in WO2005062972, WO 2005062967, WO 2008140455, or WO2023245187, the description of the anti-IL-13 antibody in each of which is incorporated herein by reference. In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody containing at least one antigen-specific binding site that specifically binds to IL-13 or IL-13R comprises HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and / or LC CDR3 of any of the anti-IL-13 antibodies or anti-IL-13R antibodies listed in Table 2. In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody containing at least one antigen-specific binding site that specifically binds to IL-13 or IL-13R comprises VH and / or VL of any of the anti-IL-13 antibodies or anti-IL-13R antibodies listed in Table 2.

[0158] In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody comprises at least one antigen-specific binding site for KLK5 / KLK7 derived from any one of KLK5 / KLK7-Dual-Ab1, KLK5 / KLK7-Dual-Ab2, KLK5 / KLK7-Dual-Ab3, and KLK5 / KLK7-Dual-Ab4, and at least one antigen-specific binding site for IL-13 derived from the aniline monoclonal antibody. In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody comprising at least one antigen-specific binding site for IL-13 comprises: heavy chain variable domains HC CDR1, HC CDR2, and HC CDR3, which comprise the amino acid sequence of SEQ ID NO: 53; and light chain variable domains LC CDR1, LC CDR2, and LC CDR3, which comprise the amino acid sequence of SEQ ID NO: 54. In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody containing at least one antigen-specific binding site that specifically binds to IL-13 comprises: HC CDR1, which contains the amino acid sequence of SEQ ID NO: 55; HC CDR2, which contains the amino acid sequence of SEQ ID NO: 56; HC CDR3, which contains the amino acid sequence of SEQ ID NO: 57; LC CDR1, which contains the amino acid sequence of SEQ ID NO: 58; LC CDR2, which contains the amino acid sequence of SEQ ID NO: 59; and LC CDR3, which contains the amino acid sequence of SEQ ID NO: 60. In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody containing at least one antigen-specific binding site that specifically binds to IL-13 comprises: HC CDR1, which contains the amino acid sequence of SEQ ID NO: 61; HC CDR2, which contains the amino acid sequence of SEQ ID NO: 62; HC CDR3, which contains the amino acid sequence of SEQ ID NO: 63; LC CDR1, which contains the amino acid sequence of SEQ ID NO: 64; LC CDR2, which contains the amino acid sequence of RAS; and LC CDR3, which contains the amino acid sequence of SEQ ID NO: 66.In some embodiments, the anti-KLK5 / KLK7+IL-13 bispecific antibody containing at least one antigen-specific binding site that specifically binds to IL-13 comprises: HC CDR1, which contains the amino acid sequence of SEQ ID NO: 67; HC CDR2, which contains the amino acid sequence of SEQ ID NO: 68; HC CDR3, which contains the amino acid sequence of SEQ ID NO: 69; LC CDR1, which contains the amino acid sequence of SEQ ID NO: 70; LC CDR2, which contains the amino acid sequence of SEQ ID NO: 71; and LC CDR3, which contains the amino acid sequence of SEQ ID NO: 72. In some embodiments, the anti-KLK5 / KLK7+IL-13 bispecific antibody containing at least one antigen-specific binding site that specifically binds to IL-13 comprises: VH, which contains the amino acid sequence of SEQ ID NO: 53; and VL, which contains the amino acid sequence of SEQ ID NO: 54.

[0159] In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody comprises at least one antigen-specific binding site for KLK5 / KLK7 derived from any one of KLK5 / KLK7-Dual-Ab1, KLK5 / KLK7-Dual-Ab2, KLK5 / KLK7-Dual-Ab3, and KLK5 / KLK7-Dual-Ab4, and at least one antigen-specific binding site for IL-13 derived from lerechizumab. In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody comprising at least one antigen-specific binding site for IL-13 comprises: heavy chain variable domains HC CDR1, HC CDR2, and HC CDR3, which comprise the amino acid sequence of SEQ ID NO: 32; and light chain variable domains LC CDR1, LC CDR2, and LC CDR3, which comprise the amino acid sequence of SEQ ID NO: 33. In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody containing at least one antigen-specific binding site that specifically binds to IL-13 comprises: HC CDR1, which contains the amino acid sequence of SEQ ID NO: 34; HC CDR2, which contains the amino acid sequence of SEQ ID NO: 35; HC CDR3, which contains the amino acid sequence of SEQ ID NO: 36; LC CDR1, which contains the amino acid sequence of SEQ ID NO: 37; LC CDR2, which contains the amino acid sequence of SEQ ID NO: 38; and LC CDR3, which contains the amino acid sequence of SEQ ID NO: 39. In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody containing at least one antigen-specific binding site that specifically binds to IL-13 comprises: HC CDR1, which contains the amino acid sequence of SEQ ID NO: 40; HC CDR2, which contains the amino acid sequence of SEQ ID NO: 41; HC CDR3, which contains the amino acid sequence of SEQ ID NO: 42; LC CDR1, which contains the amino acid sequence of SEQ ID NO: 43; LC CDR2, which contains the amino acid sequence of LAS; and LC CDR3, which contains the amino acid sequence of SEQ ID NO: 45.In some embodiments, the anti-KLK5 / KLK7+IL-13 bispecific antibody containing at least one antigen-specific binding site that specifically binds to IL-13 comprises: HC CDR1, which contains the amino acid sequence of SEQ ID NO: 46; HC CDR2, which contains the amino acid sequence of SEQ ID NO: 47; HC CDR3, which contains the amino acid sequence of SEQ ID NO: 48; LC CDR1, which contains the amino acid sequence of SEQ ID NO: 49; LC CDR2, which contains the amino acid sequence of SEQ ID NO: 50; and LC CDR3, which contains the amino acid sequence of SEQ ID NO: 51. In some embodiments, the anti-KLK5 / KLK7+IL-13 bispecific antibody containing at least one antigen-specific binding site that specifically binds to IL-13 comprises: VH, which contains the amino acid sequence of SEQ ID NO: 32; and VL, which contains the amino acid sequence of SEQ ID NO: 33.

[0160] In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody comprises at least one antigen-specific binding site specifically binding to KLK5 / KLK7 derived from KLK5 / KLK7-Dual-Ab4, and at least one antigen-specific binding site specifically binding to IL-13 derived from lerechizumab. In some embodiments, the KLK5 / KLK7 + IL-13 bispecific antibody comprises: at least one antigen-specific binding site that specifically binds to KLK5 / KLK7, the antigen-specific binding site comprising: HC CDR1, HC CDR2, and HC CDR3 of the heavy chain variable domain, which comprises the amino acid sequence of SEQ ID NO: 21; and LC CDR1, LC CDR2, and LC CDR3 of the light chain variable domain, which comprises the amino acid sequence of SEQ ID NO: 14; and at least one antigen-specific binding site that specifically binds to IL-13, the antigen-specific binding site comprising: HC CDR1, HC CDR2, and HC CDR3 of the heavy chain variable domain, which comprises the amino acid sequence of SEQ ID NO: 32; and LC CDR1, LC CDR2, and LC CDR3 of the light chain variable domain, which comprises the amino acid sequence of SEQ ID NO: 33. In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody comprises at least one antigen-specific binding site that specifically binds to KLK5 / KLK7, the antigen-specific binding site comprising: HC CDR1, comprising the amino acid sequence of SEQ ID NO: 18; HC CDR2, comprising the amino acid sequence of SEQ ID NO: 19; HC CDR3, comprising the amino acid sequence of SEQ ID NO: 20; LC CDR1, comprising the amino acid sequence of SEQ ID NO: 4; LC CDR2, comprising the amino acid sequence of SEQ ID NO: 5; and LC CDR3, comprising the amino acid sequence of SEQ ID NO: 12, and at least one antigen-specific binding site that specifically binds to IL-13 comprises: HC CDR1, comprising the amino acid sequence of SEQ ID NO: 34; HC CDR2, comprising the amino acid sequence of SEQ ID NO: 35; HC CDR3, comprising the amino acid sequence of SEQ ID NO: 36; LC CDR1, comprising the amino acid sequence of SEQ ID NO: 37; LC CDR3, comprising the amino acid sequence of SEQ ID NO: 37; LC CDR3, comprising the amino acid sequence of SEQ ID NO: 34; LC ...2, comprising the amino acid sequence of SEQ CDR2, which contains the amino acid sequence of SEQ ID NO: 38; and LC CDR3, which contains the amino acid sequence of SEQ ID NO: 39.In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody comprises at least one antigen-specific binding site that specifically binds to KLK5 / KLK7, the antigen-specific binding site comprising: HC CDR1, comprising the amino acid sequence of SEQ ID NO: 18; HCCDR2, comprising the amino acid sequence of SEQ ID NO: 19; HC CDR3, comprising the amino acid sequence of SEQ ID NO: 20; LC CDR1, comprising the amino acid sequence of SEQ ID NO: 4; LC CDR2, comprising the amino acid sequence of SEQ ID NO: 5; and LC CDR3, comprising the amino acid sequence of SEQ ID NO: 12, and at least one antigen-specific binding site that specifically binds to IL-13 comprises: HC CDR1, comprising the amino acid sequence of SEQ ID NO: 40; HC CDR2, comprising the amino acid sequence of SEQ ID NO: 41; HC CDR3, comprising the amino acid sequence of SEQ ID NO: 42; LC CDR1, comprising the amino acid sequence of SEQ ID NO: 43; LC CDR3, comprising the amino acid sequence of SEQ ID NO: 43; LC CDR4, comprising the amino acid sequence of SEQ ID NO: 43; LC CDR4, comprising the amino acid sequence of SEQ ID NO: 44; LC CDR3, comprising the amino acid sequence of SEQ ID NO: 44; LC CDR4, comprising the amino acid sequence of SEQ ID NO: 45 ... CDR2, which contains the amino acid sequence of SEQ ID NO: 44; and LCCDR3, which contains the amino acid sequence of SEQ ID NO: 45. In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody comprises at least one antigen-specific binding site that specifically binds to KLK5 / KLK7, the antigen-specific binding site comprising: HC CDR1, comprising the amino acid sequence of SEQ ID NO: 18; HC CDR2, comprising the amino acid sequence of SEQ ID NO: 19; HC CDR3, comprising the amino acid sequence of SEQ ID NO: 20; LC CDR1, comprising the amino acid sequence of SEQ ID NO: 4; LC CDR2, comprising the amino acid sequence of SEQ ID NO: 5; and LC CDR3, comprising the amino acid sequence of SEQ ID NO: 12, and at least one antigen-specific binding site that specifically binds to IL-13 comprises: HCCDR1, comprising the amino acid sequence of SEQ ID NO: 46; HC CDR2, comprising the amino acid sequence of SEQ ID NO: 47; HC CDR3, comprising the amino acid sequence of SEQ ID NO: 48; LC CDR1, comprising the amino acid sequence of SEQ ID NO: 49; LC CDR3 ... CDR2, which contains the amino acid sequence of SEQ ID NO: 50; and LC CDR3, which contains the amino acid sequence of SEQ ID NO: 50.In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody comprises: at least one antigen-specific binding site that specifically binds to KLK5 / KLK7, the antigen-specific binding site comprising: VH, which comprises the amino acid sequence of SEQ ID NO: 21; and VL, which comprises the amino acid sequence of SEQ ID NO: 14; and at least one antigen-specific binding site that specifically binds to IL-13, the antigen-specific binding site comprising: VH, which comprises the amino acid sequence of SEQ ID NO: 32; and VL, which comprises the amino acid sequence of SEQ ID NO: 33.

[0161] In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody comprises at least one antigen-specific binding site for KLK5 / KLK7 derived from any one of KLK5 / KLK7-Dual-Ab1, KLK5 / KLK7-Dual-Ab2, KLK5 / KLK7-Dual-Ab3, and KLK5 / KLK7-Dual-Ab4, and at least one antigen-specific binding site for IL-13 derived from APG777 (i.e., construct 133 as described in WO2023245187). In some embodiments, the anti-KLK5 / KLK7+IL-13 bispecific antibody containing at least one antigen-specific binding site that specifically binds to IL-13 comprises: HC CDR1, HC CDR2, and HC CDR3 of heavy chain variable domains, which contain the amino acid sequence of SEQ ID NO: 657; and LC CDR1, LC CDR2, and LC CDR3 of light chain variable domains, which contain the amino acid sequence of SEQ ID NO: 658. In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody containing at least one antigen-specific binding site that specifically binds to IL-13 comprises: HC CDR1, which contains the amino acid sequence of SEQ ID NO: 34; HC CDR2, which contains the amino acid sequence of SEQ ID NO: 35; HC CDR3, which contains the amino acid sequence of SEQ ID NO: 36; LC CDR1, which contains the amino acid sequence of SEQ ID NO: 37; LC CDR2, which contains the amino acid sequence of SEQ ID NO: 38; and LC CDR3, which contains the amino acid sequence of SEQ ID NO: 39. In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody containing at least one antigen-specific binding site that specifically binds to IL-13 comprises: HC CDR1, which contains the amino acid sequence of SEQ ID NO: 655; HC CDR2, which contains the amino acid sequence of SEQ ID NO: 41; HC CDR3, which contains the amino acid sequence of SEQ ID NO: 42; LC CDR1, which contains the amino acid sequence of SEQ ID NO: 43; LC CDR2, which contains the amino acid sequence of LAS; and LC CDR3, which contains the amino acid sequence of SEQ ID NO: 45.In some embodiments, the anti-KLK5 / KLK7+IL-13 bispecific antibody containing at least one antigen-specific binding site that specifically binds to IL-13 comprises: HC CDR1, which contains the amino acid sequence of SEQ ID NO: 656; HC CDR2, which contains the amino acid sequence of SEQ ID NO: 47; HC CDR3, which contains the amino acid sequence of SEQ ID NO: 48; LC CDR1, which contains the amino acid sequence of SEQ ID NO: 49; LC CDR2, which contains the amino acid sequence of SEQ ID NO: 50; and LC CDR3, which contains the amino acid sequence of SEQ ID NO: 51. In some embodiments, the anti-KLK5 / KLK7+IL-13 bispecific antibody containing at least one antigen-specific binding site that specifically binds to IL-13 comprises: VH, which contains the amino acid sequence of SEQ ID NO: 657; and VL, which contains the amino acid sequence of SEQ ID NO: 658.

