IL4 / IL13 receptor molecules for veterinary use

Abstract

Provided are multiple embodiments involving IL13R / IL4R contiguous polypeptides that are derived from a companion animal species and bind IL13 and / or IL4. Such contiguous polypeptides can be used in methods of treating IL13 and / or IL4-induced conditions in companion animals such as dogs, cats, and horses.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of priority to U.S. Provisional Application No. 62 / 488,509, filed on April 21, 2018, which is incorporated herein by reference in its entirety for any purpose. Technical Field

[0003] The present disclosure relates to a continuous polypeptide comprising an interleukin 4 receptor and an interleukin 13 receptor fragment derived from a companion animal species, which continuous polypeptide binds IL4 and / or IL13 of the companion animal species, for example, canine IL4 and canine IL13. The present invention also relates to methods of using the continuous polypeptide, for example, in companion animals (such as dogs, cats and horses), for example, for treating IL4 and / or IL13-induced conditions or reducing IL4 and / or IL13 signaling activity in cells. Background Art

[0004] Interleukin 4 (IL4) is a cytokine that promotes the differentiation of naive helper T cells into Th2 cells. Interleukin 13 (IL13) has a similar effect on immune cells. Both IL4 and IL13 play an important role in T cell-mediated immune responses directly related to allergic reactions (e.g., atopic dermatitis and asthma). It is generally understood that IL4 can form a signaling complex with heterodimeric receptors IL4 receptor subunit α (IL4R) and γc or IL4R and IL13 receptor subunit α-1 (IL13R). IL13 can form a signaling complex with heterodimeric receptors IL4Ra and IL13Ra1. The extracellular domain of IL4Ra or IL13Ra1 can bind to IL4 and / or IL13 and reduce the free concentration of cytokines, thereby reducing clinical signs and symptoms associated with dermatitis, asthma and other conditions.

[0005] Companion species animals, such as cats, dogs, and horses, suffer from many allergic diseases similar to human allergic diseases, including atopic dermatitis and asthma. Therefore, there remains a need for methods and compounds that can be specifically used to bind companion animal IL4 and / or IL13 to treat IL4 / IL13-induced conditions and reduce IL4 / IL13 signaling activity.

[0006] Overview

[0007] In some embodiments, an IL13R / IL4R contiguous polypeptide is provided that comprises an extracellular domain of an IL13R polypeptide and an extracellular domain of an IL4R polypeptide, wherein the IL13R and IL4R polypeptides are derived from a companion animal species.

[0008] In some embodiments, the IL13R / IL4R contiguous polypeptide comprises formula (I) IL13R-L1-IL4R-L2-FP or formula (II) IL4R-L1-IL13R-L2-FP, wherein:

[0009] a. IL13R is an extracellular domain of an IL13R polypeptide derived from said companion animal species,

[0010] b. IL4R is an extracellular domain of an IL4R polypeptide derived from said companion animal species,

[0011] c.L1 is the first optional linker,

[0012] d. L2 is the second optional linker, and

[0013] e. FP is the fusion partner.

[0014] In some embodiments, the IL13R / IL4R contiguous polypeptide is present at less than 5 x 10 -6 M, less than 1x 10 -6 M, less than 5x 10 -7 M, less than 1x 10 -7 M, less than 5x 10 -8 M, less than 1x 10 -8 M, less than 5x 10 -9 M, less than 1x 10 -9 M, less than 5x 10 -10 M, less than 1x 10 -10 M, less than 5x 10 -11 M, less than 1x 10 -11 M, less than 5x 10 -12 M or less than 1x 10 -12 Dissociation constant (Kd) of M binding to IL13 from companion animal species.

[0015] In some embodiments, the IL13R / IL4R contiguous polypeptide is present at less than 5 x 10 -6 M, less than 1x 10 -6 M, less than 5x 10 -7 M, less than 1x 10 -7 M, less than 5x 10 -8 M, less than 1x 10 -8 M, less than 5x 10 -9 M, less than 1x 10 -9 M, less than 5x 10 -10 M, less than 1x 10 -10 M, less than 5x 10-11 M, less than 1x 10 -11 M, less than 5x 10 -12 M or less than 1x 10 -12 Dissociation constant (Kd) of M binding to IL4 from companion animal species.

[0016] In some embodiments, the IL13R / IL4R contiguous polypeptide reduces IL13 and / or IL4 signaling in the companion animal species.

[0017] In some embodiments, the companion animal species is canine, feline, or equine.

[0018] In some embodiments, the extracellular domain of the IL13R polypeptide has at least 85% identity to the amino acid sequence of SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 32, SEQ ID NO: 34, or SEQ ID NO: 36. In some embodiments, the extracellular domain of the IL13R polypeptide has at least 90% identity to the amino acid sequence of SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 32, SEQ ID NO: 34, or SEQ ID NO: 36. In some embodiments, the extracellular domain of the IL13R polypeptide has at least 95% identity to the amino acid sequence of SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 32, SEQ ID NO: 34, or SEQ ID NO: 36. In some embodiments, the extracellular domain of the IL13R polypeptide is at least 98% identical to the amino acid sequence of SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:32, SEQ ID NO:34, or SEQ ID NO:36.

[0019] In some embodiments, the extracellular domain of the IL13R polypeptide comprises a cysteine ​​at a position corresponding to SEQ ID NO: 22, at position 18 of SEQ ID NO: 24, or at position 18 of SEQ ID NO: 26. In some embodiments, the extracellular domain of the IL13R polypeptide comprises a cysteine ​​at position 18 of SEQ ID NO: 22, at position 18 of SEQ ID NO: 24, at position 18 of SEQ ID NO: 26, at position 15 of SEQ ID NO: 32, at position 15 of SEQ ID NO: 34, or at position 15 of SEQ ID NO: 36. In some embodiments, the extracellular domain of the IL13R polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 32, SEQ ID NO: 34, and SEQ ID NO: 36. In some embodiments, the extracellular domain of the IL13R polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 22, SEQ ID NO: 24, and SEQ ID NO: 26.

[0020] In some embodiments, the extracellular domain of the IL4R polypeptide has at least 85% identity to the amino acid sequence of SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 33, SEQ ID NO: 35, or SEQ ID NO: 37. In some embodiments, the extracellular domain of the IL4R polypeptide has at least 90% identity to the amino acid sequence of SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 33, SEQ ID NO: 35, or SEQ ID NO: 37. In some embodiments, the extracellular domain of the IL4R polypeptide has at least 95% identity to the amino acid sequence of SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 33, SEQ ID NO: 35, or SEQ ID NO: 37. In some embodiments, the extracellular domain of the IL4R polypeptide has at least 98% identity to the amino acid sequence of SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 33, SEQ ID NO: 35, or SEQ ID NO: 37. In some embodiments, the extracellular domain of the IL4R polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 33, SEQ ID NO: 35, and SEQ ID NO: 37. In some embodiments, the extracellular domain of the IL4R polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 23, SEQ ID NO: 25, and SEQ ID NO: 27.

[0021] In some embodiments, L1 and L2, if present, each independently comprises an amino acid sequence selected from the group consisting of G, GG, GGG, S, SS, SSS, GS, GSGS (SEQ ID NO:38), GSGSGS (SEQ ID NO:39), GGS, GGSGGS (SEQ ID NO:40), GGSGGSGGS (SEQ ID NO:41), GGGS (SEQ ID NO:42), GGGSGGGS (SEQ ID NO:43), GGGSGGGSGGGS (SEQ ID NO:44), GSS, GSSGSS (SEQ ID NO:45), GSSGSSGSS (SEQ ID NO:46), GGSS (SEQ ID NO:47), GGSSGGSS (SEQ ID NO:48), and GGSSGGSSGGSS (SEQ ID NO:49).

[0022] In some embodiments, the fusion partner is selected from Fc, albumin and albumin binding fragment. In some embodiments, the Fc is (a) a canine heavy chain constant region selected from IgG-A, IgG-B, IgG-C and IgG-D constant regions; (b) a cat heavy chain constant region selected from IgG1, IgG2a and IgG2b constant regions; or (c) a horse heavy chain constant region selected from IgG1, IgG2, IgG3, IgG4, IgG5, IgG6 and IgG7 constant regions.

[0023] In some embodiments, the IL13R / IL4R contiguous polypeptide comprises a sequence selected from the group consisting of SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, and SEQ ID NO: 31.

[0024] In some embodiments, an isolated nucleic acid is provided, encoding the IL13R / IL4R continuous polypeptide described above. In some embodiments, a host cell is provided, comprising a nucleic acid encoding the IL13R / IL4R continuous polypeptide described above. In some embodiments, a method of producing an IL13R / IL4R continuous polypeptide is provided, comprising culturing such a host cell comprising a nucleic acid encoding the IL13R / IL4R continuous polypeptide described above and isolating the continuous polypeptide. In some embodiments, a pharmaceutical composition is provided, comprising the IL13R / IL4R continuous polypeptide described above and a pharmaceutically acceptable carrier.

[0025] In some embodiments, a method of treating a companion animal species having an IL13 and / or IL4-induced condition is provided, comprising administering to the companion animal species a therapeutically effective amount of an IL13R / IL4R contiguous polypeptide described herein or a pharmaceutical composition comprising an IL13R / IL4R contiguous polypeptide described herein. In some embodiments, the companion animal species is a dog, a cat, or a horse. In some embodiments, the IL13 and / or IL4-induced condition is an itch condition or an allergic condition, such as atopic dermatitis, pruritus, asthma, psoriasis, scleroderma, or eczema.

[0026] In some embodiments, the IL13R / IL4R continuous polypeptide or the pharmaceutical composition is administered parenterally. In some embodiments, the IL13R / IL4R continuous polypeptide or the pharmaceutical composition is administered by intramuscular route, intraperitoneal route, intracerebrospinal route, subcutaneous route, intraarterial route, intrasynovial route, intrathecal route or inhalation route.

[0027] In some embodiments, the method further comprises administering a Jak inhibitor, a PI3K inhibitor, an AKT inhibitor or a MAPK inhibitor. In some embodiments, the method further comprises administering one or more antibodies selected from anti-IL17 antibodies, anti-IL31 antibodies, anti-TNFα antibodies, anti-CD20 antibodies, anti-CD19 antibodies, anti-CD25 antibodies, anti-IL4 antibodies, anti-IL13 antibodies, anti-IL23 antibodies, anti-IgE antibodies, anti-CD11α antibodies, anti-IL6R antibodies, anti-α4-integrin antibodies, anti-IL12 antibodies, anti-IL1β antibodies and anti-BlyS antibodies.

[0028] In some embodiments, a method of reducing IL13 and / or IL4 signaling activity in a cell is provided, comprising exposing the cell to an IL13R / IL4R continuous polypeptide or pharmaceutical composition as described herein, under conditions that permit binding of the continuous polypeptide to IL13 and / or IL4, thereby (a) reducing the binding of IL / 4 and / or IL-13 to a native IL13 receptor and / or a native IL-4 receptor and reducing IL13 and / or IL-4 mediated signaling. In some embodiments, the cell is exposed to the IL13R / IL4R continuous polypeptide or pharmaceutical composition ex vivo. In some embodiments, the cell is exposed to the continuous polypeptide or pharmaceutical composition in vivo. In some embodiments, the cell is a canine cell, a cat cell, or a horse cell.

