Humanized Anti-il17br antibody

Abstract

To provide humanized versions of the murine anti-IL17BR antibody D9.2 comprising mutations of certain residues within the heavy and / or light chain variable domains, which exhibit improved expression without adversely affecting specificity or affinity.SOLUTION: Provided is an antibody comprising a heavy chain variable domain and a light chain variable domain, wherein: a) the heavy chain variable domain comprises a VHCDR1, VHCDR2, and VHCDR3 each having a specific sequence, and b) the light chain variable domain (VK domain) comprises a VLCDR1, VLCDR2, and VLCDR3 each having a specific sequence.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to a humanized antibody that binds to interleukin-17 receptor B (IL17BR). [Background technology]

[0002] The IL-17B receptor (IL-25 receptor) is expressed on innate lymphoid cells (ILC2s) and plays a key role in initiating type 2 immune responses and has recently been associated with a complex role in the transition from innate to adaptive immunity. ILC2s provide an important source of type 2 cytokines that stimulate effector responses and cause inflammation.

[0003] The mouse anti-IL17BR antibody D9.2 was originally raised against a mouse IL-17Br / Fc fusion protein (Non-Patent Document 1, Patent Document 1) and has been shown to slow ongoing inflammation in a colitis model (Non-Patent Document 2).

[0004] Humanized versions of D9.2 are useful in clinical applications, such as for the treatment of allergic and inflammatory conditions, such as colitis. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] U.S. Patent No. 8,852,589 [Non-patent literature]

[0006] [Non-Patent Document 1] Neill DR, et al. Nature. 2010 464:1367-1370 [Non-Patent Document 2] Camelo et al J Gastroenterol. 2012 47(11): 1198-1211 Summary of the Invention

[0007] The inventors have unexpectedly discovered that expression of a humanized version of the D9.2 murine anti-IL17BR antibody can be improved by mutation of certain residues within the heavy and / or light chain variable domains without compromising specificity or affinity, which may be useful, for example, for development candidate molecules for clinical use.

[0008] A first aspect of the invention provides a humanized antibody molecule comprising a heavy chain variable domain and a light chain variable domain, a) the heavy chain variable domain (VH) comprises a VHCDR1 of SEQ ID NO: 1, a VHCDR2 of SEQ ID NO: 2 and a VHCDR3 of SEQ ID NO: 3; b) a humanized antibody molecule, wherein the light chain variable domain (VL domain) comprises a VLCDR1 of SEQ ID NO: 4, a VLCDR2 of SEQ ID NO: 5 and a VLCDR3 of SEQ ID NO: 6.

[0009] The antibody molecule may specifically bind to interleukin-17 receptor B (IL17BR), preferably to the extracellular domain of IL17BR.

[0010] The antibody molecule of the first aspect comprises a VH domain of SEQ ID NO: 8, and may optionally have up to four additional framework substitutions. For example, the antibody may comprise a VH domain of SEQ ID NO: 9, SEQ ID NO: 10 or SEQ ID NO: 11, and may optionally have up to four additional framework substitutions. The antibody of the first aspect comprises a VL domain of SEQ ID NO: 13, and may optionally have up to four additional framework substitutions.

[0011] In some preferred embodiments, the antibody molecule of the first aspect may comprise a VH domain of SEQ ID NO:9 and a VL domain of SEQ ID NO:13.

[0012] A second aspect described herein provides a pharmaceutical composition comprising the antibody molecule of the first aspect and a pharma- ceutically acceptable carrier.

[0013] A third aspect described herein provides a nucleic acid encoding an antibody molecule of the first aspect or a heavy chain variable domain and / or a light chain variable domain thereof.

[0014] A fourth aspect described herein provides a vector comprising the nucleic acid of the third aspect.

[0015] A fifth aspect described herein provides a host cell comprising the nucleic acid of the third aspect or the vector of the fourth aspect.

[0016] A sixth aspect described herein provides a method of making an antibody molecule according to the first aspect, comprising expressing a vector according to the fourth aspect in a host cell culture to produce said antibody, and recovering the antibody molecule from the cell culture.

[0017] A seventh aspect described herein provides a method of treating an allergic or inflammatory condition by administering to an individual in need of treatment an effective amount of an antibody molecule according to the first aspect or a pharmaceutical composition according to the second aspect.

[0018] An eighth aspect described herein provides an antibody molecule according to the first aspect or a pharmaceutical composition according to the second aspect for use as a medicament or for use in a method of treatment of the human or animal body.

[0019] A ninth aspect described herein provides an antibody molecule according to the first aspect or a pharmaceutical composition according to the second aspect for use in a method of treating an allergic or inflammatory condition, or a cancer condition in an individual.

[0020] These and other aspects and embodiments described herein are described in more detail below. [Brief description of the drawings]

[0021] [Figure 1] FIG. 1 shows binding of chimeric cD9001 antibodies to IL-17BR for human, mouse and cynomolgus monkey. [Diagram 2] FIG. 1 shows the lack of binding of IL-17B to IL-17BR in the presence of one of the humanized antibodies hD9030. [Diagram 3] FIG. 1 shows the ability of IL-25 to bind to IL-17BR in the presence of one of the humanized antibodies hD9030. [Figure 4] FIG. 1 shows the binding of humanized antibodies hD9040-42 to human IL-17BR. [Diagram 5] FIG. 1 shows the kinetics of humanized antibodies hD9040-42 using human IL-17BR. [Figure 6] FIG. 1 shows the thermal stability of humanized antibodies hD9040-42. [Figure 7] FIG. 1 shows thermal shift analysis of humanized antibodies hD9040-42. [Figure 8] FIG. 1 shows aggregation analysis of humanized antibodies hD9040-42. [Figure 9] FIG. 1 shows non-specific protein-protein interactions (interaction chromatography) of humanized antibodies hD9040 to 42. [Figure 10] FIG. 1 shows purified humanized antibodies hD9040-42 evaluated for solubility. [Figure 11] FIG. 11 shows freeze / thaw stress analysis of humanized antibodies hD9040 (11A), hD9041 (11B) and hD9042 (11C). [Figure 12] FIG. 12 shows heat-induced stress analysis of humanized antibodies hD9040 (12A), hD9041 (12B) and hD9042 (12C). [Figure 13] FIG. 1 shows the self-association analysis of humanized antibodies hD9040-42 by DLS. [Figure 14] FIG. 1 shows the results of an ELISA assay to determine the binding specificity of the lead candidate hD9042 for human IL17BR compared to human IL17RA, IL17RC and IL17RD. [Figure 15]Figure 15 shows the results of assaying induction of human IL-5 (Figure 15A) and IL-13 (Figure 15B) by IL-25 and IL-2 in the presence of hD9042. [Figure 16] FIG. 1 shows the results of an assay for induction of human IL-8 by IL-25 and TNFα in the presence of hD9042 or hD9043. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0022] The present invention relates to the discovery that the expression of a humanized version of the murine anti-IL17BR antibody D9.2 is significantly improved by mutation of certain residues within the variable domains, without adversely affecting specificity or affinity.

[0023] The antibody molecules described herein may specifically bind to interleukin-17 receptor B (IL17BR). Preferably, the antibody molecules described herein specifically bind to the extracellular domain of IL17BR (residues 18-292 of human IL17BR).

[0024] The antibody molecules described herein may bind to human IL17BR, mouse IL17BR and / or cynomolgus IL17BR. For example, the antibody molecule may bind to human IL17BR and show no or substantially no binding to mouse IL17BR and / or cynomolgus IL17BR. The antibody molecules of the present invention are preferably cross-reactive with human, mouse and cynomolgus IL17BR. For example, the cross-reactive antibody molecule may bind to human IL17BR, mouse IL17BR and cynomolgus IL17BR. Typically, specificity can be determined by a binding assay such as ELISA using a panel of antigens.

[0025] The antibody molecules described herein are generally specific for IL17BR. In other words, the antibody molecules are capable of binding to IL17BR, but show no or substantially no binding to other members of the IL-17R family. Preferably, antibody molecules specific for IL17BR bind to IL17BR, but show no or substantially no binding to IL-17RA, IL-17RC and / or IL-17RD.

[0026] The interleukin-17B receptor, variously known as IL-25R, IL17BR, IL-17RB, or IL-17RH1, was first identified in expressed sequence tag databases by its homology to the IL-17A receptor (IL-17RA) (Tian et al., 2000 Oncogene 19, 2098-2109). IL17BR was subsequently shown to bind both IL17B and IL25 (Lee, J., et al., J Biol Chem 276, 1660-1664 (2001); Shi, Y., et al., J Biol Chem 275, 19167-19176 (2000); Tian et al., 2000 (supra) IL25 binds to IL17BR with stronger affinity (1.4 nM) than IL-17B (7.6 nM). Mouse IL17BR (gene ID 50905: nucleic acids: NM_019583.3 GI:142368701; NP_062529.2 GI:83025064) is described in Tian et al., 2000 (supra). Human IL17BR (gene ID 50905: nucleic acids: NM_019583.3 GI:142368701; NP_062529.2 GI:83025064) is described in Tian et al., 2000 (supra). D:55540, Nucleic acid:NM_018725.3 GI:112382255; Protein NP_061195.2 GI:27477074) is Shi, Y., et al., 2000 (above) This is described in the above.

[0027] IL17BR is available as an Fc fusion protein from commercial sources (e.g., R&D Systems, MN, USA) or can be cloned or synthesized using sequences of IL17BR available in the art. For the generation of antibodies or use in immunoassays, fragments of recombinant IL17BR may be used, particularly fragments containing the extracellular domain (e.g., residues 18-292 of the human IL17BR sequence of NP_061195.2).

[0028] The antibody molecules described herein preferably inhibit or block the binding or functional interaction of IL17BR with a ligand such as IL25 or IL17B. For example, the antibody molecule may inhibit the functional interaction of IL17BR with IL25 and / or may block the binding of IL17BR to IL17B.

[0029] The antibody molecules described herein preferably reduce or inhibit at least one of IL-25 mediated airway hyperresponsiveness (AHR), IL-13 production, IL-25 mediated IL-5 production, IL-25 mediated IL-8 production, and γδ T cell proliferation and infiltration.

[0030] In some embodiments, the binding of the antibody molecules described herein to IL17BR may be abolished by competition with recombinant ligands such as IL25 or IL17B.

[0031] The anti-IL17BR antibodies described herein may comprise a heavy chain variable (VH) domain and a light chain variable (VL) domain. The VH domain may comprise the VHCDR1, VHCDR2, and VHCDR3 sequences of SEQ ID NOs: 1-3, respectively. In some embodiments, the VHCDR2 may have the sequence of SEQ ID NO: 14 or SEQ ID NO: 15. The VL domain may comprise the VLCDR1, VLCDR2, and VLCDR3 sequences of SEQ ID NOs: 4-6, respectively.

[0032] In some embodiments, the VH domain of the antibody molecules described herein has the following Kabat position 94 with a T to R substitution, and optionally at Kabat positions 1, 24, 60, 64, 65, and 69 For example, the VH domain of an antibody molecule described herein may comprise the amino acid sequence of SEQ ID NO: 7 with N60A, K64Q, D65G and T94R substitutions, or with E1Q, T24A, L69M and T94R substitutions (Kabat numbering) and optionally with one, two, three or all six substitutions in the framework regions. may comprise the amino acid sequence of SEQ ID NO:7 with four additional substitutions.

[0033] The VH domain of the antibody molecules described herein may comprise the amino acid sequence of SEQ ID NO: 8, or the amino acid sequence of SEQ ID NO: 8 with one or more, e.g., two, three or four or more, amino acid substitutions, deletions or insertions independently in the framework regions. The substitutions may be conservative substitutions. For example, the anti-IL17BR antibodies described herein may comprise a VH domain of SEQ ID NO: 9, 10 or 11, optionally with one, two, three or four amino acid substitutions in the framework regions.

[0034] In some embodiments, the VL domain of the antibody molecules described herein has the following Kabat position It may comprise the amino acid sequence of SEQ ID NO:12 having an R to G substitution at position 93 and, optionally, one, two, three or four additional amino acid substitutions in the framework regions.

[0035] The VL domain of the antibody molecules described herein may have the amino acid sequence of SEQ ID NO: 13, or may have, independently, one or more, e.g., two, three or four, amino acid residues in the framework regions. The anti-IL17BR antibodies described herein may have the amino acid sequence of SEQ ID NO: 13 with one, two, three or four amino acid substitutions, deletions or insertions. The substitutions may be conservative substitutions. For example, the anti-IL17BR antibodies described herein may comprise the VL domain of SEQ ID NO: 13, optionally with one, two, three or four amino acid substitutions in the framework regions.

[0036] A suitable anti-IL17BR antibody may comprise (i) a VH domain of SEQ ID NO: 9 and a VL domain of SEQ ID NO: 13, (ii) a VH domain of SEQ ID NO: 10 and a VL domain of SEQ ID NO: 13, or (iii) a VH domain of SEQ ID NO: 11 and a VL domain of SEQ ID NO: 13. Some preferred anti-IL17BR antibodies may comprise a VH domain of SEQ ID NO: 9 and a VL domain of SEQ ID NO: 13.