[0162] In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody comprises at least one antigen-specific binding site that specifically binds to KLK5 / KLK7 derived from KLK5 / KLK7-Dual-Ab4, and at least one antigen-specific binding site that specifically binds to IL-13 derived from APG777. In some embodiments, the KLK5 / KLK7 + IL-13 bispecific antibody comprises: at least one antigen-specific binding site that specifically binds to KLK5 / KLK7, the antigen-specific binding site comprising: HC CDR1, HC CDR2, and HC CDR3 of the heavy chain variable domain, which comprises the amino acid sequence of SEQ ID NO:21; and LC CDR1, LC CDR2, and LC CDR3 of the light chain variable domain, which comprises the amino acid sequence of SEQ ID NO:14; and at least one antigen-specific binding site that specifically binds to IL-13, the antigen-specific binding site comprising: HC CDR1, HC CDR2, and HC CDR3 of the heavy chain variable domain, which comprises the amino acid sequence of SEQ ID NO:657; and LC CDR1, LC CDR2, and LC CDR3 of the light chain variable domain, which comprises the amino acid sequence of SEQ ID NO:658. In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody comprises at least one antigen-specific binding site that specifically binds to KLK5 / KLK7, the antigen-specific binding site comprising: HC CDR1, comprising the amino acid sequence of SEQ ID NO: 18; HC CDR2, comprising the amino acid sequence of SEQ ID NO: 19; HC CDR3, comprising the amino acid sequence of SEQ ID NO: 20; LC CDR1, comprising the amino acid sequence of SEQ ID NO: 4; LC CDR2, comprising the amino acid sequence of SEQ ID NO: 5; and LC CDR3, comprising the amino acid sequence of SEQ ID NO: 12, and at least one antigen-specific binding site that specifically binds to IL-13 comprises: HC CDR1, comprising the amino acid sequence of SEQ ID NO: 34; HC CDR2, comprising the amino acid sequence of SEQ ID NO: 35; HC CDR3, comprising the amino acid sequence of SEQ ID NO: 36; LC CDR1, comprising the amino acid sequence of SEQ ID NO: 37; LC CDR3, comprising the amino acid sequence of SEQ ID NO: 37; LC CDR3, comprising the amino acid sequence of SEQ ID NO: 36; LC CDR1, comprising the amino acid sequence of SEQ ID NO: 37; LC CDR3, comprising the amino acid sequence of SEQ ID NO: 34; LC CDR2, comprising the amino acid sequence of SEQ ID NO: 35; HC ...3, comprising the amino acid sequence of SEQ CDR2, which contains the amino acid sequence of SEQ ID NO:38; and LC CDR3, which contains the amino acid sequence of SEQ ID NO:39.In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody comprises at least one antigen-specific binding site that specifically binds to KLK5 / KLK7, the antigen-specific binding site comprising: HC CDR1, comprising the amino acid sequence of SEQ ID NO: 18; HCCDR2, comprising the amino acid sequence of SEQ ID NO: 19; HC CDR3, comprising the amino acid sequence of SEQ ID NO: 20; LC CDR1, comprising the amino acid sequence of SEQ ID NO: 4; LC CDR2, comprising the amino acid sequence of SEQ ID NO: 5; and LC CDR3, comprising the amino acid sequence of SEQ ID NO: 12, and at least one antigen-specific binding site that specifically binds to IL-13 comprises: HC CDR1, comprising the amino acid sequence of SEQ ID NO: 655; HC CDR2, comprising the amino acid sequence of SEQ ID NO: 41; HC CDR3, comprising the amino acid sequence of SEQ ID NO: 42; LC CDR1, comprising the amino acid sequence of SEQ ID NO: 43; LC CDR3, comprising the amino acid sequence of SEQ ID NO: 43; LC CDR3, comprising the amino acid sequence of SEQ ID NO: 42; LC CDR1, comprising the amino acid sequence of SEQ ID NO: 43; LC CDR3, comprising the amino acid sequence of SEQ ID NO: 42; LC CDR3 ...2; LC CDR3, comprising the amino acid sequence of SEQ ID NO: 42; LC CDR3, comprising the amino acid sequence of SEQ CDR2, which contains the amino acid sequence of SEQ ID NO: 44; and LC CDR3, which contains the amino acid sequence of SEQ ID NO: 45. In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody comprises at least one antigen-specific binding site that specifically binds to KLK5 / KLK7, the antigen-specific binding site comprising: HC CDR1, comprising the amino acid sequence of SEQ ID NO: 18; HC CDR2, comprising the amino acid sequence of SEQ ID NO: 19; HC CDR3, comprising the amino acid sequence of SEQ ID NO: 20; LC CDR1, comprising the amino acid sequence of SEQ ID NO: 4; LC CDR2, comprising the amino acid sequence of SEQ ID NO: 5; and LC CDR3, comprising the amino acid sequence of SEQ ID NO: 12, and at least one antigen-specific binding site that specifically binds to IL-13 comprises: HC CDR1, comprising the amino acid sequence of SEQ ID NO: 656; HC CDR2, comprising the amino acid sequence of SEQ ID NO: 47; HC CDR3, comprising the amino acid sequence of SEQ ID NO: 48; LC CDR1, comprising the amino acid sequence of SEQ ID NO: 49; LC CDR3 ... CDR2, which contains the amino acid sequence of SEQ ID NO: 50; and LC CDR3, which contains the amino acid sequence of SEQ ID NO: 51.In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody comprises: at least one antigen-specific binding site that specifically binds to KLK5 / KLK7, the antigen-specific binding site comprising: VH, which comprises the amino acid sequence of SEQ ID NO: 21; and VL, which comprises the amino acid sequence of SEQ ID NO: 14; and at least one antigen-specific binding site that specifically binds to IL-13, the antigen-specific binding site comprising: VH, which comprises the amino acid sequence of SEQ ID NO: 657; and VL, which comprises the amino acid sequence of SEQ ID NO: 658.

[0163] In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody comprises at least one antigen-specific binding site for KLK5 / KLK7 derived from any one of KLK5 / KLK7-Dual-Ab1, KLK5 / KLK7-Dual-Ab2, KLK5 / KLK7-Dual-Ab3, and KLK5 / KLK7-Dual-Ab4, and at least one antigen-specific binding site for IL-13 derived from trorocalumab. In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody comprising at least one antigen-specific binding site for IL-13 comprises: heavy chain variable domains HC CDR1, HC CDR2, and HC CDR3, which comprise the amino acid sequence of SEQ ID NO: 74; and light chain variable domains LC CDR1, LC CDR2, and LC CDR3, which comprise the amino acid sequence of SEQ ID NO: 75. In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody containing at least one antigen-specific binding site that specifically binds to IL-13 comprises: HC CDR1, which contains the amino acid sequence of SEQ ID NO: 76; HC CDR2, which contains the amino acid sequence of SEQ ID NO: 77; HC CDR3, which contains the amino acid sequence of SEQ ID NO: 78; LC CDR1, which contains the amino acid sequence of SEQ ID NO: 79; LC CDR2, which contains the amino acid sequence of SEQ ID NO: 80; and LC CDR3, which contains the amino acid sequence of SEQ ID NO: 81. In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody containing at least one antigen-specific binding site that specifically binds to IL-13 comprises: HC CDR1 containing the amino acid sequence of SEQ ID NO: 82; HC CDR2 containing the amino acid sequence of SEQ ID NO: 83; HC CDR3 containing the amino acid sequence of SEQ ID NO: 84; LC CDR1 containing the amino acid sequence of SEQ ID NO: 85; LC CDR2 containing the amino acid sequence of DDG; and LC CDR3 containing the amino acid sequence of SEQ ID NO: 87.In some embodiments, the anti-KLK5 / KLK7+IL-13 bispecific antibody containing at least one antigen-specific binding site that specifically binds to IL-13 comprises: HC CDR1, which contains the amino acid sequence of SEQ ID NO: 88; HC CDR2, which contains the amino acid sequence of SEQ ID NO: 89; HC CDR3, which contains the amino acid sequence of SEQ ID NO: 90; LC CDR1, which contains the amino acid sequence of SEQ ID NO: 91; LC CDR2, which contains the amino acid sequence of SEQ ID NO: 92; and LC CDR3, which contains the amino acid sequence of SEQ ID NO: 93. In some embodiments, the anti-KLK5 / KLK7+IL-13 bispecific antibody containing at least one antigen-specific binding site that specifically binds to IL-13 comprises: VH, which contains the amino acid sequence of SEQ ID NO: 74; and VL, which contains the amino acid sequence of SEQ ID NO: 75.

[0164] In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody comprises at least one antigen-specific binding site for KLK5 / KLK7 derived from any one of KLK5 / KLK7-Dual-Ab1, KLK5 / KLK7-Dual-Ab2, KLK5 / KLK7-Dual-Ab3, and KLK5 / KLK7-Dual-Ab4, and at least one antigen-specific binding site for IL-13R derived from ixazolidinyl. In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody comprising at least one antigen-specific binding site for IL-13R comprises: heavy chain variable domains HC CDR1, HC CDR2, and HC CDR3, comprising the amino acid sequence of SEQ ID NO: 94; and light chain variable domains LC CDR1, LC CDR2, and LC CDR3, comprising the amino acid sequence of SEQ ID NO: 95. In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody containing at least one antigen-specific binding site that specifically binds to IL-13R comprises: HC CDR1, which contains the amino acid sequence of SEQ ID NO: 96; HC CDR2, which contains the amino acid sequence of SEQ ID NO: 97; HC CDR3, which contains the amino acid sequence of SEQ ID NO: 98; LC CDR1, which contains the amino acid sequence of SEQ ID NO: 99; LC CDR2, which contains the amino acid sequence of SEQ ID NO: 100; and LC CDR3, which contains the amino acid sequence of SEQ ID NO: 101. In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody containing at least one antigen-specific binding site that specifically binds to IL-13R comprises: HC CDR1 containing the amino acid sequence of SEQ ID NO: 102; HC CDR2 containing the amino acid sequence of SEQ ID NO: 103; HC CDR3 containing the amino acid sequence of SEQ ID NO: 104; LC CDR1 containing the amino acid sequence of SEQ ID NO: 105; LCCDR2 containing the amino acid sequence of GAS; and LC CDR3 containing the amino acid sequence of SEQ ID NO: 107.In some embodiments, the anti-KLK5 / KLK7+IL-13 bispecific antibody containing at least one antigen-specific binding site that specifically binds to IL-13R comprises: HC CDR1, which contains the amino acid sequence of SEQ ID NO: 108; HC CDR2, which contains the amino acid sequence of SEQ ID NO: 109; HC CDR3, which contains the amino acid sequence of SEQ ID NO: 110; LC CDR1, which contains the amino acid sequence of SEQ ID NO: 111; LC CDR2, which contains the amino acid sequence of SEQ ID NO: 112; and LC CDR3, which contains the amino acid sequence of SEQ ID NO: 113. In some embodiments, the anti-KLK5 / KLK7+IL-13 bispecific antibody containing at least one antigen-specific binding site that specifically binds to IL-13R comprises: VH, which contains the amino acid sequence of SEQ ID NO: 94; and VL, which contains the amino acid sequence of SEQ ID NO: 95.

[0165] In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody comprises at least one antigen-specific binding site for KLK5 / KLK7 derived from any one of KLK5 / KLK7-Dual-Ab1, KLK5 / KLK7-Dual-Ab2, KLK5 / KLK7-Dual-Ab3, and KLK5 / KLK7-Dual-Ab4, and at least one antigen-specific binding site for IL-13 derived from sendazol. In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody comprising at least one antigen-specific binding site for IL-13 comprises: heavy chain variable domains HC CDR1, HC CDR2, and HC CDR3, comprising the amino acid sequence of SEQ ID NO: 114; and light chain variable domains LC CDR1, LC CDR2, and LC CDR3, comprising the amino acid sequence of SEQ ID NO: 115. In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody containing at least one antigen-specific binding site that specifically binds to IL-13 comprises: HC CDR1, which contains the amino acid sequence of SEQ ID NO: 116; HC CDR2, which contains the amino acid sequence of SEQ ID NO: 117; HC CDR3, which contains the amino acid sequence of SEQ ID NO: 118; LC CDR1, which contains the amino acid sequence of SEQ ID NO: 119; LC CDR2, which contains the amino acid sequence of SEQ ID NO: 120; and LCCDR3, which contains the amino acid sequence of SEQ ID NO: 121. In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody containing at least one antigen-specific binding site that specifically binds to IL-13 comprises: HC CDR1, which contains the amino acid sequence of SEQ ID NO: 122; HC CDR2, which contains the amino acid sequence of SEQ ID NO: 123; HC CDR3, which contains the amino acid sequence of SEQ ID NO: 124; LC CDR1, which contains the amino acid sequence of SEQ ID NO: 125; LCCDR2, which contains the amino acid sequence of YTS; and LC CDR3, which contains the amino acid sequence of SEQ ID NO: 127.In some embodiments, the anti-KLK5 / KLK7+IL-13 bispecific antibody containing at least one antigen-specific binding site that specifically binds to IL-13 comprises: HC CDR1, which contains the amino acid sequence of SEQ ID NO: 128; HC CDR2, which contains the amino acid sequence of SEQ ID NO: 129; HC CDR3, which contains the amino acid sequence of SEQ ID NO: 130; LC CDR1, which contains the amino acid sequence of SEQ ID NO: 131; LC CDR2, which contains the amino acid sequence of SEQ ID NO: 132; and LC CDR3, which contains the amino acid sequence of SEQ ID NO: 133. In some embodiments, the anti-KLK5 / KLK7+IL-13 bispecific antibody containing at least one antigen-specific binding site that specifically binds to IL-13 comprises: VH, which contains the amino acid sequence of SEQ ID NO: 114; and VL, which contains the amino acid sequence of SEQ ID NO: 115.

[0166] In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody comprises at least one antigen-specific binding site for KLK5 / KLK7 derived from any one of KLK5 / KLK7-Dual-Ab1, KLK5 / KLK7-Dual-Ab2, KLK5 / KLK7-Dual-Ab3, and KLK5 / KLK7-Dual-Ab4, and at least one antigen-specific binding site for IL-13 derived from detkumab. In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody comprising at least one antigen-specific binding site for IL-13 comprises: heavy chain variable domains HC CDR1, HC CDR2, and HC CDR3, comprising the amino acid sequence of SEQ ID NO: 134; and light chain variable domains LC CDR1, LC CDR2, and LC CDR3, comprising the amino acid sequence of SEQ ID NO: 135. In some embodiments, the Th2-targeting antibody contains an antigen-specific binding site for IL-13 or its receptor. In some embodiments, the anti-KLK5 / KLK7+IL-13 bispecific antibody containing at least one antigen-specific binding site for IL-13 comprises: HC CDR1 containing the amino acid sequence of SEQ ID NO: 136; HC CDR2 containing the amino acid sequence of SEQ ID NO: 137; HC CDR3 containing the amino acid sequence of SEQ ID NO: 138; LC CDR1 containing the amino acid sequence of SEQ ID NO: 139; LC CDR2 containing the amino acid sequence of SEQ ID NO: 140; and LC CDR3 containing the amino acid sequence of SEQ ID NO: 141. In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody containing at least one antigen-specific binding site that specifically binds to IL-13 comprises: HC CDR1, which contains the amino acid sequence of SEQ ID NO: 142; HC CDR2, which contains the amino acid sequence of SEQ ID NO: 143; HC CDR3, which contains the amino acid sequence of SEQ ID NO: 144; LC CDR1, which contains the amino acid sequence of SEQ ID NO: 145; LC CDR2, which contains the amino acid sequence of DAS; and LC CDR3, which contains the amino acid sequence of SEQ ID NO: 147.In some embodiments, the anti-KLK5 / KLK7+IL-13 bispecific antibody comprising at least one antigen-specific binding site that specifically binds to IL-13 comprises: HC CDR1, comprising the amino acid sequence of SEQ ID NO: 148; HC CDR2, comprising the amino acid sequence of SEQ ID NO: 149; HC CDR3, comprising the amino acid sequence of SEQ ID NO: 150; LC CDR1, comprising the amino acid sequence of SEQ ID NO: 151; LC CDR2, comprising the amino acid sequence of SEQ ID NO: 152; and LC CDR3, comprising the amino acid sequence of SEQ ID NO: 153. In some embodiments, the anti-KLK5 / KLK7+IL-13 bispecific antibody comprising at least one antigen-specific binding site that specifically binds to IL-13 comprises: VH, comprising the amino acid sequence of SEQ ID NO: 134; and VL, comprising the amino acid sequence of SEQ ID NO: 135.

[0167] Due to variations in size, valency, structure, arrangement, flexibility, geometry, heavy / light chain pairing, etc., bispecific antibodies are known to not retain the same binding properties to each antigen as the parent antibodies from which they are derived (see, for example, Brinkmanna et al., The making of bispecific antibodies, MAbs February to March 2017; 9(2): 182-212).

[0168] In some embodiments, relative to the binding affinity of KLK5 / KLK7-Dual-Ab4 to KLK5 / KLK7 and lerechizumab to IL-13, the binding affinity includes CDR and / or VH / VL derived from or corresponding to KLK5 / KLK7-Dual-Ab4 or the same CDR and / or VH / VL thereas, and CDR and / or VH / VL derived from or corresponding to anti-IL-13 antibodies such as lerechizumab or the same CDR and / or VH / VL thereas. The IL-13 bispecific antibody retains a similar range of binding affinity (e.g., no more than 10%, no more than 20%, no more than 30%, no more than 40%, no more than 50%, no more than 10%, less than 10-fold difference, less than 9-fold difference, less than 8-fold difference, less than 7-fold difference, less than 6-fold difference, less than 5-fold difference, less than 4-fold difference, less than 3-fold difference, less than 2-fold difference, or less than 1-fold difference). Therefore, in some embodiments, anti-KLK5 / KLK7+IL-13 bispecific antibodies containing CDR and / or VH / VL derived from or corresponding to KLK5 / KLK7-Dual-Ab4 or the same CDR and / or VH / VL as thereon, and anti-IL-13 antibodies containing CDR and / or VH / VL derived from or corresponding to anti-IL-13 antibodies (such as lerechizumab) or the same CDR and / or VH / VL as thereon, retain similar ranges of binding affinity relative to KLK5 / KLK7-Dual-Ab4 (e.g., no more than 10%, no more than 20%, no more than 30%, no more than 40%, no more than 50%, no more than 10%, less than 10-fold difference, less than 9-fold difference, less than 8-fold difference, less than 7-fold difference, less than 6-fold difference, less than 5-fold difference, less than 4-fold difference, less than 3-fold difference, less than 2-fold difference, or less than 1-fold difference).Therefore, in some embodiments, anti-IL-13 bispecific antibodies containing CDR and / or VH / VL derived from or corresponding to KLK5 / KLK7-Dual-Ab4 or the same CDR and / or VH / VL as thereon, and anti-IL-13 bispecific antibodies containing CDR and / or VH / VL derived from or corresponding to anti-IL-13 antibodies (such as lerechizumab) or the same CDR and / or VH / VL as thereon, retain similar ranges of binding affinity relative to the binding affinity of the bispecific antibody derived from or corresponding to it (such as lerechizumab). (e.g., no more than 10% difference, no more than 20% difference, no more than 30% difference, no more than 40% difference, no more than 50% difference, no more than 10% difference, less than 10-fold difference, less than 9-fold difference, less than 8-fold difference, less than 7-fold difference, less than 6-fold difference, less than 5-fold difference, less than 4-fold difference, less than 3-fold difference, less than 2-fold difference, or less than 1-fold difference.) The binding affinity between a bispecific antibody and its parent antibody can be measured by any suitable method (e.g., BIAcore, ELISA, etc.).