[0029] In some embodiments, a method for detecting IL13 or IL4 in a sample from a companion animal species is provided, comprising contacting the sample with an IL13R / IL4R contiguous polypeptide or pharmaceutical composition as described herein, under conditions that permit binding of the contiguous polypeptide to IL13 and / or IL4, and detecting whether a complex is formed between the contiguous polypeptide and IL13 and / or IL4 in the sample. In some embodiments, the sample is a biological sample obtained from a dog, cat, or horse. Sequence Listing <110> Elanco USA <120> IL4 / IL13 receptor molecules for veterinary use <130> 01157-0008-00PCT <150> US 62 / 488,509 <151> April 21, 2017 <160> 49 <170> PatentIn version 3.5 <210> 1 <211> 131 <212> PRT <213> Wolf <400> 1 Met Gly Leu Thr Ser Gln Leu Ile Pro Thr Leu Val Cys Leu Leu Ala 1 5 10 15 Leu Thr Ser Thr Phe Val His Gly His Asn Phe Asn Ile Thr Ile Lys 20 25 30 Glu Ile Ile Lys Met Leu Asn Ile Leu Thr Ala Arg Asn Asp Ser Cys 35 40 45 Met Glu Leu Thr Val Asp Val Phe Thr Ala Pro Lys Asn Thr Ser Asp 50 55 60 Lys Glu Ile Phe Cys Arg Ala Ala Thr Val Leu Arg Gln Ile Tyr Thr 65 70 75 80 His Asn Cys Ser Asn Arg Tyr Leu Arg Gly Leu Tyr Arg Asn Leu Ser 85 90 95 Ser Met Ala Asn Lys Thr Cys Ser Met Asn Glu Ile Lys Lys Ser Thr 100 105 110 Leu Lys Asp Phe Leu Glu Arg Leu Lys Val Ile Met Gln Lys Lys Tyr 115 120 125 Tyr Arg His 130 <210> 2 <211> 132 <212> PRT <213> Domestic cat <400> 2 Met Asp Leu Thr Ser Gln Leu Ile Pro Ala Leu Val Cys Leu Leu Ala 1 5 10 15 Phe Thr Ser Thr Phe Val His Gly Gln Asn Phe Asn Asn Thr Leu Lys 20 25 30 Glu Ile Ile Lys Thr Leu Asn Ile Leu Thr Ala Arg Asn Asp Ser Cys 35 40 45 Met Glu Leu Thr Met Asp Val Leu Ala Ala Pro Lys Asn Thr Ser Asp 50 55 60 Lys Glu Ile Phe Cys Arg Ala Thr Thr Val Leu Arg Gln Ile Tyr Thr 65 70 75 80 His His Asn Cys Ser Thr Lys Phe Leu Lys Gly Leu Asp Arg Asn Leu 85 90 95 Ser Ser Met Ala Asn Arg Thr Cys Ser Val Asn Glu Val Lys Lys Cys 100 105 110 Thr Leu Lys Asp Phe Leu Glu Arg Leu Lys Ala Ile Met Gln Lys Lys 115 120 125 Tyr Ser Lys His 130 <210> 3 <211> 137 <212> PRT <213> Horse <400> 3 Met Gly Leu Thr Tyr Gln Leu Ile Pro Ala Leu Val Cys Leu Leu Ala 1 5 10 15 Cys Thr Ser Asn Phe Ile Gln Gly Cys Lys Tyr Asp Ile Thr Leu Gln 20 25 30 Glu Ile Ile Lys Thr Leu Asn Asn Leu Thr Asp Gly Lys Gly Lys Asn 35 40 45 Ser Cys Met Glu Leu Thr Val Ala Asp Ala Phe Ala Gly Pro Lys Asn 50 55 60 Thr Asp Gly Lys Glu Ile Cys Arg Ala Ala Lys Val Leu Gln Gln Leu 65 70 75 80 Tyr Lys Arg His Asp Arg Ser Leu Ile Lys Glu Cys Leu Ser Gly Leu 85 90 95 Asp Arg Asn Leu Lys Gly Met Ala Asn Gly Thr Cys Cys Thr Val Asn 100 105 110 Glu Ala Lys Lys Ser Thr Leu Lys Asp Phe Leu Glu Arg Leu Lys Thr 115 120 125 Ile Met Lys Glu Lys Tyr Ser Lys Cys 130 135 <210> 4 <211> 131 <212> PRT <213> Wolf <400> 4 Met Ala Leu Trp Leu Thr Val Val Ile Ala Leu Thr Cys Leu Gly Gly 1 5 10 15 Leu Ala Ser Pro Ser Pro Val Thr Pro Ser Pro Thr Leu Lys Glu Leu 20 25 30 Ile Glu Glu Leu Val Asn Ile Thr Gln Asn Gln Ala Ser Leu Cys Asn 35 40 45 Gly Ser Met Val Trp Ser Val Asn Leu Thr Ala Gly Met Tyr Cys Ala 50 55 60 Ala Leu Glu Ser Leu Ile Asn Val Ser Asp Cys Ser Ala Ile Gln Arg 65 70 75 80 Thr Gln Arg Met Leu Lys Ala Leu Cys Ser Gln Lys Pro Ala Ala Gly 85 90 95 Gln Ile Ser Ser Glu Arg Ser Arg Asp Thr Lys Ile Glu Val Ile Gln 100 105 110 Leu Val Lys Asn Leu Leu Thr Tyr Val Arg Gly Val Tyr Arg His Gly 115 120 125 Asn Phe Arg 130 <210> 5 <211> 183 <212> PRT <213> Domestic cat <400> 5 Met Trp Phe Leu Asp Ser Thr Arg Gln Ser Gly Asp Gln Gly Gly Arg 1 5 10 15 Arg His Thr Trp Pro Ile Lys Ala Thr Ala Arg Gly Gln Gly His Lys 20 25 30 Pro Leu Ser Leu Gly Gln Pro Thr Cys Pro Leu Leu Ala Pro Pro Val 35 40 45 Leu Ala Leu Gly Ser Met Ala Leu Trp Leu Thr Val Val Ile Ala Leu 50 55 60 Thr Cys Leu Gly Gly Leu Ala Ser Pro Gly Pro His Ser Arg Arg Glu 65 70 75 80 Leu Lys Glu Leu Ile Glu Glu Leu Val Asn Ile Thr Gln Asn Gln Val 85 90 95 Ser Leu Cys Asn Gly Ser Met Val Trp Ser Val Asn Leu Thr Thr Gly 100 105 110 Met Tyr Cys Ala Ala Leu Glu Ser Leu Ile Asn Val Ser Asp Cys Thr 115 120 125 Ala Ile Gln Arg Thr Gln Arg Met Leu Lys Ala Leu Cys Thr Gln Lys 130 135 140 Pro Ser Ala Gly Gln Thr Ala Ser Glu Arg Ser Arg Asp Thr Lys Ile 145 150 155 160 Glu Val Ile Gln Leu Val Lys Asn Leu Leu Asn His Leu Arg Arg Asn 165 170 175 Phe Arg His Gly Asn Phe Lys 180 <210> 6 <211> 133 <212> PRT <213> horse <400> 6 Met Ala Leu Trp Leu Thr Ala Val Ile Ala Leu Ala Cys Leu Gly Gly 1 5 10 15 Leu Ala Ser Pro Ala Pro Leu Pro Ser Ser Met Ala Leu Lys Glu Leu 20 25 30 Ile Lys Glu Leu Val Asn Ile Thr Gln Asn Gln Ala Pro Leu Cys Asn 35 40 45 Gly Ser Met Val Trp Ser Val Asn Leu Thr Ala Asp Thr Tyr Cys Arg 50 55 60 Ala Leu Glu Ser Leu Ser Asn Val Ser Thr Cys Ser Ala Ile Gln Asn 65 70 75 80 Thr Arg Lys Met Leu Thr Lys Leu Cys Pro His Gln Leu Ser Ala Gly 85 90 95 Gln Val Ser Ser Glu Arg Ala Arg Asp Thr Lys Ile Glu Val Ile Val 100 105 110 Leu Val Lys Asp Leu Leu Lys Asn Leu Arg Lys Ile Phe His Gly Gly 115 120 125 Lys His Val Asp Ala 130 <210> 7 <211> 823 <212> PRT <213> Wolf <400> 7 Met Gly Arg Leu Cys Ser Gly Leu Thr Phe Pro Val Ser Cys Leu Val 1 5 10 15 Leu Val Trp Val Ala Ser Ser Gly Ser Val Lys Val Leu His Glu Pro 20 25 30 Ser Cys Phe Ser Asp Tyr Ile Ser Thr Ser Val Cys Gln Trp Lys Met 35 40 45 Asp His Pro Thr Asn Cys Ser Ala Glu Leu Arg Leu Ser Tyr Gln Leu 50 55 60 Asp Phe Met Gly Ser Glu Asn His Thr Cys Val Pro Glu Asn Arg Glu 65 70 75 80 Asp Ser Val Cys Val Cys Ser Met Pro Ile Asp Asp Ala Val Glu Ala 85 90 95 Asp Val Tyr Gln Leu Asp Leu Trp Ala Gly Gln Gln Leu Leu Trp Ser 100 105 110 Gly Ser Phe Gln Pro Ser Lys His Val Lys Pro Arg Thr Pro Gly Asn 115 120 125 Leu Thr Val His Pro Asn Ile Ser His Thr Trp Leu Leu Met Trp Thr 130 135 140 Asn Pro Tyr Pro Thr Glu Asn His Leu His Ser Glu Leu Thr Tyr Met 145 150 155 160 Val Asn Val Ser Asn Asp Asn Asp Pro Glu Asp Phe Lys Val Tyr Asn 165 170 175 Val Thr Tyr Met Gly Pro Thr Leu Arg Leu Ala Ala Ser Thr Leu Lys 180 185 190 Ser Gly Ala Ser Tyr Ser Ala Arg Val Arg Ala Trp Ala Gln Thr Tyr 195 200 205 Asn Ser Thr Trp Ser Asp Trp Ser Pro Ser Thr Thr Trp Leu Asn Tyr 210 215 220 Tyr Glu Pro Trp Glu Gln His Leu Pro Leu Gly Val Ser Ile Ser Cys 225 230 235 240 Leu Val Ile Leu Ala Ile Cys Leu Ser Cys Tyr Phe Ser Ile Ile Lys 245 250 255 Ile Lys Lys Gly Trp Trp Asp Gln Ile Pro Asn Pro Ala His Ser Pro 260 265 270 Leu Val Ala Ile Val Ile Gln Asp Ser Gln Val Ser Leu Trp Gly Lys 275 280 285 Arg Ser Arg Gly Gln Glu Pro Ala Lys Cys Pro His Trp Lys Thr Cys 290 295 300 Leu Thr Lys Leu Leu Pro Cys Leu Leu Glu His Gly Leu Gly Arg Glu 305 310 315 320 Glu Glu Ser Pro Lys Thr Ala Lys Asn Gly Pro Leu Gln Gly Pro Gly 325 330 335 Lys Pro Ala Trp Cys Pro Val Glu Val Ser Lys Thr Ile Leu Trp Pro 340 345 350 Glu Ser Ile Ser Val Val Gln Cys Val Glu Leu Ser Glu Ala Pro Val 355 360 365 Asp Asn Glu Glu Glu Glu Glu Val Glu Glu Asp Lys Arg Ser Leu Cys 370 375 380 Pro Ser Leu Glu Gly Ser Gly Gly Ser Phe Gln Glu Gly Arg Glu Gly 385 390 395 400 Ile Val Ala Arg Leu Thr Glu Ser Leu Phe Leu Asp Leu Leu Gly Gly 405 410 415 Glu Asn Gly Gly Phe Cys Pro Gln Gly Leu Glu Glu Ser Cys Leu Pro 420 425 430 Pro Pro Ser Gly Ser Val Gly Ala Gln Met Pro Trp Ala Gln Phe Pro 435 440 445 Arg Ala Gly Pro Arg Ala Ala Pro Glu Gly Pro Glu Gln Pro Arg Arg 450 455 460 Pro Glu Ser Ala Leu Gln Ala Ser Pro Thr Gln Ser Ala Gly Ser Ser 465 470 475 480 Ala Phe Pro Glu Pro Pro Pro Val Val Thr Asp Asn Pro Ala Tyr Arg 485 490 495 Ser Phe Gly Ser Phe Leu Gly Gln Ser Ser Asp Pro Gly Asp Gly Asp 500 505 510 Ser Asp Pro Glu Leu Ala Asp Arg Pro Gly Glu Ala Asp Pro Gly Ile 515 520 525 Pro Ser Ala Pro Gln Pro Pro Glu Pro Pro Ala Ala Leu Gln Pro Glu 530 535 540 Pro Glu Ser Trp Glu Gln Ile Leu Arg Gln Ser Val Leu Gln His Arg 545 550 555 560 Ala Ala Pro Ala Pro Gly Pro Gly Pro Gly Ser Gly Tyr Arg Glu Phe 565 570 575 Thr Cys Ala Val Lys Gln Gly Ser Ala Pro Asp Ala Gly Gly Pro Gly 580 585 590 Phe Gly Pro Ser Gly Glu Ala Gly Tyr Lys Ala Phe Cys Ser Leu Leu 595 600 605 Pro Gly Gly Ala Thr Cys Pro Gly Thr Ser Gly Gly Glu Ala Gly Ser 610 615 620 Gly Glu Gly Gly Tyr Lys Pro Phe Gln Ser Leu Thr Pro Gly Cys Pro 625 630 635 640 Gly Ala Pro Thr Pro Val Pro Val Pro Leu Phe Thr Phe Gly Leu Asp 645 650 655 Thr Glu Pro Pro Gly Ser Pro Gln Asp Ser Leu Gly Ala Gly Ser Ser 660 665 670 Pro Glu His Leu Gly Val Glu Pro Ala Gly Lys Glu Glu Asp Ser Arg 675 680 685 Lys Thr Leu Leu Ala Pro Glu Gln Ala Thr Asp Pro Leu Arg Asp Asp 690 695 700 Leu Ala Ser Ser Ile Val Tyr Ser Ala Leu Thr Cys His Leu