[0037] The terms "immunoglobulin" and "antibody" can be used interchangeably to refer to any protein containing an antibody antigen-binding site that has the capacity to specifically bind to one or more antigens.

[0038] An "antigen" is an entity (e.g., a proteinaceous entity or peptide) to which an immunoglobulin or antibody (or an antigen-binding fragment thereof) specifically binds. The antigen of the anti-IL17BR antibodies described herein may include IL17BR or a fragment thereof, such as a fragment comprising the extracellular domain of IL17BR or an immunogenic portion thereof.

[0039] Native antibodies are usually heterotetrameric glycoproteins of about 150,000 daltons, composed of two identical light (L) chains and two identical heavy (H) chains. Each light chain is linked to a heavy chain by one covalent disulfide bond, with a variable number of disulfide linkages between the heavy chains of different antibody isotypes. Each heavy and light chain also has regularly spaced intrachain disulfide bridges.

[0040] Antibodies contain globular regions of heavy or light polypeptide chains called "domains." A domain may contain, for example, peptide loops, usually three to four loops, stabilized by β-pleated sheets and / or intrachain disulfide bonds. Domains are generally designated "constant" or "variable" based on the relative lack of sequence variation within the domain of different class members, in the case of "constant" domains, or the pronounced variation within the domain of different class members, in the case of "variable" domains. Antibody or polypeptide "domains" are often referred to interchangeably in the art as antibody or polypeptide "regions."

[0041] The "constant" domains of an antibody light chain can be referred to as "light chain constant regions", "light chain constant domains", "CL" regions or "CL" domains. The "constant" domains of an antibody heavy chain can be referred to as "heavy chain constant regions", "heavy chain constant domains", "CH" regions or "CH" domains. The constant domain of the light chain aligns with the first constant domain of the heavy chain.

[0042] The constant domain of the heavy chain containing the tail region of an antibody is referred to herein as the fragment crystallizable (Fc) domain or Fc region. The Fc region interacts with cell surface Fc receptors and the complement system. The Fc region contains three heavy chain constant domains in each polypeptide chain.

[0043] In some embodiments, the VH domain of the antibody molecules described herein may be fused to an antibody constant region, for example an IgG4 constant region, such as SEQ ID NO: 16. The amino acid sequence of the VH domain fused to the IgG4 constant region may comprise SEQ ID NO: 17.

[0044] The "variable" domain of an antibody light chain is called a "light chain variable region", "light chain variable domain", or "VL". The "variable" domains of an antibody heavy chain can be referred to as "VL" domains (where "L" refers to the "light" rather than the light chain isotype "lambda"). These may be referred to as "heavy chain variable region," "heavy chain variable domain," "VH" region or "VH" domain. An intact light chain, for example, has two domains (VL and CL), and an intact heavy chain, for example, has four or five domains (VH, CH1, CH2 and CH3).

[0045] The light and heavy chain variable domains contain "hypervariable regions" (HVR or HV), also known as "complementarity determining regions" (CDRs), which are hypervariable in sequence and can form structurally defined loops. Generally, antibodies contain six hypervariable regions, three in the heavy chain (H1, H2, H3) and three in the light chain (L1, L2, L3), interspersed among relatively conserved framework regions (FR). The amino acid sequences of the variable domains in the antibodies described herein are shown below. The CDRs can be readily identified in these sequences using standard techniques.

[0046] The variable regions of each light / heavy chain pair form an antigen-binding site. The term "antigen-binding site" refers to the site that specifically binds (immunoreacts with) an antigen. The antibodies described herein contain at least one antigen-binding site, and preferably contain two antigen-binding sites. The antigen-binding site is formed by the heavy and light chain CDRs aligned by the framework regions, thereby allowing binding to a specific epitope. An "antigen-binding region" or "antigen-binding domain" is an antibody region or domain that comprises the antibody-binding site. The antibodies described herein have at least one antigen-binding site that recognizes IL17BR.

[0047] Naturally occurring or recombinantly produced antibody chains can be expressed with leader sequences that are removed during cellular processing to yield mature chains that may be recombinantly produced containing non-naturally occurring leader sequences, for example to enhance secretion or alter processing of the particular chain of interest.

[0048] The constant regions of antibody heavy and light chains can exhibit phenotypic variation. Antibody light chains are classified as either kappa (κ) or lambda (λ) based on the amino acid sequence of the light chain constant region and are approximately 230 residues long. The antibodies described herein contain a kappa light chain (the variable domain of the kappa light chain is referred to herein as VK). Heavy chains from humans and higher mammals are classified as gamma (γ), mu (μ), alpha (α), delta (δ) or epsilon (ε), are approximately 450 to 600 residues long, and define the antibody isotype as IgG, IgM, IgA, IgD, and IgE, respectively. There are two subclasses of IgM (H and L), three subclasses of IgA (IgA1, IgA2, and secretory IgA), and four subclasses of IgG (IgG1, IgG2, IgG3, and IgG4). The antibodies described herein are preferably immunoglobulin G (IgG) antibodies, and most preferably IgG1 antibodies.

[0049] The antibodies described herein can comprise a heavy chain belonging to any of the immunoglobulin isotypes described herein. The antibodies described herein may also comprise sequences from more than one class or isotype.

[0050] The anti-IL17BR antibodies described herein can exhibit cytotoxic activity. In such antibodies, the constant domain is usually a complement fixing constant domain, and the class is usually IgG1. Human isotypes IgG1 and IgG4 are exemplary.

[0051] The antibody molecules described herein may include fragments of whole antibodies. The term "fragment" refers to a portion or part of an antibody or antibody chain that contains fewer amino acid residues than an intact or complete antibody or antibody chain, which portion is not normally present in an intact antibody. Preferably, the fragment retains at least one, and preferably most or all, of the functions associated with that portion when present. Fragments can be obtained by chemical or enzymatic treatment of an intact or complete antibody or antibody chain. Fragments can also be obtained by recombinant means.

[0052] Fragments of the antibodies described herein can bind to an antigen or compete for antigen binding (i.e., specific binding) with the intact antibody (i.e., the intact antibody from which the fragment is derived). The antibodies described herein bind to IL17BR. Binding fragments are produced by recombinant DNA methods or by enzymatic or chemical cleavage of intact immunoglobulins.

[0053] The antibodies described herein may exist as binding fragments, including but not limited to, Fab, Fab', F(ab')2, chemically linked F(ab')2, monospecific Fab2, bispecific Fab2, trispecific Fab2, monovalent IgG, scFv (single chain variable fragment), di-scFv (bivalent scFv), bispecific diabody, trispecific triabody, scFv-Fc, minibody or sdAb (single domain antibody), and retain the ability to bind to IL17BR.

[0054] The antibody molecules described herein may be part of a bispecific or trispecific antibody. A bispecific antibody is an artificial hybrid antibody having two different heavy / light chain pairs and two different antigen binding sites, and a trispecific antibody is an artificial hybrid antibody having three different heavy / light chain pairs and three different antigen binding sites. Bispecific and trispecific antibodies can be produced by a variety of methods including fusion of hybridomas or linking of Fab' fragments. See, e.g., Songsivilai & Lachmann, Clin. Exp. Immunol. 79:315-321 (1990); Kostelny et al., J. Immunol. 148, 1547-1553 (1992). Exemplary antibodies described herein may be bispecific antibodies, which comprise at least two different antigen-binding sites.

[0055] Specific binding refers to the situation where an antibody binds to an epitope on an antigen and shows little binding to molecules other than the specific epitope on the antigen. This term also applies, for example, when an antigen-binding domain is specific to an epitope carried by multiple antigens, in which case an antibody carrying the antigen-binding domain can bind to various antigens carrying the epitope.

[0056] The anti-IL17BR antibody molecules described herein or nucleic acids encoding such antibody molecules are in an isolated state. The antibody molecules and nucleic acids are free or substantially free of other polypeptides or nucleic acids found with the substances with which they are naturally associated, such as their natural environment, or in the environment in which they are prepared (e.g. cell cultures) if the preparation is by recombinant DNA techniques carried out in vitro or in vivo. The antibody molecules and nucleic acids can be formulated with a diluent or adjuvant and still be isolated for practical purposes. For example, antibodies are usually mixed with gelatin or other carriers when used to coat microtiter plates used in immunoassays, or with pharma- ceutically acceptable carriers or diluents when used in diagnostics or therapeutics.

[0057] Another aspect of the invention provides a nucleic acid encoding an antibody disclosed herein, or a light chain, a heavy chain, a VH domain or a VL domain thereof. The nucleic acid may for example encode a heavy chain variable domain (VH domain) comprising SEQ ID NO:8, as described above, a VH domain comprising, for example, SEQ ID NO:9, SEQ ID NO:10 or SEQ ID NO:11, and / or a light chain variable domain (VL domain) comprising SEQ ID NO:13. Optionally, the encoded VH domain and / or VL domain may have up to four additional amino acid mutations in the framework regions.

[0058] Nucleic acids may include DNA and RNA sequences in which thymine nucleobases are substituted with uracil.

[0059] The anti-IL17BR antibody molecules described herein can be produced by any suitable technique, including those described herein and other techniques known in the art. The antibodies described herein can be produced on a commercial scale using methods established in the art for large-scale production of antibodies. For example, recombinant expression systems as described herein can be used.

[0060] The antibody molecules described herein can be produced by recombinant expression. Nucleic acids as described above encoding light and heavy chain variable regions, optionally linked to constant regions, can be inserted into an expression vector. A vector containing a nucleic acid encoding an antibody described herein is itself an aspect of the invention. The light and heavy chains can be cloned into the same or different expression vectors. The nucleic acids encoding the antibody chains described herein can be operably linked to one or more control sequences in the expression vector(s) that ensure the expression of the antibody chain. Expression control sequences include, but are not limited to, promoters (e.g., naturally associated or heterologous promoters), signal sequences, enhancer elements, and transcription termination sequences. Preferably, the expression control sequences are eukaryotic promoter systems in vectors capable of transforming or transfecting eukaryotic host cells (e.g., COS, CHO, or Expi293 cells). Such vectors can be incorporated into a suitable host, which is maintained under conditions suitable for high level expression of the nucleotide sequences, and for harvesting and purifying the antibody.

[0061] Aspects of the invention provide a nucleic acid encoding an anti-IL17BR antibody molecule described herein, or a VH or VL domain thereof, a vector, preferably an expression vector, comprising one or more nucleic acids encoding an antibody described herein, and a vector comprising one or more nucleic acids encoding an antibody molecule described herein, or a VH or VL domain thereof, operably linked to a promoter. Exemplary expression vectors are pHuK and pHuG1, which comprise a nucleotide sequence encoding an antibody described herein, in combination with a nucleic acid disclosed herein. Other vectors providing nucleotide sequences encoding the constant regions of the antibody light and heavy chains may also be used.

[0062] The expression vectors used as described herein are typically replicable in the host organism, either as episomes or as an integral part of the host chromosomal DNA. Generally, the expression vector contains a selection marker (e.g., ampicillin resistance, hygromycin resistance, tetracycline resistance, kanamycin resistance, or neomycin resistance) that allows detection of cells transformed with the desired DNA sequence (see, e.g., U.S. Pat. No. 4,704,362).

[0063] The vectors described herein for use in eukaryotic host cells may encode a signal sequence or other polypeptide that has a specific cleavage site at the N-terminus of the mature antibody chain or polypeptide. Suitable signal sequences may be heterologous and may be one that is recognized and processed (i.e., cleaved by a signal peptidase) by the host cell. In mammalian cell expression, mammalian signal sequences as well as viral secretory leaders, such as the herpes simplex gD signal, are available.

[0064] Alternatively, sequences encoding the antibodies described herein can be incorporated into a transgene for introduction into the genome of a transgenic animal and subsequent expression in the milk of the transgenic animal (see, e.g., U.S. Pat. No. 5,741,957; U.S. Pat. No. 5,304,433). (See U.S. Patent No. 5,849,992 and U.S. Patent No. 5,849,992.) Suitable transgenes include light and / or heavy chain coding sequences operably linked to a promoter and enhancer derived from a mammary gland-specific gene such as casein or beta-lactoglobulin.

[0065] The vectors described herein containing the polynucleotide sequences of interest (e.g., sequences encoding heavy and light chains and expression control sequences) can be transferred into host cells by known methods depending on the type of cellular host. For example, calcium chloride transfection is commonly utilized for prokaryotic cells, while calcium phosphate treatment, electroporation, lipofection, biolistics or viral-based transfection can be used for other cellular hosts (see generally, Green and Sambrook, Molecular Cloning: A Laboratory Manual (Cold Spring Harbor Press, 4th ed., 2012)). Other methods used for transformation of mammalian cells include the use of polybrene, protoplast fusion, liposomes, electroporation and microinjection (see generally, Sambrook et al., supra). For the generation of transgenic animals, Thus, transgenes can be microinjected into fertilized oocytes or integrated into the genome of embryonic stem cells and the nuclei of such cells transferred into enucleated oocytes.