[0169] In some embodiments, the anti-KLK5 / KLK7 + IL-13 bispecific antibody retains a similar range of inhibitory activity against each antigen (e.g., KLK5 / KLK7 and the Th2 targets described herein) relative to the inhibitory activity of antibodies derived from or corresponding to or identical to each antigen-specific binding site in the bispecific antibody’s antigen-specific binding site (e.g., no more than 10%, no more than 20%, no more than 30%, no more than 40%, no more than 50%, no more than 10%, less than 10-fold difference, less than 9-fold difference, less than 8-fold difference, less than 7-fold difference, less than 6-fold difference, less than 5-fold difference, less than 4-fold difference, less than 3-fold difference, less than 2-fold difference, or less than 1-fold difference). In some embodiments, relative to the KLK5 / KLK7-Dual-Ab4's activity against the KLK5 / KLK7 protease and the inhibitory activity of lerechizumab against IL-13 signaling, the anti-KLK5 / KLK7+ antibody comprises a CDR and / or VH / VL derived from or corresponding to KLK5 / KLK7-Dual-Ab4 or the same CDR and / or VH / VL thereof, and an anti-IL-13 antibody derived from or corresponding to the CDR and / or VH / VL thereof (such as lerechizumab) or the same CDR and / or VH / VL thereof. The IL-13 bispecific antibodies retained similar ranges of inhibitory activity (e.g., differences not exceeding 10%, 20%, 30%, 40%, 50%, 10%, less than 10-fold, less than 9-fold, less than 8-fold, less than 7-fold, less than 6-fold, less than 5-fold, less than 4-fold, less than 3-fold, less than 2-fold, or less than 1-fold). The inhibitory activity of both the bispecific and parental antibodies can be measured by any suitable method (e.g., KLK5 / KLK7 protease activity against BOC-Val-Pro-Arg-AMC and / or KHLF-AMC, RT-PCR, Western blotting, etc.).

[0170] In some embodiments, one or more anti-KLK5 / KLK7 antibodies may be combined with any other suitable anti-KLK7 antibody to produce multispecific or bispecific anti-KLK5 / KLK7 antibodies. For example, the anti-KLK5 / KLK7 antibodies described herein (Tables 1a and 1b) may be combined with any other suitable anti-KLK7 antibody to produce bispecific antibodies. Non-limiting examples of suitable anti-KLK7 antibodies include 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 “RECOMBINANTHUMAN ANTIBODIES FOR INHIBITING HUMAN TISSUE KALLIKREIN 7 (KLK7) AND USE INDISEASES RELATED TO THE PROCESS OF SKIN DESQUAMATION”, published November 18, 2021; and International Patent Application Publication No. WO2021226695, entitled “DIAGNOSTICS AND THERAPEUTICS FOR DISEASES ASSOCIATED WITH The contents of the international patent application publication number WO2005075667 for “KALLIKREIN 7 (KLK7)” are incorporated herein by reference.

[0171] In some embodiments, one or more anti-KLK5 / KLK7 antibodies can be combined with any suitable anti-KLK5 antibody to produce multispecific or bispecific anti-KLK5 / KLK7 antibodies. For example, the KLK5 / KLK7 antibodies described herein (Tables 1a and 1b) can be combined with any suitable anti-KLK5 antibody. Non-limiting examples of anti-KLK5 antibodies include 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; and others. The contents of the following patent applications are incorporated herein by reference: International Patent Application Publication No. WO2021156170, entitled “ANTI-KLK5 ANTIBODIES AND METHODS OF USE”, published on September 30, 2021; U.S. Patent Application Publication No. 2021-0301032, entitled “ANTI-KLK5 ANTIBODIES AND METHODS OF USE”, published on May 6, 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”.

[0172] In some embodiments, a 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 targeting different antigens and / or unique antigen-specific binding sites targeting different regions of the same antigen. In some embodiments, the multispecific antibody comprises at least one antigen-specific binding site targeting a first antigen and at least one antigen-specific binding site targeting a second antigen. In some embodiments, the multispecific antibody comprises two or more antigen-specific binding sites targeting different regions of a first antigen and / or two or more antigen-specific binding sites targeting different regions of a second antigen.

[0173] 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).

[0174] In some embodiments, the multispecific antibody lacks Fc-mediated effector functions such as antibody-dependent cell-mediated cytotoxicity (ADCC), antibody-dependent phagocytosis (ADCP), complement fixation, and FcRn-mediated recycling. However, in some embodiments, the multispecific antibody includes one or more Fc regions that support Fc-mediated effector functions such as antibody-dependent cell-mediated cytotoxicity (ADCC), antibody-dependent phagocytosis (ADCP), complement fixation, and FcRn-mediated recycling.

[0175] In some embodiments, the antibodies provided herein are bispecific antibodies. In some embodiments, the bispecific antibody comprises at least two distinct Fv regions. In some embodiments, the bispecific antibody comprises two distinct heavy chains and two distinct light chains. In some embodiments, the bispecific antibody comprises one or more IgG molecules. In some embodiments, the bispecific antibody comprises one or more IgG molecules containing additional antigen-specific binding sites, for example, IgG molecules containing additional or modified Ig-like structures.

[0176] In some embodiments, the bispecific antibody comprises two single-chain variable fragments (scFv) linked by a linker. In some embodiments, the bispecific antibody comprises two single-domain antibodies, such as VH or VL domains, VHH, VNAR, or nanobodies linked by a linker (e.g., a glycine-rich flexible linker, such as (G4S)3-linker). In some embodiments, the bispecific antibody is in the form of a biantibody, such as described in P Holliger, T Prospero, and G Winter, "Diabodies": small bivalent and bispecific antibody fragments, Proc Natl AcadSci US A. July 15, 1993; 90(14): 6444–6448, the entire contents of which are incorporated herein by reference. In some embodiments, the bispecific antibody is a Fab fusion protein, such as a Fab-Fab fusion protein, a Fab-scFv fusion protein, or a Fab-Fv fusion protein. In some implementations, bispecific antibodies contain an antigen-binding site, such as scFv, which is modified to include a second and unique antigen-specific binding site as a component of the antibody, such as scFv.

[0177] In some embodiments, the bispecific antibody is in fragment-based, symmetrical, or asymmetrical form. In some embodiments, the fragment-based bispecific antibody does not contain an Fc region. In some embodiments, the bispecific antibody is in the form of tandem VHH, tandem scFv, DART, bifunctional antibody, F(ab)2, scFv-Fab, tandem VHH, (scFv)2-Fab, or tandem bifunctional antibody. In some embodiments, the bispecific antibody is in asymmetrical form selected from the following: rat-mouse hybrid IgG, heterologous H HL exchange and / or assembled IgG, heterologous H forced HL IgG, cH IgG, heterologous HCrossMab, scFv-Fab IgG, DART-Fc, LP-DART, CODV-Fab-TL, HLE-BiTE, and F(ab)3CrossMab. In some implementations, the bispecific antibody is in a symmetrical form selected from the following: 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.

[0178] In some embodiments, bispecific antibodies are engineered to facilitate formation via a knock-in-holes technique, for example, to promote heterodimerization. In some embodiments, the knock-in-holes technique can be used to produce bispecific IgG molecules, trivalent Ig-like antibodies, bispecific Fc and CH3 fusion proteins, and other forms, as discussed below: 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 ahomodimer 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.

[0179] In some embodiments, the bispecific antibody lacks Fc-mediated effector functions such as antibody-dependent cell-mediated cytotoxicity (ADCC), antibody-dependent phagocytosis (ADCP), complement fixation, and / or FcRn-mediated recycling. However, in some embodiments, the bispecific antibody includes one or more Fc regions that support Fc-mediated effector functions such as antibody-dependent cell-mediated cytotoxicity (ADCC), antibody-dependent phagocytosis (ADCP), complement fixation, and FcRn-mediated recycling.

[0180] III. Antibody Preparation The antibodies described herein can be prepared by any method known in the art. See, for example, Harlow and Lane, (1998) Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, New York.

[0181] In some embodiments, antibodies specific to target antigens (e.g., KLK5 and / or KLK7) can be prepared using conventional hybridoma techniques. Full-length target antigens or fragments thereof, optionally conjugated to a carrier protein (such as KLH), can be used to immunize a host animal to produce antibodies that bind to that antigen. The immunization pathway and schedule for the host animal are generally consistent with established and conventional techniques for antibody stimulation and production, as further described herein. General techniques for producing mouse antibodies, humanized antibodies, and human antibodies are known in the art and described herein. It is conceivable that any mammalian subject (including humans) or cells from which antibodies are produced can be manipulated as a basis for the production of mammalian (including human) hybridoma cell lines. Typically, the host animal is inoculated with a predetermined amount of immunogen (including the immunogens described herein) via intraperitoneal, intramuscular, oral, subcutaneous, plantar, and / or intradermal administration.

[0182] In some implementations, bispecific antibodies targeting two different antigens (e.g., KLK5 / KLK7 and IL-13 or their receptors) can be prepared by suitable methods, for example, as described in Nature Reviews Drug Discovery, Vol. 18, pp. 585-608 (2019) and Brinkmann U and Kontermann EE. The making of bispecific antibodiesAs described in MAbs. 2017 Feb / Mar;9(2):182-212. In some embodiments, anti-KLK5 / KLK7+ IL-13 cytokines or their receptors can be generated by expressing VH and / or VL on one arm and VH and / or VL on the other arm in one or more host cells. In some embodiments, arm-specific coding sequences for KLK5 / KLK7 are described herein. In some embodiments, arm-specific coding sequences for IL-13 or their receptors can be developed to express the amino acid sequences of the IL-13-targeting antibodies described herein. In some embodiments, different chains of bispecific antibodies can be assembled using any suitable method to form bispecific antibodies.

[0183] If desired, the antibody of interest (monoclonal or polyclonal) (e.g., generated by a hybridoma) can be sequenced, and the polynucleotide sequence can then be cloned into a vector for expression or propagation. The sequence encoding the antibody of interest can be held in a vector within host cells, which can then be amplified and frozen for future use. Alternatively, the polynucleotide sequence can be genetically manipulated to “humanize” the antibody or to improve affinity (affinity maturation) or other characteristics of the antibody. For example, if the antibody is used in clinical trials and treatments in humans, the constant region can be engineered to be more similar to the human constant region to avoid an immune response. Genetic manipulation of the antibody sequence may be necessary to obtain higher affinity and greater potency for the target antigen. It will be apparent to those skilled in the art that one or more polynucleotide alterations can be made to an antibody while still maintaining its binding specificity to the target antigen.

[0184] In other embodiments, fully human antibodies can be obtained using commercially available mice that have been engineered to express specific human immunoglobulins. Transgenic animals designed to produce a more desirable (e.g., fully human antibody) or more robust immune response can also be used to generate humanized or human antibodies. Examples of such technologies are XenomouseRTM from Amgen, Inc. (Fremont, CA) and HuMAb-MouseRTM and TC MouseTM from Medarex, Inc. (Princeton, NJ), or H2L2 mice from Harbour Antibodies BV (Holland). In another alternative, antibodies can be recombinantly prepared using phage display or yeast technology. See, for example, U.S. Patent Nos. 5,565,332, 5,580,717, 5,733,743, and 6,265,150; and Winter et al., (1994) Annu. Rev. Immunol. 12:433-455. Alternatively, phage display technology (McCafferty et al., (1990) Nature 348:552-553) can be used to generate human antibodies and antibody fragments in vitro from a gene library of immunoglobulin variable (V) domains from non-immunized donors.

[0185] Antigen-binding fragments of complete antibodies (full-length antibodies) can be prepared using conventional methods. For example, the F(ab')2 fragment can be produced by pepsin digestion of antibody molecules, and the Fab fragment can be produced by reducing the disulfide bonds of the F(ab')2 fragment. Genetically engineered antibodies, such as humanized antibodies, chimeric antibodies, single-chain antibodies, and bispecific antibodies, can be produced using, for example, conventional recombinant techniques. In one instance, DNA encoding a monoclonal antibody specific to a target antigen can be readily isolated and sequenced using conventional methods (e.g., by using oligonucleotide probes capable of specifically binding to genes encoding the heavy and light chains of the monoclonal antibody). Hybridoma cells are a preferred source of such DNA. Once isolated, the DNA can be placed in one or more expression vectors and then transfected into host cells that do not additionally produce immunoglobulins, such as *E. coli* cells, simian COS cells, Chinese hamster ovary (CHO) cells, human HEK293 cells, or myeloma cells, to synthesize monoclonal antibodies in recombinant host cells. See, for example, PCT Publication No. WO 87 / 04462. DNA can then be modified, for example, by replacing the homologous mouse sequences with coding sequences that replace the constant domains of the human heavy and light chains (Morrison et al., (1984) Proc. Nat. Acad. Sci. 81:6851), or by covalently linking all or part of the coding sequence of a non-immunoglobulin polypeptide to an immunoglobulin coding sequence. In this way, genetically engineered antibodies with target antigen binding specificity (such as “chimeric” or “hybrid” antibodies) can be prepared.

[0186] Single-chain antibodies can be prepared using recombinant technology by linking nucleotide sequences encoding the variable region of the heavy chain and the variable region of the light chain. Preferably, a flexible linker is incorporated between the two variable regions.

[0187] Antibodies obtained according to methods known in the art and described herein can be characterized using methods well-known in the art. For example, one approach is to identify antigen-binding epitopes, or “epitope mapping.” Numerous methods are known in the art for locating and characterizing epitopes on proteins, including resolving the crystal structure of antibody-antigen complexes, competitive assays, gene fragment 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, NY, 1999. In one instance, epitope localization can be accomplished using H / D-Ex (hydrogen-deuterium exchange) combined with proteolysis and mass spectrometry. In another embodiment, epitope localization can be used to determine the sequence that binds to the antibody. Epitopes can be linear epitopes, i.e., contained within a single amino acid segment, or conformational epitopes formed by the three-dimensional interactions of amino acids, which are not necessarily contained within a single segment (a linear sequence of primary structure). Peptides of varying lengths (e.g., at least 4-6 amino acids long) can be isolated or synthesized (e.g., recombined) and used for antibody binding assays. In another example, the epitope to which an antibody binds can be identified in a systematic screening process by using overlapping peptides derived from the target antigen sequence and determining antibody binding. Based on gene fragment expression assays, open reading frames encoding the target antigen are randomly or fragmented via specific gene constructions, and the reactivity of the antigen expression fragment with the test antibody is determined. Gene fragments can be generated, for example, by PCR, and then transcribed and translated into proteins in vitro in the presence of radioactive amino acids. The binding of the antibody to the radiolabeled antigen fragment is then determined by immunoprecipitation and gel electrophoresis. Certain epitopes can also be identified using a large library (phage library) of random peptide sequences displayed on the surface of phage particles. Alternatively, the binding of a defined library of overlapping peptide fragments to the test antibody can be tested in a simple binding assay. In another example, mutagenesis of the antigen-binding domain, domain exchange assays, and alanine scan mutagenesis can be performed to identify sufficient and / or essential residues required for epitope binding. Alternatively, a competitive assay can be performed using other antibodies known to bind to the same antigen to determine whether an antibody binds to the same epitope as other antibodies. Competitive assays are well known to those skilled in the art.

[0188] In some instances, the antibodies described herein are prepared using recombinant techniques exemplified below. Nucleic acids encoding the heavy and light chains of the antibodies described herein can be cloned into an expression vector, with each nucleotide sequence operatively linked to a suitable promoter. In one instance, each nucleotide sequence encoding the heavy and light chains is operatively linked to a unique promoter. Alternatively, the nucleotide sequences encoding the heavy and light chains can be operatively linked to a single promoter, such that both the heavy and light chains are expressed by the same promoter. If necessary, an internal ribosome entry site (IRES) can be inserted between the heavy chain coding sequence and the light chain coding sequence.

[0189] In some instances, 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 of them can be isolated from the host cell expressing them, and the separated heavy and light chains can be mixed and incubated under suitable conditions that allow antibody formation.