Cys Gly 705 710 715 720 His Leu Lys Gln Trp His Asp Gln Glu Glu Arg Gly Lys Ala His Ile 725 730 735 Val Pro Ser Pro Cys Cys Gly Cys Cys Cys Gly Asp Arg Ser Ser Leu 740 745 750 Leu Leu Ser Pro Leu Arg Ala Pro Asn Val Leu Pro Gly Gly Val Leu 755 760 765 Leu Glu Ala Ser Leu Ser Pro Ala Ser Leu Val Pro Ser Gly Val Ser 770 775 780 Lys Glu Gly Lys Ser Ser Pro Phe Ser Gln Pro Ala Ser Ser Ser Ala 785 790 795 800 Gln Ser Ser Ser Gln Thr Pro Lys Lys Leu Ala Val Leu Ser Thr Glu 805 810 815 Pro Thr Cys Met Ser Ala Ser 820 <210> 8 <211> 881 <212> PRT <213> Domestic cat <400> 8 Met Gly Arg Leu Cys Ser Gly Leu Thr Phe Pro Val Ser Cys Leu Ile 1 5 10 15 Leu Met Trp Ala Ala Gly Ser Gly Ser Val Lys Val Leu Arg Ala Pro 20 25 30 Thr Cys Phe Ser Asp Tyr Phe Ser Thr Ser Val Cys Gln Trp Asn Met 35 40 45 Asp Ala Pro Thr Asn Cys Ser Ala Glu Leu Arg Leu Ser Tyr Gln Leu 50 55 60 Asn Phe Met Gly Ser Glu Asn Arg Thr Cys Val Pro Glu Asn Gly Glu 65 70 75 80 Gly Ala Ala Cys Ala Cys Ser Met Leu Met Asp Asp Phe Val Glu Ala 85 90 95 Asp Val Tyr Gln Leu His Leu Trp Ala Gly Thr Gln Leu Leu Trp Ser 100 105 110 Gly Ser Phe Lys Pro Ser Ser His Val Lys Pro Arg Ala Pro Gly Asn 115 120 125 Leu Thr Val His Pro Asn Val Ser His Thr Trp Leu Leu Arg Trp Ser 130 135 140 Asn Pro Tyr Pro Pro Glu Asn His Leu His Ala Glu Leu Thr Tyr Met 145 150 155 160 Val Asn Ile Ser Ser Glu Asp Asp Pro Thr Asp Val Ser Val Cys Ala 165 170 175 Ser Gly Phe Leu Cys His Leu Leu Gly Leu Arg Arg Val Glu Thr Gly 180 185 190 Ala Pro Gly Ala Arg Leu Pro Pro Trp Leu Cys Ala Pro Arg Pro Arg 195 200 205 Arg Val Pro Gly Ser Gln Cys Ala Val Ile Ser Cys Cys Arg Trp Val 210 215 220 Leu Ile Ala Leu Thr Ser Arg Gly Gly Arg Trp Arg Leu Thr Pro Gly 225 230 235 240 Leu Arg Ser Gln Thr Arg Tyr Val Ser Val Ala Glu Gly Leu Phe Gly 245 250 255 Ala Thr Pro Arg Val Leu Cys Pro Gly Thr Gln Ala Gly Leu Ala Ser 260 265 270 Ala Ala Arg Glu Gln Met Ser Pro Asp Pro Ser Ala Phe His Ser Ile 275 280 285 Asp Tyr Glu Pro Trp Glu Gln His Leu Pro Leu Gly Val Ser Ile Ser 290 295 300 Cys Leu Val Ile Leu Ala Val Cys Leu Ser Cys Tyr Leu Ser Val Ile 305 310 315 320 Lys Ile Lys Lys Glu Trp Trp Asp Gln Ile Pro Asn Pro Ala His Ser 325 330 335 His Leu Val Ala Ile Val Ile Gln Asp Pro Gln Val Ser Leu Trp Gly 340 345 350 Lys Arg Ser Arg Gly Gln Glu Pro Ala Lys Cys Pro His Trp Lys Thr 355 360 365 Cys Leu Arg Lys Leu Leu Pro Cys Leu Leu Glu His Gly Met Glu Arg 370 375 380 Lys Glu Asp Pro Ser Lys Ile Ala Arg Asn Gly Pro Ser Gln Cys Ser 385 390 395 400 Gly Lys Ser Ala Trp Cys Pro Val Glu Val Ser Lys Thr Ile Leu Trp 405 410 415 Pro Glu Ser Ile Ser Val Val Arg Cys Val Glu Leu Leu Glu Ala Pro 420 425 430 Val Glu Ser Glu Glu Glu Glu Glu Glu Glu Glu Asp Lys Gly Ser Phe 435 440 445 Cys Pro Ser Pro Val Asn Leu Glu Asp Ser Phe Gln Glu Gly Arg Glu 450 455 460 Gly Ile Ala Ala Arg Leu Thr Glu Ser Leu Phe Met Asp Leu Leu Gly 465 470 475 480 Val Glu Lys Gly Gly Phe Gly Pro Gln Gly Ser Leu Glu Ser Trp Phe 485 490 495 Pro Pro Pro Ser Gly Ser Ala Gly Ala Gln Met Pro Trp Ala Glu Phe 500 505 510 Pro Gly Pro Gly Pro Gln Glu Ala Ser Pro Gln Gly Lys Glu Gln Pro 515 520 525 Phe Asp Pro Arg Ser Asp Pro Leu Ala Thr Leu Pro Gln Ser Pro Ala 530 535 540 Ser Pro Thr Phe Pro Glu Thr Pro Pro Val Val Thr Asp Asn Pro Ala 545 550 555 560 Tyr Arg Ser Phe Gly Thr Phe Gln Gly Arg Ser Ser Gly Pro Gly Glu 565 570 575 Cys Asp Ser Gly Pro Glu Leu Ala Gly Arg Leu Gly Glu Ala Asp Pro 580 585 590 Gly Ile Pro Ala Ala Pro Gln Pro Ser Glu Pro Pro Ser Ala Leu Gln 595 600 605 Pro Glu Ala Glu Thr Trp Glu Gln Ile Leu Arg Gln Arg Val Leu Gln 610 615 620 His Arg Gly Ala Pro Ala Pro Ala Pro Gly Ser Gly Tyr Arg Glu Phe 625 630 635 640 Val Cys Ala Val Arg Gln Gly Ser Thr Gln Asp Ser Gly Val Gly Asp 645 650 655 Phe Gly Pro Ser Glu Glu Ala Gly Tyr Lys Ala Phe Ser Ser Leu Leu 660 665 670 Thr Ser Gly Ala Val Cys Pro Glu Ser Gly Gly Glu Ala Gly Ser Gly 675 680 685 Asp Gly Gly Tyr Lys Pro Phe Gln Ser Leu Thr Pro Gly Cys Pro Gly 690 695 700 Ala Pro Ala Pro Val Pro Val Pro Leu Phe Thr Phe Gly Leu Asp Ala 705 710 715 720 Glu Pro Pro His Cys Pro Gln Asp Ser Pro Leu Pro Gly Ser Ser Pro 725 730 735 Glu Pro Ala Gly Lys Ala Gln Asp Ser His Lys Thr Pro Pro Ala Pro 740 745 750 Glu Gln Ala Ala Asp Pro Leu Arg Asp Asp Leu Ala Ser Gly Ile Val 755 760 765 Tyr Ser Ala Leu Thr Cys His Leu Cys Gly His Leu Lys Gln Cys His 770 775 780 Gly Gln Glu Glu Gly Gly Glu Ala His Pro Val Ala Ser Pro Cys Cys 785 790 795 800 Gly Cys Cys Cys Gly Asp Arg Ser Ser Pro Leu Val Ser Pro Leu Arg 805 810 815 Ala Pro Asp Pro Leu Pro Gly Gly Val Pro Leu Glu Ala Ser Leu Ser 820 825 830 Pro Ala Ser Pro Ala Pro Leu Ala Val Ser Glu Glu Gly Pro Pro Ser 835 840 845 Leu Cys Phe Gln Pro Ala Leu Ser His Ala His Ser Ser Ser Gln Thr 850 855 860 Pro Lys Lys Val Ala Met Leu Ser Pro Glu Pro Thr Cys Thr Met Ala 865 870 875 880 Serum <210> 9 <211> 809 <212> PRT <213> horse <400> 9 Met Gly Cys Leu Cys Pro Gly Leu Thr Leu Pro Val Ser Cys Leu Ile 1 5 10 15 Leu Val Trp Ala Ala Gly Ser Gly Ser Val Lys Val Leu His Leu Thr 20 25 30 Ala Cys Phe Ser Asp Tyr Ile Ser Ala Ser Thr Cys Glu Trp Lys Met 35 40 45 Asp Arg Pro Thr Asn Cys Ser Ala Gln Leu Arg Leu Ser Tyr Gln Leu 50 55 60 Asn Asp Glu Phe Ser Asp Asn Leu Thr Cys Ile Pro Glu Asn Arg Glu 65 70 75 80 Asp Glu Val Cys Val Cys Arg Met Leu Met Asp Asn Ile Val Ser Glu 85 90 95 Asp Val Tyr Glu Leu Asp Leu Trp Ala Gly Asn Gln Leu Leu Trp Asn 100 105 110 Ser Ser Phe Lys Pro Ser Arg His Val Lys Pro Arg Ala Pro Gln Asn 115 120 125 Leu Thr Val His Ala Ile Ser His Thr Trp Leu Leu Thr Trp Ser Asn 130 135 140 Pro Tyr Pro Leu Lys Asn His Leu Trp Ser Glu Leu Thr Tyr Leu Val 145 150 155 160 Asn Ile Ser Lys Glu Asp Asp Pro Thr Asp Phe Lys Ile Tyr Asn Val 165 170 175 Thr Tyr Met Asp Pro Thr Leu Arg Val Thr Ala Ser Thr Leu Lys Ser 180 185 190 Arg Ala Thr Tyr Ser Ala Arg Val Lys Ala Arg Ala Gln Asn Tyr Asn 195 200 205 Ser Thr Trp Ser Glu Trp Ser Pro Ser Thr Thr Trp His Asn Tyr Tyr 210 215 220 Glu Gln Pro Leu Glu Gln Arg Leu Pro Leu Gly Val Ser Ile Ser Cys 225 230 235 240 Val Val Ile Leu Ala Ile Cys Leu Ser Cys Tyr Phe Ser Ile Ile Lys 245 250 255 Ile Lys Lys Glu Trp Trp Asp Gln Ile Pro Asn Pro Ala His Ser Pro 260 265 270 Leu Val Ala Ile Val Leu Gln Asp Ser Gln Val Ser Leu Trp Gly Lys 275 280 285 Gln Ser Arg Gly Gln Glu Pro Ala Lys Cys Pro Arg Trp Lys Thr Cys 290 295 300 Leu Thr Lys Leu Leu Pro Cys Leu Leu Glu His Gly Leu Gln Lys Glu 305 310 315 320 Glu Asp Ser Ser Lys Thr Val Arg Asn Gly Pro Phe Gln Ser Pro Gly 325 330 335 Lys Ser Ala Trp His Thr Val Glu Val Asn His Thr Ile Leu Arg Pro 340 345 350 Glu Ile Ile Ser Val Val Pro Cys Val Glu Leu Cys Glu Ala Gln Val 355 360 365 Glu Ser Glu Glu Glu Glu Val Glu Glu Asp Arg Gly Ser Phe Cys Pro 370 375 380 Ser Pro Glu Ser Ser Gly Ser Gly Phe Gln Glu Gly Arg Glu Gly Val 385 390 395 400 Ala Ala Arg Leu Thr Glu Ser Leu Phe Leu Gly Leu Leu Gly Ala Glu 405 410 415 Asn Gly Ala Leu Gly Glu Ser Cys Leu Leu Pro Pro Leu Gly Ser Ala 420 425 430 His Met Pro Trp Ala Arg Ile Ser Ser Ala Gly Pro Gln Glu Ala Ala 435 440 445 Ser Gln Gly Glu Glu Gln Pro Leu Asn Pro Glu Ser Asn Pro Leu Ala 450 455 460 Thr Leu Thr Gln Ser Pro Gly Ser Leu Ala Phe Thr Glu Ala Pro Ala 465 470 475 480 Val Val Ala Asp Asn Pro Ala Tyr Arg Ser Phe Ser Asn Ser Leu Ser 485 490 495 Gln Pro Arg Gly Pro Gly Glu Leu Asp Ser Asp Pro Gln Leu Ala Glu 500 505 510 His Leu Gly Gln Val Asp Pro Ser Ile Pro Ser Ala Pro Gln Pro Ser 515 520 525 Glu Pro Pro Thr Ala Leu Gln Pro Glu Pro Glu Thr Trp Glu Gln Met 530 535 540 Leu Arg Gln Ser Val Leu Gln Gln Gly Ala Ala Pro Ala Pro Ala Ser 545 550 555 560 Ala Pro Thr Gly Gly Tyr Arg Glu Phe Ala Gln Ala Val Lys Gln Gly 565 570 575 Gly Gly Ala Ala Gly Ser Gly Pro Ser Gly Glu Ala Gly Tyr Lys Ala 580 585 590 Phe Ser Ser Leu Leu Ala Gly Ser Ala Val Cys Pro Gly Gln Ser Gly 595 600 605 Val Glu Ala Ser Ser Gly Glu Gly Gly Tyr Arg Pro Tyr Glu Ser Pro 610 615 620 Asp Pro Gly Ala Pro Ala Pro Val Pro Val Pro Leu Phe Thr Phe Gly 625 630 635 640 Leu Asp Val Glu Pro Pro His Ser Pro Gln Asn Ser Leu Leu Pro Gly 645 650 655 Gly Ser Pro Glu Leu Pro Gly Pro Glu Pro Thr Val Lys Gly Glu Asp 660 665 670 Pro Arg Lys Pro Leu Leu Ser Ala Gln Gln Ala Thr Asp Ser Leu Arg 675 680 685 Asp Asp Leu Gly Ser Gly Ile Val Tyr Ser Ala Leu Thr Cys His Leu 690 695 700 Cys Gly His Leu Lys Gln Cys His Gly Gln Glu Glu His Gly Glu Ala 705 710 715 720 His Thr Val Ala Ser Pro Cys Cys Gly Cys Cys Cys Gly Asp Arg Ser 725 730 735 Ser Pro Pro Val Ser Pro Val Arg Ala Leu Asp Pro Pro Pro Gly Gly 740 745 750 Val Pro Leu Glu