[0066] Host cells can be transformed with the expression vector and cultured in conventional nutrient media suitable for inducing promoters, selecting transformants and / or amplifying the genes encoding the desired sequences. Host cells containing the above-described nucleic acids or vectors are provided as one aspect of the invention.

[0067] Another aspect of the present invention provides a method for producing an anti-IL17BR antibody molecule as described herein, comprising expressing a vector as described herein in a host cell culture to produce said antibody, and recovering said antibody from the cell culture. The method may comprise introducing a vector comprising one or more nucleic acids encoding an anti-IL17BR antibody molecule or its antibody chains as described above into a host cell as described herein, growing the host cell culture under conditions allowing expression of the nucleic acid(s), and recovering the expressed anti-IL17BR antibody molecule. Any suitable method known in the art may be used.

[0068] Microbial host organisms suitable for use in cloning and expressing the nucleic acids and vectors described herein include prokaryotic hosts; bacilli such as Escherichia coli, Bacillus subtilis, and other species such as Salmonella, Serratia, and various Pseudomonas species. Enterobacteriaceae. In these prokaryotic hosts, expression vectors can also be made that typically contain expression control sequences (e.g., origin of replication) compatible with the host cell. In addition, there are a number of different known promoters, such as the lactose promoter system, the tryptophan (trp) promoter system, the β-lactamase promoter system, or the promoter system derived from phage lambda. The promoter typically controls expression, optionally with an operator sequence, and has ribosome binding site sequences, etc., for initiation and termination of transcription and translation. Vectors used in prokaryotic cells may also require an origin of replication component.

[0069] Other microorganisms, such as yeast, can also be used to express the nucleic acids or vectors described herein. Saccharomyces is a preferred yeast host, with suitable vectors containing expression control sequences (e.g., promoters), an origin of replication, termination sequences, etc. as desired. Typical promoters include 3-phosphoglycerate kinase and other glycolytic enzymes. Inducible yeast promoters include those derived from alcohol dehydrogenase, isocytochrome C, and enzymes involved in maltose and galactose utilization, among others. Examples of promoters include

[0070] In addition to microorganisms, mammalian tissue cell cultures can be used to express the nucleic acids or vectors described herein and produce antibody polypeptides (e.g., polypeptides encoding antibodies or fragments thereof). The nucleic acids or vectors described herein may also be used to produce heterologous proteins (nucleotides) (see, e.g., Winnacker, From Genes to Clones, VCH Publishers, NY 1987). Eukaryotic or mammalian cell hosts comprising the nucleic acids or vectors described herein are themselves an aspect of the invention. In practice, eukaryotic cells are preferred, as many suitable host cell lines capable of secreting heterologous proteins (e.g., intact antibodies) have been developed in the art, including CHO cell lines, various COS cell lines, HeLa cells, Expi293 cells, ExpiCHO cells, myeloma cell lines, or transformed B cells or hybridomas. The cells may be human or non-human, e.g., non-human mammalian cells. In some preferred embodiments, the cells are Expi293 human cells. The antibodies described herein can be produced in cell lines engineered to produce afucosylated proteins, such as the Potelligent™ CHOK1SV cell line (BioWa / Lonza), GlymaxX™ engineered cells (ProBioGen) or the duck embryonic stem cell line EB66 (Valneva). Expression vectors for mammalian cells generally include, but are not limited to, one or more of the following: a signal sequence, one or more marker genes, an enhancer element, a promoter, and necessary processing information sites, such as ribosome binding sites, RNA splice sites, polyadenylation sites, and transcription terminator sequences. Preferred expression control sequences are promoters derived from immunoglobulin genes, SV40, adenovirus, bovine papilloma virus, cytomegalovirus, etc. (see, e.g., Co et al., J. Immunol. 148:1149 1992).

[0071] If the heavy and light chains are cloned on separate expression vectors, the vectors are co-transfected to achieve expression and assembly of the intact antibodies described herein. After expression, whole antibodies, dimers thereof, individual light and heavy chains, or other immunoglobulin forms described herein can be purified according to standard procedures in the art, including ammonium sulfate precipitation, affinity columns, column chromatography, HPLC purification, gel electrophoresis, and the like (see generally, Scopes, Protein Purification (Springer-Verlag, NY, (1982)). (See, for example, US Pat. No. 5,393,963, which is incorporated herein by reference). Substantially pure anti-IL17BR antibody molecules of at least about 90%-95% homogeneity are preferred, with 98%-99% or more homogeneity being most preferred for the pharmaceutical uses described herein. Standard protein purification techniques known in the art can be used. The following procedures are illustrative of suitable protein purification procedures: fractionation on immunoaffinity or ion exchange columns, ethanol precipitation, reverse phase HPLC, chromatography on cation exchange resins such as silica or DEAE, chromatofocusing, SDS-PAGE, ammonium sulfate precipitation, and gel filtration.

[0072] The anti-IL17BR antibody molecules described herein are capable of specifically binding to human IL17BR. The antibodies described herein may also exhibit desirable structural, physical, biophysical and chemical properties as described below and with reference to the Examples.

[0073] The affinity of the anti-IL17BR antibody molecules described herein is the degree or strength of binding of the antibody to an epitope or antigen. The dissociation constant K D and affinity constant K A is a quantitative measure of affinity. K D is the antibody dissociation rate (k off ) and the antibody association rate (k on) The binding of an antibody to its antigen is a reversible process, and the rate of the binding reaction is proportional to the concentrations of the reactants. At equilibrium, the rate of (antibody)(antigen) complex formation is equal to the rate of dissociation into its components (antibody) + (antigen). Measurement of the reaction rate constants can be used to determine the equilibrium or affinity constant, K A (K A =1 / K D ) can be specified. D The lower the value, the greater the affinity of the antibody for its target. Most antibodies have low micromolar (10 -6 ) to nanomolar (10 -7 ~10 -9 ) range K D High affinity antibodies generally have a molecular weight in the low nanomolar range (10 -9 ), whereas ultra-high affinity antibodies are considered to have a molecular weight of picomolar (10 -12 ) range.

[0074] The anti-IL17BR antibody molecules described herein are at least 2×10 2 M -1 s -1 , at least 5 × 10 2 M -1 s -1 , at least 10 3 M -1 s -1 Or at least 5 x 10 3 M -1 s -1 , at least 10 4 M -1 s -1 , at least 5 × 10 4 M -1 s -1 , at least 10 5 M -1 s -1 , at least 5 × 10 5 M -1 s -1 Or at least 10 6 M -1 s -1 The association rate constant (k on For example, the anti-IL17BR antibody molecule may have a concentration of 3×10 6 M -1 s -1~5×10 6 M -1 s -1 may have an association rate of

[0075] The anti-IL17BR antibody molecules described herein are administered in a concentration of 5×10 -1 s -1 Less than 10 -1 s -1 Less than 5×10 -2 s -1 Less than 10 -2 s -1 Less than or 5 x 10 -3 s -1 Less than 10 -3 s -1 Less than 5×10 -4 s -1 Less than 10 -4 s -1 Less than or 5 x 10 -5 s -1 Antibody dissociation (k off For example, an anti-IL17BR antibody molecule may have a 6×10 -5 s -1 ~9×10 -5 s -1 may have a dissociation rate of

[0076] In some embodiments, the anti-IL17BR antibody molecules described herein are administered in amounts of 5×10 6 M -1 That's it, 10 6 M -1 That's it, 5 x 10 7 M -1 That's it, 10 7 M -1 That's it, 5 x 10 8 M -1 , 10 -8 M -1 That's it, 5 x 10 9 M -1 That's it, 10 9 M -1 That's it, 5 x 10 10 M -1 That's it, 10 10 M -1 or more or 5 x 10 11 M -1 The equilibrium association constant, i.e., K AThe anti-IL17BR antibody molecules described herein may bind to IL17BR with an affinity substantially similar to that of D9.2, for example, 90% to 110% of the binding affinity of D9.2.

[0077] The anti-IL17BR antibody molecules described herein are administered in a concentration of 5×10 -6 Less than M, 10 -6 Less than M, 5×10 -7 Less than M, 10 -7 Less than M, 5×10 -8 Less than M, 10 -8 Less than M, 5×10 -9 Less than M, 10 -9 Less than M, 5×10 -10 Less than M, 10 -10 Less than M or 5 x 10 -11 The equilibrium dissociation constant from IL17BR, i.e., K, is less than M D For example, the anti-IL17BR antibody molecule may have a concentration of 1×10 -11 M~2×10 -11 The equilibrium dissociation constant of M, i.e., K D may have:

[0078] Specific binding of an anti-IL17BR antibody molecule means that the antibody exhibits considerable affinity for a particular antigen or epitope and generally exhibits little cross-reactivity. An antibody that exhibits "little cross-reactivity" is one that does not appreciably bind to undesired entities (e.g., undesired proteinaceous entities). An antibody specific for a particular epitope will not cross-react with distant epitopes, for example, on the same protein or peptide. The specific binding of an antibody as described herein, i.e., k off , k on , K A and K D can be determined according to any art-recognized means of determining such binding.

[0079] The anti-IL17BR antibody molecules described herein can be thermally stable, i.e., the antibodies described herein can bind to IL17BR at temperatures between 30° C. and 85° C., specifically up to 75° C. The antibodies described herein can have a melting temperature between 50° C. and 100° C., specifically between 60° C. and 80° C., more specifically around 66° C. to 67° C.

[0080] The anti-IL17BR antibody molecules described herein can have a low tendency to aggregate. The tendency to aggregate can be analyzed using standard techniques such as multi-angle light scattering or dynamic light scattering as described herein. The antibodies described herein can have a low tendency to non-specific protein-protein interactions and good solubility.

[0081] The anti-IL17BR antibody molecules described herein can have a low tendency to aggregate when concentrated. The formulations described herein can be concentrated at concentrations of 50 mg / ml to 200 mg / ml without forming soluble aggregates in aqueous solutions maintained at physiological pH, e.g., with Dulbecco's PBS. The antibody may be concentrated to a concentration of 0.00 mg / ml, for example 75 mg / ml to 150 mg / ml, preferably 80 mg / ml to 120 mg / ml, and more preferably 90 mg / ml to 110 mg / ml (the preferred concentration is about 100 mg / ml).

[0082] The anti-IL17BR antibody molecules described herein can have a low tendency to aggregate when subjected to repeated freeze-thaw cycles or to temperatures above normal body temperature for extended periods of time, e.g., 50° C. for 30 days in Dulbecco's PBS.

[0083] The anti-IL17BR antibody molecules described herein may have an isoelectric point (pI) between pH 7 and pH 9, preferably between pH 7.4 and pH 8.2.

[0084] The anti-IL17BR antibody molecules described herein can retain binding ability to IL17BR after incubation in serum from mouse, human and / or cynomolgus primates at 37° C. For example, the antibodies described herein can retain binding ability to IL17BR after incubation in mouse, human and / or cynomolgus serum for 10 to 50 days, preferably 20 to 40 days, more preferably 30 days. Antibodies that retain binding ability can exhibit the same or substantially the same binding ability at 37° C. as observed in antibodies that have not been incubated in serum or that have been incubated in a control solution.

[0085] The anti-IL17BR antibody molecules disclosed herein may be aglycosylated. The Fc region of IgG antibodies possesses a highly conserved N-glycosylation site, and glycosylation of the Fc fragment is essential for Fc receptor-mediated activity. The N-gly carbohydrate moiety attached to this site is predominantly a complex, core-fucosylated diantennary structure. In addition, a small proportion of these N-glycans also carry bisecting GlcNAc and α-2,6-linked sialic acid residues. An aglycosylated antibody may lack one or more carbohydrate moieties, for example, by chemical or enzymatic processes, the absence or mutation of one or more glycosylation sites, or expression in bacteria. An aglycosylated antibody described herein may be a deglycosylated antibody in which the Fc carbohydrate has been removed, for example, chemically or enzymatically. Alternatively, an aglycosylated antibody described herein may be a nonglycosylated or unglycosylated antibody expressed without an Fc carbohydrate moiety, for example, by mutation of one or more residues that code for a glycosylation pattern, or by expression in an organism that does not add carbohydrate to proteins, for example, bacteria.

[0086] The anti-IL17BR antibody molecules described herein may be afucosylated, i.e. modified so that the carbohydrate moiety in the Fc region of the antibody does not have any fucose sugar units. Alternatively, the anti-Aβ antibodies disclosed herein may have a reduced number of fucose sugar units. Afucosylated antibodies are more effective in antibody-dependent cell-mediated cytotoxicity (see below). The afucosylated antibodies described herein can be produced in cell lines modified to produce afucosylated proteins, such as the Potelligent™ CHOK1SV cell line (BioWa / Lonza), GlymaxX™ modified cells (ProBioGen) or the duck embryonic stem cell line EB66 (Valneva).