[0190] Typically, methods known in the art can be used to clone a nucleic acid sequence encoding one or all strands of an antibody into a suitable expression vector operatively linked to a suitable promoter. For example, the nucleotide sequence and the vector can be contacted with a restriction enzyme under suitable conditions to generate complementary ends on each molecule, which can then be paired with each other and ligated together with a ligase. Alternatively, synthetic nucleic acid adapters can be ligated to the ends of the gene. These synthetic adapters contain nucleic acid sequences corresponding 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.

[0191] A variety of promoters can be used for the expression of the antibodies described herein, including but not limited to the cytomegalovirus (CMV) intermediate early promoter, viral LTRs (such as Rous sarcoma virus LTR, HIV-LTR, HTLV-1 LTR), simian virus 40 (SV40) early promoter, Escherichia coli lac UV promoter and herpes simplex virus tk promoter.

[0192] Adjustable promoters can also be used. Such adjustable promoters include those that use lac repressor from E. coli as a transcription regulator to regulate transcription from mammalian cell promoters carrying lac operons [[Brown, M. et al., Cell, 49:603-612 (1987)]], and those that use 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 the FK506 dimer using estradiol, RU486, diphenol murislerone, or rapamycin, VP16, or p65. Inducible systems are available from Invitrogen, Clontech, and Ariad, among others.

[0193] Adjustable promoters including repressor factors with operons can be used. In one embodiment, the lac repressor factor from *E. coli* can act as a transcription regulator to regulate transcription from a mammalian cell promoter carrying a lac operon, which combines a tetracycline repressor factor (tetR) with a transcription activator (VP16) to produce the tetR-mammalian cell transcription activator fusion protein tTa (tetR-VP16) [[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)]], wherein tetR carries a minimal promoter derived from a human cytomegalovirus (hCMV) promoter to produce the tetR-tet operon system to control gene expression in mammalian cells. In one embodiment, a tetracycline-inducible switch is used. When the tetracycline operon is appropriately located downstream of the TATA element of the CMVIE promoter, the tetracycline repressor (tetR) alone, rather than a tetR-mammalian cell transcription factor fusion derivative, can act as an effective trans-regulator to modulate gene expression in mammalian cells (Yao et al., Human Gene Therapy). A particular advantage of this tetracycline-inducible switch is that it does not require the use of tetracycline repressor-mammalian cell trans-activator or repressor fusion proteins, which in some cases may be cellularly toxic (Gossen et al., Natl. Acad. Sci. USA, 89:5547-5551 (1992); Shockett et al., Proc. Natl. Acad. Sci. USA, 92:6522-6526 (1995)) to achieve its modulated effect.

[0194] In addition, the vector may contain some or all of the following: selection marker genes, such as the neomycin gene for selecting stable or transient transfectants in mammalian cells; enhancer / promoter sequences from the immediate early gene of human CMV for high-level transcription; transcription termination and RNA processing signals for mRNA stability from SV40; ColE1 for SV40 multitumor origin of replication and appropriate appendage replication; internal ribosome binding sites (IRESes), multifunctional multiple cloning sites; and T7 and SP6 RNA promoters for in vitro transcription of sense and antisense RNA. Suitable vectors and methods for generating vectors containing transgenes are well known and available in the art. Examples of polyadenylation signals that can be used to implement the methods described herein include, but are not limited to, human type I collagen polyadenylation signals, human type II collagen polyadenylation signals, and SV40 polyadenylation signals.

[0195] One or more vectors (e.g., expression vectors) containing a nucleic acid encoding any of the antibodies (e.g., nucleic acid coding sequences listed in Table 3) can be introduced into suitable host cells to produce antibodies. 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 (NSO) cells, human embryonic retina (PER.C6) cells, Sp2 / O cells, young hamster kidney (BHK) cells, Martin-Darby canine kidney (MDCK) cells, Martin-Darby bovine kidney (MDBK) cells, and monkey kidney CV1 lines transformed from SV40 (COS) cells. In some embodiments, the host cell expressing the antibody described herein is a CHO cell. The host cell can be cultured under suitable conditions for expressing the antibody or any polypeptide chain thereof. Such antibodies or polypeptide chains thereof can be recovered from cultured cells (e.g., from cells or culture supernatant) by conventional methods (e.g., affinity purification). If desired, the antibody polypeptide chain can be incubated under suitable conditions for a suitable time, thereby allowing antibody production. In some embodiments, the host cell contains nucleic acid encoding the antibody heavy chain described herein. In some embodiments, the host cell contains nucleic acid encoding the antibody light chain described herein. In some embodiments, the host cell contains nucleic acid encoding both the heavy chain and the light chain.

[0196] In some embodiments, the method for preparing the antibodies described herein involves a recombinant expression vector encoding the heavy and light chains of the antibodies described herein, as also described herein. The recombinant expression vector can be introduced into suitable host cells (e.g., dhfr-CHO cells) using conventional methods (e.g., calcium phosphate-mediated transfection). Positive transformant host cells can be selected and cultured under suitable conditions that allow for the expression of both polypeptide chains that form the antibody, which can then 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 antibody formation.

[0197] In one example, 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 suitable host cells (e.g., dhfr-CHO cells) using conventional methods (e.g., calcium phosphate-mediated transfection).

[0198] 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 the expression of the antibody polypeptide chain. When both expression vectors are introduced into the same host cell, the antibodies produced therein can be recovered from the host cell or culture medium. If necessary, the polypeptide chain can be recovered from the host cell or culture medium and then incubated under suitable conditions that allow for antibody formation. When the two expression vectors are introduced into different host cells, each of them can be recovered from the corresponding host cell or corresponding culture medium. The two polypeptide chains can then be incubated under suitable conditions to form antibodies.

[0199] Standard molecular biology techniques are used to prepare recombinant expression vectors, transfect host cells, select transformants, culture host cells, and recover antibodies from culture media. For example, some antibodies can be separated by affinity chromatography using a matrix conjugated with protein A or protein G.

[0200] Any nucleic acid of the heavy chain, light chain, or both that encodes the antibodies described herein (e.g., as provided in Table 3), a vector containing such nucleic acid (e.g., an expression vector), and a host cell containing the vector are all within the scope of this disclosure.

[0201] Table 3: Nucleic acid sequences of VH / VL for the anti-KLK5 / KLK7 antibodies listed in Table 1a

[0202] In some embodiments, this disclosure provides an isolated nucleic acid comprising a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with any of SEQ ID NO: 22, 24, 26, or 27. In some embodiments, this disclosure provides an isolated nucleic acid comprising a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with any of SEQ ID NO: 23, 25, 659, or 660. In some embodiments, this disclosure provides an isolated nucleic acid comprising a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with any of SEQ ID NO: 22-27 or 659-660.

[0203] In some embodiments, this disclosure provides an expression vector encoding the anti-KLK5 / KLK7 antibody described herein. In some embodiments, the expression vector comprises isolated nucleic acids having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with any of SEQ ID NO: 22, 24, 26, or 27. In some embodiments, the expression vector comprises isolated nucleic acids having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with any of SEQ ID NO: 23, 25, 659, or 660. In some embodiments, the expression vector contains isolated nucleic acids having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with any of SEQ ID NO: 22-27 or 659-660.

[0204] In some embodiments, the anti-KLK5 / KLK7 antibody described herein is generated by expression in recombinant cells of: (i) isolated nucleic acid having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with any of SEQ ID NO: 22, 24, 26, or 27, and / or (ii) isolated nucleic acid having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with any of SEQ ID NO: 23, 25, 659, or 660.

[0205] In some embodiments, the anti-KLK5 / KLK7 antibody described herein is generated by expressing an expression vector in recombinant cells, the expression vector comprising: (i) isolated nucleic acids having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with any of SEQ ID NO: 22, 24, 26, and 27, and / or (ii) isolated nucleic acids having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with any of SEQ ID NO: 23, 25, 659, or 660.

[0206] 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, protein, nucleic acid, oligonucleotide, or any molecular entity capable of modulating the activity or function of a gene, protein, and / or nucleic acid in a cell.

[0207] In some embodiments, the multispecific antibody comprises a direct fusion or linking of different antigen-specific binding sites. In some embodiments, the multispecific antibody comprises an immunoglobulin-derived heteropolymerized domain to generate the multispecific antibody. In some embodiments, the multispecific antibody can be formed by co-expression of different heavy chains and two different light chains. In some embodiments, the multispecific antibody can be formed by co-expression of different heavy chains and a common light chain. In some embodiments, the multispecific antibody comprises an engineered CH1 domain (the first constant Ig domain of the heavy chain) that facilitates proper heavy-light chain pairing, for example in such co-expression systems as disclosed in International Patent Application Publication No. WO2021067404, published April 8, 2021, entitled “CH1 DOMAIN VARIANTS ENGINEERED FOR PREFERENTIALLIGHT CHAIN ​​PAIRING AND MULTISPECIFIC ANTIBODIES COMPRISING THE SAME,” the contents of which are incorporated herein by reference.

[0208] In some embodiments, the multispecific antibody comprises a variant CH1 domain that pairs (e.g., preferably pairs) with a specific variant CL domain. For example, in some embodiments, the multispecific antibody comprises a heavy chain that comprises a variant CH1 domain that preferentially pairs with a variant CLK or CLλ domain. Non-limiting examples of such multispecific antibody configurations are provided in International Patent Application Publication No. WO2022150787, published July 14, 2022, entitled “VARIANT CH1 DOMAINS AND VARIANT CL DOMAINS ENGINEERED FOR PREFERENTIAL CHAIN ​​PAIRING AND MULTI-SPECIFIC ANTIBODIES COMPRISING THE SAME,” the contents of which are incorporated herein by reference in their entirety.

[0209] In some embodiments, the multispecific antibody includes a variant CH3 domain that preferentially forms a CH3-CH3 heterodimer rather than a CH3-CH3 homodimer. The incorporation of such a variant CH3 domain promotes, for example, heterodimerization of different antibodies to form a multispecific antibody. Non-limiting examples of such multispecific antibody configurations are provided in International Patent Application Publication No. WO2022150785, published July 14, 2022, entitled “VARIANT CH3 DOMAINS ENGINEERED FORPREFERENTIAL CH3 HETERODIMERIZATION, MULTI-SPECIFIC ANTIBODIES COMPRISING THESAME, AND METHODS OF MAKING THEREOF,” the contents of which are incorporated herein by reference in their entirety.

[0210] In some embodiments, the bispecific antibody comprises a direct fusion or linking 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 a common light chain. In some embodiments, the fusion of two antibody-producing cell lines allows for the combination of heavy and light chains of two different antibodies, such that the resulting bispecific antibody comprises the heavy and light chains of a first antibody and the heavy and light chains of a second antibody. In some embodiments, the heavy chain constant region and the light chain constant region are the same isotype. In some embodiments, the heavy chain constant region and the light chain constant region are different isotypes.

[0211] In some embodiments, the bispecific antibody comprises a modified heavy chain and / or light chain that compels proper assembly between the two heavy chains and the homologous heavy and light chains, or facilitates the purification of a properly assembled bispecific antibody (see, for example, Figures 3 and 4 and Table 1 in Brinkmann U and Kontermann EE, The making of bispecific antibodies, MAbs. 2017 Feb / Mar;9(2):182-212). In some embodiments, the bispecific antibody is formed using a mortar and pestle technique, for example, to facilitate heterodimerization. In some implementations, the mortar and pestle technique can be used to generate bispecific IgG molecules, trivalent Ig-like antibodies, bispecific Fc and CH3 fusion proteins, and other forms, as discussed below: 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.

[0212] In some embodiments, the bispecific antibody comprises an engineered CH1 domain (the first constant Ig domain of the heavy chain) that facilitates proper heavy-light chain pairing, for example in such co-expression systems as disclosed in International Patent Application Publication No. WO2021067404, published April 8, 2021, entitled “CH1 DOMAIN VARIANTS ENGINEERED FORPREFERENTIAL LIGHT CHAIN ​​PAIRING AND MULTISPECIFIC ANTIBODIES COMPRISING THESAME”, the contents of which are incorporated herein by reference.

[0213] In some embodiments, the bispecific antibody comprises a variant CH1 domain that pairs (e.g., preferably pairs) with a specific variant CL domain. For example, in some embodiments, the bispecific antibody comprises a heavy chain that comprises a variant CH1 domain that preferentially pairs with a variant CLK or CLλ domain. Non-limiting examples of such bispecific antibody configurations are provided in International Patent Application Publication No. WO2022150787, published July 14, 2022, entitled “VARIANT CH1 DOMAINS AND VARIANT CL DOMAINS ENGINEERED FOR PREFERENTIAL CHAIN ​​PAIRING AND MULTI-SPECIFIC ANTIBODIES COMPRISING THE SAME,” the contents of which are incorporated herein by reference in their entirety.

[0214] In some embodiments, the bispecific antibody comprises a variant CH3 domain that preferentially forms a CH3-CH3 heterodimer rather than a CH3-CH3 homodimer. The incorporation of such a variant CH3 domain promotes, for example, heterodimerization of different antibodies to form a bispecific antibody. A non-limiting example of such bispecific antibody conformation is provided in International Patent Application Publication No. WO2022150785, published July 14, 2022, entitled “VARIANT CH3 DOMAINS ENGINEERED FORPREFERENTIAL CH3 HETERODIMERIZATION, MULTI-SPECIFIC ANTIBODIES COMPRISING THESAME, AND METHODS OF MAKING THEREOF”, the contents of which are incorporated herein by reference in their entirety.

[0215] 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 docking-locking (DNL) assembly of heterodimers utilizing the regulatory subunit of cAMP-dependent protein kinase (PKA) and the anchoring domain (AD) of kinase-anchored protein (AKAP). In some embodiments, non-immunoglobulin heterodimerization modules can be used to combine different antigen-specific binding sites (such as the barnase-barstar system, a linker / docking tag module based on a mutated RNase I fragment, and a SNARE module based on the interaction of three protein synaptic fusion proteins, synaptic vesicle proteins, and SNAP25) to form bispecific antibodies.

[0216] IV. Pharmaceutical Compositions Antibodies as described herein, as well as those encoding nucleic acids or nucleic acid sequences, vectors containing them, or host cells containing vectors, can be mixed with pharmaceutically acceptable excipients to form pharmaceutical compositions for treating a target disease. "Acceptable" means that the vector must be compatible with (and preferably, capable of stabilizing) the active ingredient of the composition and harmless to the subject to be treated. Pharmaceutically acceptable excipients include buffers well known in the art. See, for example, Remington: The Science and Practice of Pharmacy, 20th Edition (2000), Lippincott Williams and Wilkins, eds. KE Hoover.

[0217] The pharmaceutical compositions disclosed herein containing an anti-KLK5 / KLK7 + IL-13 bispecific antibody having at least one antigen-specific binding site may also contain a suitable buffer. A buffer is a weak acid or weak base used to maintain the pH of a solution near a selected value after the addition of another acid or base. In some embodiments, the buffers disclosed herein may be buffers that maintain physiological pH despite changes in carbon dioxide concentration (generated by cellular respiration). Exemplary buffers include, but are not limited to, HEPES (4-(2-hydroxyethyl)-1-piperazine ethanesulfonic acid) buffer, Dürr-Becker phosphate-buffered saline (DPBS) buffer, or phosphate-buffered saline (PBS) buffer. Such buffers may contain disodium hydrogen phosphate and sodium chloride, or potassium dihydrogen phosphate and potassium chloride.

[0218] 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, DA; West, DM; Holler, JF; Crouch, SR (2004). Chapters 14-16. Fundamentals of Analytical Chemistry (8th Edition)). Salts consist of a relevant number of cations (positively charged ions) and anions (negative ions), and therefore the product is electrically neutral (without a net charge).

[0219] In some embodiments, the pharmaceutical composition may comprise a pharmaceutically acceptable carrier, excipient, or stabilizer in the form of a lyophilized formulation or an aqueous solution. (Remington: The Science and Practice of Pharmacy, 20th Edition (2000), Lippincott Williams and Wilkins, eds. KE Hoover). In some embodiments, the pharmaceutical composition may be formulated for intravenous injection. In some embodiments, the pharmaceutical composition may be formulated for subcutaneous injection.

[0220] Pharmaceutical compositions intended for in vivo administration must be sterile. This can be readily achieved, for example, by filtration through a sterile filter membrane. Therapeutic antibody compositions are typically placed in containers with sterile access ports, such as intravenous or subcutaneous solution bags or vials with stoppers that can be punctured by a hypodermic needle.