Ala Gly Leu Ser Leu Ala Ser Leu Gly Ser Leu Gly 755 760 765 Leu Ser Glu Glu Arg Lys Pro Ser Leu Phe Phe Gln Pro Ala Pro Gly 770 775 780 Asn Ala Gln Ser Ser Ser Gln Thr Pro Leu Thr Val Ala Met Leu Ser 785 790 795 800 Thr Gly Pro Thr Cys Thr Ser Ala Ser 805 <210> 10 <211> 427 <212> PRT <213> Wolf <400> 10 Met Glu Arg Pro Ala Arg Leu Cys Gly Leu Trp Ala Leu Leu Leu Cys 1 5 10 15 Ala Ala Gly Gly Arg Gly Gly Gly Val Ala Ala Pro Thr Glu Thr Gln 20 25 30 Pro Pro Val Thr Asn Leu Ser Val Ser Val Glu Asn Leu Cys Thr Val 35 40 45 Ile Trp Thr Trp Asp Pro Pro Glu Gly Ala Ser Pro Asn Cys Thr Leu 50 55 60 Arg Tyr Phe Ser His Phe Asp Asn Lys Gln Asp Lys Lys Ile Ala Pro 65 70 75 80 Glu Thr His Arg Ser Lys Glu Val Pro Leu Asn Glu Arg Ile Cys Leu 85 90 95 Gln Val Gly Ser Gln Cys Ser Thr Asn Glu Ser Asp Asn Pro Ser Ile 100 105 110 Leu Val Glu Lys Cys Thr Pro Pro Pro Glu Gly Asp Pro Glu Ser Ala 115 120 125 Val Thr Glu Leu Gln Cys Val Trp His Asn Leu Ser Tyr Met Lys Cys 130 135 140 Thr Trp Leu Pro Gly Arg Asn Thr Ser Pro Asp Thr Asn Tyr Thr Leu 145 150 155 160 Tyr Tyr Trp His Ser Ser Leu Gly Lys Ile Leu Gln Cys Glu Asp Ile 165 170 175 Tyr Arg Glu Gly Gln His Ile Gly Cys Ser Phe Ala Leu Thr Asn Leu 180 185 190 Lys Asp Ser Ser Phe Glu Gln His Ser Val Gln Ile Val Val Lys Asp 195 200 205 Asn Ala Gly Lys Ile Arg Pro Ser Phe Asn Ile Val Pro Leu Thr Ser 210 215 220 His Val Lys Pro Asp Pro Pro His Ile Lys Arg Leu Phe Phe Gln Asn 225 230 235 240 Gly Asn Leu Tyr Val Gln Trp Lys Asn Pro Gln Asn Phe Tyr Ser Arg 245 250 255 Cys Leu Ser Tyr Gln Val Glu Val Asn Asn Ser Gln Thr Glu Thr Asn 260 265 270 Asp Ile Phe Tyr Val Glu Glu Ala Lys Cys Gln Asn Ser Glu Phe Glu 275 280 285 Gly Asn Leu Glu Gly Thr Ile Cys Phe Met Val Pro Gly Val Leu Pro 290 295 300 Asp Thr Leu Asn Thr Val Arg Ile Arg Val Arg Thr Asn Lys Leu Cys 305 310 315 320 Tyr Glu Asp Asp Lys Leu Trp Ser Asn Trp Ser Gln Ala Met Ser Ile 325 330 335 Gly Glu Asn Thr Asp Pro Thr Phe Tyr Ile Thr Met Leu Leu Ala Thr 340 345 350 Pro Val Ile Val Ala Gly Ala Ile Ile Val Leu Leu Leu Tyr Leu Lys 355 360 365 Arg Leu Lys Ile Ile Ile Phe Pro Pro Ile Pro Asp Pro Gly Lys Ile 370 375 380 Phe Lys Glu Met Phe Gly Asp Gln Asn Asp Asp Thr Leu His Trp Arg 385 390 395 400 Lys Tyr Asp Ile Tyr Glu Lys Gln Thr Lys Glu Glu Thr Asp Ser Val 405 410 415 Val Leu Ile Glu Asn Leu Lys Lys Ala Ser Gln 420 425 <210> 11 <211> 479 <212> PRT <213> Domestic cat <400> 11 Met Met Thr Lys Cys Ser Ser Asp Arg Asn Val Phe Lys Arg Lys Trp 1 5 10 15 Phe Leu Phe Pro Ala Ser Gln Tyr Thr Phe Arg Pro Ile His Gln Ala 20 25 30 Arg Pro Cys Glu Val Pro Ala Val His Leu Glu Pro Ser Pro Pro Trp 35 40 45 Glu Val Gly Leu Gly Leu Leu Asn Leu Glu Ser Glu Phe Arg Lys Leu 50 55 60 Gly Leu Arg Gly Arg Arg Leu Ala Ala Ala Pro Pro Asp Ser Arg Ala 65 70 75 80 Glu Ala Ala Ser Gln Thr Gln Pro Pro Val Thr Asn Leu Ser Val Ser 85 90 95 Val Glu Asn Leu Cys Thr Val Ile Trp Thr Trp Asp Pro Pro Glu Gly 100 105 110 Ala Ser Pro Asn Cys Thr Leu Arg Tyr Phe Ser His Phe Asp Asn Lys 115 120 125 Gln Asp Lys Lys Ile Ala Pro Glu Thr His Arg Ser Lys Glu Val Pro 130 135 140 Leu Asn Glu Arg Ile Cys Leu Gln Val Gly Ser Gln Cys Ser Thr Asn 145 150 155 160 Glu Ser Asp Asn Pro Ser Ile Leu Val Glu Lys Cys Thr Pro Pro Pro 165 170 175 Glu Gly Asp Pro Glu Ser Ala Val Thr Glu Leu Gln Cys Val Trp His 180 185 190 Asn Leu Ser Tyr Met Lys Cys Thr Trp Leu Pro Gly Arg Asn Thr Ser 195 200 205 Pro Asp Thr Asn Tyr Thr Leu Tyr Tyr Trp His Ser Ser Leu Gly Lys 210 215 220 Ile Leu Gln Cys Glu Asn Ile Tyr Arg Glu Gly Gln His Ile Gly Cys 225 230 235 240 Ser Phe Ala Leu Thr Asn Leu Lys Asp Ser Ser Phe Glu Gln His Ser 245 250 255 Val Gln Ile Val Val Lys Asp Asn Ala Gly Lys Ile Arg Pro Ser Phe 260 265 270 Asn Ile Val Pro Leu Thr Ser His Val Lys Pro Asp Pro Pro His Ile 275 280 285 Lys Arg Leu Phe Phe Gln Asn Gly Asn Leu Tyr Val Gln Trp Lys Asn 290 295 300 Pro Gln Asn Phe Tyr Ser Arg Cys Leu Ser Tyr Gln Val Glu Val Asn 305 310 315 320 Asn Ser Gln Thr Glu Thr His Asp Ile Phe Tyr Val Glu Glu Ala Lys 325 330 335 Cys Gln Asn Ser Glu Phe Glu Gly Asn Leu Glu Gly Thr Ile Cys Phe 340 345 350 Met Val Pro Gly Ile Leu Pro Asp Thr Leu Asn Thr Val Arg Ile Arg 355 360 365 Val Arg Thr Asn Lys Leu Cys Tyr Glu Asp Asp Arg Leu Trp Ser Asn 370 375 380 Trp Ser Gln Ala Met Ser Ile Gly Glu Asn Thr Asp Pro Thr Phe Tyr 385 390 395 400 Ile Thr Met Leu Leu Ala Thr Pro Val Ile Val Ala Gly Ala Ile Ile 405 410 415 Val Leu Leu Leu Tyr Leu Lys Arg Leu Lys Ile Ile Ile Phe Pro Pro 420 425 430 Ile Pro Asp Pro Gly Lys Ile Phe Lys Glu Met Phe Gly Asp Gln Asn 435 440 445 Asp Asp Ser Leu His Trp Lys Lys Tyr Asp Ile Tyr Glu Lys Gln Thr 450 455 460 Lys Glu Glu Thr Asp Ser Val Val Leu Ile Glu Asn Ala Ser Gln 465 470 475 <210> 12 <211> 407 <212> PRT <213> Horse <400> 12 Met Tyr Phe Leu Cys Leu Ile Trp Thr Glu Ser Gln Pro Pro Val Thr 1 5 10 15 Asn Leu Ser Val Ser Val Glu Asn Leu Cys Thr Val Ile Trp Thr Trp 20 25 30 Asn Pro Pro Glu Gly Val Ser Pro Asn Cys Ser Leu Trp Tyr Phe Ser 35 40 45 His Phe Gly Asn Lys Gln Asp Lys Lys Ile Ala Pro Glu Thr His Arg 50 55 60 Ser Lys Glu Val Pro Leu Asn Glu Arg Ile Cys Leu Gln Val Gly Ser 65 70 75 80 Gln Cys Ser Thr Asn Glu Ser Asp Asn Pro Ser Ile Leu Val Glu Lys 85 90 95 Cys Ile Ser Pro Pro Glu Gly Asp Pro Glu Ser Ala Val Thr Glu Leu 100 105 110 Gln Cys Val Trp His Asn Leu Ser Tyr Met Lys Cys Thr Trp Leu Pro 115 120 125 Gly Lys Asn Ala Ser Pro Asp Thr Asn Tyr Thr Leu Tyr Tyr Trp His 130 135 140 Ser Ser Leu Gly Lys Ile Leu Gln Cys Glu Asp Ile Tyr Arg Glu Gly 145 150 155 160 Gln His Ile Gly Cys Ser Phe Ala Leu Thr Glu Val Lys Asp Ser Ile 165 170 175 Phe Glu Gln His Ser Val Gln Ile Met Val Lys Asp Asn Ala Gly Lys 180 185 190 Ile Arg Pro Phe Phe Asn Ile Val Pro Leu Thr Ser His Val Lys Pro 195 200 205 Asp Pro Pro His Ile Lys Lys Leu Phe Phe Gln Asn Gly Asp Leu Tyr 210 215 220 Val Gln Trp Lys Asn Pro Gln Asn Phe Tyr Ser Arg Cys Leu Ser Tyr 225 230 235 240 Gln Val Glu Val Asn Asn Ser Gln Thr Glu Thr Arg Asp Ile Phe Ser 245 250 255 Val Glu Glu Ala Lys Cys Gln Asn Pro Glu Phe Glu Gly Asp Leu Glu 260 265 270 Gly Thr Ile Cys Phe Met Val Pro Gly Val Leu Pro Asp Thr Val Asn 275 280 285 Thr Val Arg Ile Arg Val Lys Thr Asn Lys Leu Cys Tyr Glu Asp Asp 290 295 300 Lys Leu Trp Ser Asn Trp Ser Gln Ala Met Ser Ile Gly Lys Lys Ala 305 310 315 320 Asp Pro Thr Phe Tyr Ile Ala Met Leu Leu Ile Ile Pro Val Ile Val 325 330 335 Ala Gly Ala Ile Ile Val Leu Leu Leu Tyr Leu Lys Arg Leu Lys Ile 340 345 350 Ile Met Phe Pro Pro Ile Pro Asp Pro Gly Lys Ile Phe Lys Glu Met 355 360 365 Phe Gly Asp Gln Asn Asp Asp Thr Leu His Trp Lys Lys Tyr Asp Ile 370 375 380 Tyr Glu Lys Gln Thr Lys Glu Glu Thr Asp Ser Val Val Leu Ile Glu 385 390 395 400 Asn Leu Lys Arg Ala Ser Gln 405 <210> 13 <211> 761 <212> PRT <213> Artificial Sequence <220> <223> Wolf IL13R-IL4R-Fc (without signal sequence) <400> 13 Thr Glu Thr Gln Pro Pro Val Thr Asn Leu Ser Val Ser Val Glu Asn 1 5 10 15 Leu Cys Thr Val Ile Trp Thr Trp Asp Pro Pro Glu Gly Ala Ser Pro 20 25 30 Asn Cys Thr Leu Arg Tyr Phe Ser His Phe Asp Asn Lys Gln Asp Lys 35 40 45 Lys Ile Ala Pro Glu Thr His Arg Ser Lys Glu Val Pro Leu Asn Glu 50 55 60 Arg Ile Cys Leu Gln Val Gly Ser Gln Cys Ser Thr Asn Glu Ser Asp 65 70 75 80 Asn Pro Ser Ile Leu Val Glu Lys Cys Thr Pro Pro Pro Glu Gly Asp 85 90 95 Pro Glu Ser Ala Val Thr Glu Leu Gln Cys Val Trp His Asn Leu Ser 100 105 110 Tyr Met Lys Cys Thr Trp Leu Pro Gly Arg Asn Thr Ser Pro Asp Thr 115 120 125 Asn Tyr Thr Leu Tyr Tyr Trp His Ser Ser Leu Gly Lys Ile Leu Gln 130 135 140 Cys Glu Asp Ile Tyr Arg Glu Gly Gln His Ile Gly Cys Ser Phe Ala 145 150 155 160 Leu Thr Asn Leu Lys Asp Ser Ser Phe Glu Gln His Ser Val Gln Ile 165 170 175 Val Val Lys Asp Asn Ala Gly Lys Ile Arg Pro Ser Phe Asn Ile Val 180 185 190 Pro Leu Thr Ser His Val Lys Pro Asp Pro Pro His Ile Lys Arg Leu 195 200 205 Phe Phe Gln Asn Gly Asn Leu Tyr Val Gln Trp Lys Asn Pro Gln Asn 210 215 220 Phe Tyr Ser Arg Cys Leu Ser Tyr Gln Val Glu Val Asn Asn Ser Gln 225 230 235 240 Thr Glu Thr Asn Asp Ile Phe Tyr Val Glu Glu Ala Lys Cys Gln Asn 245 250 255 Ser Glu Phe Glu Gly Asn Leu Glu Gly Thr Ile Cys Phe Met Val Pro 260 265 270 Gly Val Leu Pro Asp Thr Leu Asn Thr Val Arg Ile Arg Val Arg Thr 275 280 285 Asn Lys Leu Cys Tyr Glu Asp Asp Lys Leu Trp Ser Asn Trp Ser Gln 290 295 300 Ala Met Ser Ile Gly Glu Asn Thr Asp Pro Thr Gly Gly Gly Ser Gly 305 310 315 320 Ser Gly Ser Val Lys Val Leu His Glu Pro Ser Cys Phe Ser Asp Tyr 325 330 335 Ile Ser Thr Ser Val Cys Gln Trp Lys Met Asp His Pro Thr Asn Cys 340 345 