[0087] The anti-IL17BR antibody molecules described herein can be modified to enhance their antibody-dependent cell-mediated cytotoxicity (ADCC). ADCC is a cell-mediated reaction in which non-specific cytotoxic cells (e.g., natural killer (NK) cells, neutrophils, and macrophages) recognize bound antibodies on target cells, which subsequently cause lysis of the target cells. Such cells can be human cells. ADCC activity of an antibody is generally believed to require binding of the Fc region of the antibody to antibody receptors present on the surface of effector cells, such as killer cells, natural killer cells, and activated macrophages. The carbohydrate structure of the humanized antibody By altering (e.g., reducing or eliminating) the fucosylation of the antibody's structure (i.e., in the Fc region), the ADCC activity of the antibody can be enhanced in vitro, for example 10-fold or 20-fold or 30-fold or 40-fold or 50-fold or 100-fold or 500-fold or 600-fold or 700-fold or 1000-fold relative to an unmodified humanized antibody. Because of the increased ADCC activity, such modified antibodies can be used at lower dosages than their unmodified counterparts and generally have fewer or reduced side effects in patients.

[0088] The anti-IL17BR antibodies described herein can be used in complement-dependent cytotoxicity (CDC). CDC involves the central innate complement system, which acts as an effector of adaptive immunity. The classical CDC pathway is triggered by antibody molecules binding to antigens on target cells and is initiated by the binding of C1q protein to the Fc domain of the bound antibody. The resulting complement cascade activates the membrane attack pathway, resulting in the formation of a membrane attack complex that induces lysis of the target cell. The antibodies described herein can be modified to enhance their ability to induce CDC by any method known in the art, including, but not limited to, modification of the protein backbone, such as containing amino acid residue substitutions in the constant domain of the antibody heavy chain. For examples of IgG1 amino acid substitution combinations used to enhance CDC activity, see Moore et al., mAbs, 2(2), 181-189 (2010). The CDC activity of the modified antibodies described herein can be enhanced, for example, 10-fold or 20-fold or 30-fold or 40-fold or 50-fold or 100-fold or 500-fold or 600-fold or 700-fold or 1000-fold relative to an unmodified humanized antibody.

[0089] The anti-IL17BR antibody molecules can be further modified by chemical modification, such as PEGylation or incorporation into liposomes, to improve their pharmaceutical properties, for example by increasing their in vivo half-life.

[0090] The anti-IL17BR antibody molecules described herein can be formulated and / or administered as pharmaceutical compositions comprising an active therapeutic antibody agent and various other pharma- ceutically acceptable components. See Remington: The Science and Practice of Pharmacy (22nd ed., Pharmaceutical Press, London, Pa. (2013)). The preferred form depends on the intended method of administration and therapeutic application. The composition may also contain pharma- ceutically acceptable non-toxic carriers or diluents, defined as vehicles commonly used to formulate pharmaceutical compositions for administration to animals or humans, depending on the desired formulation. The diluents are selected so as not to affect the biological activity of the combination. Examples of such diluents are distilled water, phosphate-buffered saline, Ringer's solution, dextrose solution, and Hank's solution. In addition, the pharmaceutical composition or formulation may also contain other carriers, adjuvants, or non-toxic non-therapeutic non-immunogenic stabilizers, and the like.

[0091] Pharmaceutical compositions containing the anti-IL17BR antibody molecules described herein may also include large slowly metabolized macromolecules such as proteins, polysaccharides such as chitosan, polylactic acids, polyglycolic acids and copolymers (e.g., latex-functionalized Sepharose™, agarose, cellulose, etc.), polymeric amino acids, amino acid copolymers, and lipid aggregates (such as oil droplets or liposomes). Additionally, these carriers can function as immunostimulants (i.e., adjuvants).

[0092] For parenteral administration, the antibodies or compositions described herein can be administered as injectable dosages of a solution or suspension of the substance in a physiologically acceptable diluent, including a pharmaceutical carrier, which can be a sterile liquid, such as water, an oil, saline, glycerol, or ethanol. Additionally, auxiliary substances, such as wetting or emulsifying agents, surfactants, pH buffering substances, and the like, may be present in the composition. Other components of pharmaceutical compositions are those of petroleum, animal, vegetable, or synthetic origin, for example, peanut oil, soybean oil, and mineral oil. Generally, propylene glycol is used. Glycols, such as glycols or polyethylene glycol, are preferred liquid carriers, particularly for injectable solutions. The antibodies can be administered in the form of a depot injection or implant preparation which can be formulated so as to permit a sustained release of the active ingredient.

[0093] The term "parenteral" as used herein includes subcutaneous, intravenous, intradermal, intramuscular, intraperitoneal and intrathecal administration of the antibody or composition described herein. The anti-IL17BR antibody or composition described herein can also be administered by intranasal or intragastric methods. Typically, the compositions are prepared as injectables, either as liquid solutions or suspensions, although solid forms suitable for solution or suspension in liquid vehicles prior to injection can also be prepared. The preparations can also be emulsified or encapsulated in liposomes or microparticles, such as polylactides, polyglycolides or copolymers for enhanced adjuvant effect as discussed above (see Langer, Science 249: 1527 (1990) and Hanes, Advanced Drug Delivery Reviews 28:97 (1997)). The antibody molecules of the present invention can be administered in the form of depot injection or implant preparations, which can be formulated to allow sustained or pulsatile release of the active ingredient.

[0094] Additional formulations suitable for other modes of administration include oral, intranasal and pulmonary formulations, suppositories and transdermal applications. Oral formulations contain excipients such as pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose and magnesium carbonate. These compositions take the form of solutions, suspensions, tablets, pills, capsules, sustained release formulations or powders and contain 10%-95% of the active ingredient, preferably 25%-70%.

[0095] Therapeutic formulations of anti-IL17BR antibody molecules can be prepared in the form of a lyophilized powder or aqueous solution by mixing the antibody molecule having the desired purity for storage with any physiologically acceptable carriers, excipients or stabilizers (see, e.g., "Remington: The Science and Practice of Pharmacy", 20th Edition, 2000, pub. Lippincott, Williams & Wilkins.). Acceptable carriers, excipients, or stabilizers are non-toxic to recipients at the dosages and concentrations employed, and include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid; low molecular weight (less than about 10 residues) polypeptides; proteins such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugar alcohols such as mannitol or sorbitol; salt-forming counterions such as sodium; and / or non-ionic surfactants such as Tween, Pluronic, or polyethylene glycol (PEG).

[0096] For anti-IL17BR antibody molecules to be used for in vivo administration, they must be sterile, which is readily accomplished by filtration through sterile filtration membranes, prior to or following lyophilization and reconstitution. The antibody molecule is typically stored in lyophilized form or in a solution.

[0097] Preferably, the anti-IL17BR antibody molecules described herein or compositions comprising the anti-IL17BR antibodies described herein can be administered intravenously (IV), intramuscularly (IM) or topically, e.g., by topical solutions such as aerosols, lotions, ointments, or drops.

[0098] Topical application can result in transdermal or intradermal delivery. Topical administration can be facilitated by coadministration of the agent with cholera toxin, or a detoxified derivative or subunit thereof, or other similar bacterial toxins (see Glenn et al., Nature 391, 851 (1998)). Simultaneous administration can be achieved by using the components as a mixture, or as a linked molecule obtained by chemical cross-linking or expression as a fusion protein. Alternatively, transdermal delivery can be achieved using skin patches or using transferosomes (Paul et al., Eur. J. Immunol. 25:3521 (1995); Cevc et al., J. Immunol. 25:3521 (1995); et al., Biochem. Biophys. Acta 1368:201-15 (1998)).

[0099] The composition may comprise an anti-IL17BR antibody molecule as described herein, a pharma- ceutically acceptable carrier as described herein, and other therapeutic agents, particularly prophylactic or therapeutic agents, useful for the prevention, management or treatment of allergic or inflammatory conditions. Such therapeutic agents may include analgesics, anti-inflammatories, antiviral drugs, drugs that reverse fever or elevated body temperature, and / or therapeutic compounds designed to relieve pain, such as mouthwashes or sprays that can relieve mouth pain. The compositions described herein may additionally comprise compositions for rehydrating a subject, for example, by intravenous therapy.

[0100] The compositions described herein can include nucleic acids, i.e., DNA or RNA, encoding the anti-IL17BR antibody molecules described herein, and any method of delivery of such nucleic acids, with or without any of the other components of the composition discussed above. The compositions can also include vectors, such as, but not limited to, expression vectors described herein, which themselves contain a nucleic acid described herein.

[0101] The compositions described herein may include viral vectors used as a delivery system for nucleic acid to cells.Suitable viral vector nucleic acid delivery systems include retroviral systems, adenoviral vectors, viral vectors of the family Poxviridae, including vaccinia virus and avian poxvirus, and viral vectors of the genus Alphavirus.The nucleic acid encoding the antibody described herein or the vector containing it can be packaged into liposomes for delivery to individuals or cells, which can be incorporated into compositions as described.Vector and nucleic acid encoding the antibody can also be adsorbed or associated with particle carriers.

[0102] The compositions described herein may include gene therapy vectors containing nucleotide sequences encoding the antibodies described herein, or naked antibody polypeptide chains according to the invention. The compositions may include such vectors or polypeptides in combination with the antibodies described herein and any other compositional components described above.

[0103] The anti-IL17BR antibody molecules described herein can be used in kits. The term "kit" is used in reference to a combination of reagents and other materials that facilitate sample analysis. In some embodiments, the immunoassay kits described herein include a suitable antigen, a binding agent that includes a detectable moiety, and a detection reagent. A system for amplifying the signal generated by the detectable moiety may or may not be included in the kit. Additionally, in other embodiments, the kit includes components such as, but not limited to, devices for sample collection, sample tubes, holders, trays, racks, dishes, plates, instructions for the user of the kit, solutions or other chemical reagents, and samples used for standardization, normalization, and / or control samples.

[0104] The kit may include at least one antibody described herein. The kit may include a composition described herein in one or more containers, optionally with one or more other prophylactic or therapeutic agents useful for the diagnosis, prevention, management, or treatment of an allergic or inflammatory condition, or a cancer condition. A device capable of delivering the components of the kit by a route of administration, such as a syringe, may be included. The kit may further include instructions for use to prevent, treat, manage, or ameliorate an allergic or inflammatory condition, as well as information on side effects and dosages for administration methods. .

[0105] The present invention also provides diagnostic kits. The antibodies described herein are useful for monitoring, diagnosing or providing a prognosis for the development or progression of an allergic or inflammatory condition, or a cancer condition, and can be used in a kit suitable for such purposes. The antibodies described herein can be used in diagnostic kits for detecting the presence of IL17BR in a sample of bodily fluid taken from an individual, which can be a human or a mammal, such as a non-human primate, or an experimental animal, including mice, rats and rabbits. A sample of bodily fluid, such as, but not limited to, blood or serum, is taken from the individual and tested for the presence of IL17BR using the antibodies described herein. Measuring the level of IL17BR in the blood of an individual using the antibodies described herein can provide information about susceptibility, risk of developing, diagnosis or prognosis of an allergic or inflammatory condition, or a cancer condition in the individual, or about a suitable administration schedule or dosage of the antibodies or compositions described herein for the treatment of the individual. The diagnostic method is generally performed in vitro. A method for detecting the presence of IL17BR in a sample from an individual can include contacting the sample with an anti-IL17BR antibody described herein and determining binding of the antibody to IL17BR in the sample.

[0106] Kits useful for the above diagnostics may include various enzymes used in assays including EIA and ELISA, such as, but not limited to, horseradish peroxidase, alkaline phosphatase, β-galactosidase, or acetylcholinesterase; prosthetic groups, such as, but not limited to, streptavidin / biotin and avidin / biotin; particles, such as latex beads or bacteria, used in agglutination tests; fluorescent substances, such as, but not limited to, umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride, or phycoerythrin; luminescent substances, such as, but not limited to, luminol; bioluminescent substances, such as, but not limited to, luciferase, luciferin, and aequorin; iodine ( 131 I, 125 I, 123 I, 121 I), Carbon ( 14 C), sulfur ( 35 S), tritium ( 3 H), Indium ( 115 In, 113 In, 112 In, 111 In) and technetium ( 99 Tc), thallium ( 201 Ti), Gallium ( 68 Ga, 67 Ga), palladium ( 103 Pd), Molybdenum ( 99 Mo), Xenon ( 133 Xe), fluorine ( 18 F), 153 Sm, 177 Lu, 159 Gd, 149 Pm, 140 La, 175 Yb, 166 Ho, 90 Y, 47 Sc, 186 Re, 188 Re, 142 Pr, 105 Rh, 97 Ru, 68 Ge, 57 Co, 65Zn, 85 Sr, 32 P, 153 Gd, 169 Yb, 51 Cr, 54 Mn, 75 Se, 113 Sn and 117 The antibody described herein may be conjugated to a detectable substance, including, but not limited to, radioactive substances such as Sn; positron-emitting metals using various positron emission tomography, non-radioactive paramagnetic metal ions, and molecules that are radiolabeled or conjugated to specific radioisotopes. Any detectable label that can be easily measured can be conjugated to the antibody described herein and used to diagnose the diseases described herein. The detectable substance can be directly bound or conjugated to the antibody using techniques known in the art, or indirectly through an intermediate (e.g., a linker known in the art, etc.). Metal ions that can be conjugated to antibodies used as diagnostic agents are known in the art (see, for example, U.S. Pat. No. 474,900).