[0221] V. Usage Instructions In some aspects, this disclosure provides methods and related compositions for treating disorders associated with KLK5 and KLK7 dysregulation, including, for example, Natherton's syndrome, atopic dermatitis (with and without filaggrin mutations), eosinophilic esophagitis, nodular prurigo, chronic pruritus of unknown cause (CPUO), dry skin, asthma (e.g., particularly KLK5-related asthma), ichthyosis vulgaris, and pruritus. In some embodiments, Th2 cytokines (e.g., IL-13, IL-4, IL-5, IL-6, IL-9, IL-31, and IL-17E (IL-25)) are also associated with the KLK5 and KLK7 dysregulation disorders described herein. Therefore, in some aspects, this disclosure provides methods and compositions for inhibiting KLK5 and KLK7, as well as IL-13. In some embodiments, this disclosure provides bispecific antibodies for inhibiting KLK5 and KLK7, as well as IL-13, for the treatment of the disorders described herein.

[0222] This disclosure relates to methods and compositions (e.g., anti-KLK5 / KLK7 dual inhibitor antibodies or anti-KLK5 / KLK7 + IL-13 bispecific antibodies) for promoting proper barrier function (e.g., epidermal barrier function). Overactive kallikrein 5 / 7 causes hereditary and spontaneous disruption of epidermal barrier function and is associated with related conditions such as Natherton's syndrome, eosinophilic esophagitis, and atopic dermatitis. Therefore, in some embodiments, the methods provided herein include administering to a subject an effective amount of one or more anti-KLK5 / KLK7 antibodies or one or more anti-KLK5 / KLK7 + IL-13 bispecific antibodies provided herein for the purpose of restoring the epithelial barrier in a subject with this need. In other aspects, methods for addressing one or more aspects of barrier function alterations are provided. For example, in some embodiments, the methods provided herein include administering an effective amount of one or more anti-KLK5 / KLK7 antibodies or one or more anti-KLK5 / KLK7 + IL-13 bispecific antibodies provided herein to a subject for the purpose of reducing dermal infiltration in a subject with this need. In some embodiments, the methods provided herein include administering an effective amount of one or more anti-KLK5 / KLK7 antibodies or one or more anti-KLK5 / KLK7 + IL-13 bispecific antibodies provided herein to a subject for the purpose of alleviating epithelial inflammation in a subject with this need. In some embodiments, the methods provided herein include administering an effective amount of one or more anti-KLK5 / KLK7 antibodies or one or more anti-KLK5 / KLK7 + IL-13 bispecific antibodies provided herein to a subject for the purpose of reducing epithelial permeability in a subject with this need. In some embodiments, the methods provided herein include administering an effective amount of the presently provided anti-KLK5 / KLK7 antibody or one or more anti-KLK5 / KLK7 + IL-13 bispecific antibodies to a subject for the purpose of reducing parakeratosis in a subject with this need. In some embodiments, the methods provided herein include administering an effective amount of the presently provided anti-KLK5 / KLK7 antibody or one or more anti-KLK5 / KLK7 + IL-13 bispecific antibodies to a subject for the purpose of reducing inflammatory cytokines in a subject with this need. In some embodiments, the methods provided herein include administering an effective amount of the presently provided anti-KLK5 / KLK7 antibody or one or more anti-KLK5 / KLK7 + IL-13 bispecific antibodies to a subject for the purpose of reducing transepidermal water loss in a subject with this need.

[0223] Other aspects of this disclosure relate to methods and compositions (e.g., anti-KLK5 / KLK7 antibodies or anti-KLK5 / KLK7 + IL-13 bispecific antibodies) for treating atopic dermatitis. Atopic dermatitis (AD) (also known as eczema) is a common chronic, itchy, inflammatory skin condition. In some embodiments, atopic dermatitis begins in the subject's infancy or early childhood (e.g., 2 years of age or about 2 years of age). Therefore, in some embodiments, the methods provided herein can be used to treat subjects aged 2 years or older with atopic dermatitis. Atopic dermatitis may be associated with elevated total serum IgE concentrations. Therefore, in some embodiments, the methods provided herein can be used to treat subjects with atopic dermatitis who have elevated total serum IgE levels (e.g., compared to normal IgE levels in subjects who do not have atopic dermatitis or related conditions). In some embodiments, atopic dermatitis is associated with chronic relapsing skin inflammation, epidermal barrier dysfunction (e.g., ultimately leading to dry skin), and / or IgE-mediated sensitization to allergens such as food and environmental allergens. Therefore, in some embodiments, this document provides a method for treating a subject with atopic dermatitis, comprising administering to the subject an effective amount of an anti-KLK5 / KLK7 antibody or one or more anti-KLK5 / KLK7 + IL-13 bispecific antibodies. Furthermore, in some embodiments, the subject to be treated exhibits chronic relapsing skin inflammation, epidermal barrier dysfunction, and / or IgE-mediated sensitization to allergens.

[0224] In some implementations, administration of the anti-KLK5 / KLK7 + IL-13 bispecific antibody described herein to subjects with AD resulted in a reduction in ear thickness of more than 10%, more than 20%, more than 30%, more than 40%, more than 50%, more than 60%, more than 70%, more than 80%, more than 90%, or more than 100%, relative to subjects receiving either of the parental antibodies (e.g., parental anti-KLK5 / KLK7 antibody or parental anti-IL-13 or anti-IL-13R antibody).

[0225] In some implementations, administration of the anti-KLK5 / KLK7 + IL-13 bispecific antibody described herein to a subject with AD resulted in a reduction of skin erythema / bleeding by more than 10%, more than 20%, more than 30%, more than 40%, more than 50%, more than 60%, more than 70%, more than 80%, more than 90%, or more than 100% in the subject, relative to a subject receiving either of the parental antibodies (e.g., parental anti-KLK5 / KLK7 antibody or parental anti-IL-13 or anti-IL-13R antibody).

[0226] In some implementations, administration of the anti-KLK5 / KLK7 + IL-13 bispecific antibody described herein to subjects with AD resulted in a reduction of skin peeling / erosion by more than 10%, more than 20%, more than 30%, more than 40%, more than 50%, more than 60%, more than 70%, more than 80%, more than 90%, or more than 100% in the subject, relative to a subject receiving either of the parental antibodies (e.g., parental anti-KLK5 / KLK7 antibody or parental anti-IL-13 or anti-IL-13R antibody).

[0227] In some implementations, administration of the anti-KLK5 / KLK7 + IL-13 bispecific antibody described herein to subjects with AD resulted in a reduction of skin edema by more than 10%, more than 20%, more than 30%, more than 40%, more than 50%, more than 60%, more than 70%, more than 80%, more than 90%, or more than 100%, relative to subjects receiving either of the parent antibodies of the bispecific antibody (e.g., parent anti-KLK5 / KLK7 antibody or parent anti-IL-13 or anti-IL-13R antibody).

[0228] In some implementations, administration of the anti-KLK5 / KLK7 + IL-13 bispecific antibody described herein to subjects with AD resulted in a reduction of skin desquamation / dryness by more than 10%, more than 20%, more than 30%, more than 40%, more than 50%, more than 60%, more than 70%, more than 80%, more than 90%, or more than 100% in the subject, relative to a subject receiving either of the parental antibodies (e.g., parental anti-KLK5 / KLK7 antibody or anti-IL-13 or anti-IL-13R antibody).

[0229] In some implementations, administration of the anti-KLK5 / KLK7 + IL-13 bispecific antibody described herein to subjects with AD resulted in a reduction in the severity of skin lesions of more than 10%, more than 20%, more than 30%, more than 40%, more than 50%, more than 60%, more than 70%, more than 80%, more than 90%, or more than 100%, relative to subjects receiving either of the parental antibodies (e.g., parental anti-KLK5 / KLK7 antibody or parental anti-IL-13 or anti-IL-13R antibody). The severity of skin lesions (four parameters: erythema / hemorrhagic, edema, exfoliation / erosion, and desquamation / dryness) was assessed using signs of the skin on the ears, neck, and back. The total clinical skin severity score was defined as the sum of the individual scores (0: none; 1: mild; 2: moderate; 3: severe).

[0230] Other aspects of this disclosure relate to methods and compositions (e.g., anti-KLK5 / KLK7 antibodies or anti-KLK5 / KLK7 + IL-13 bispecific antibodies) for treating subjects with Natherton syndrome. In some embodiments, this document provides a method for treating a subject with Natherton syndrome comprising administering to the subject an effective amount of one or more anti-KLK5 / KLK7 antibodies or one or more anti-KLK5 / KLK7 + IL-13 bispecific antibodies. Natherton syndrome is a rare and severe autosomal recessive skin disorder. In some embodiments, Natherton syndrome is associated with atopic manifestations such as congenital erythroderma, specific hair shaft abnormalities, and / or elevated IgE levels (e.g., compared to normal IgE levels in subjects without Natherton syndrome or related disorders). In some embodiments, subjects with Natherton's syndrome exhibit atopic manifestations, including eczematous rash, atopic dermatitis, pruritus, hay fever, angioedema, urticaria, high levels of IgE in serum, and / or eosinophilia. In some embodiments, Natherton's syndrome is caused by a mutation in the serine protease inhibitor Kazal-5 (SPINK5) gene, which encodes a protease inhibitor associated with Kazal-5 lymphoepithelial protease. In some embodiments, a deficiency of this protease inhibitor leads to keratinization secondary to excessive activity of epidermal proteases. Therefore, in some embodiments, the methods provided herein include administering to a subject an effective amount of one or more anti-KLK5 / KLK7 antibodies or one or more anti-KLK5 / KLK7 + IL-13 bispecific antibodies provided herein to alleviate one or more aspects or symptoms associated with Natherton's syndrome (e.g., atopic manifestations such as rash desquamation, keratinization).

[0231] Other aspects of this disclosure relate to methods and compositions (e.g., anti-KLK5 / KLK7 antibodies or anti-KLK5 / KLK7 + IL-13 bispecific antibodies) for treating eosinophilic esophagitis. In some embodiments, symptoms of eosinophilic esophagitis include dysphagia, malnutrition, vomiting, epigastric or chest pain, dysphagia, and food impaction. In some embodiments, the subject with eosinophilic esophagitis is a young male with a relatively high susceptibility to atopic diseases. In some embodiments, the subject with eosinophilic esophagitis is diagnosed by endoscopic examination and / or biopsy of eosinophils isolated from the esophagus. In some embodiments, eosinophilic esophagitis is histologically defined by the presence of proliferative changes, which in some embodiments include thickening of the basal epithelium and / or papillary elongation, at least 24 eosinophils per high-power field in the distal esophagus, and / or the absence of eosinophilia in any other evaluated intestinal segment. In some embodiments, subjects with eosinophilic esophagitis exhibit low levels or a deficiency of certain serine protease inhibitors belonging to the Kazal-type inhibitory protein family of lymphoepithelial cells, such as SPINK7, for example, in esophageal biopsies. In some embodiments, eosinophilic esophagitis is differentiated from reflux esophagitis based on the magnitude of mucosal eosinophilia and the lack of response to acid inhibition. In some embodiments, this document provides a method for treating subjects with eosinophilic esophagitis comprising administering to the subject an effective amount of one or more anti-KLK5 / KLK7 antibodies or one or more anti-KLK5 / KLK7 + IL-13 bispecific antibodies.

[0232] Other aspects of this disclosure relate to methods and compositions (e.g., anti-KLK5 / KLK7 antibodies or anti-KLK5 / KLK7 + IL-13 bispecific antibodies) for treating nodular prurigo. Nodular prurigo is a chronic inflammatory skin disease, the most common symptom of which is an intensely itchy, symmetrically distributed rash on the arms, legs, upper back, and / or abdomen. In some embodiments, nodular prurigo occurs alone. 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, changes in the function of the immune system and nerves in the skin are thought to be associated with increased itching (pruritus) leading to frequent scratching; thus, such frequent scratching and pinching of the skin leads to further thickening and formation of lesions. Therefore, in some embodiments, this document provides a method for treating a subject suffering from nodular prurigo, comprising administering to the subject an effective amount of one or more anti-KLK5 / KLK7 antibodies or one or more anti-KLK5 / KLK7 + IL-13 bispecific antibodies.

[0233] Other aspects of this disclosure relate to methods and compositions (e.g., anti-KLK5 / KLK7 antibodies or anti-KLK5 / KLK7 + IL-13 bispecific antibodies) for treating chronic pruritus. In some embodiments, chronic pruritus is associated with itching lasting longer than 6 weeks (e.g., up to 3 months, up to 6 months, up to 1 year, or longer). In some embodiments, chronic pruritus is associated with underlying unrelated diseases, including chronic kidney disease, hepatobiliary disease, and neuropathic diseases such as brachioradialis pruritus and paresthetic low back pain. In some embodiments, chronic pruritus of unknown cause (CPUO) is established when the underlying cause of the pruritus cannot be determined. In some embodiments, chronic pruritus is associated with severe itching and significant scratching lesions. In some embodiments, this document provides a method for treating a subject suffering from chronic pruritus (including CPUO), comprising administering to the subject an effective amount of one or more anti-KLK5 / KLK7 antibodies or one or more anti-KLK5 / KLK7 + IL-13 bispecific antibodies.

[0234] Other aspects of this disclosure relate to methods and compositions for treating ichthyosis vulgaris (e.g., anti-KLK5 / KLK7 antibodies or anti-KLK5 / KLK7 + IL-13 bispecific antibodies). In some embodiments, ichthyosis vulgaris is caused by a heterozygous mutation in the filaggrin gene. In some embodiments, subjects with homozygous or compound heterozygous mutations in this gene have a more severe phenotype. In some embodiments, the histological features of ichthyosis vulgaris are the absence or reduction of keratinocytes and mild hyperkeratosis in the epidermis. Keratinocytes contain histidine-rich proteins and are a precursor form of filaggrin (a keratin filament aggregate). In some embodiments, filaggrin precursors and filaggrin are reduced or absent in subjects with ichthyosis vulgaris. In some embodiments, ichthyosis vulgaris includes increased palmar creases, keratosis pilaris, and fine peeling most prominent on the lower abdomen, arms, and legs. In some embodiments, subjects may exhibit significant desquamation. In some implementations, subjects may exhibit increased palmar creases, keratosis pilaris, and in some cases, fine scaling. Therefore, in some implementations, this document provides a method for treating subjects with ichthyosis vulgaris or one or more of its symptoms or phenotypic features, comprising administering to the subject an effective amount of one or more anti-KLK5 / KLK7 antibodies or one or more anti-KLK5 / KLK7 + IL-13 bispecific antibodies.

[0235] Other aspects of this disclosure relate to methods and compositions for treating psoriasis (e.g., anti-KLK5 / KLK7 antibodies or anti-KLK5 / KLK7 + IL-13 bispecific antibodies). Psoriasis (or psoriasis vulgaris) is a chronic inflammatory skin disease. In some embodiments, psoriasis is characterized by red, scaly plaques of skin 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, a subject experiences psoriasis onset 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, significant psoriatic inflammatory infiltration at the dermal-epidermal junction may include activated T cells and antigen-presenting cells (APCs). In some embodiments, the presence of activated T cells and APCs in such infiltration precedes the development of epidermal hyperproliferation. In some embodiments, increased levels of inflammatory cytokines can be detected in the diseased 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, comprising administering to the subject an effective amount of one or more anti-KLK5 / KLK7 antibodies or one or more anti-KLK5 / KLK7 + IL-13 bispecific antibodies. In some embodiments, the subject is treated prior to the development of epidermal hyperproliferation. However, in some embodiments, the subject is treated after epidermal hyperproliferation and accelerated differentiation of keratinocytes.

[0236] Other aspects of this disclosure relate to methods and compositions (e.g., anti-KLK5 / KLK7 antibodies or anti-KLK5 / KLK7 + IL-13 bispecific antibodies) for treating rosacea. Rosacea is an inflammatory disease characterized by erythema, papular 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 cathelicidin peptide found in rosacea is proteolytically processed differently from that present in normal subjects. In some embodiments, a method is provided herein for treating a subject with rosacea, comprising administering to the subject an effective amount of one or more anti-KLK5 / KLK7 antibodies or one or more anti-KLK5 / KLK7 + IL-13 bispecific antibodies.