[0107] Detection of the antigen using any of the above methods or detectable substances, using the antibodies described herein in the kits described herein, can produce a positive result for the presence of IL17BR and diagnose an individual as having an allergic or inflammatory condition, or a cancer condition, or determine whether an individual has an allergic or inflammatory condition, or a cancer condition. or at risk for an allergic or inflammatory or cancerous condition, which may subsequently require and / or receive treatment for the allergic or inflammatory or cancerous condition described herein.

[0108] Aspects of the invention also relate to methods of treatment, including prevention of an allergic or inflammatory condition, or a cancerous condition, by administering an effective amount of an antibody or composition described herein to an individual in need of treatment. An antibody or composition described herein, preferably a pharmaceutical composition (e.g., a composition comprising an antibody described herein, a pharma- ceutically acceptable excipient, and optionally an additional therapeutic agent), can be used as a medicament, for example, in a method of treatment of the human or animal body. An antibody or composition described herein, preferably a pharmaceutical composition, can be used in a method of treatment of the human or animal body, where the treatment is a therapeutic or prophylactic treatment of an allergic or inflammatory condition, or a cancerous condition in an individual.

[0109] Allergic or inflammatory conditions can include IL25-mediated diseases such as allergies; inflammatory bowel disease (IBD); inflammation of the colon; asthma, including allergic asthma and rhinovirus-exacerbated asthma; airway hyperresponsiveness (AHR); fibrosis, including idiopathic pulmonary fibrosis; colitis, including chronic colitis, ulcerative colitis, and Crohn's disease; and inflammatory eye diseases such as keratoconjunctivitis sicca (dry eye syndrome).

[0110] Cancerous conditions can include pancreatic, breast, liver and thyroid cancer.

[0111] The anti-IL17BR antibody molecules may be useful, for example, in preventing or alleviating AHR in a subject. A method of preventing or alleviating AHR in a subject (e.g., a human) in need thereof may comprise administering to the subject an anti-IL17BR antibody molecule described herein.

[0112] The anti-IL17BR antibody molecule may be useful, for example, in preventing or reducing inflammation of the colon. A method of preventing or reducing inflammation of the colon in a subject (e.g., a human) in need thereof may comprise administering to the subject an antibody molecule that binds to IL17BR as described above. For example, the anti-IL17BR antibody molecule may be useful, for example, in preventing, reducing or treating IBD. A method of preventing, reducing or treating IBD may comprise administering to the subject an antibody molecule that binds to IL17BR as described above.

[0113] The above-mentioned methods of treatment can include administration to an individual of an antibody or composition described herein (e.g., a composition comprising an antibody described herein, a pharma- ceutically acceptable excipient, and optionally an additional therapeutic agent) under conditions that result in a therapeutic response in the individual that is beneficial for, e.g., the prevention or treatment of an allergic or inflammatory condition.

[0114] Individuals suitable for the treatments described herein may suffer from an allergic or inflammatory condition, or a cancer condition. The treatment methods described herein may be used for both asymptomatic patients and patients currently showing symptoms of an allergic or inflammatory condition, or a cancer condition. The antibodies described herein may be administered prophylactically to individuals who do not have an allergic or inflammatory condition, or a cancer condition. The antibodies described herein may be administered to individuals who do not have or are not showing symptoms of an allergic or inflammatory condition, or a cancer condition. The antibodies described herein may be administered to individuals who have or are likely to have an allergic or inflammatory condition, or a cancer condition. Individuals suitable for treatment include individuals who are at risk or susceptible to an allergic or inflammatory condition but are not showing symptoms, and individuals who are suspected of having an allergic or inflammatory condition, or a cancer condition, as well as individuals who are currently showing symptoms. The antibodies described herein may be administered to individuals who are at risk or susceptible to an allergic or inflammatory condition but are not showing symptoms, as well as individuals who are suspected of having an allergic or inflammatory condition, or a cancer condition, as well as individuals who are currently showing symptoms. It can be administered prophylactically to the general population without the need for risk assessment.

[0115] In some embodiments, an individual suitable for the treatment described herein may exhibit elevated levels of biomarkers associated with a Th2 response compared to control individuals, e.g., healthy members of a population, or compared to levels previously determined in the individual. Biomarkers associated with a Th2 response may include exhaled nitric oxide, blood eosinophils, or serum biomarkers, e.g., IgE, IL-13, or chemokines, e.g., eotaxin-3, and thymus and activation regulated chemokine (TARC / CCL17). The method may include obtaining a sample from the individual, determining the level of one or more biomarkers in the sample, and administering an antibody described herein if the level of one or more biomarkers associated with a Th2 response is elevated.

[0116] The terms "treat", "treating" or "treatment" (or grammatical equivalents) mean that the severity of an individual's condition is reduced or at least partially ameliorated or ameliorated, and / or some relief, alleviation or diminution of at least one clinical symptom is achieved, and / or the progression of the condition is inhibited or slowed and / or the onset of a disease or disorder is prevented or delayed.

[0117] The anti-IL17BR antibody molecules described herein that can be used in the methods of treatment of an allergic or inflammatory condition, or a cancer condition, can be antibodies of any of the sequences and formats described above. The antibody molecules used in the methods of treatment described herein can also be fragments, such as antigen-binding fragments, of the anti-IL17BR antibodies described herein.

[0118] The anti-IL17BR antibody molecule described herein can be administered to an individual in need of treatment with a pharmaceutical carrier or pharmaceutical composition, or in any of the compositions described herein. Alternatively, the anti-IL17BR antibody molecule can be administered to an individual by administering a polynucleotide encoding at least one antibody chain. The polynucleotide is expressed to produce the antibody chain in the patient. Optionally, the polynucleotide encodes the heavy and light chains of the anti-IL17BR antibody molecule. The polynucleotide is expressed to produce the heavy and light chains in the individual.

[0119] The terms "patient," "individual," or "subject" include humans and other mammalian subjects who receive either prophylactic or therapeutic treatment with one or more of the agents (e.g., immunotherapeutic agents or antibodies) described herein. Mammalian subjects include primates, e.g., non-human primates. Mammalian subjects also include laboratory animals commonly used in research, such as, but not limited to, rabbits, and rodents such as rats and mice.

[0120] The anti-IL17BR antibody molecules described herein can be used in a method of preventing or treating an allergic or inflammatory condition, or a cancerous condition, comprising administering to a patient an effective dose of an antibody described herein. As used herein, an "effective amount" or "effective dosage" or "sufficient amount" of a therapeutic antibody described herein (or grammatically equivalent terms) refers to an amount of an antibody or composition described herein that is effective to produce a desired effect, which is optionally a therapeutic or prophylactic effect (i.e., by administering a therapeutically effective amount). For example, an "effective amount" or "effective dosage" or "sufficient amount" can be an amount such that the severity of an individual's condition, e.g., an allergic or inflammatory condition, or cancerous condition, is reduced or at least partially ameliorated or ameliorated, and / or some relief, alleviation or reduction of at least one clinical symptom is achieved, and / or the progression of the allergic or inflammatory condition, or cancerous condition is inhibited or delayed, and / or the onset of the allergic or inflammatory condition, or cancerous condition is prevented or delayed.

[0121] The dosage of the compositions described herein that is effective in treating the above conditions will vary depending on many different factors, including the means of administration, the target site, the physiological state of the patient, whether the patient is human or animal, other drugs administered, and whether the treatment is prophylactic or therapeutic. Typically, the patient is a human, but non-human mammals, including transgenic mammals, such as non-human primates, rabbits, rats, and mice, can also be treated. Treatment dosages should be titrated to optimize safety and efficacy.

[0122] For passive immunization with the antibodies described herein, the dosage is in the range of about 0.0001 mg / kg to 100 mg / kg of host body weight, more usually 0.01 mg / kg to 5 mg / kg (e.g., 0.02 mg / kg, 0.25 mg / kg, 0.5 mg / kg, 0.75 mg / kg, 1 mg / kg, 2 mg / kg, etc.). For example, the dosage can be 1 mg / kg body weight or 10 mg / kg body weight, or in the range of 1 mg / kg to 10 mg / kg, preferably at least 1 mg / kg. In another example, the dosage can be 0.5 mg / kg body weight or 15 mg / kg body weight, or in the range of 0.5 mg / kg to 15 mg / kg, preferably at least 1 mg / kg. In another example, the dosage can be 0.5 mg / kg body weight or 20 mg / kg body weight, or in the range of 0.5 mg / kg to 20 mg / kg, preferably at least 1 mg / kg. In another example, the dosage may be 0.5 mg / kg (body weight) or 30 mg / kg (body weight), or within the range of 0.5 mg / kg to 30 mg / kg, preferably at least 1 mg / kg. In a preferred example, the dosage may be about 30 mg / kg.

[0123] The methods described herein may include administration of the antibody to the subject as a single dose, two doses, or multiple doses. The dose of the antibody may be about 100 μg / kg to 100 mg / kg of the patient's body weight, about 300 μg / kg to 30 mg / kg of the patient's body weight, or about 1 mg / kg to 10 mg / kg of the patient's body weight. The subject may be administered such doses daily, every other day, weekly, or according to any other schedule determined by empirical analysis. Treatment may include administration in multiple doses over a long period of time, for example, at least six months. Additional treatment regimens may include administration once every two weeks or once a month or once every three to six months. Exemplary administration schedules include 1 mg / kg to 20 mg / kg or 15 mg / kg daily, 30 mg / kg every other day, or 60 mg / kg weekly.

[0124] In both prophylactic and therapeutic treatment regimes, the reagents can be administered in several doses until a sufficient immune response is achieved. The term "immune response" or "immunological response" includes the development of a humoral (antibody-mediated) and / or cellular (antigen-specific T-cells or their secreted products) response to an antigen in a recipient subject. Typically, the immune response is monitored and repeated administrations are given if the immune response begins to wane. For example, the antibodies described herein can be administered multiple times. The intervals between single doses can be weekly, monthly or yearly. The intervals can be irregular as specified by measuring blood levels of anti-IL17BR antibodies in the patient. In some methods, the dosage is adjusted to achieve a plasma antibody concentration of 1 μg / ml to 1000 μg / ml, and in some methods 25 μg / ml to 300 μg / ml. Alternatively, the antibodies described herein can be administered as a sustained release formulation, in which case less frequent administration is required. The dosage and frequency vary depending on the half-life of the antibody in the patient. Generally, humanized antibodies exhibit a longer half-life than chimeric and non-human antibodies.

[0125] The dosage and frequency of administration can vary depending on whether the treatment is prophylactic or therapeutic. In prophylactic applications, compositions containing the antibodies or cocktails thereof described herein are administered to patients not already in a disease state to enhance the patient's resistance. Such an amount is defined as a "prophylactically effective dose." Again, in this application, the exact amount will vary depending on the patient's state of health and general immunity, but will generally be between 0.1 mg and 25 mg per administration. , particularly in the range of 0.5 mg to 2.5 mg per dose. Relatively low doses are administered at relatively infrequent intervals over a long period of time.

[0126] Doses of nucleic acids encoding the antibodies described herein range from about 10 ng-1 g, 100 ng-100 mg, 1 μg-10 mg, or 30 μg-300 μg of DNA per patient. Doses of infectious viral vectors range from 10-100 or more virions per administration.

[0127] The antibodies and compositions described herein can be administered by parenteral, topical, intravenous, oral, intragastric, subcutaneous, intraarterial, intracranial, intraperitoneal, intranasal or intramuscular methods as described herein for therapeutic and / or prophylactic treatment. Intramuscular injection or intravenous infusion is preferred for administration of the antibodies.

[0128] Other aspects and embodiments described herein provide for the aspects and embodiments described above in which the term "comprising" is replaced with the term "consisting of," as well as the aspects and embodiments described above in which the term "comprising" is replaced with the term "consisting essentially of."

[0129] It is to be understood that the present application discloses all combinations of any of the above aspects and embodiments unless the context requires otherwise. Similarly, the present application discloses all combinations of preferred and / or optional features alone or in combination with any of the other aspects unless the context requires otherwise.

[0130] Modifications of the above-described embodiments, further embodiments and modifications thereof will be apparent to those of skill in the art upon reading this disclosure, and therefore are encompassed within the scope described herein.

[0131] All publications and sequence database entries referred to herein are incorporated by reference in their entirety for all purposes.

[0132] The antibody residue positions described herein are numbered according to the scheme described in Kabat, EA, Wu, TT, Perry, HM, Gottesmann, KS & Foeller, C. (1991). Sequences of Proteins of Immunological Interest, 5th edit., NIH Publication no. 91-3242. US Department of Health and Human Services. Where appropriate, the positions of substitutions are numbered according to the scheme described in Kabat, EA, Wu, TT, Perry, HM, Gottesmann, KS & Foeller, C. (1991). Sequences of Proteins of Immunological Interest, 5th edit., NIH Publication no. 91-3242. US Department of Health and Human Services. The Kabat numbering can be written for residues that are invariant in immunoglobulin sequences. An alternative antibody numbering scheme is described in Honegger, A and Plueckthun A. (2001). J. It is described in Mol. Biol 309, 657-67.

[0133] "And / or," as used herein, shall be construed as a specific disclosure of each of the two specified features or components with or without the other. For example, "A and / or B" shall be construed as a specific disclosure of (i) A, (ii) B, and (iii) each of A and B, as if each were individually set forth herein. EXAMPLES

[0134] experiment method Biacore kinetics for human IL-17BR-His Using a Human Antibody Capture kit, anti-human IgG (Fc) antibody (in 10 mM sodium acetate (pH 5.0) immobilization buffer) was coated onto channels 2 to 4 of a CM5 chip. Channel 1 was blocked and used as a blank control. This anti-human IgG-coated chip was then used to immobilize humanized antibody mutants. A concentration series of 20 nM to 1.25 nM of human IL17BR-His was used using a 2-fold dilution series. , HBS-EP + The purified antibody samples were prepared using HBS-EP buffer. + The samples were diluted to 1 μg / ml with buffer and analyzed by a Biacore T200 (GE Healthcare). The assay was performed in one coated channel on the chip at 10 μl / min for 2 min. The association / dissociation kinetics was analyzed using a 1:1 model in the BIAevaluation software with an association time of 8 min and a dissociation time of 90 min at a flow rate of 30 μl / min.

[0135] Biacore competition assay for IL-17BR First, a Protein A biacore chip was prepared by capturing 10 μg / ml biotinylated Protein A (Sigma) onto an SA sensor chip. The chip was then inserted into a Biacore 2000 instrument. Anti-IL-17BR antibody was injected at 10 μg / ml at a flow rate of 20 μl / min. This was followed by an injection of 10 μl of IL-17B / IL-25 to confirm non-specific binding of the antibody to the ligand. IL-17B receptor was then injected over the chip at 10 μg / ml, followed by a second injection of IL-17B / IL-25 at the same concentration. Regeneration was performed with 10 μl of 10 mM glycine-HCl (pH 2.2).

[0136] Comparison of thermal stability Fully humanized antibodies and chimeric controls were diluted to 1 μg / mL in PBS / 0.2% Tween and aliquoted into PCR tubes in volumes appropriate for EC80 concentration. Volumes were brought up to 100 μl with the same buffer. Each tube was heated separately for 10 minutes at temperatures between 30° C. and 85° C. at 5° C. intervals and cooled to 4° C. Binding assays were performed against human IL-17BR-His extracellular domain using 100 μl of each antibody per well in 96-well plates (each temperature assayed in duplicate).

[0137] Thermal shift comparison Samples were prepared directly in a 96-well white PCR plate in a final volume of 25 μL (1 μM and 2 μM purified antibody final concentrations). Sypro Orange stock was diluted 1:100 in PBS buffer and then added 1:10 to the final sample (e.g., 2.5 μL in 25 μL). Samples were loaded into the qPCR instrument and amplified using MxPro software, SYBR Green method (filter=FRROX, no reference dye) with a thermal profile setting of 71 cycles with 1°C increments. Results were plotted and Tm determined.

[0138] SEC-MALS (freeze-thaw and heat-induced stress testing) Size exclusion chromatography-multi-angle light scattering (SEC-MALS) analysis was performed on the "unstressed" and "stressed" samples to detect the presence and / or induction of aggregates that could potentially cause downstream manufacturing problems. 10 μl of each sample (1 mg / mL) was injected onto a SEC column (Acquity UPLC BEH200 SEC, 4.6×150 mm, 1.7 μm) followed by three consecutive detectors: UV (Agilent's 1260 Infinity HPLC system with a thermostatted column compartment), light scattering (Wyatt Technology's DAWN HELEOS) and differential refractometer (Wyatt Technology's Opt Detection was performed by a HPLC-ilab TRex. A constant flow rate of 0.4 mL / min was applied with a mobile phase of Dulbecco's PBS (Sigma's D8537 containing 0.01% sodium azide). All experiments were performed at 25°C. Data were analyzed using ASTRA software from Wyatt Technology (version 6.1.2.83) with the refractive index increment (dn / dc) set to 0.185 (i.e., for protein analysis). All samples were stored at 4°C prior to analysis by SEC-MALS.

[0139] Complementary Interaction Chromatography (CIC) Complementary interaction chromatography (CIC) analysis was performed to assess the propensity for non-specific protein-protein interactions and to provide an indication of any solubility issues that may result in downstream manufacturing problems. First, 1 ml of 1000 μg of human polyclonal IgG (Sigma 14506) was conjugated to the L of NHS-activated resin (GE Healthcare), and a 1 mL blank column as a control. The assay was carried out by two separate 20 μl injections (0.5 mg / mL) onto combined NHS-activated resin. The mobile phase was Dulbecco's PBS containing 0.01% sodium azide (Sigma). All experiments were performed at 25°C. Eluting samples were detected by UV absorbance (Agilent HPLC with thermostatted column compartment). 1260 Infinity HPLC system, and the data was analyzed using ASTRA software from Wyatt Technology (version 6.1.2.83) to determine sample peak retention times. These were then used to determine the retention factor k':

number

[0140] Evaluation of solubility by solvent absorption Vivapore™ solvent absorption concentrators were loaded with 3.5-5.0 ml of antibody at 1 mg / ml in PBS at time 0. Antibody concentration was monitored once every 10 min by sampling a small aliquot and measuring with a Nanodrop 2000 (ε=1.4) and continuing until the concentrated volume reached a dead volume of approximately 30 μl-50 μl. Concentration values ​​(mg / ml) were plotted against the corresponding time points to generate a concentration profile.

[0141] Dynamic Light Scattering (DLS) Dynamic light scattering was used as a complementary technique (i.e. to static light scattering such as SEC-MALS) for the detection of soluble aggregates. When performed, 30 μl of sample (Dulbecco's PBS; D8537 from Sigma) was loaded into a 348-well polypropylene plate (Greiner bio-one) and data was recorded on a Zetasizer APS (Malvern). All values were recorded in triplicate and processed using the associated Zetasizer software (version 7.10). Cumulant analysis was performed to obtain the mean particle size (z-average) and polydispersity index (PDI).

[0142] ELISA (enzyme-linked immunosorbent assay) ELISA plates were coated with 2μg / ml human IL17RA-Fc fusion protein (Sino Biological, 10895-H03H), human IL17BR-his tag protein (Sino Biological, 13091-H08H), human IL17RC-his tag protein (11747-H08B), human IL17RD-his tag (10507-H08H) at 4℃ overnight. The next day, the plates were blocked with 1% BSA in PBS, followed by the addition of hD9042. The binding specificity of hD9042 was measured as absorbance at 450nm using anti-human IgG F(ab')2 peroxidase antibody (Jackson ImmunoResearch Inc).

[0143] In vitro human IL-5 and IL-13 cytokine production assay in human peripheral blood mononuclear cells (PBMCs) Fresh human PBMCs were resuspended in RPMI 1640 (Sigma Aldrich) containing 10% heat-inactivated human AB serum and then plated in a total of 200 μL of culture medium in a 96-deep-well plate (Nunc). 7.5×10 in culture medium 4 Cells were seeded as above at a density of 100 cells / well. For induction of IL-5 and IL-13 expression, 10 U / mL human IL-2 (Peprotech, 200-02) combined with 2 ng / ml or 10 ng / ml human IL-25 (R&D systems, 1258-IL / CF) was added to each well. For inhibition studies, PBMCs were incubated with 2 μg / mL hD9 After treatment with 042 or isotype control (ChromPure Human IgG, whole molecule, Jackson ImmunoResearch Inc) for 1 hour, IL-2 and IL-25 were added. Supernatants were collected 7 days after cytokine and antibody treatment and analyzed by IL-5 and IL-13 ELISA kits (SAB biotech).

[0144] In vitro human IL-8 cytokine production assay in human renal cell carcinoma cell line TK-10 TK-10 cells were maintained in RPMI 1640 medium containing 10% fetal bovine serum at 37 °C in a humidified atmosphere with 5% CO. For IL-8 stimulation, TK-10 cells were cultured at 2.5 × 10 in OptiMEM serum-reduced medium (Thermofisher Scientific) in a total volume of 100 μL. 4 Cells were seeded at a density of 10 cells / well, followed by addition of 10 ng / ml human TNFα (Peprotech, 30 0-01A) and 100ng / ml human IL-25 (R&D systems, 1258-IL / C For inhibition studies, TK-10 cells were treated with 100 ng / mL hD9042, hD9043 or isotype control (ChromPure Human IgG, whole molecule, Jackson Immuno Research Inc) for 1 hour, followed by TNFα and IL-1. 25 was added. The supernatants collected after cytokine and antibody treatments were analyzed by human IL-8 ELISA kit (R&D systems, D8000C).

[0145] result Sequencing of D9.2 antibody Cloning and sequencing of mouse D9.2 was performed as described in US Pat. No. 5,399,633.

[0146] Generation of a chimeric version of the D9.2 antibody 1. Construction of chimeric D9.2 expression vector Construction of chimeric expression vectors involves the addition of suitable leader sequence DNA to the VH and VK genes following the HindIII restriction site. Leader sequences are listed in the Kabat database. The most similar sequence in the VK gene was selected. Furthermore, the construction of the chimeric heavy chain expression vector involved the introduction of a 5'-fragment of the human IgG1 heavy chain constant region gene up to the native ApaI restriction site adjacent to the 3' end of the J region of the VH gene. For the construction of the chimeric kappa light chain expression vector, a 3'-splice donor site and a 5'BamHI restriction site were added to the VK gene. The splice donor sequence is important for the correct in-frame addition of the variable region gene to its appropriate constant region gene and excision of the V:C intron. The appropriate constant region gene is encoded downstream of the inserted variable region sequence. The 400 bp (approximately) PCR product of the cD9.2 VK Advantage HF2 (Clontech) reaction was cloned into the pCR2.1 vector using the TOPO-TA cloning kit and the manufacturer's protocol. DNA plasmid minipreps of selected clones with the correct fragment size were cut with HindIII and BamHI and ligated into the vector pKN100. Clones were isolated after transformation of chemically-competent DH5α bacteria with the ligation products. Overnight cultures (5 ml) of four clones were processed using the QIAprep Spin Miniprep Kit according to the manufacturer's protocol to generate DNA plasmid minipreps, which were sequenced by GATC-Biotech using primer Hu-K3 and named D9.2 cVK.pKN100. The 450 bp (approx.) PCR product of the cD9.2 VH Advantage HF2 (Clontech) reaction was treated with DpnI, cut with HindIII and ApaI, and directly ligated into the expression vector pG1D200. Clones were isolated after transformation of chemically-competent DH5α bacteria with the ligation products and screened by PCR using primers HCMVi (F0951) and chD9.2VH REV. Overnight cultures (5 ml) of five clones that produced PCR products of the correct size were purified using QIAprep Processed using the Spin Miniprep Kit according to the manufacturer's protocol. A DNA plasmid miniprep was generated and sequenced by GATC-Biotech using primer HCMVi and designated D9.2 cVH.pG1D200.

[0147] 2. Generation of chimeric D9.2 antibody Plasmid preparations of D9.2cVH.pG1D200 and D9.2cVK.pKN100 were prepared using Promega's Maxiprep System from overnight cultures of transformed DH5α bacteria. The plasmids were purified using TEM and the manufacturer's protocol. These two plasmids were used to co-transfect 293T cells. Conditioned medium was harvested 3-4 days later. IgG1 kappa antibody concentrations in conditioned medium from 293T transfections measured by quantitative ELISA were 1862.78ng / ml and 3573.13ng / ml in two sets of triplicate transfections for D9.2.

[0148] 3. IL-17BR binding activity of chimeric cD9001 antibody Recombinant human IL-17BR-His (R&D Systems), mouse IL-17BR-(Fc ) (Sino Biological) and the extracellular domain of cynomolgus monkey IL-17BR-monoFc The antibody binding activity to the IL-17BR protein was measured by binding ELISA. Figure 1 shows that the chimeric antibody cD9001 has similar EC 50 The binding is indicated by the value.

[0149] D9.2 Design of humanized antibody variants 1. Human VH and VK cDNA database International Immunogenetics Database 2009 (Lefranc, 2015) and Kabat Database Release 5 of Sequences of Proteins of Immunological Interest (last updated in 1999) Human and mouse immunoglobulin proteins from the 1990s (Kabat et al. 1991) A database of human immunoglobulin sequences in a Kabat alignment using quality sequences. Our database contains 10406 ​​VH and 2894 VK sequences.

[0150] 2. Molecular model of D9.2 Homology models of the murine D9.2 antibody variable regions were calculated using Discovery Studio 3.5 program running in automatic mode. The atomic coordinates of 3rkd_C.pdb and 1mj8_H.pdb were the highest scoring sequence templates for VL and VH, respectively, as determined by Blast analysis of the Accelrys antibody pdb structural database, and the atomic coordinates of 4aeh_LH.pdb were the highest scoring global (interface) sequence template. These templates were used to generate 30 initial models. The top scoring models were refined by modeling each CDR loop with its five best loop templates. The 20 final models were used to determine the identity of residues within 4 Å of the CDR loops.