[0237] Other aspects of this disclosure relate to methods for treating a subject with asthma, including administering to the subject an effective amount of one or more antibodies disclosed herein (e.g., anti-KLK5 / KLK7 antibody or anti-KLK5 / KLK7 + IL-13 bispecific antibody). In some embodiments, this document provides a method for treating a subject with asthma, including administering an effective amount of one or more anti-KLK5 / KLK7 antibodies or one or more anti-KLK5 / KLK7 + IL-13 bispecific antibodies. In some embodiments, the subject has asthma (e.g., a subject to be treated with a KLK5-targeting antibody provided herein). In some implementations, asthma is selected from: allergic asthma, aspirin-sensitive / exacerbated asthma, smoking-induced asthma, asthma uncontrolled by corticosteroids or other chronic asthma control medications, atopic asthma, bronchial obstruction-related asthma, pathogenesis-related asthma, chronic asthma, corticosteroid-induced asthma, corticosteroid-resistant asthma, corticosteroid-resistant asthma, eosinophilic hyperasthma, eosinophilic hypoasthma, exercise-induced asthma, mild asthma, moderate to severe asthma, Natherton syndrome asthma, newly diagnosed and / or untreated asthma, non-allergic asthma, non-Th2 driven asthma, hyperperiosteal asthma, hypoperiosteal asthma, Th2 hypoasthma, type 2 (T2) driven asthma, and type 2 hypoinflammatory asthma. In some implementations, the subject has atopic asthma or allergic asthma. In some implementations, the subject has aspirin-sensitive or aspirin-exacerbated asthma. In some implementations, the subject has asthma associated with nonsteroidal anti-inflammatory drugs (NSAIDs). Therefore, in some embodiments, the subject suffers from asthma triggered by aspirin or a similar NSAID (e.g., recently ingested aspirin or a similar NSAID). In some embodiments, the subject suffers from bronchospasm, which may or may not be characterized asthma. For example, in some embodiments, the subject suffers from exercise-induced bronchospasm.

[0238] 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 high eosinophilic asthma (e.g., at least about 150, 200, 250, 300, 350, or 400 eosinophil counts / ml of blood). In some embodiments, the subject has low eosinophilic asthma (e.g., less than about 150 eosinophil counts / μl of blood or less than about 100 eosinophil counts / μl of blood). These and further examples of asthma-related disorders that can be treated with the compositions provided herein (e.g., anti-KLK5 / KLK7 antibodies) are disclosed in WO2015 / 061441, METHODS OFDIAGNOSING AND TREATING EOSINOPHILIC DISORDERS, published on April 30, 2015, the contents of which are incorporated herein by reference.

[0239] In some implementations, the subject has exercise-induced asthma, intermittent or exercise-induced asthma, mild asthma, mild or corticosteroid-induced asthma, moderate to severe asthma, Netherton syndrome asthma, newly diagnosed asthma, untreated asthma, or severe asthma. In some implementations, the subject has asthma that previously did not require or involve the use (e.g., long-term use) of inhaled topical or systemic steroids to control or manage symptoms (e.g., symptoms such as cough, wheezing, shortness of breath / dyspnea, and chest pain). In some embodiments, the subject has low periosteal protein asthma (e.g., periosteal protein levels below about 20 ng / mL serum). In some embodiments, the subject has high periosteal protein asthma (e.g., periosteal protein levels of at least about 20 ng / mL, 25 ng / mL, or 50 ng / mL serum). In some embodiments, the subject has non-allergic asthma (e.g., which may or may not be triggered by infection (e.g., respiratory viruses (e.g., influenza, coronavirus, parainfluenza, rhinovirus, human metapneumovirus, and respiratory syncytial virus)) or inhalation of irritants (air pollutants, smoke, combustion particles (e.g., diesel particles), volatile chemicals, indoor or outdoor gases) or relatively cold, dry air). In some embodiments, the subject has asthma due to acute or chronic primary or secondary exposure to smoke (cigarettes, cigars, pipes, or other combustion products) or due to inhalation or evaporation (nicotine, cannabis, or other similar substances). In some embodiments, the subject has persistent chronic severe asthma with acute events of potentially life-threatening symptom exacerbation (exacerbation or flare-up).

[0240] In some embodiments, the subject has type 2 (Th2) T helper lymphocytes or type 2 (Th2) hyperasthmatic disorder. In some embodiments, the subject has Th2-induced asthma. For example, in some embodiments, Th2 cells and / or their secreted effector molecules mediate an immune response to an allergen and are triggered by exposure to a specific allergen that causes the subject's allergic asthma. In some embodiments, the subject has activated Th2 cell-mediated asthma, which may be partially caused by the secretion of interleukins (e.g., IL-13). Therefore, in some embodiments, a KLK5 antibody (or other antibody) may be used alone or in combination with one or more antibodies targeting IL-13 and allergy-related targets (such as IgE) as a multispecific antibody. Examples of such antibodies for the treatment of asthma include, but are not limited to, omalizumab (XOLAIR®) (targeting soluble IgE); lerekizumab (targeting IL-13); and quilizumab (targeting membrane-bound IgE).

[0241] In addition, in other embodiments, the antibodies described herein are used to treat, but are not limited to, the following diseases: inflammatory conditions, infectious diseases, allergic diseases, and autoimmune diseases. In some embodiments, inflammatory conditions are selected from, but are not limited to, rosacea, nodular prurigo, Crohn's disease, ankylosing spondylitis, ulcerative colitis, hidradenitis suppurativa, and uveitis, or one or more of their barrier-related symptoms. In some embodiments, allergic diseases are selected from, but are not limited to, eczema, atopic dermatitis, asthma, sinusitis, and eosinophilic esophagitis. In some embodiments, autoimmune diseases are selected from, but are not limited to, rheumatoid arthritis, psoriatic arthritis, juvenile idiopathic arthritis, Behçet's disease, and plaque psoriasis, or one or more of their barrier-related symptoms.

[0242] Based on the teachings provided herein, it will be apparent to those skilled in the art that determining whether an antibody (e.g., an anti-KLK5 / KLK7 antibody or an anti-KLK5 / KLK7 + IL-13 bispecific antibody) achieves a therapeutic effect is readily apparent. As will be appreciated by those skilled in the art, effective amounts vary depending on the specific disease being treated, the severity of the disease, individual patient parameters (including age, physical condition, body size, sex, and weight), duration of treatment, the nature of concomitant therapies (if any), the specific route of administration, and similar factors within the knowledge and expertise of the healthcare practitioner. The specific dosage regimens (i.e., dosage, time, and repetition) used in the methods described herein will depend on the specific subject and that subject's medical history, as discussed herein.

[0243] Empirical considerations (such as time to reach maximum effect, half-life, and / or time exceeding a specific concentration) often aid in dosage determination. For example, antibodies compatible with the human immune system (such as humanized or fully human antibodies) can be used to prolong the antibody's half-life and prevent it from being attacked by the host immune system. Other reasons for dosage adjustment include differences in pharmacokinetic or pharmacodynamic responses driven by sex, age, individual response, antibody target, and / or receptor polymorphisms involved in antibody clearance. The frequency of administration can be determined and adjusted during treatment and is generally, but not necessarily, based on the treatment and / or inhibition and / or mitigation and / or delay of the target disease / symptom. Alternatively, a sustained-release formulation of the antibody may be suitable. Various formulations and devices for achieving sustained release are known in the art.

[0244] The frequency of administration may vary depending on the method of protection required. In some embodiments, the composition may be administered once. In some embodiments, the composition may be administered multiple times. In some embodiments, the frequency of administration is weekly, 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 monthly, every 2 months, or every 3 months, or longer. In some embodiments, the composition may be administered daily, every two weeks, weekly, every two months, monthly, or at any time interval that provides appropriate (e.g., maximum) efficacy while minimizing safety risks to the subject. Typically, efficacy and treatment and safety risks can be monitored throughout the course of treatment.

[0245] In some embodiments, the composition provided herein (e.g., anti-KLK5 / KLK7 antibody or anti-KLK5 / KLK7 + IL-13 bispecific antibody) may be administered to the subject at one or more intervals during a defined time period. In some cases, the time periods during which the composition is administered to the subject at one or more intervals may be separated by time periods during which the composition is not administered to the subject. In some embodiments, the relative duration of the respective time periods may depend on the subject's response to treatment or the severity of the disease, or both, and / or may be determined based on the judgment of the treating physician.

[0246] In some implementations, the antibody can be administered parenterally. For example, the parenterally administered composition can be applied locally or via mucosal means through subcutaneous, intradermal, intravenous, intraperitoneal, intratumoral, intramuscular, intra-articular, intra-arterial, or infusion techniques.

[0247] In some embodiments, antibodies (e.g., anti-KLK5 / KLK7 antibodies or anti-KLK5 / KLK7 + IL-13 bispecific antibodies) are administered intravenously. In some embodiments, antibodies (e.g., anti-KLK5 / KLK7 antibodies or anti-KLK5 / KLK7 + IL-13 bispecific antibodies) are administered subcutaneously or locally.

[0248] For intravenous administration, water-soluble antibodies can be administered via infusion, thereby infusing a pharmaceutical preparation containing the antibody and a physiologically acceptable excipient. Physiologically acceptable excipients may include, for example, 5% glucose, 0.9% saline, Ringer's solution, or other suitable excipients. Other injectable compositions may contain various carriers, such as vegetable oils, dimethylacetamide, dimethylformamide, ethyl lactate, ethyl carbonate, isopropyl myristate, ethanol, and polyols (glycerol, propylene glycol, liquid polyethylene glycol, etc.). In some cases, the preparation (e.g., a sterile preparation in the form of a suitable soluble salt of the antibody) may be dissolved and administered in a pharmaceutical excipient (such as water for injection, 0.9% saline, or 5% glucose solution).

[0249] In one implementation, the antibody is administered via a site-specific or targeted local delivery technique. Examples of site-specific or targeted local delivery techniques include various implantable, percutaneous, or transmucosal antibody storage sources or local delivery systems.

[0250] Anti-KLK5 / KLK7 antibodies or one or more anti-KLK5 / KLK7 + IL-13 bispecific antibodies, along with treatments such as those described in this disclosure, may be used in combination with other types of therapies targeting the disease or condition described herein. In this case, the antibody composition and the therapeutic agent may be administered simultaneously or sequentially. These therapies may be administered simultaneously or sequentially (in any order) with treatments according to this disclosure.

[0251] Therefore, aspects of this disclosure relate to methods and compositions (e.g., anti-KLK5 / KLK7 antibodies or anti-KLK5 / KLK7 + IL-13 bispecific antibodies). In some embodiments, the antibodies described herein can be administered as a combination therapy (concurrent or sequential, e.g., over time). In some embodiments, the combination therapy comprises administering one or more of the antibodies described herein (e.g., anti-KLK5 / KLK7 antibodies or anti-KLK5 / KLK7 + IL-13 bispecific antibodies) and at least one additional therapeutic agent (e.g., one, two, three, four, five, six, or seven therapeutic agents). In some embodiments, one or more of the antibodies described herein and at least one additional therapeutic agent (e.g., one, two, three, four, five, six, or seven therapeutic agents) are administered together. In some embodiments, one or more of the antibodies described herein and at least one additional therapeutic agent (e.g., one, two, three, four, five, six, or seven therapeutic agents) are administered separately.

[0252] In some implementations, the additional treatment is an anti-inflammatory agent. In some implementations, 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, certolizumab pegol, etanercept, adalimumab, infliximab, golimumab, ustekinumab, secukinumab, ixekizumab, brolinumab, abatacept, tetrajizumab, lecithinumab, and gusezimab. 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., intravenous, subcutaneous, or intramuscular).

[0253] In some embodiments, the treatment combination comprises one or more of the antibodies described herein (e.g., anti-KLK5 / KLK7 antibody or anti-KLK5 / KLK7 + IL-13 bispecific antibody) delivered together with one or more additional antibodies or fragments thereof (e.g., one, two, three, four, five, six, or seven antibodies). In some embodiments, the treatment combination comprising one or more of the antibodies described herein and one or more additional antibodies or fragments thereof (e.g., one, two, three, four, five, six, or seven antibodies) is delivered separately. In some embodiments, the treatment combination comprising one or more of the antibodies described herein and one or more additional antibodies or fragments thereof (e.g., one, two, three, four, five, six, or seven antibodies) is delivered together.

[0254] In some embodiments, a therapeutic combination comprising one or more of the antibodies 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-specific binding site, and one or more additional unique antigen-specific binding sites from one or more additional antibodies. In some embodiments, the multispecific antibody comprises a direct fusion or linking of different antigen-specific binding sites. In some implementations, the additional antibody or fragment thereof is selected from, but not limited to: anti-IL4R antibody, anti-IL-13 antibody, anti-TNF antibody, anti-IL-12 / 23 antibody, anti-IL-17 antibody, doxycycline, dupilumab, pecelilizumab, etanercept, adalimumab, infliximab, golimumab, ustekinumab, secukinumab, ixekizumab, brolinumab, abatacept, tetragercizumab, levozizumab, and / or gusezizumab.

[0255] In some embodiments, in non-limiting examples, in the treatment of rosacea, additional therapeutic agents may be selected from topical steroids; oral methotrexate; oral cyclosporine; and / or TNF inhibitors. In some embodiments, in non-limiting examples, in the treatment of atopic dermatitis, additional therapeutic agents may be selected from subcutaneous dupilumab and / or topical steroids.

[0256] Any anti-KLK5 / KLK7 antibodies disclosed herein can also be used for the in vitro or in vivo detection of the presence of KLK5 and / or KLK7. Results obtained from such assays can be used for diagnostic purposes (e.g., diagnosing diseases associated with KLK5 and / or KLK7) or for scientific research purposes (e.g., identifying novel KLK5-secreting cell types, studying the biological activity and / or regulation of secreted KLK5 and / or KLK7). For assay applications (such as diagnostic applications), anti-KLK5 / KLK7 antibodies or anti-KLK5 / KLK7 + IL-13 bispecific antibodies as described herein can be conjugated to detectable markers (e.g., imaging agents, such as contrast agents) for the in vivo or in vitro detection of the presence of KLK5 and / or KLK7.

[0257] As used herein, “attachment” or “connection” means the association of two entities, preferably with sufficient affinity to realize the therapeutic / diagnostic benefits of the association between the two entities. The association between the two entities can be direct or through a joint, such as a polymer joint. Attachment or connection can include covalent or non-covalent bonding as well as other forms of association, such as entrapment, for example, one entity on or within another entity, or one or two entities on or within a third entity (such as micelles).

[0258] In other embodiments, the anti-KLK5 / KLK7 antibody, as described herein, can be linked to a detectable label, which is a compound capable of directly or indirectly releasing a detectable signal, enabling the aptamer to be detected, measured, and / or qualitatively analyzed in vitro or in vivo. Examples of such "detectable labels" include, but are not limited to, fluorescent labels, chemiluminescent labels, colorimetric labels, enzyme markers, radioisotopes, and affinity labels (such as biotin). Such labels can be conjugated to the aptamer directly or indirectly using conventional methods.

[0259] Reporter agents can also be dyes, such as fluorophores, which can be used to detect diseases mediated by cells expressing KLK5 and / or KLK7 in tissue samples, respectively.

[0260] For in vitro diagnostic assays, anti-KLK5 / KLK7 antibodies can be contacted with samples suspected of containing KLK5 and / or KLK7 (e.g., cells expressing KLK5 in the disease microenvironment or soluble KLK5). The antibody and sample can be incubated under appropriate conditions for a suitable time to allow the antibody to bind to the KLK5 antigen. This interaction can then be detected using conventional methods (e.g., ELISA, histological staining, or FACS). For in vivo diagnostic assays, an appropriate amount of anti-KLK5 / KLK7 antibody conjugated to a label (e.g., an imaging agent or contrast agent) can be administered to the subject requiring examination. The presence of the labeled antibody can be detected using conventional methods based on the signal released from the label.

[0261] For scientific research purposes, anti-KLK5 / KLK7 antibodies 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 function of KLK5. For example, an appropriate amount of anti-KLK5 / KLK7 can be contacted with a sample suspected of producing KLK5 and / or KLK7 (e.g., a new cell type not previously identified as producing KLK5 and / or KLK7). Cells are permeabilized before contact with the anti-KLK5 / KLK7 antibody. The antibody and sample can be incubated under suitable conditions for an appropriate time to allow the antibody to bind to the KLK5 antigen. Such interactions can then be detected using conventional methods (e.g., ELISA, histological staining, or FACS).

[0262] VI. Kits for therapeutic and diagnostic applications This disclosure also provides kits for the therapeutic or diagnostic applications disclosed herein. Such kits may include one or more containers containing antibodies (e.g., any of the antibodies described herein).

[0263] In some embodiments, the kit may include instructions for use according to any of the methods described herein. The included instructions may include a description of administering the antibody to treat, delay the onset of, or alleviate a target disease as described herein. The kit may also include a description of selecting suitable individuals for treatment based on the identification of whether an individual has the target disease. In other embodiments, the instructions include a description of administering the antibody to an individual at risk of the target disease.

[0264] Instructions for use related to the antibody typically include information about dosage, dosing regimen, and route of administration for the intended treatment. Containers may be unit dose, bulk packaging (e.g., multi-dose packaging), or subunit dose. Instructions provided with the kits of this invention are typically written instructions on a label or packaging insert (e.g., a slip of paper included in the kit), but machine-readable instructions (e.g., instructions carried on a magnetic or optical storage disk) are also acceptable.

[0265] The label or packaging insert indicates that the composition is used to treat, delay the onset of, and / or alleviate a disease or condition. Instructions for use in carrying out any of the methods described herein may be provided.

[0266] The kit of the present invention is packaged in suitable packaging. Suitable packaging includes, but is not limited to, vials, bottles, wide-mouth bottles, flexible packaging (e.g., sealed polyester film (Mylar) or plastic bags), etc.

[0267] Packaging for use in conjunction with specific devices, such as infusion devices or micropumps, is also envisioned. The kit may have a sterile access port (e.g., the container may be an intravenous infusion bag or a vial with a stopper that can be pierced by a hypodermic needle). The container may also have a sterile access port (e.g., the container may be an intravenous infusion bag or a vial with a stopper that can be pierced by a hypodermic needle). At least one active agent in the composition is an antibody as described herein.

[0268] The kit may optionally provide additional components, such as buffer solutions and explanatory information. Typically, the kit includes a container and a label or packaging insert on or associated with the container. In some embodiments, the invention provides an article of manufacture containing the contents of the aforementioned kit.

[0269] This document also provides kits for detecting target proteins (e.g., KLK5 and / or KLK7, or KLK5 and KLK7 plus IL-13) in samples. Such kits may contain any of the antibodies described herein. In some cases, antibodies may be conjugated to detectable markers as described herein. As used herein, “conjugation” or “linkage” means the association of two entities, preferably with sufficient affinity to realize the therapeutic / diagnostic benefits of association between the two entities. Association between two entities can be direct or through a linker, such as a polymer linker. Conjugation or linkage can include covalent or non-covalent binding as well as other forms of association, such as entrapment, for example, one entity on or within another entity, or one or two entities on or within a third entity (such as micelles).

[0270] Alternatively, the kit may contain a secondary antibody capable of binding to the antibody described herein. The kit may also include instructions for using the antibody to detect the target protein (e.g., KLK5 and / or KLK7, or KLK5 and KLK7 plus IL-13).

[0271] Example Example 1: Generation and selection of dual-inhibitor anti-KLK5 / KLK7 antibodies (i) Affinity Antibody binding kinetics assays were performed using Biacore to screen for anti-KLK5 / KLK7 antibodies with high affinity for the corresponding targets. All screening assays were performed at 25°C. The run buffer used was 20 mM HEPES, 300 mM NaCl, 0.01% Tween-20, pH 7.5. Antibodies (1 μg / mL) were captured on Fc2-4 of the Cytiva S-series protein A sensor chip at a flow rate of 10 μL / mL for 30 sec. Kinetic measurements were performed using a series of 4–5 concentrations in single-cycle kinetic mode, with the highest concentration being 25 nM–100 nM and serial dilutions of 4-fold. The contact time was typically 300 sec, and the dissociation time was 1800 sec–3600 sec. A flow rate of 30 μL / mL was typically used. The sensor chip was regenerated with 10 mM glycine (pH 1.5) at a contact time of 30 sec and a flow rate of 50 μL / mL. The results showed that the dual inhibitor antibodies described in Table 1a exhibited binding specificity for both KLK5 and KLK7. Table 4 below provides a summary of the binding kinetics of the anti-KLK5 antibody.

[0272] Table 4. Affinity and titer of anti-KLK5 / KLK7 antibodies

[0273] (ii) Protease inhibition assay To evaluate the ability of the antibodies described in Table 1a to inhibit the activity of KLK5 and / or KLK7 proteases, 1.5 nM human or mouse KLK5 or 0.5 nM human or mouse KLK7 was prepared in assay buffer (1 M NaH2PO4 pH 7.5 for KLK5, and 50 mM Tris, 150 mM NaCl pH 7.5 for KLK7). Antibodies diluted to the test concentration in PBS were added. Then, substrates were added (50 μM BOC-Val-Pro-Arg-AMC for KLK5, and 30 μM KHLF-AMC for KLK7). Fluorescence was then measured at 30 min per minute at room temperature for 30 minutes.

[0274] The results showed that the anti-KLK5 / KLK7 antibody could inhibit the activity of KLK5 and KLK7 proteases, as described in Table 4.

[0275] Example 2: Generation of anti-KLK5 / KLK7+ Th2 antibodies targeting bispecific antibodies Experiments showed that in the Nc / Nga mouse model, inhibition of KLK5 / KLK7 using a dual-inhibitor anti-KLK7 antibody had a superior effect compared to anti-IL4R antibodies. Atopic dermatitis was induced in Nc / Nga mice (a strain with an intrinsic barrier defect due to reduced ceramide expression) by topical application of house dust mite (HDM) allergen. Eight to ten-week-old animals received Biostir-AD ointment (120 mg / mouse) containing a house dust mite allergen, twice weekly for two weeks, for a total of 480 mg Biostir-AD per mouse, on the skin areas of the ears and back (including the neck). Two hours prior to the second treatment (day 4), 150 µL of SDS was applied. Starting on day -6, the mediator or test product (control IgG, KLK5-KLK7-Dual Ab4, KLK5 / 7 inhibitor, or anti-IL-4R inhibitor antibody) was administered intraperitoneally (IP) three times weekly at a dose of 30 mg / kg for a total of 9 doses. The anti-IL-4R inhibitor antibody inhibited IL-4 signaling. The reference compound (tacrolimus ointment) was applied topically once daily (QD) starting on day -6 for a total of 21 days (QD×21), one (1) hour after the Biostir-AD administration on the Biostir-AD challenge day. On day 15, the model underwent gross assessment of skin lesions and ear thickness measurement. Ear thickness was measured using a Dyer micrometer. The severity of skin lesions (four parameters: erythema / hemoptysis, edema, exfoliation / erosion, and desquamation / dryness) was assessed by signs of skin on the ears, neck, and back. The overall clinical skin severity score was defined as the sum of individual scores (0: none; 1: mild; 2: moderate; 3: severe).

[0276] The results showed that, compared with the anti-IL4R antibody, inhibiting KLK5 / KLK7 had a more significant effect in reducing ear thickness and clinical skin scores.

[0277] Therefore, it is advantageous to generate bispecific antibodies that inhibit KLK5 / KLK7 activity and Th2 signaling in a single molecule. For example, bispecific antibodies having one arm of an antigen-specific binding site for anti-KLK5 / KLK7 antibodies as described in Table 1a and another arm of an antigen-specific binding site for Th2-targeting antibodies as described in Table 2 are suitable for inhibiting KLK5 / KLK7 and Th2 signaling by a single molecule.

[0278] In one example, the bispecific antibody inhibiting KLK5 / KLK7 and IL-13 was constructed from one arm having an antigen-binding site for KLK5 / KLK7 derived from KLK5 / KLK7-Dual-Ab4 and another arm having an antigen-binding site for an anti-IL-13 antibody (designated K13-0004) containing a VH with the amino acid sequence of SEQ ID NO: 32 and a VL with the amino acid sequence of SEQ ID NO: 33. The binding affinity of the anti-KLK5 / KLK7 + IL-13 bispecific antibody was evaluated and compared with that of monospecific monoclonal antibodies having the same antigen-binding site for the corresponding antigens, as shown in Tables 5 and 7, thus demonstrating that such bispecific antibodies inhibit both pathways in a single molecule (see also...). Figures 4A to 4D Monospecific monoclonal antibodies are bivalent to their corresponding antigens. Bispecific antibodies are monovalent to each antigen.

[0279] Table 5. Affinity and antibody D (KD) of anti-KLK5 / KLK7 + IL-13 bispecific antibodies.

[0280] Table 7. Affinity and titer (IC50) of anti-KLK5 / KLK7 + IL-13 binding antibodies

[0281] Figures 4A to 4C It showed binding to KLK5 with the anti-KLK5 / KLK7+IL-13 bispecific antibody. Figure 4A ), KLK7 ( Figure 4B ) and IL13 ( Figure 4C The K corresponding to Tables 5 and 7) D Data. Binding was achieved by capturing bispecific antibodies (1 μg / mL) on the surface of the protein A chip. The flow rate during the capture step was 10 μL / min for 30 seconds. Then, different concentrations of hKLK5, hKLK7, or hIL-13 were applied to the chip using a single-cycle kinetic approach. The concentrations of hKLK5 and hKLK7 included 0.156 nM, 0.625 nM, 2.5 nM, 10 nM, and 40 nM. The concentrations of hIL-13 included 0.39 nM, 1.56 nM, 6.25 nM, 25 nM, and 100 nM. The flow rate during sample injection was 50 μL / min, the association time was 300 seconds, and the dissociation time was 3600 seconds. The chip was regenerated using 10 mM glycine at pH 1.5. The flow rate during the regeneration step was 50 μL / min for 30 seconds. The anti-KLK5 / KLK7 + IL-13 bispecific antibody binds to all targets with sub-nanomolar affinity.

[0282] To evaluate the ability of bispecific antibodies to inhibit the activity of hKLK5 and hKLK7 proteases, 1.5 nM of hKLK5 or hKLK7 was prepared in assay buffer (0.1 M Tris-HCl, 0.1% (v / v) Triton X-100, pH 8.3). Antibodies diluted to the test concentration in assay buffer were added and incubated with the proteases for 15 min. Then, substrates were added (500 uM BOC-Val-Pro-Arg-AMC for hKLK5 and 50 uM KHLF-AMC for hKLK7). Fluorescence was then measured at room temperature for 1 hour at a rate of 1 minute. Anti-KLK5 / KLK7 + IL-13 bispecific antibodies inhibited KLK5, KLK7, and IL-13, as did anti-KLK5 / KLK7 bispecific antibodies (KLK5 and KLK7). Figures 4D to 4G ) or anti-IL13 and anti-IL4 antibodies (IL-13, Figure 4H This indicates that configuring anti-KLK5 / KLK7 antibodies as bispecific antibodies does not interfere with the anti-KLK5 / KLK7 activity of anti-KLK5 / KLK7 bispecific antibodies or anti-IL-13 antibodies. Compared with treatment with anti-IL-13 antibodies or anti-IL-4 antibodies, the concentration of periosteal protein (an extracellular matrix protein (ECM) that is increased in some allergic diseases) decreased to a comparable level after treatment with anti-KLK5 / KLK7 + IL-13 bispecific antibodies, further indicating that the bispecific form of anti-KLK5 / anti-KLK7 + IL-13 targeting antibodies does not alter efficacy.

[0283] Example 3: Anti-KLK5 / KLK7 + IL-13 bispecific antibody inhibits primary human keratinocyte barrier disruption. Corning Transwell inserts were treated with 100 μL of 50 μg / mL rat tail type I collagen for 1 hour, followed by washing with 1X PBS. Primary human keratinocytes were seeded at a density of 50,000 cells / well on the collagen-treated inserts. After 24 hours, the medium was replaced with medium containing 1.5 mM calcium chloride to promote differentiation, and the cells were incubated for 72 hours. After 72 hours, the medium was replaced with 0.1 mM calcium chloride, and the apical and basal sides of the differentiated cells were treated with a mixture of 125 nM KLK5 and 125 nM KLK7 containing and without inhibitory antibodies ranging from 30 nM to 500 nM. After 24 hours, transepithelial resistance of the cell barrier was measured using an Applied Biophysics TEER24 instrument. After the transepithelial resistance measurement, the medium was replaced with fresh medium, and 1 mg / mL FITC-dextran dye was added to the apical side of the transwell inserts. After 24 hours, the culture medium was collected from the bottom side, and the amount of migrated dye was measured by fluorescence under 487 nm excitation and 528 nm emission.

[0284] Activation of KLK5 and KLK7 leads to disruption of the epithelial barrier, thereby reducing transepithelial resistance (TEER) and increasing FITC-dextran dye migration. This barrier disruption is inhibited by both the anti-KLK5 / KLK7 + IL-13 bispecific antibody (K13-0004) and the anti-KLK5 / KLK7 dual inhibitory antibody (KLK5 / KLK7-Dual-Ab4), which results in a dose-dependent increase in transepithelial resistance. Figures 5A to 5B and the reduction of dye migration ( Figures 5C to 5D This has been confirmed.

[0285] Other implementation plans All features disclosed in this specification can be combined in any combination. Each feature disclosed in this specification can be used to replace alternative features for the same, equivalent, or similar purposes. Therefore, unless otherwise expressly stated, each disclosed feature is merely one example of a general series of equivalent or similar features.

[0286] Based on the above description, those skilled in the art can readily determine the basic features of the present invention, and various changes and modifications can be made to the present invention to suit various uses and conditions without departing from its spirit and scope.

[0287] Equivalent schemes and scope In the claims, unless otherwise indicated or clearly apparent from the context, articles such as “a,” “an,” and “the” may mean one or more. Unless otherwise indicated or clearly apparent from the context, a claim or description including “or” among one or more members of the group is considered satisfied if one, more than one, or all members of the group are present in, used in, or otherwise associated with a given product or method. The invention includes embodiments in which exactly one member of the group is present in, used in, or otherwise associated with a given product or method. The invention includes embodiments in which more than one or all members of the group are present in, used in, or otherwise associated with a given product or method.

[0288] Furthermore, this invention encompasses all variations, combinations, and arrangements in which one or more limitations, elements, clauses, and descriptive terms from one or more of the listed claims are incorporated into another claim. For example, any claim dependent on another claim may be modified to incorporate one or more limitations seen in any other claim dependent on the same basic claim. When elements are presented as a list, for example in Markush group format, each subgroup of said elements is also disclosed, and any one or more elements may be removed from said group. It should be understood that, generally, when the invention or multiple aspects of the invention are referred to as containing specific elements and / or features, certain embodiments of the invention or multiple aspects of the invention consist of, or are substantially consist of, such elements and / or features. For the sake of brevity, those embodiments are not specifically listed herein in their original wording.

[0289] As used in this specification and claims, the phrase “and / or” should be understood to mean “any one or both” of the elements so combined, that is, the elements are combined in some cases and separate in others. Multiple elements listed using “and / or” should be interpreted in the same way, that is, “one or more” of the elements so combined. Other elements may be optional, whether related to or unrelated to those explicitly identified by the “and / or” clause. Therefore, as a non-limiting embodiment, when used in conjunction with open-ended language (e.g., “comprising”), the reference to “A and / or B” may, in one embodiment, refer only to A (optionally including elements other than B); in another embodiment, refer only to B (optionally including elements other than A); in yet another embodiment, refer to both A and B (optionally including other elements), and so on.

[0290] 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 separatin...

Claims

1. A bispecific antibody comprising an antigen-specific binding site that specifically binds to KLK5 and KLK7 and an antigen-specific binding site that specifically binds to IL-13 or its receptor.

2. The bispecific antibody of claim 1, wherein the antigen-specific binding sites that specifically bind to KLK5 and KLK7 bind to the active sites of KLK5 and KLK7 and inhibit enzyme activity.

3. The bispecific antibody of claim 1 or 2, wherein the antigen-specific binding sites that specifically bind to KLK5 and KLK7 comprise HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2 and / or LC CDR3 of any of the antibodies listed in Tables 1a and 1b.

4. The bispecific antibody according to any one of claims 1 to 3, wherein the antigen-specific binding sites that specifically bind to KLK5 and KLK7 comprise: (a) HC CDR1, HC CDR2, and HC CDR3 of heavy chain variable domains having the amino acid sequence of SEQ ID NO: 7, and LC CDR1, LC CDR2, and LC CDR3 of light chain variable domains having the amino acid sequence of SEQ ID NO:

8. (b) HC CDR1, HC CDR2 and HC CDR3 of heavy chain variable domains having the amino acid sequence of SEQ ID NO: 13, and LC CDR1, LC CDR2 and LC CDR3 of light chain variable domains having the amino acid sequence of SEQ ID NO:

14. (c) HC CDR1, HC CDR2, and HC CDR3 of the heavy chain variable domain, having the amino acid sequence of SEQ ID NO: 17, and LC CDR1, LC CDR2, and LC CDR3 of the light chain variable domain, having the amino acid sequence of SEQ ID NO: 14; or (d) HC CDR1, HC CDR2 and HC CDR3 of heavy chain variable domains having the amino acid sequence of SEQ ID NO: 21, and LC CDR1, LC CDR2 and LC CDR3 of light chain variable domains having the amino acid sequence of SEQ ID NO:

14.

5. The bispecific antibody according to any one of claims 1 to 4, wherein the antigen-specific binding sites that specifically bind to KLK5 and KLK7 comprise: (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.

6. The bispecific antibody according to any one of claims 1 to 5, wherein the antigen-specific binding sites specifically binding to KLK5 and KLK7 are VH and / or VL of any of the antibodies listed in Table 1a.