[0151] 3. Human Framework Selection Humanization requires the identification of suitable human V regions. Human VH and VK databases were matched against the D9.2 VH and VK protein sequences using various selection criteria. FW residues within 4 Å (Kabat definition) of the CDR residues in the structure of the murine D9.2 antibody were identified and selected as "4 The sequences were designated "4 Å adjacent residues". Humanized and incomplete sequences were excluded from the analysis. Sequence EF178110 was chosen as the human heavy chain donor candidate. This sequence has a very high sequence identity and similarity score and has six 4 Å adjacent residue changes, which was the minimum number of changes obtained (Table 1).

[0152] Similarly, the sequence Y14869 was chosen as a candidate human kappa light chain donor. 9.2 The sequence identity and similarity scores to VK are very high, and IGKV1-NL1 * 01 has no somatic mutations from the germline. It has two potential 4 Å adjacent residue changes (Table 2).

[0153] 4. Design of D9 HA and HB With suitable human frameworks identified, synthetic protein and DNA sequences can be designed. The initial design of a humanized version of D9.2 was to graft CDRs 1, 2 and 3 from the D9.2 VH into the acceptor FW of EF178110, resulting in the mutant D9 HA (SEQ ID NO: 7). The humanized version D9 was then Six 4 Å adjacent residues at positions 52, 74-76, 79, and 105 were backmutated to the equivalent murine residues in HB and mutated one at a time in subsequent variants. The sequences were computationally assembled and designated D9 HC through D9 HH. Table 1 compares the murine and humanized versions of the D9 VH protein sequence.

[0154] 5. Design of D9 KA and D9 KB The framework derived from Y14869 was used to design the DNA and protein for the humanized construct. CDR1, 2 and 3 from D9 VK were grafted into the acceptor FW of Y14869 to generate the initial version of humanized D9 (KA SEQ ID NO: 12). Two mismatched 4 Å adjacent residues 54 and 76 were backmutated to the equivalent mouse residues in humanized version D9 KB and mutated one at a time in subsequent variants. The sequences were computationally assembled and designated D9 KC and KD (Table 2).

[0155] Generation and Characterization of a Humanized Version of the D9.2 Antibody 1. Generation of D9 humanized antibody Construction of humanized expression vectors involves cloning the amplified variable regions into IgG / kappa vectors (pHuK and pHuG1) using ligase-independent cloning (LIC). After digestion of the vectors (pCMV modified) with BfuA1 (BspM1), compatible overhangs are generated using T4 DNA polymerase 3'-5' exonuclease activity (+dATP).

[0156] The D9 HA, HB, KA and KB variable region genes were synthesized by GenScript. The native human framework sequences EF178110 and Y14869, heavy and light chains, respectively, and the native mouse CDR sequences were computationally assembled and designated D9 HA to D9 HH and D9 KA to D9 KE. The sequences of HA / B and KA / B were optimized by silent mutagenesis and synthesized to use codons preferentially utilized by human cells. The KA / B and HA / B constructs were PCR amplified with specific primers for the expression vector plus insert. First, the variable region was amplified by PCR using a primer containing either the 3' end of the leader sequence (most of the sequence is in the vector) - forward primer - or the beginning of the constant region (IgG1 or kappa) - reverse primer - followed by the beginning of the variable region (in each orientation). Complementary overhangs were generated in the PCR products by T4 DNA polymerase + dTTP treatment. The vectors and inserts were incubated at room temperature, transformed into chemically competent TOP10 bacteria, and plated on kanamycin plates. Several clones were isolated and colonies were screened by PCR using primers HCMVi and HuG1 LIC Rev for VH or HuK LIC Rev for VK. Clones generating PCR products of the correct size were selected, miniprepped using a QIAGEN kit and sequenced using the same primers. Versions HA and KA were subsequently modified by PCR mutagenesis to obtain other humanized variants, as annotated in Tables 1 and 2, respectively. Clones were sequenced and plasmid DNA was purified using QIAGEN's Plasmid Miniprep Kit or Promega's Plasmid Maxipre Kit. Expression plasmid preparations encoding (humanized or chimeric) VH, HA, KA and VK were used to transfect Expi293 cells and cultured in serum-free medium for 5-7 days, at which point conditioned medium containing the secreted antibodies was harvested.

[0157] 2. Antibody Expression The concentration of IgG1κ antibody in Expi293 cell conditioned medium was measured by HTRF and is shown in Table 3. This was determined by combining XL665-labeled IgG and cryptate-labeled Mab anti-human F c This is a competitive immunoassay based on the HTRF technique (homogeneous time-resolved fluorescence) for the quantitative determination of human IgG capable of displacing the binding between F and F in samples or standards. c The expression level is inversely proportional to the concentration of KM. The chimeric antibody (cD9001) showed good expression similar to other antibodies that expressed chimeric light chains (D9 HAcK and D9 HBcK). However, all of the antibodies containing KA light chains (cHKA, D9 HAKA and D9 HBKA) had very low levels of expression. Therefore, a model light chain named KM shown in Table 2 was used to determine the heavy chain to be used. Most of the antibodies that expressed KM as the light chain showed higher expression levels (hD9002, hD9006, hD9010, hD9014, hD9018, hD9022, hD9026 and hD9030).

[0158] 3. Antigen binding by early versions of the humanized D9 antibody Off-rate ranking was performed using Biacore for the heavy chain variants with KM to determine the heavy chain version with the slowest off-rate with human IL-17BR. The data was fitted using association and dissociation, and the K offThe values ​​were used to rank the data (Table 4). Heavy chain version H produced the slowest off-rate (hD9030) and was therefore chosen as the lead heavy chain version at this stage.

[0159] 4. Biacore competition assay with IL-17B or IL-25 To determine whether the humanized antibodies directly compete with IL-17B or IL-25 for binding to IL-17BR, a Biacore competition assay was performed. First, the humanized antibody hD9030 was loaded onto a pre-coated Protein A chip. IL-17B or IL-25 were injected to confirm binding to the antibody alone. This was followed by injection of IL-17BR, where binding to the antibody could be observed as shown in Figures 2 and 3. IL-17B / IL-25 were then injected again to see if the natural ligands could bind to the receptor in the presence of the humanized antibody. In the case of IL-17B, the latter was unable to bind to IL-17BR in the presence of the humanized antibody (Figure 2), suggesting that their epitopes may overlap. However, for IL-25, binding to IL-17BR could be observed after binding of the humanized antibody to the receptor (Figure 3), suggesting that they do not directly compete for the same site.

[0160] 5. Reengineering of heavy and light chain sequences Given the data indicating that the KA light chain mutant was poorly expressed, the light chain sequence was analyzed further: site-directed mutagenesis was used to mutate Arg99 in D9 KA to Gly to generate version KE (Table 2).

[0161] It was also desired to modify the heavy chain sequence to have a higher identity percentage (>85%) to the human germline. The identity of the KE light chain to the human germline is 86% and no modifications were necessary in this construct. For the heavy chain, the current lead heavy chain H has an identity of 80.4% to the human germline and a large number of mutations were required to reach >85%. The sequence was examined by molecular modeling and two promising versions were considered: mutation of three residues in the framework regions that appear to be non-essential for binding or mutation of three residues in CDRH2. Both of these heavy chain versions were modified using site-directed mutations. Generate D9 HI and D9 HI clones using heterotransferase induction (section 8.8) as shown in Table 1. We named him HJ.

[0162] 6. Reengineered Antibodies; Expression and Purification The expression levels of antibodies containing D9 HH-HJ heavy chains with either the original KA or modified KE light chains (transfected with Expi293 cells) were quantified using the Octet quantification assay. Again, as shown in Table 3, antibodies containing D9 KA as the light chain (hD9035, hD9036 and hD9037) all had low expression levels, while antibodies containing D9 KE as the light chain (hD9040, hD9041 and hD9042) showed moderate expression levels. Antibodies hD9040, hD9041 and hD9042 were selected for further characterization as three lead humanized antibody candidates and were purified using affinity and size exclusion chromatography.

[0163] 7. Antigen binding by humanized D9 antibody Binding of human IL-17BR to antibody versions hD9040, hD9041 and hD9042 was tested by ELISA. All three humanized variants showed very similar EC 50 It bound to human IL-17BR-His at 100 mM levels (Figure 4).

[0164] 8. Kinetics of binding of humanized hD9040, hD9041 and hD9042 antibodies to IL-17BR To further characterize the binding of the lead humanized candidate hD9040, hD9041, and hD9042 antibodies to human IL-17BR-His, kinetic experiments were performed using a Biacore T200 instrument. The three humanized antibodies showed good kinetics with affinities in the low picomolar range (Figure 5). Although these stringent affinities reach the limits of the Biacore instrument, it is clear that all three antibodies have affinities comparable to the chimeric antibody.

[0165] 9. Thermostability of humanized hD9040, hD9041 and hD9042 antibodies at high temperatures The purpose of this experiment is to measure the EC of each candidate by exposing them to various high temperatures from 30°C to 85°C for 10 minutes, then cooling them to 4°C. 80 The aim of this study was to test the thermal stability of the humanized antibodies when used in an ELISA assay at various concentrations. Humanized candidates hD9040 and hD9041 retain their binding ability to human IL-17BR-His up to 73° C., and hD9042 retains binding up to 75° C. (FIG. 6).

[0166] 10. Determination of Tm (melting temperature) of humanized hD9040, hD9041 and hD9042 antibodies To determine the melting temperatures of the lead humanized antibodies hD9040, hD9041 and hD9042, they were tested in a thermal shift assay. Samples were incubated with a fluorescent dye (Sypro Orange) in a qPCR thermal cycler for 71 cycles with an increase of 1°C per cycle. The Tm values ​​of the humanized antibodies were calculated to be 72°C-74°C (Figure 7, Table 5).

[0167] 11. Aggregation of humanized hD9040, hD9041 and hD9042 antibodies Samples were injected at 0.4 mL / min onto a size exclusion column in an HPLC system and analyzed by multi-angle light scattering to determine absolute molar mass and confirm aggregation. For humanized candidates hD9040, hD9041 and hD9042, the profiles show no signs of aggregation (Figure 8, Table 6) and the average molecular weight is between 136 kDa and 132 kDa (Table 6), which is the expected range for IgG monomers in this analytical setup. The antibodies are monodisperse (Mw / Mn<1.05). The mass recovery was 100% (calculated mass over injected mass), indicating good protein recovery and that the samples do not appear to contain insoluble aggregates that stick to the column or are retained by the guard column. Overall, the data indicates that the humanized antibodies hD9040, hD9041 and hD9042 are highly aggregated and show no signs of aggregation. This suggests that there are no aggregation problems with 041 and hD9042.

[0168] 12. Non-specific protein-protein interactions (CIC) of humanized hD9040, hD9041 and hD9042 antibodies Interaction chromatography using bulk purified human polyclonal IgG is a technique for monitoring non-specific protein-protein interactions and can be used to distinguish between soluble and insoluble antibodies. An increase in retention index (k') indicates a tendency for self-interaction and poor solubility. Antibody hD9042 has the lowest tendency for non-specific interactions, followed by hD9040 and then hD9041. All three humanized antibodies show retention indices below 0.2, indicating a generally low tendency for non-specific interactions and good solubility (Figure 9).

[0169] 13. Solubility of humanized hD9040, hD9041 and hD9042 antibodies Humanized antibodies hD9040, hD9041 and hD9042 were concentrated using a solvent absorption concentrator (MWCO 7500 kDa) and the concentration was measured at regular intervals. The antibodies were concentrated to over 65 mg / mL without obvious precipitation, demonstrating that they were soluble in PBS to high concentrations (Figure 10).

[0170] 14. Freeze / Thaw Stress Analysis of Humanized hD9040, hD9041 and hD9042 Antibodies Samples of purified candidate antibodies were subjected to 10 cycles of -80°C for 15 minutes followed by thawing at room temperature for 15 minutes. Samples were then analyzed by SEC-MALS to confirm aggregation (Figure 11). The data suggest that freeze / thaw does not cause aggregation in humanized antibodies hD9040, hD9041 and hD9042 (Figures 11A, 11B and 11C, respectively).

[0171] 15. Heat-induced stress analysis of humanized hD9040, hD9041 and hD9042 antibodies Samples of purified candidate antibodies were exposed to a) 4°C, b) 25°C, c) 37°C and d) 50°C for 33 days. Samples were then analyzed by SEC-MALS as described in section 8.19 to confirm aggregation (Figure 12). Overall, the data suggests that there are no aggregation issues with humanized antibodies hD9040, hD9041 and hD9042 (Figures 12A, 12B and 12C, respectively).

[0172] 16. pI Analysis of Humanized hD9040, hD9041, and hD9042 Antibodies The pI analysis of the humanized antibodies hD9040, hD9041 and hD9042 was carried out using capillary isoelectric focusing (cIEF). This technique allows the antibodies to be separated according to their isoelectric point (pI) using a pH gradient across a capillary. Table 7 shows the main pI isoforms of each antibody and the pI range of each antibody. The pI of hD9040 and hD9041 is 7.8 and the pI of the hD9042 antibody is 7.9.

[0173] 17. Self-association analysis of humanized hD9040, hD9041 and hD9042 antibodies by DLS A two-fold dilution series was made for each antibody, hD9040, hD9041, and hD9042, at concentrations from 10 mg / ml to 0.63 mg / ml (five samples total). The hydrodynamic diameter and diffusion constant were then determined for each sample using dynamic light scattering (DLS). As shown in Figure 13, hD9042 has the lowest tendency to self-associate, followed by hD9040 and hD9041.

[0174] 18. hD9042 Binding Specificity to the hIL17 Receptor Family Binding specificity of lead candidate hD9042 to human IL17BR was assessed by ELISA. It was determined that hD9042 specifically binds to human IL17BR, but does not cross-react with human IL17RA, IL17RC, or IL17RD (FIG. 14).

[0175] 19. In vitro human IL-5 and IL-13 cytokine production assays In the presence of IL-2, IL-25 significantly induces the production of type 2 cytokines (IL-5 and IL-13) in human PBMC. hD9042 blocked the production of IL-5 (FIG. 15A) and IL-13 (FIG. 15B) induced by IL-25. Statistical significance is expressed as NS (p>0.05). * p ≤ 0.05, ** Shown as p≦0.01 (by unpaired two-tailed Student's t test).

[0176] 20. Generation of hD9043, a hinge-stabilized IgG4 antibody To minimize the effector function of the Fc region, D9HJ was fused to the IgG4 constant region to produce D9HJ-IgG4. * To inhibit Fab arm exchange and stabilize the hinge region, a point mutation was introduced to mutate the serine at position 228 to proline (EU numbering system) (Angal, S. 1993 Mol Immunol, 30:105-108, Aalberse et al. 2002 Immunol, 105:9-19). D9HJ-IgG4* The sequence is shown in SEQ ID NO: 17. Next, D9HJ-IgG4 * The sequence of was optimized by silent mutagenesis to use codons preferentially utilized by human cells and synthesized by GenScript. To generate hD9043, D9HJ-IgG4 * was inserted into the pcDNA3.1 vector and co-transfected with D9KE into ExpiCHO cells. hD9043 was purified using affinity and size exclusion chromatography.

[0177] 21. In vitro human IL-8 cytokine production assay IL-25 in combination with TNFα was able to induce the production of IL-8 from TK-10 cells. IL-8 production could be significantly blocked by hD9042 and hD9043 (Figure 16). * p ≤ 0.05, ND, not detected.

[0178] Summary of data for hD9040-42 compared with cD9001 A summary of the binding, kinetic affinity and biophysical properties of humanized candidates hD9040, hD9041 and hD9042 is shown in Table 8. Considering all the data, hD9042 was selected as the lead humanization candidate since it had the slowest off-rate, was the most stable, had the lowest tendency to self-associate and showed the best expression level among the three candidates.

[0179] When the D9.2 antibody was humanized, the resulting antibody retained binding to IL-17BR and had good biophysical properties. The antibody was engineered and expressed as a fully humanized antibody with little loss of binding capacity. Experiments with humanized antibodies hD9040, hD9041, and hD9042 showed high affinity in the low picomolar range in binding ELISA and kinetic studies using Biacore (Figures 4 and 5). Initial experiments showed that the CDR-grafted KA light chain antibody was poorly expressed, but simple re-engineering of the light chain significantly improved expression of the antibody. The heavy chain was successfully engineered to have >85% identity to the human germline and therefore can be referred to as a "humanized antibody" in accordance with the new regulations. The hD9042 antibody was chosen as a lead candidate since it showed the best drug-like properties and excellent binding kinetics (Table 8). The combination of excellent binding, expression, thermal stability, and affinity makes hD9042 a good candidate antibody for further development.

[0180] array SYWMN SEQ ID NO:1 VHCDR1 RIDPYDSEIQYX1QKFX2X3 (X1 is N or A, and X2 is K or Q) and X3 is D or G) SEQ ID NO:2 VHCDR2 SGGFDWFAY SEQ ID NO:3 VHCDR3 RASENINSNLA SEQ ID NO:4 VLCDR1 DVTNLAD SEQ ID NO:5 VLCDR2 QHFWGPPYT SEQ ID NO:6 VLCDR3 TIFF2025041747000003.tif10170 SEQ ID NO: 7 VH sequence HA (CDRs are shaded) (Kabat positions 1, 24, 60, 64, 65 , 69 and 94 are underlined) TIFF2025041747000004.tif10170 (X1 is Q or E, X2 is A or T, X3 is A or N, X4 is Q or K, X5 is G or D, and X6 is M or L) SEQ ID NO: 8 VH sequence with modifications at Kabat positions 1, 24, 60, 64, 65 and 69 ( CDR is shaded) TIFF2025041747000005.tif10170 SEQ ID NO: 9 VH sequence HJ (CDRs are shaded) TIFF2025041747000006.tif10170 SEQ ID NO: 10 VH sequence HH (CDRs are shaded) TIFF2025041747000007.tif11170 SEQ ID NO: 11 VH sequence HI (CDRs are shaded) TIFF2025041747000008.tif10170 SEQ ID NO: 12 VL sequence KA (CDRs are shaded) (Kabat position 93 is underlined) TIFF2025041747000009.tif9170 SEQ ID NO: 13 VL sequence KE (CDRs are shaded) RIDPYDSEIQYNQKFKD SEQ ID NO:14 VHCDR2 HJ and HH RIDPYDSEIQYAQKFQG SEQ ID NO:15 VHCDR2 HI ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVE VHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: 16 Hinge stabilized IgG4 constant region QVQLVQSGAEVKKPGASVKVSCKASGYTFISYWMNWVRQA PGQGLEWMGRIDPYDSEIQYNQKFKDRVTMTRDTSISTAYMELSRLRSDDTAVYYCARSGGFDWFAYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPRE PQVYTLPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: 17 D9HJ-IgG4 * array

[0181] [Table 1]

[0182] [Table 2]

[0183] [Table 3]

[0184] [Table 4]

[0185] [Table 5]

[0186]

Table 6

[0187]

Table 7

[0188]

Table 8

Claims

1. An antibody comprising a heavy chain variable domain and a light chain variable domain, a) the heavy chain variable domain comprises a VHCDR1 of SEQ ID NO:1, a VHCDR2 of SEQ ID NO:2, and a VHCDR3 of SEQ ID NO:3; b) An antibody, wherein the light chain variable domain (VK domain) comprises a VLCDR1 of SEQ ID NO: 4, a VLCDR2 of SEQ ID NO: 5 and a VLCDR3 of SEQ ID NO:

6.

2. The antibody of claim 1, which specifically binds to interleukin-17 receptor B (IL17BR).

3. 3. The antibody of claim 2, which binds to the interleukin-17 receptor with a binding affinity that is at least 75% of the binding affinity of murine antibody D9.2 to interleukin-17 receptor B as measured by ELISA.

4. The antibody according to any one of claims 1 to 3, wherein the expression level of the antibody is at least 40% of the expression level of mouse antibody D9.2 in mammalian cells.

5. The antibody of any one of claims 1 to 4, wherein the heavy chain variable domain (VH domain) comprises SEQ ID NO:8, optionally with up to four additional framework substitutions.

6. The heavy chain variable domain (VH domain) comprises SEQ ID NO:8, ３ The antibody of claim 5 , wherein is N.

7. The heavy chain variable domain (VH domain) comprises SEQ ID NO:8, ４ The antibody of claim 5 or 6, wherein is K.

8. The heavy chain variable domain (VH domain) comprises SEQ ID NO:8, ５ The antibody according to any one of claims 5 to 7, wherein is D.

9. The heavy chain variable domain (VH domain) comprises SEQ ID NO:8, １ The antibody of any one of claims 5 to 8, wherein is Q.

10. The heavy chain variable domain (VH domain) comprises SEQ ID NO:8, ２ The antibody according to any one of claims 5 to 9, wherein is A.

11. The heavy chain variable domain (VH domain) comprises SEQ ID NO:8, ６ The antibody according to any one of claims 5 to 10, wherein is M.

12. The antibody of any one of claims 1 to 11, wherein the heavy chain variable domain (VH domain) comprises SEQ ID NO: 9 with up to four additional framework substitutions.

13. The antibody of claim 12, wherein the heavy chain variable domain (VH) comprises SEQ ID NO:

9.

14. The heavy chain variable domain (VH domain) comprises SEQ ID NO:8, １ The antibody according to any one of claims 5 to 7, wherein is E.

15. The heavy chain variable domain (VH domain) comprises SEQ ID NO:8, ２ The antibody of any one of claims 5 to 7 and 14, wherein is T.

16. The heavy chain variable domain (VH domain) comprises SEQ ID NO:8, ６ The antibody of any one of claims 5 to 7, 14 and 15, wherein is L.

17. The antibody of any one of claims 5 to 7 and 14 to 16, wherein the heavy chain variable domain (VH domain) comprises SEQ ID NO: 10 with up to four additional framework substitutions.

18. The antibody of claim 17, wherein the heavy chain variable domain (VH) comprises SEQ ID NO:

10.

19. The heavy chain variable domain (VH domain) comprises SEQ ID NO:8, ３ The antibody of claim 5, wherein is A.

20. The heavy chain variable domain (VH domain) comprises SEQ ID NO:8, ４ The antibody of claim 5 or 19, wherein is Q.

21. The heavy chain variable domain (VH domain) comprises SEQ ID NO:8, ５ The antibody of claim 5, 19 or 20, wherein is G.

22. The heavy chain variable domain (VH domain) comprises SEQ ID NO:8, １ The antibody of any one of claims 5 and 19 to 21, wherein is E.

23. The heavy chain variable domain (VH domain) comprises SEQ ID NO:8, ２ The antibody of any one of claims 5 and 19 to 22, wherein is T.

24. The heavy chain variable domain (VH domain) comprises SEQ ID NO:8, ６ The antibody of any one of claims 5 and 19 to 23, wherein is L.

25. 25. The antibody of any one of claims 5 and 19 to 24, wherein the heavy chain variable domain (VH domain) comprises SEQ ID NO: 11 with up to four additional framework substitutions.

26. 26. The antibody of claim 25, wherein the heavy chain variable domain (VH) comprises SEQ ID NO:

11.

27. The antibody of any one of claims 1 to 26, wherein the VH domain is fused to an antibody constant region comprising SEQ ID NO:

16.

28. The antibody of claim 27, wherein the amino acid sequence of the VH domain and constant region comprises SEQ ID NO:

17.

29. The antibody of any one of claims 1 to 28, wherein the light chain variable domain (VL domain) comprises SEQ ID NO: 13 with up to four additional framework substitutions.

30. The antibody of claim 29, wherein the light chain variable domain (VL domain) comprises SEQ ID NO:

13.

31. The antibody of any one of claims 1 to 30, further comprising a second heavy chain variable domain and a second light chain variable domain which form an antigen-binding site for an antigen other than IL17BR.

32. A pharmaceutical composition comprising the antibody of any one of claims 1 to 31 and a pharma- ceutically acceptable carrier.

33. A nucleic acid encoding the antibody of any one of claims 1 to 31.

34. 34. A vector comprising the nucleic acid of claim 33 operably linked to a promoter.

35. 35. A host cell comprising a nucleic acid according to claim 33 or a vector according to claim 34.

36. 32. A method of making an antibody according to any one of claims 1 to 31, comprising expressing the vector according to claim 34 in a host cell culture to produce the antibody, and recovering the antibody from the cell culture.

37. A method for treating or preventing an allergic or inflammatory condition by administering to an individual in need of such treatment an effective amount of the antibody of any one of claims 1 to 31, the pharmaceutical composition of claim 32, the nucleic acid of claim 33 or the vector of claim 34.

38. 35. A method for treating or preventing a cancerous condition by administering to an individual in need of such treatment an effective amount of an antibody according to any one of claims 1 to 31, a pharmaceutical composition according to claim 32, a nucleic acid according to claim 33 or a vector according to claim 34.

39. An antibody according to any one of claims 1 to 31, a pharmaceutical composition according to claim 32, a nucleic acid according to claim 33 or a vector according to claim 34 for use in a method for the treatment of the human or animal body.

40. An antibody according to any one of claims 1 to 31, a pharmaceutical composition according to claim 32, a nucleic acid according to claim 33 or a vector according to claim 34 for use in a method for the treatment of an allergic or inflammatory condition in an individual.

41. 41. The method of claim 37, or the antibody, pharmaceutical composition, nucleic acid or vector for use according to claim 40, wherein the allergic or inflammatory condition is asthma, ulcerative colitis or fibrosis.

42. 42. The antibody for use or method of claim 41, wherein the asthma is allergic asthma or rhinovirus exacerbated asthma.

43. 42. The antibody for use or method of claim 41 , wherein the fibrosis is idiopathic pulmonary fibrosis.

44. An antibody according to any one of claims 1 to 31, a pharmaceutical composition according to claim 32, a nucleic acid according to claim 33 or a vector according to claim 34 for use in a method for the treatment of a cancerous condition in an individual.

45. 45. The method of claim 38 or the antibody, pharmaceutical composition, nucleic acid or vector of claim 44, wherein the cancer condition is breast cancer or pancreatic cancer.

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