7. The bispecific antibody according to any one of claims 1 to 6, wherein the antigen-specific binding sites that specifically bind to KLK5 and KLK7 comprise: (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.

8. The bispecific antibody according to any one of claims 1 to 7, wherein the antigen-specific binding site that specifically binds to IL-13 or its receptor comprises any of the antibodies listed in Table 2, namely HC CDR1, HC CDR2, HCCDR3, LC CDR1, LC CDR2 and / or LC CDR3.

9. The bispecific antibody of claim 8, wherein the antigen-specific binding site that specifically binds to IL-13 or IL-13R comprises: (a) HC CDR1, HC CDR2, and HC CDR3 of heavy chain variable domains having the amino acid sequence of SEQ ID NO: 53, and LC CDR1, LC CDR2, and LC CDR3 of light chain variable domains having the amino acid sequence of SEQ ID NO:

54. (b) HC CDR1, HC CDR2 and HC CDR3 of heavy chain variable domains having the amino acid sequence of SEQ ID NO: 32, and LC CDR1, LC CDR2 and LC CDR3 of light chain variable domains having the amino acid sequence of SEQ ID NO: 33; (c) HC CDR1, HC CDR2 and HC CDR3 of heavy chain variable domains having the amino acid sequence of SEQ ID NO: 74, and LC CDR1, LC CDR2 and LC CDR3 of light chain variable domains having the amino acid sequence of SEQ ID NO:

75. (d) HC CDR1, HC CDR2 and HC CDR3 of heavy chain variable domains having the amino acid sequence of SEQ ID NO: 94, and LC CDR1, LC CDR2 and LC CDR3 of light chain variable domains having the amino acid sequence of SEQ ID NO: 95; (e) HC CDR1, HC CDR2 and HC CDR3 of heavy chain variable domains having the amino acid sequence of SEQ ID NO: 114, and LC CDR1, LC CDR2 and LC CDR3 of light chain variable domains having the amino acid sequence of SEQ ID NO:

115. (f) HC CDR1, HC CDR2, and HC CDR3 of the heavy chain variable domains, having the amino acid sequence of SEQ ID NO: 134, and LC CDR1, LC CDR2, and LC CDR3 of the light chain variable domains, having the amino acid sequence of SEQ ID NO: 135; or (g) HC CDR1, HC CDR2 and HC CDR3 of heavy chain variable domains having the amino acid sequence of SEQ ID NO: 657, and LC CDR1, LC CDR2 and LC CDR3 of light chain variable domains having the amino acid sequence of SEQ ID NO:

658.

10. The bispecific antibody of claim 8 or 9, wherein the antigen-specific binding site that specifically binds to IL-13 or IL-13R comprises: (a) HC CDR1 having the amino acid sequence of SEQ ID NO: 55; HC CDR2 having the amino acid sequence of SEQ ID NO: 56; HC CDR3 having the amino acid sequence of SEQ ID NO: 57; LC CDR1 having the amino acid sequence of SEQ ID NO: 58; LC CDR2 having the amino acid sequence of SEQ ID NO: 59; and LC CDR3 having the amino acid sequence of SEQ ID NO: 60; (b) HC CDR1 having the amino acid sequence of SEQ ID NO: 61; HC CDR2 having the amino acid sequence of SEQ ID NO: 62; HC CDR3 having the amino acid sequence of SEQ ID NO: 63; LC CDR1 having the amino acid sequence of SEQ ID NO: 64; LC CDR2 having the amino acid sequence of SEQ ID NO: 65; and LC CDR3 having the amino acid sequence of SEQ ID NO: 66; (c) HC CDR1 having the amino acid sequence of SEQ ID NO: 67; HC CDR2 having the amino acid sequence of SEQ ID NO: 68; HC CDR3 having the amino acid sequence of SEQ ID NO: 69; LC CDR1 having the amino acid sequence of SEQ ID NO: 70; LC CDR2 having the amino acid sequence of SEQ ID NO: 71; and LC CDR3 having the amino acid sequence of SEQ ID NO: 72; (d) HC CDR1 having the amino acid sequence of SEQ ID NO: 34; HC CDR2 having the amino acid sequence of SEQ ID NO: 35; HC CDR3 having the amino acid sequence of SEQ ID NO: 36; LC CDR1 having the amino acid sequence of SEQ ID NO: 37; LC CDR2 having the amino acid sequence of SEQ ID NO: 38; and LC CDR3 having the amino acid sequence of SEQ ID NO: 39; (e) HC CDR1 having the amino acid sequence of SEQ ID NO: 40; HC CDR2 having the amino acid sequence of SEQ ID NO: 41; HC CDR3 having the amino acid sequence of SEQ ID NO: 42; LC CDR1 having the amino acid sequence of SEQ ID NO: 43; LC CDR2 having the amino acid sequence of SEQ ID NO: 44; and LC CDR3 having the amino acid sequence of SEQ ID NO: 45; (f) HC CDR1 having the amino acid sequence of SEQ ID NO: 46; HC CDR2 having the amino acid sequence of SEQ ID NO: 47; HC CDR3 having the amino acid sequence of SEQ ID NO: 48; LC CDR1 having the amino acid sequence of SEQ ID NO: 49; LC CDR2 having the amino acid sequence of SEQ ID NO: 50; and LC CDR3 having the amino acid sequence of SEQ ID NO: 51; (g) HC CDR1 having the amino acid sequence of SEQ ID NO: 76; HC CDR2 having the amino acid sequence of SEQ ID NO: 77; HC CDR3 having the amino acid sequence of SEQ ID NO: 78; LC CDR1 having the amino acid sequence of SEQ ID NO: 79; LC CDR2 having the amino acid sequence of SEQ ID NO: 80; and LC CDR3 having the amino acid sequence of SEQ ID NO: 81; (h) HC CDR1 having the amino acid sequence of SEQ ID NO: 82; HC CDR2 having the amino acid sequence of SEQ ID NO: 83; HC CDR3 having the amino acid sequence of SEQ ID NO: 84; LC CDR1 having the amino acid sequence of SEQ ID NO: 85; LC CDR2 having the amino acid sequence of SEQ ID NO: 86; and LC CDR3 having the amino acid sequence of SEQ ID NO: 87; (i) HC CDR1 having the amino acid sequence of SEQ ID NO: 88; HC CDR2 having the amino acid sequence of SEQ ID NO: 89; HC CDR3 having the amino acid sequence of SEQ ID NO: 90; LC CDR1 having the amino acid sequence of SEQ ID NO: 91; LC CDR2 having the amino acid sequence of SEQ ID NO: 92; and LC CDR3 having the amino acid sequence of SEQ ID NO: 93; (j) HC CDR1 having the amino acid sequence of SEQ ID NO: 96; HC CDR2 having the amino acid sequence of SEQ ID NO: 97; HC CDR3 having the amino acid sequence of SEQ ID NO: 98; LC CDR1 having the amino acid sequence of SEQ ID NO: 99; LC CDR2 having the amino acid sequence of SEQ ID NO: 100; and LC CDR3 having the amino acid sequence of SEQ ID NO: 101; (k) HC CDR1 having the amino acid sequence of SEQ ID NO: 102; HC CDR2 having the amino acid sequence of SEQ ID NO: 103; HC CDR3 having the amino acid sequence of SEQ ID NO: 104; LC CDR1 having the amino acid sequence of SEQ ID NO: 105; LC CDR2 having the amino acid sequence of SEQ ID NO: 106; and LC CDR3 having the amino acid sequence of SEQ ID NO: 107; (l) HC CDR1 having the amino acid sequence of SEQ ID NO: 108; HC CDR2 having the amino acid sequence of SEQ ID NO: 109; HC CDR3 having the amino acid sequence of SEQ ID NO: 110; LC CDR1 having the amino acid sequence of SEQ ID NO: 111; LC CDR2 having the amino acid sequence of SEQ ID NO: 112; and LC CDR3 having the amino acid sequence of SEQ ID NO: 113; (m) HC CDR1 having the amino acid sequence of SEQ ID NO: 116; HC CDR2 having the amino acid sequence of SEQ ID NO: 117; HC CDR3 having the amino acid sequence of SEQ ID NO: 118; LC CDR1 having the amino acid sequence of SEQ ID NO: 119; LC CDR2 having the amino acid sequence of SEQ ID NO: 120; and LC CDR3 having the amino acid sequence of SEQ ID NO: 121; (n) HC CDR1 having the amino acid sequence of SEQ ID NO: 122; HC CDR2 having the amino acid sequence of SEQ ID NO: 123; HC CDR3 having the amino acid sequence of SEQ ID NO: 124; LC CDR1 having the amino acid sequence of SEQ ID NO: 125; LC CDR2 having the amino acid sequence of SEQ ID NO: 126; and LC CDR3 having the amino acid sequence of SEQ ID NO: 127; (o) HC CDR1 having the amino acid sequence of SEQ ID NO: 128; HC CDR2 having the amino acid sequence of SEQ ID NO: 129; HC CDR3 having the amino acid sequence of SEQ ID NO: 130; LC CDR1 having the amino acid sequence of SEQ ID NO: 131; LC CDR2 having the amino acid sequence of SEQ ID NO: 132; and LC CDR3 having the amino acid sequence of SEQ ID NO: 133; (p) HC CDR1 having the amino acid sequence of SEQ ID NO: 136; HC CDR2 having the amino acid sequence of SEQ ID NO: 137; HC CDR3 having the amino acid sequence of SEQ ID NO: 138; LC CDR1 having the amino acid sequence of SEQ ID NO: 139; LC CDR2 having the amino acid sequence of SEQ ID NO: 140; and LC CDR3 having the amino acid sequence of SEQ ID NO: 141; (q) HC CDR1 having the amino acid sequence of SEQ ID NO: 142; HC CDR2 having the amino acid sequence of SEQ ID NO: 143; HC CDR3 having the amino acid sequence of SEQ ID NO: 144; LC CDR1 having the amino acid sequence of SEQ ID NO: 145; LC CDR2 having the amino acid sequence of SEQ ID NO: 146; and LC CDR3 having the amino acid sequence of SEQ ID NO: 147; (r) HC CDR1 having the amino acid sequence of SEQ ID NO: 148; HC CDR2 having the amino acid sequence of SEQ ID NO: 149; HC CDR3 having the amino acid sequence of SEQ ID NO: 150; LC CDR1 having the amino acid sequence of SEQ ID NO: 151; LC CDR2 having the amino acid sequence of SEQ ID NO: 152; and LC CDR3 having the amino acid sequence of SEQ ID NO: 153; (s) HC CDR1 having the amino acid sequence of SEQ ID NO: 655; HC CDR2 having the amino acid sequence of SEQ ID NO: 41; HC CDR3 having the amino acid sequence of SEQ ID NO: 42; LC CDR1 having the amino acid sequence of SEQ ID NO: 43; LC CDR2 having the amino acid sequence of SEQ ID NO: 44; and LC CDR3 having the amino acid sequence of SEQ ID NO: 45; or (t) HC CDR1, having the amino acid sequence of SEQ ID NO: 656; HC CDR2, having the amino acid sequence of SEQ ID NO: 47; HC CDR3, having the amino acid sequence of SEQ ID NO: 48; LC CDR1, having the amino acid sequence of SEQ ID NO: 49; LC CDR2, having the amino acid sequence of SEQ ID NO: 50; and LC CDR3, having the amino acid sequence of SEQ ID NO:

51.

11. The bispecific antibody of claim 9 or 10, wherein the antigen-specific binding site that specifically binds to IL-13 or IL-13R comprises: (a) VH, which contains the amino acid sequence of SEQ ID NO: 53; and VL, which contains the amino acid sequence of SEQ ID NO: 54; (b) VH, which contains the amino acid sequence of SEQ ID NO: 32; and VL, which contains the amino acid sequence of SEQ ID NO: 33; (c) VH, which contains the amino acid sequence of SEQ ID NO: 74; and VL, which contains the amino acid sequence of SEQ ID NO: 75; (d) VH, which contains the amino acid sequence of SEQ ID NO: 94; and VL, which contains the amino acid sequence of SEQ ID NO: 95; (e) VH, which contains the amino acid sequence of SEQ ID NO: 114; and VL, which contains the amino acid sequence of SEQ ID NO: 115; (f) VH, comprising the amino acid sequence of SEQ ID NO: 134; and VL, comprising the amino acid sequence of SEQ ID NO: 135; or (g) VH, which contains the amino acid sequence of SEQ ID NO: 657; and VL, which contains the amino acid sequence of SEQ ID NO:

658.

12. The bispecific antibody according to any one of claims 1 to 11, wherein the antigen-specific binding site that specifically binds to IL-13 or its receptor has a VH and / or VL of any of the antibodies in Table 2.

13. The bispecific antibody according to any one of claims 1 to 12, wherein the binding affinity of the bispecific antibody for KLK5 and KLK7 is not more than 20% different from that of an anti-KLK5 / KLK7 antibody comprising the same CDR and / or VH / VL as the bispecific antibody.

14. The bispecific antibody according to any one of claims 1 to 13, wherein the binding affinity of the bispecific antibody for IL-13 signaling is not more than 20% different from that of an anti-IL-13 antibody comprising the same CDR and / or VH / VL as the bispecific antibody.

15. The bispecific antibody according to any one of claims 1 to 14, wherein the bispecific antibody retains at least 80% inhibitory activity against KLK5 and KLK7 relative to an anti-KLK5 / KLK7 antibody comprising the same CDR and / or VH / VL as the bispecific antibody.

16. The bispecific antibody according to any one of claims 1 to 15, wherein the bispecific antibody retains at least 80% inhibitory activity against IL-13 signaling relative to an anti-IL-13 antibody comprising the same CDR and / or VH / VL as the bispecific antibody.

17. A composition comprising a bispecific antibody as described in any one of claims 1 to 16, and an acceptable carrier.

18. A method comprising administering to a subject a bispecific antibody as described in any one of claims 1 to 16 or a composition as described in claim 17.

19. The method of claim 18, wherein the subject has a skin barrier defect.

20. A method for treating skin barrier defects, the method comprising administering to a subject an effective amount of a bispecific antibody as described in any one of claims 1 to 16 or a composition as described in claim 17.

21. The method of claim 19 or 20, wherein the skin barrier defect is associated with Natherton syndrome, atopic dermatitis, eosinophilic esophagitis, nodular prurigo, chronic pruritus of unknown cause (CPUO), dry skin, asthma (especially KLK5), ichthyosis vulgaris, or itching or chronic pruritus.

22. The method of any one of claims 18 to 21, wherein the subject suffers from atopic dermatitis.

23. The method of claims 18 to 22, wherein the application reduces ear thickness by more than 30% relative to a subject receiving an anti-KLK5 / KLK7 antibody comprising the same CDR and / or VH / VL as the bispecific antibody.

24. The method of any one of claims 18 to 23, wherein the application reduces ear thickness by more than 30% relative to the administration of an anti-IL-13 antibody comprising the same CDR and / or VH / VL as the bispecific antibody to a subject.

25. The method of any one of claims 18 to 24, wherein the administration reduces skin erythema / bleeding by more than 30% relative to the administration of an anti-KLK5 / KLK7 antibody comprising the same CDR and / or VH / VL as the bispecific antibody to a subject.

26. The method of any one of claims 18 to 25, wherein the administration reduces skin erythema / bleeding by more than 30% relative to the administration of an anti-IL-13 antibody comprising the same CDR and / or VH / VL as the bispecific antibody to a subject.

27. The method of any one of claims 18 to 26, wherein the administration reduces skin erythema / bleeding by more than 30% relative to the administration of an anti-KLK5 / KLK7 antibody comprising the same CDR and / or VH / VL as the bispecific antibody to a subject.

28. The method of any one of claims 18 to 27, wherein the application reduces skin abrasion / erosion by more than 30% relative to the administration of an anti-IL-13 antibody comprising the same CDR and / or VH / VL as the bispecific antibody to a subject.

29. The method of any one of claims 18 to 28, wherein the administration reduces skin desquamation / dryness by more than 30% relative to the administration of an anti-KLK5 / KLK7 antibody comprising the same CDR and / or VH / VL as the bispecific antibody to a subject.

30. The method of any one of claims 18 to 29, wherein the application reduces skin desquamation / dryness by more than 30% relative to the administration of an anti-IL-13 antibody comprising the same CDR and / or VH / VL as the bispecific antibody to a subject.

31. The method of any one of claims 18 to 30, wherein the administration reduces skin edema by more than 30% relative to a subject receiving an anti-KLK5 / KLK7 antibody comprising the same CDR and / or VH / VL as the bispecific antibody.

32. The method of any one of claims 18 to 31, wherein the administration reduces skin edema by more than 30% relative to the administration of an anti-IL-13 antibody comprising the same CDR and / or VH / VL as the bispecific antibody to a subject.

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