350 Ser Ala Glu Leu Arg Leu Ser Tyr Gln Leu Asp Phe Met Gly Ser Glu 355 360 365 Asn His Thr Cys Val Pro Glu Asn Arg Glu Asp Ser Val Cys Val Cys 370 375 380 Ser Met Pro Ile Asp Asp Ala Val Glu Ala Asp Val Tyr Gln Leu Asp 385 390 395 400 Leu Trp Ala Gly Gln Gln Leu Leu Trp Ser Gly Ser Phe Gln Pro Ser 405 410 415 Lys His Val Lys Pro Arg Thr Pro Gly Asn Leu Thr Val His Pro Asn 420 425 430 Ile Ser His Thr Trp Leu Leu Met Trp Thr Asn Pro Tyr Pro Thr Glu 435 440 445 Asn His Leu His Ser Glu Leu Thr Tyr Met Val Asn Val Ser Asn Asp 450 455 460 Asn Asp Pro Glu Asp Phe Lys Val Tyr Asn Val Thr Tyr Met Gly Pro 465 470 475 480 Thr Leu Arg Leu Ala Ala Ser Thr Leu Lys Ser Gly Ala Ser Tyr Ser 485 490 495 Ala Arg Val Arg Ala Trp Ala Gln Thr Tyr Asn Ser Thr Trp Ser Asp 500 505 510 Trp Ser Pro Ser Thr Thr Trp Leu Asn Tyr Tyr Glu Pro Lys Arg Glu 515 520 525 Asn Gly Arg Val Pro Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala Pro 530 535 540 Glu Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys 545 550 555 560 Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val Val 565 570 575 Asp Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp 580 585 590 Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln Phe 595 600 605 Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Gln Asp 610 615 620 Trp Leu Lys Gly Lys Gln Phe Thr Cys Lys Val Asn Asn Lys Ala Leu 625 630 635 640 Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala His 645 650 655 Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser Lys 660 665 670 Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro Asp 675 680 685 Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser Lys 690 695 700 Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu 705 710 715 720 Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp Thr 725 730 735 Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 740 745 750 Glu Ser Leu Ser His Ser Pro Gly Lys 755 760 <210> 14 <211> 758 <212> PRT <213> Artificial Sequence <220> <223> Wolf IL4R - IL13R - IgGA (without signal sequence) <400> 14 Ser Gly Ser Val Lys Val Leu His Glu Pro Ser Cys Phe Ser Asp Tyr 1 5 10 15 Ile Ser Thr Ser Val Cys Gln Trp Lys Met Asp His Pro Thr Asn Cys 20 25 30 Ser Ala Glu Leu Arg Leu Ser Tyr Gln Leu Asp Phe Met Gly Ser Glu 35 40 45 Asn His Thr Cys Val Pro Glu Asn Arg Glu Asp Ser Val Cys Val Cys 50 55 60 Ser Met Pro Ile Asp Asp Ala Val Glu Ala Asp Val Tyr Gln Leu Asp 65 70 75 80 Leu Trp Ala Gly Gln Gln Leu Leu Trp Ser Gly Ser Phe Gln Pro Ser 85 90 95 Lys His Val Lys Pro Arg Thr Pro Gly Asn Leu Thr Val His Pro Asn 100 105 110 Ile Ser His Thr Trp Leu Leu Met Trp Thr Asn Pro Tyr Pro Thr Glu 115 120 125 Asn His Leu His Ser Glu Leu Thr Tyr Met Val Asn Val Ser Asn Asp 130 135 140 Asn Asp Pro Glu Asp Phe Lys Val Tyr Asn Val Thr Tyr Met Gly Pro 145 150 155 160 Thr Leu Arg Leu Ala Ala Ser Thr Leu Lys Ser Gly Ala Ser Tyr Ser 165 170 175 Ala Arg Val Arg Ala Trp Ala Gln Thr Tyr Asn Ser Thr Trp Ser Asp 180 185 190 Trp Ser Pro Ser Thr Thr Trp Leu Asn Tyr Tyr Glu Pro Gly Gly Gly 195 200 205 Ser Gly Thr Glu Thr Gln Pro Pro Val Thr Asn Leu Ser Val Ser Val 210 215 220 Glu Asn Leu Cys Thr Val Ile Trp Thr Trp Asp Pro Pro Glu Gly Ala 225 230 235 240 Ser Pro Asn Cys Thr Leu Arg Tyr Phe Ser His Phe Asp Asn Lys Gln 245 250 255 Asp Lys Lys Ile Ala Pro Glu Thr His Arg Ser Lys Glu Val Pro Leu 260 265 270 Asn Glu Arg Ile Cys Leu Gln Val Gly Ser Gln Cys Ser Thr Asn Glu 275 280 285 Ser Asp Asn Pro Ser Ile Leu Val Glu Lys Cys Thr Pro Pro Pro Glu 290 295 300 Gly Asp Pro Glu Ser Ala Val Thr Glu Leu Gln Cys Val Trp His Asn 305 310 315 320 Leu Ser Tyr Met Lys Cys Thr Trp Leu Pro Gly Arg Asn Thr Ser Pro 325 330 335 Asp Thr Asn Tyr Thr Leu Tyr Tyr Trp His Ser Ser Leu Gly Lys Ile 340 345 350 Leu Gln Cys Glu Asp Ile Tyr Arg Glu Gly Gln His Ile Gly Cys Ser 355 360 365 Phe Ala Leu Thr Asn Leu Lys Asp Ser Ser Phe Glu Gln His Ser Val 370 375 380 Gln Ile Val Val Lys Asp Asn Ala Gly Lys Ile Arg Pro Ser Phe Asn 385 390 395 400 Ile Val Pro Leu Thr Ser His Val Lys Pro Asp Pro Pro His Ile Lys 405 410 415 Arg Leu Phe Phe Gln Asn Gly Asn Leu Tyr Val Gln Trp Lys Asn Pro 420 425 430 Gln Asn Phe Tyr Ser Arg Cys Leu Ser Tyr Gln Val Glu Val Asn Asn 435 440 445 Ser Gln Thr Glu Thr Asn Asp Ile Phe Tyr Val Glu Glu Ala Lys Cys 450 455 460 Gln Asn Ser Glu Phe Glu Gly Asn Leu Glu Gly Thr Ile Cys Phe Met 465 470 475 480 Val Pro Gly Val Leu Pro Asp Thr Leu Asn Thr Val Arg Ile Arg Val 485 490 495 Arg Thr Asn Lys Leu Cys Tyr Glu Asp Asp Lys Leu Trp Ser Asn Trp 500 505 510 Ser Gln Ala Met Ser Ile Gly Glu Asn Thr Asp Pro Thr Phe Asn Glu 515 520 525 Cys Arg Cys Thr Asp Thr Pro Pro Cys Pro Val Pro Glu Pro Leu Gly 530 535 540 Gly Pro Ser Val Leu Ile Phe Pro Pro Lys Pro Lys Asp Ile Leu Arg 545 550 555 560 Ile Thr Arg Thr Pro Glu Val Thr Cys Val Val Leu Asp Leu Gly Arg 565 570 575 Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Glu Val 580 585 590 His Thr Ala Lys Thr Gln Ser Arg Glu Gln Gln Phe Asn Gly Thr Tyr 595 600 605 Arg Val Val Ser Val Leu Pro Ile Glu His Gln Asp Trp Leu Thr Gly 610 615 620 Lys Glu Phe Lys Cys Arg Val Asn His Ile Asp Leu Pro Ser Pro Ile 625 630 635 640 Glu Arg Thr Ile Ser Lys Ala Arg Gly Arg Ala His Lys Pro Ser Val 645 650 655 Tyr Val Leu Pro Pro Ser Pro Lys Glu Leu Ser Ser Ser Asp Thr Val 660 665 670 Ser Ile Thr Cys Leu Ile Lys Asp Phe Tyr Pro Pro Asp Ile Asp Val 675 680 685 Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Arg Lys His Arg Met 690 695 700 Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys 705 710 715 720 Leu Ser Val Asp Lys Ser Arg Trp Gln Gln Gly Asp Pro Phe Thr Cys 725 730 735 Ala Val Met His Glu Thr Leu Gln Asn His Tyr Thr Asp Leu Ser Leu 740 745 750 Ser His Ser Pro Gly Lys 755 <210> 15 <211> 762 <212> PRT <213> Artificial sequence <220> <223> Wolf IL4R-IL13R-IgGB (without signal sequence) <400> 15 Ser Gly Ser Val Lys Val Leu His Glu Pro Ser Cys Phe Ser Asp Tyr 1 5 10 15 Ile Ser Thr Ser Val Cys Gln Trp Lys Met Asp His Pro Thr Asn Cys 20 25 30 Ser Ala Glu Leu Arg Leu Ser Tyr Gln Leu Asp Phe Met Gly Ser Glu 35 40 45 Asn His Thr Cys Val Pro Glu Asn Arg Glu Asp Ser Val Cys Val Cys 50 55 60 Ser Met Pro Ile Asp Asp Ala Val Glu Ala Asp Val Tyr Gln Leu Asp 65 70 75 80 Leu Trp Ala Gly Gln Gln Leu Leu Trp Ser Gly Ser Phe Gln Pro Ser 85 90 95 Lys His Val Lys Pro Arg Thr Pro Gly Asn Leu Thr Val His Pro Asn 100 105 110 Ile Ser His Thr Trp Leu Leu Met Trp Thr Asn Pro Tyr Pro Thr Glu 115 120 125 Asn His Leu His Ser Glu Leu Thr Tyr Met Val Asn Val Ser Asn Asp 130 135 140 Asn Asp Pro Glu Asp Phe Lys Val Tyr Asn Val Thr Tyr Met Gly Pro 145 150 155 160 Thr Leu Arg Leu Ala Ala Ser Thr Leu Lys Ser Gly Ala Ser Tyr Ser 165 170 175 Ala Arg Val Arg Ala Trp Ala Gln Thr Tyr Asn Ser Thr Trp Ser Asp 180 185 190 Trp Ser Pro Ser Thr Thr Trp Leu Asn Tyr Tyr Glu Pro Gly Gly Gly 195 200 205 Ser Gly Thr Glu Thr Gln Pro Pro Val Thr Asn Leu Ser Val Ser Val 210 215 220 Glu Asn Leu Cys Thr Val Ile Trp Thr Trp Asp Pro Pro Glu Gly Ala 225 230 235 240 Ser Pro Asn Cys Thr Leu Arg Tyr Phe Ser His Phe Asp Asn Lys Gln 245 250 255 Asp Lys Lys Ile Ala Pro Glu Thr His Arg Ser Lys Glu Val Pro Leu 260 265 270 Asn Glu Arg Ile Cys Leu Gln Val Gly Ser Gln Cys Ser Thr Asn Glu 275 280 285 Ser Asp Asn Pro Ser Ile Leu Val Glu Lys Cys Thr Pro Pro Pro Glu 290 295 300 Gly Asp Pro Glu Ser Ala Val Thr Glu Leu Gln Cys Val Trp His Asn 305 310 315 320 Leu Ser Tyr Met Lys Cys Thr Trp Leu Pro Gly Arg Asn Thr Ser Pro 325 330 335 Asp Thr Asn Tyr Thr Leu Tyr Tyr Trp His Ser Ser Leu Gly Lys Ile 340 345 350 Leu Gln Cys Glu Asp Ile Tyr Arg Glu Gly Gln His Ile Gly Cys Ser 355 360 365 Phe Ala Leu Thr Asn Leu Lys Asp Ser Ser Phe Glu Gln His Ser Val 370 375 380 Gln Ile Val Val Lys Asp Asn Ala Gly Lys Ile Arg Pro Ser Phe Asn 385 390 395 400 Ile Val Pro Leu Thr Ser His Val Lys Pro Asp Pro Pro His Ile Lys 405 410 415 Arg Leu Phe Phe Gln Asn Gly Asn Leu Tyr Val Gln Trp Lys Asn Pro 420 425 430 Gln Asn Phe Tyr Ser Arg Cys Leu Ser Tyr Gln Val Glu Val Asn Asn 435 440 445 Ser Gln Thr Glu Thr Asn Asp Ile Phe Tyr Val Glu Glu Ala Lys Cys 450 455 460 Gln Asn Ser Glu Phe Glu Gly Asn Leu Glu Gly Thr Ile Cys Phe Met 465 470 475 480 Val Pro Gly Val Leu Pro Asp Thr Leu Asn Thr Val Arg Ile Arg Val 485 490 495 Arg Thr Asn Lys Leu Cys Tyr Glu Asp Asp Lys Leu Trp Ser Asn Trp 500 505 510 Ser Gln Ala Met Ser Ile Gly Glu Asn Thr Asp Pro Thr Pro Lys Arg 515 520 525 Glu Asn Gly Arg Val Pro Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala 530 535 540 Pro Glu Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro 545 550 555 560 Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val 565 570 575 Val Asp Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val 580 585 590 Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln 595 600 605 Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Gln 610 615 620 Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Lys Val Asn Asn Lys Ala 625 630 635 640 Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala 645 650 655 His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser 660 665 670 Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro 675 680 685 Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser 690 695 700 Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe 705 710 715 720 Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp 725 730 735 Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His Tyr Thr 740 745 750 Gln Glu Ser Leu Ser His Ser Pro Gly Lys 755 760 <210> 16 <211> 760 <212> PRT <213> Artificial Sequence <220> <223> Wolf IL4R-IL13R-IgGC (without signal sequence) <400> 16 Ser Gly Ser Val Lys Val Leu His Glu Pro Ser Cys Phe Ser Asp Tyr 1 5 10 15 Ile Ser Thr Ser Val Cys Gln Trp Lys Met Asp His Pro Thr Asn Cys 20 25 30 Ser Ala Glu Leu Arg Leu Ser Tyr Gln Leu Asp Phe Met Gly Ser Glu 35 40 45 Asn His Thr Cys Val Pro Glu Asn Arg Glu Asp Ser Val Cys Val Cys 50 55 60 Ser Met Pro Ile Asp Asp Ala Val Glu Ala Asp Val Tyr Gln Leu Asp 65 70 75 80 Leu Trp Ala Gly Gln Gln Leu Leu Trp Ser Gly Ser Phe Gln Pro Ser 85 90 95 Lys His Val Lys Pro Arg Thr Pro Gly Asn Leu Thr Val His Pro Asn 100 105 110 Ile Ser His Thr Trp Leu Leu Met Trp Thr Asn Pro Tyr Pro Thr Glu 115 120 125 Asn His Leu His Ser Glu Leu Thr Tyr Met Val Asn Val Ser Asn Asp 130 135 140 Asn Asp Pro Glu Asp Phe Lys Val Tyr Asn Val Thr Tyr Met Gly Pro 145 150 155 160 Thr Leu Arg Leu Ala Ala Ser Thr Leu Lys Ser Gly Ala Ser Tyr Ser 165 170 175 Ala Arg Val Arg Ala Trp Ala Gln Thr Tyr Asn Ser Thr Trp Ser Asp 180 185 190 Trp Ser Pro Ser Thr Thr Trp Leu Asn Tyr Tyr Glu Pro Gly Gly Gly 195 200 205 Ser Gly Thr Glu Thr Gln Pro Pro Val Thr Asn Leu Ser Val Ser Val 210 215 220 Glu Asn Leu Cys Thr Val Ile Trp Thr Trp Asp Pro Pro Glu Gly Ala 225 230 235 240 Ser Pro Asn Cys Thr Leu Arg Tyr Phe Ser His Phe Asp Asn Lys Gln 245 250 255 Asp Lys Lys Ile Ala Pro Glu Thr His Arg Ser Lys Glu Val Pro Leu 260 265 270 Asn Glu Arg Ile Cys Leu Gln Val Gly Ser Gln Cys Ser Thr Asn Glu 275 280 285 Ser Asp Asn Pro Ser Ile Leu Val Glu Lys Cys Thr Pro Pro Pro Glu 290 295 300 Gly Asp Pro Glu Ser Ala Val Thr Glu Leu Gln Cys Val Trp His Asn 305 310 315 320 Leu Ser Tyr Met Lys Cys Thr Trp Leu Pro Gly Arg Asn Thr Ser Pro 325 330 335 Asp Thr Asn Tyr Thr Leu Tyr Tyr Trp His Ser Ser Leu Gly Lys Ile 340 345 350 Leu Gln Cys Glu Asp Ile Tyr Arg Glu Gly Gln His Ile Gly Cys Ser 355 360 365 Phe Ala Leu Thr Asn Leu Lys Asp Ser Ser Phe Glu Gln His Ser Val 370 375 380 Gln Ile Val Val Lys Asp Asn Ala Gly Lys Ile Arg Pro Ser Phe Asn 385 390 395 400 Ile Val Pro Leu Thr Ser His Val Lys Pro Asp Pro Pro His Ile Lys 405 410 415 Arg Leu Phe Phe Gln Asn Gly Asn Leu Tyr Val Gln Trp Lys Asn Pro 420 425 430 Gln Asn Phe Tyr Ser Arg Cys Leu Ser Tyr Gln Val Glu Val Asn Asn 435 440 445 Ser Gln Thr Glu Thr Asn Asp Ile Phe Tyr Val Glu Glu Ala Lys Cys 450 455 460 Gln Asn Ser Glu Phe Glu Gly Asn Leu Glu Gly Thr Ile Cys Phe Met 465 470 475 480 Val Pro Gly Val Leu Pro Asp Thr Leu Asn Thr Val Arg Ile Arg Val 485 490 495 Arg Thr Asn Lys Leu Cys Tyr Glu Asp Asp Lys Leu Trp Ser Asn Trp 500 505 510 Ser Gln Ala Met Ser Ile Gly Glu Asn Thr Asp Pro Thr Ala Lys Glu 515 520 525 Cys Glu Cys Lys Cys Asn Cys Asn Asn Cys Pro Cys Pro Gly Cys Gly 530 535 540 Leu Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp 545 550 555 560 Ile Leu Val Thr Ala Arg Thr Pro Thr Val Thr Cys Val Val Val Asp 565 570 575 Leu Asp Pro Glu Asn Pro Glu Val Gln Ile Ser Trp Phe Val Asp Ser 580 585 590 Lys Gln Val Gln Thr Ala Asn Thr Gln Pro Arg Glu Glu Gln Ser Asn 595 600 605 Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Gln Asp Trp 610 615 620 Leu Ser Gly Lys Gln Phe Lys Cys Lys Val Asn Asn Lys Ala Leu Pro 625 630 635 640 Ser Pro Ile Glu Glu Ile Ile Ser Lys Thr Pro Gly Gln Ala His Gln 645 650 655 Pro Asn Val Tyr Val Leu Pro Pro Ser Arg Asp Glu Met Ser Lys Asn 660 665 670 Thr Val Thr Leu Thr Cys Leu Val Lys Asp Phe Phe Pro Pro Glu Ile 675 680 685 Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr 690 695 700 Arg Met Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr 705 710 715 720 Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe 725 730 735 Ile Cys Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Ile 740 745 750 Ser Leu Ser His Ser Pro Gly Lys 755 760 <210> 17 <211> 758 <212> PRT <213> Artificial sequence <220> <223> Wolf IL4R-IL13R-IgGD (without signal sequence) <400> 17 Ser Gly Ser Val Lys Val Leu His Glu Pro Ser Cys Phe Ser Asp Tyr 1 5 10 15 Ile Ser Thr Ser Val Cys Gln Trp Lys Met Asp His Pro Thr Asn Cys 20 25 30 Ser Ala Glu Leu Arg Leu Ser Tyr Gln Leu Asp Phe Met Gly Ser Glu 35 40 45 Asn His Thr Cys Val Pro Glu Asn Arg Glu Asp Ser Val Cys Val Cys 50 55 60 Ser Met Pro Ile Asp Asp Ala Val Glu Ala Asp Val Tyr Gln Leu Asp 65 70 75 80 Leu Trp Ala Gly Gln Gln Leu Leu Trp Ser Gly Ser Phe Gln Pro Ser 85 90 95 Lys His Val Lys Pro Arg Thr Pro Gly Asn Leu Thr Val His Pro Asn 100 105 110 Ile Ser His Thr Trp Leu Leu Met Trp Thr Asn Pro Tyr Pro Thr Glu 115 120 125 Asn His Leu His Ser Glu Leu Thr Tyr Met Val Asn Val Ser Asn Asp 130 135 140 Asn Asp Pro Glu Asp Phe Lys Val Tyr Asn Val Thr Tyr Met Gly Pro 145 150 155 160 Thr Leu Arg Leu Ala Ala Ser Thr Leu Lys Ser Gly Ala Ser Tyr Ser 165 170 175 Ala Arg Val Arg Ala Trp Ala Gln Thr Tyr Asn Ser Thr Trp Ser Asp 180 185 190 Trp Ser Pro Ser Thr Thr Trp Leu Asn Tyr Tyr Glu Pro Gly Gly Gly 195 200 205 Ser Gly Thr Glu Thr Gln Pro Pro Val Thr Asn Leu Ser Val Ser Val 210 215 220 Glu Asn Leu Cys Thr Val Ile Trp Thr Trp Asp Pro Pro Glu Gly Ala 225 230 235 240 Ser Pro Asn Cys Thr Leu Arg Tyr Phe Ser His Phe Asp Asn Lys Gln 245 250 255 Asp Lys Lys Ile Ala Pro Glu Thr His Arg Ser Lys Glu Val Pro Leu 260 265 270 Asn Glu Arg Ile Cys Leu Gln Val Gly Ser Gln Cys Ser Thr Asn Glu 275 280 285 Ser Asp Asn Pro Ser Ile Leu Val Glu Lys Cys Thr Pro Pro Pro Glu 290 295 300 Gly Asp Pro Glu Ser Ala Val Thr Glu Leu Gln Cys Val Trp His Asn 305 310 315 320 Leu Ser Tyr Met Lys Cys Thr Trp Leu Pro Gly Arg Asn Thr Ser Pro 325 330 335 Asp Thr Asn Tyr Thr Leu Tyr Tyr Trp His Ser Ser Leu Gly Lys Ile 340 345 350 Leu Gln Cys Glu Asp Ile Tyr Arg Glu Gly Gln His Ile Gly Cys Ser 355 360 365 Phe Ala Leu Thr Asn Leu Lys Asp Ser Ser Phe Glu Gln His Ser Val 370 375 380 Gln Ile Val Val Lys Asp Asn Ala Gly Lys Ile Arg Pro Ser Phe Asn 385 390 395 400 Ile Val Pro Leu Thr Ser His Val Lys Pro Asp Pro Pro His Ile Lys 405 410 415 Arg Leu Phe Phe Gln Asn Gly Asn Leu Tyr Val Gln Trp Lys Asn Pro 420 425 430 Gln Asn Phe Tyr Ser Arg Cys Leu Ser Tyr Gln Val Glu Val Asn Asn 435 440 445 Ser Gln Thr Glu Thr Asn Asp Ile Phe Tyr Val Glu Glu Ala Lys Cys 450 455 460 Gln Asn Ser Glu Phe Glu Gly Asn Leu Glu Gly Thr Ile Cys Phe Met 465 470 475 480 Val Pro Gly Val Leu Pro Asp Thr Leu Asn Thr Val Arg Ile Arg Val 485 490 495 Arg Thr Asn Lys Leu Cys Tyr Glu Asp Asp Lys Leu Trp Ser Asn Trp 500 505 510 Ser Gln Ala Met Ser Ile Gly Glu Asn Thr Asp Pro Thr Pro Lys Glu 515 520 525 Ser Thr Cys Lys Cys Ile Ser Pro Cys Pro Val Pro Glu Ser Leu Gly 530 535 540 Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Ile Leu Arg 545 550 555 560 Ile Thr Arg Thr Pro Glu Ile Thr Cys Val Val Leu Asp Leu Gly Arg 565 570 575 Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Glu Val 580 585 590 His Thr Ala Lys Thr Gln Pro Arg Glu Gln Gln Phe Asn Ser Thr Tyr 595 600 605 Arg Val Val Ser Val Leu Pro Ile Glu His Gln Asp Trp Leu Thr Gly 610 615 620 Lys Glu Phe Lys Cys Arg Val Asn His Ile Gly Leu Pro Ser Pro Ile 625 630 635 640 Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val 645 650 655 Tyr Val Leu Pro Pro Ser Pro Lys Glu Leu Ser Ser Ser Asp Thr Val 660 665 670 Thr Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro Glu Ile Asp Val 675 680 685 Glu Trp Gln Ser Asn Gly Gln Pro Glu Pro Glu Ser Lys Tyr His Thr 690 695 700 Thr Ala Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys 705 710 715 720 Leu Ser Val Asp Lys Ser Arg Trp Gln Gln Gly Asp Thr Phe Thr Cys 725 730 735 Ala Val Met His Glu Ala Leu Gln Asn His Tyr Thr Asp Leu Ser Leu 740 745 750 Ser His Ser Pro Gly Lys 755 <210> 18 <211> 590 <212> PRT <213> Artificial Sequence <220> <223> Feline IL4R-IL13R (without signal sequence) <400> 18 Ser Gly Ser Val Lys Val Leu Arg Ala Pro Thr Cys Phe Ser Asp Tyr 1 5 10 15 Phe Ser Thr Ser Val Cys Gln Trp Asn Met Asp Ala Pro Thr Asn Cys 20 25 30 Ser Ala Glu Leu Arg Leu Ser Tyr Gln Leu Asn Phe Met Gly Ser Glu 35 40 45 Asn Arg Thr Cys Val Pro Glu Asn Gly Glu Gly Ala Ala Cys Ala Cys 50 55 60 Ser Met Leu Met Asp Asp Phe Val Glu Ala Asp Val Tyr Gln Leu His 65 70 75 80 Leu Trp Ala Gly Thr Gln Leu Leu Trp Ser Gly Ser Phe Lys Pro Ser 85 90 95 Ser His Val Lys Pro Arg Ala Pro Gly Asn Leu Thr Val His Pro Asn 100 105 110 Val Ser His Thr Trp Leu Leu Arg Trp Ser Asn Pro Tyr Pro Pro Glu 115 120 125 Asn His Leu His Ala Glu Leu Thr Tyr Met Val Asn Ile Ser Ser Glu 130 135 140 Asp Asp Pro Thr Asp Val Ser Val Cys Ala Ser Gly Phe Leu Cys His 145 150 155 160 Leu Leu Gly Leu Arg Arg Val Glu Thr Gly Ala Pro Gly Ala Arg Leu 165 170 175 Pro Pro Trp Leu Cys Ala Pro Arg Pro Arg Arg Val Pro Gly Ser Gln 180 185 190 Cys Ala Val Ile Ser Cys Cys Arg Trp Val Leu Ile Ala Leu Thr Ser 195 200 205 Arg Gly Gly Arg Trp Arg Leu Thr Pro Gly Leu Arg Ser Gln Thr Arg 210 215 220 Tyr Val Ser Val Ala Glu Gly Leu Phe Gly Ala Thr Pro Arg Val Leu 225 230 235 240 Cys Pro Gly Thr Gln Ala Gly Leu Ala Ser Ala Ala Arg Glu Gln Met 245 250 255 Ser Pro Asp Pro Ser Ala Phe His Ser Ile Asp Tyr Glu Pro Gly Gly 260 265 270 Gly Ser Gly Ser Gln Thr Gln Pro Pro Val Thr Asn Leu Ser Val Ser 275 280 285 Val Glu Asn Leu Cys Thr Val Ile Trp Thr Trp Asp Pro Pro Glu Gly 290 295 300 Ala Ser Pro Asn Cys Thr Leu Arg Tyr Phe Ser His Phe Asp Asn Lys 305 310 315 320 Gln Asp Lys Lys Ile Ala Pro Glu Thr His Arg Ser Lys Glu Val Pro 325 330 335 Leu Asn Glu Arg Ile Cys Leu Gln Val Gly Ser Gln Cys Ser Thr Asn 340 345 350 Glu Ser Asp Asn Pro Ser Ile Leu Val Glu Lys Cys Thr Pro Pro Pro 355 360 365 Glu Gly Asp Pro Glu Ser Ala Val Thr Glu Leu Gln Cys Val Trp His 370 375 380 Asn Leu Ser Tyr Met Lys Cys Thr Trp Leu Pro Gly Arg Asn Thr Ser 385 390 395 400 Pro Asp Thr Asn Tyr Thr Leu Tyr Tyr Trp His Ser Ser Leu Gly Lys 405 410 415 Ile Leu Gln Cys Glu Asn Ile Tyr Arg Glu Gly Gln His Ile Gly Cys 420 425 430 Ser Phe Ala Leu Thr Asn Leu Lys Asp Ser Ser Phe Glu Gln His Ser 435 440 445 Val Gln Ile Val Val Lys Asp Asn Ala Gly Lys Ile Arg Pro Ser Phe 450 455 460 Asn Ile Val Pro Leu Thr Ser His Val Lys Pro Asp Pro Pro His Ile 465 470 475 480 Lys Arg Leu Phe Phe Gln Asn Gly Asn Leu Tyr Val Gln Trp Lys Asn 485 490 495 Pro Gln Asn Phe Tyr Ser Arg Cys Leu Ser Tyr Gln Val Glu Val Asn 500 505 510 Asn Ser Gln Thr Glu Thr His Asp Ile Phe Tyr Val Glu Glu Ala Lys 515 520 525 Cys Gln Asn Ser Glu Phe Glu Gly Asn Leu Glu Gly Thr Ile Cys Phe 530 535 540 Met Val Pro Gly Ile Leu Pro Asp Thr Leu Asn Thr Val Arg Ile Arg 545 550 555 560 Val Arg Thr Asn Lys Leu Cys Tyr Glu Asp Asp Arg Leu Trp Ser Asn 565 570 575 Trp Ser Gln Ala Met Ser Ile Gly Glu Asn Thr Asp Pro Thr 580 585 590 <210> 19 <211> 525 <212> PRT <213> Artificial sequence <220> <223> Horse IL4R-IL13R (without signal sequence) <400> 19 Ser Gly Ser Val Lys Val Leu His Leu Thr Ala Cys Phe Ser Asp Tyr 1 5 10 15 Ile Ser Ala Ser Thr Cys Glu Trp Lys Met Asp Arg Pro Thr Asn Cys 20 25 30 Ser Ala Gln Leu Arg Leu Ser Tyr Gln Leu Asn Asp Glu Phe Ser Asp 35 40 45 Asn Leu Thr Cys Ile Pro Glu Asn Arg Glu Asp Glu Val Cys Val Cys 50 55 60 Arg Met Leu Met Asp Asn Ile Val Ser Glu Asp Val Tyr Glu Leu Asp 65 70 75 80 Leu Trp Ala Gly Asn Gln Leu Leu Trp Asn Ser Ser Phe Lys Pro Ser 85 90 95 Arg His Val Lys Pro Arg Ala Pro Gln Asn Leu Thr Val His Ala Ile 100 105 110 Ser His Thr Trp Leu Leu Thr Trp Ser Asn Pro Tyr Pro Leu Lys Asn 115 120 125 His Leu Trp Ser Glu Leu Thr Tyr Leu Val Asn Ile Ser Lys Glu Asp 130 135 140 Asp Pro Thr Asp Phe Lys Ile Tyr Asn Val Thr Tyr Met Asp Pro Thr 145 150 155 160 Leu Arg Val Thr Ala Ser Thr Leu Lys Ser Arg Ala Thr Tyr Ser Ala 165 170 175 Arg Val Lys Ala Arg Ala Gln Asn Tyr Asn Ser Thr Trp Ser Glu Trp 180 185 190 Ser Pro Ser Thr Thr Trp His Asn Tyr Tyr Glu Gln Pro Gly Gly Gly 195 200 205 Ser Gly Thr Glu Ser Gln Pro Pro Val Thr Asn Leu Ser Val Ser Val 210 215 220 Glu Asn Leu Cys Thr Val Ile Trp Thr Trp Asn Pro Pro Glu Gly Val 225 230 235 240 Ser Pro Asn Cys Ser Leu Trp Tyr Phe Ser His Phe Gly Asn Lys Gln 245 250 255 Asp Lys Lys Ile Ala Pro Glu Thr His Arg Ser Lys Glu Val Pro Leu 260 265 270 Asn Glu Arg Ile Cys Leu Gln Val Gly Ser Gln Cys Ser Thr Asn Glu 275 280 285 Ser Asp Asn Pro Ser Ile Leu Val Glu Lys Cys Ile Ser Pro Pro Glu 290 295 300 Gly Asp Pro Glu Ser Ala Val Thr Glu Leu Gln Cys Val Trp His Asn 305 310 315 320 Leu Ser Tyr Met Lys Cys Thr Trp Leu Pro Gly Lys Asn Ala Ser Pro 325 330 335 Asp Thr Asn Tyr Thr Leu Tyr Tyr Trp His Ser Ser Leu Gly Lys Ile 340 345 350 Leu Gln Cys Glu Asp Ile Tyr Arg Glu Gly Gln His Ile Gly Cys Ser 355 360 365 Phe Ala Leu Thr Glu Val Lys Asp Ser Ile Phe Glu Gln His Ser Val 370 375 380 Gln Ile Met Val Lys Asp Asn Ala Gly Lys Ile Arg Pro Phe Phe Asn 385 390 395 400 Ile Val Pro Leu Thr Ser His Val Lys Pro Asp Pro Pro His Ile Lys 405 410 415 Lys Leu Phe Phe Gln Asn Gly Asp Leu Tyr Val Gln Trp Lys Asn Pro 420 425 430 Gln Asn Phe Tyr Ser Arg Cys Leu Ser Tyr Gln Val Glu Val Asn Asn 435 440 445 Ser Gln Thr Glu Thr Arg Asp Ile Phe Ser Val Glu Glu Ala Lys Cys 450 455 460 Gln Asn Pro Glu Phe Glu Gly Asp Leu Glu Gly Thr Ile Cys Phe Met 465 470 475 480 Val Pro Gly Val Leu Pro Asp Thr Val Asn Thr Val Arg Ile Arg Val 485 490 495 Lys Thr Asn Lys Leu Cys Tyr Glu Asp Asp Lys Leu Trp Ser Asn Trp 500 505 510 Ser Gln Ala Met Ser Ile Gly Lys Lys Ala Asp Pro Thr 515 520 525 <210> 20 <211> 780 <212> PRT <213> Artificial sequence <220> <223> Wolf IL13R-IL4R-IgGB (with signal sequence) <400> 20 Met Ala Val Leu Gly Leu Leu Phe Cys Leu Val Thr Phe Pro Ser Cys 1 5 10 15 Val Leu Ser Thr Glu Thr Gln Pro Pro Val Thr Asn Leu Ser Val Ser 20 25 30 Val Glu Asn Leu Cys Thr Val Ile Trp Thr Trp Asp Pro Pro Glu Gly 35 40 45 Ala Ser Pro Asn Cys Thr Leu Arg Tyr Phe Ser His Phe Asp Asn Lys 50 55 60 Gln Asp Lys Lys Ile Ala Pro Glu Thr His Arg Ser Lys Glu Val Pro 65 70 75 80 Leu Asn Glu Arg Ile Cys Leu Gln Val Gly Ser Gln Cys Ser Thr Asn 85 90 95 Glu Ser Asp Asn Pro Ser Ile Leu Val Glu Lys Cys Thr Pro Pro Pro 100 105 110 Glu Gly Asp Pro Glu Ser Ala Val Thr Glu Leu Gln Cys Val Trp His 115 120 125 Asn Leu Ser Tyr Met Lys Cys Thr Trp Leu Pro Gly Arg Asn Thr Ser 130 135 140 Pro Asp Thr Asn Tyr Thr Leu Tyr Tyr Trp His Ser Ser Leu Gly Lys 145 150 155 160 Ile Leu Gln Cys Glu Asp Ile Tyr Arg Glu Gly Gln His Ile Gly Cys 165 170 175 Ser Phe Ala Leu Thr Asn Leu Lys Asp Ser Ser Phe Glu Gln His Ser 180 185 190 Val Gln Ile Val Val Lys Asp Asn Ala Gly Lys Ile Arg Pro Ser Phe 195 200 205 Asn Ile Val Pro Leu Thr Ser His Val Lys Pro Asp Pro Pro His Ile 210 215 220 Lys Arg Leu Phe Phe Gln Asn Gly Asn Leu Tyr Val Gln Trp Lys Asn 225 230 235 240 Pro Gln Asn Phe Tyr Ser Arg Cys Leu Ser Tyr Gln Val Glu Val Asn 245 250 255 Asn Ser Gln Thr Glu Thr Asn Asp Ile Phe Tyr Val Glu Glu Ala Lys 260 265 270 Cys Gln Asn Ser Glu Phe Glu Gly Asn Leu Glu Gly Thr Ile Cys Phe 275 280 285 Met Val Pro Gly Val Leu Pro Asp Thr Leu Asn Thr Val Arg Ile Arg 290 295 300 Val Arg Thr Asn Lys Leu Cys Tyr Glu Asp Asp Lys Leu Trp Ser Asn 305 310 315 320 Trp Ser Gln Ala Met Ser Ile Gly Glu Asn Thr Asp Pro Thr Gly Gly 325 330 335 Gly Ser Gly Ser Gly Ser Val Lys Val Leu His Glu Pro Ser Cys Phe 340 345 350 Ser Asp Tyr Ile Ser Thr Ser Val Cys Gln Trp Lys Met Asp His Pro 355 360 365 Thr Asn Cys Ser Ala Glu Leu Arg Leu Ser Tyr Gln Leu Asp Phe Met 370 375 380 Gly Ser Glu Asn His Thr Cys Val Pro Glu Asn Arg Glu Asp Ser Val 385 390 395 400 Cys Val Cys Ser Met Pro Ile Asp Asp Ala Val Glu Ala Asp Val Tyr 405 410 415 Gln Leu Asp Leu Trp Ala Gly Gln Gln Leu Leu Trp Ser Gly Ser Phe 420 425 430 Gln Pro Ser Lys His Val Lys Pro Arg Thr Pro Gly Asn Leu Thr Val 435 440 445 His Pro Asn Ile Ser His Thr Trp Leu Leu Met Trp Thr Asn Pro Tyr 450 455 460 Pro Thr Glu Asn His Leu His Ser Glu Leu Thr Tyr Met Val Asn Val 465 470 475 480 Ser Asn Asp Asn Asp Pro Glu Asp Phe Lys Val Tyr Asn Val Thr Tyr 485 490 495 Met Gly Pro Thr Leu Arg Leu Ala Ala Ser Thr Leu Lys Ser Gly Ala 500 505 510 Ser Tyr Ser Ala Arg Val Arg Ala Trp Ala Gln Thr Tyr Asn Ser Thr 515 520 525 Trp Ser Asp Trp Ser Pro Ser Thr Thr Trp Leu Asn Tyr Tyr Glu Pro 530 535 540 Lys Arg Glu Asn Gly Arg Val Pro Arg Pro Pro Asp Cys Pro Lys Cys 545 550 555 560 Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro 565 570 575 Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys 580 585 590 Val Val Val Asp Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser Trp 595 600 605 Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu 610 615 620 Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly 625 630 635 640 His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Lys Val Asn Asn 645 650 655 Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly 660 665 670 Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu 675 680 685 Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe 690 695 700 Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro 705 710 715 720 Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser 725 730 735 Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg 740 745 750 Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His 755 760 765 Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys 770 775 780 <210> 21 <211> 781 <212> PRT <213> Artificial Sequence <220> <223> Wolf IL4R-IL13R-IgGB (with signal sequence) <400> 21 Met Ala Val Leu Gly Leu Leu Phe Cys Leu Val Thr Phe Pro Ser Cys 1 5 10 15 Val Leu Ser Ser Gly Ser Val Lys Val Leu His Glu Pro Ser Cys Phe 20 25 30 Ser Asp Tyr Ile Ser Thr Ser Val Cys Gln Trp Lys Met Asp His Pro 35 40 45 Thr Asn Cys Ser Ala Glu Leu Arg Leu Ser Tyr Gln Leu Asp Phe Met 50 55 60 Gly Ser Glu Asn His Thr Cys Val Pro Glu Asn Arg Glu Asp Ser Val 65 70 75 80 Cys Val Cys Ser Met Pro Ile Asp Asp Ala Val Glu Ala Asp Val Tyr 85 90 95 Gln Leu Asp Leu Trp Ala Gly Gln Gln Leu Leu Trp Ser Gly Ser Phe 100 105 110 Gln Pro Ser Lys His Val Lys Pro Arg Thr Pro Gly Asn Leu Thr Val 115 120 125 His Pro Asn Ile Ser His Thr Trp Leu Leu Met Trp Thr Asn Pro Tyr 130 135 140 Pro Thr Glu Asn His Leu His Ser Glu Leu Thr Tyr Met Val Asn Val 145 150 155 160 Ser Asn Asp Asn Asp Pro Glu Asp Phe Lys Val Tyr Asn Val Thr Tyr 165 170 175 Met Gly Pro Thr Leu Arg Leu Ala Ala Ser Thr Leu Lys Ser Gly Ala 180 185 190 Ser Tyr Ser Ala Arg Val Arg Ala Trp Ala Gln Thr Tyr Asn Ser Thr 195 200 205 Trp Ser Asp Trp Ser Pro Ser Thr Thr Trp Leu Asn Tyr Tyr Glu Pro 210 215 220 Gly Gly Gly Ser Gly Thr Glu Thr Gln Pro Pro Val Thr Asn Leu Ser 225 230 235 240 Val Ser Val Glu Asn Leu Cys Thr Val Ile Trp Thr Trp Asp Pro Pro 245 250 255 Glu Gly Ala Ser Pro Asn Cys Thr Leu Arg Tyr Phe Ser His Phe Asp 260 265 270 Asn Lys Gln Asp Lys Lys Ile Ala Pro Glu Thr His Arg Ser Lys Glu 275 280 285 Val Pro Leu Asn Glu Arg Ile Cys Leu Gln Val Gly Ser Gln Cys Ser 290 295 300 Thr Asn Glu Ser Asp Asn Pro Ser Ile Leu Val Glu Lys Cys Thr Pro 305 310 315 320 Pro Pro Glu Gly Asp Pro Glu Ser Ala Val Thr Glu Leu Gln Cys Val 325 330 335 Trp His Asn Leu Ser Tyr Met Lys Cys Thr Trp Leu Pro Gly Arg Asn 340 345 350 Thr Ser Pro Asp Thr Asn Tyr Thr Leu Tyr Tyr Trp His Ser Ser Leu 355 360 365 Gly Lys Ile Leu Gln Cys Glu Asp Ile Tyr Arg Glu Gly Gln His Ile 370 375 380 Gly Cys Ser Phe Ala Leu Thr Asn Leu Lys Asp Ser Ser Phe Glu Gln 385 390 395 400 His Ser Val Gln Ile Val Val Lys Asp Asn Ala Gly Lys Ile Arg Pro 405 410 415 Ser Phe Asn Ile Val Pro Leu Thr Ser His Val Lys Pro Asp Pro Pro 420 425 430 His Ile Lys Arg Leu Phe Phe Gln Asn Gly Asn Leu Tyr Val Gln Trp 435 440 445 Lys Asn Pro Gln Asn Phe Tyr Ser Arg Cys Leu Ser Tyr Gln Val Glu 450 455 460 Val Asn Asn Ser Gln Thr Glu Thr Asn Asp Ile Phe Tyr Val Glu Glu 465 470 475 480 Ala Lys Cys Gln Asn Ser Glu Phe Glu Gly Asn Leu Glu Gly Thr Ile 485 490 495 Cys Phe Met Val Pro Gly Val Leu Pro Asp Thr Leu Asn Thr Val Arg 500 505 510 Ile Arg Val Arg Thr Asn Lys Leu Cys Tyr Glu Asp Asp Lys Leu Trp 515 520 525 Ser Asn Trp Ser Gln Ala Met Ser Ile Gly Glu Asn Thr Asp Pro Thr 530 535 540 Pro Lys Arg Glu Asn Gly Arg Val Pro Arg Pro Pro Asp Cys Pro Lys 545 550 555 560 Cys Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro 565 570 575 Pro Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr 580 585 590 Cys Val Val Val Asp Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser 595 600 605 Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg 610 615 620 Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile 625 630 635 640 Gly His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Lys Val Asn 645 650 655 Asn Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg 660 665 670 Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu 675 680 685 Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe 690 695 700 Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu 705 710 715 720 Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly 725 730 735 Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln 740 745 750 Arg Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn 755 760 765 His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys 770 775 780 <210> 22 <211> 315 <212> PRT <213> Artificial sequence <220> <223> Wolf IL13R extracellular domain (without signal sequence) <400> twenty two Thr Glu Thr Gln Pro Pro Val Thr Asn Leu Ser Val Ser Val Glu Asn 1 5 10 15 Leu Cys Thr Val Ile Trp Thr Trp Asp Pro Pro Glu Gly Ala Ser Pro 20 25 30 Asn Cys Thr Leu Arg Tyr Phe Ser His Phe Asp Asn Lys Gln Asp Lys 35 40 45 Lys Ile Ala Pro Glu Thr His Arg Ser Lys Glu Val Pro Leu Asn Glu 50 55 60 Arg Ile Cys Leu Gln Val Gly Ser Gln Cys Ser Thr Asn Glu Ser Asp 65 70 75 80 Asn Pro Ser Ile Leu Val Glu Lys Cys Thr Pro Pro Pro Glu Gly Asp 85 90 95 Pro Glu Ser Ala Val Thr Glu Leu Gln Cys Val Trp His Asn Leu Ser 100 105 110 Tyr Met Lys Cys Thr Trp Leu Pro Gly Arg Asn Thr Ser Pro Asp Thr 115 120 125 Asn Tyr Thr Leu Tyr Tyr Trp His Ser Ser Leu Gly Lys Ile Leu Gln 130 135 140 Cys Glu Asp Ile Tyr Arg Glu Gly Gln His Ile Gly Cys Ser Phe Ala 145 150 155 160 Leu Thr Asn Leu Lys Asp Ser Ser Phe Glu Gln His Ser Val Gln Ile 165 170 175 Val Val Lys Asp Asn Ala Gly Lys Ile Arg Pro Ser Phe Asn Ile Val 180 185 190 Pro Leu Thr Ser His Val Lys Pro Asp Pro Pro His Ile Lys Arg Leu 195 200 205 Phe Phe Gln Asn Gly Asn Leu Tyr Val Gln Trp Lys Asn Pro Gln Asn 210 215 220 Phe Tyr Ser Arg Cys Leu Ser Tyr Gln Val Glu Val Asn Asn Ser Gln 225 230 235 240 Thr Glu Thr Asn Asp Ile Phe Tyr Val Glu Glu Ala Lys Cys Gln Asn 245 250 255 Ser Glu Phe Glu Gly Asn Leu Glu Gly Thr Ile Cys Phe Met Val Pro 260 265 270 Gly Val Leu Pro Asp Thr Leu Asn Thr Val Arg Ile Arg Val Arg Thr 275 280 285 Asn Lys Leu Cys Tyr Glu Asp Asp Lys Leu Trp Ser Asn Trp Ser Gln 290 295 300 Ala Met Ser Ile Gly Glu Asn Thr Asp Pro Thr 305 310 315 <210> twenty three <211> 207 <212> PRT <213> Artificial sequence <220> <223> Wolf IL4R extracellular domain (without signal sequence) <400> twenty three Ser Gly Ser Gly Ser Val Lys Val Leu His Glu Pro Ser Cys Phe Ser 1 5 10 15 Asp Tyr Ile Ser Thr Ser Val Cys Gln Trp Lys Met Asp His Pro Thr 20 25 30 Asn Cys Ser Ala Glu Leu Arg Leu Ser Tyr Gln Leu Asp Phe Met Gly 35 40 45 Ser Glu Asn His Thr Cys Val Pro Glu Asn Arg Glu Asp Ser Val Cys 50 55 60 Val Cys Ser Met Pro Ile Asp Asp Ala Val Glu Ala Asp Val Tyr Gln 65 70 75 80 Leu Asp Leu Trp Ala Gly Gln Gln Leu Leu Trp Ser Gly Ser Phe Gln 85 90 95 Pro Ser Lys His Val Lys Pro Arg Thr Pro Gly Asn Leu Thr Val His 100 105 110 Pro Asn Ile Ser His Thr Trp Leu Leu Met Trp Thr Asn Pro Tyr Pro 115 120 125 Thr Glu Asn His Leu His Ser Glu Leu Thr Tyr Met Val Asn Val Ser 130 135 140 Asn Asp Asn Asp Pro Glu Asp Phe Lys Val Tyr Asn Val Thr Tyr Met 145 150 155 160 Gly Pro Thr Leu Arg Leu Ala Ala Ser Thr Leu Lys Ser Gly Ala Ser 165 170 175 Tyr Ser Ala Arg Val Arg Ala Trp Ala Gln Thr Tyr Asn Ser Thr Trp 180 185 190 Ser Asp Trp Ser Pro Ser Thr Thr Trp Leu Asn Tyr Tyr Glu Pro 195 200 205 <210> twenty four <211> 315 <212> PRT <213> Artificial sequence <220> <223> Domestic cat IL13R extracellular domain (without signal sequence) <400> twenty four Ser Gln Thr Gln Pro Pro Val Thr Asn Leu Ser Val Ser Val Glu Asn 1 5 10 15 Leu Cys Thr Val Ile Trp Thr Trp Asp Pro Pro Glu Gly Ala Ser Pro 20 25 30 Asn Cys Thr Leu Arg Tyr Phe Ser His Phe Asp Asn Lys Gln Asp Lys 35 40 45 Lys Ile Ala Pro Glu Thr His Arg Ser Lys Glu Val Pro Leu Asn Glu 50 55 60 Arg Ile Cys Leu Gln Val Gly Ser Gln Cys Ser Thr Asn Glu Ser Asp 65 70 75 80 Asn Pro Ser Ile Leu Val Glu Lys Cys Thr Pro Pro Pro Glu Gly Asp 85 90 95 Pro Glu Ser Ala Val Thr Glu Leu Gln Cys Val Trp His Asn Leu Ser 100 105 110 Tyr Met Lys Cys Thr Trp Leu Pro Gly Arg Asn Thr Ser Pro Asp Thr 115 120 125 Asn Tyr Thr Leu Tyr Tyr Trp His Ser Ser Leu Gly Lys Ile Leu Gln 130 135 140 Cys Glu Asn Ile Tyr Arg Glu Gly Gln His Ile Gly Cys Ser Phe Ala 145 150 155 160 Leu Thr Asn Leu Lys Asp Ser Ser Phe Glu Gln His Ser Val Gln Ile 165 170 175 Val Val Lys Asp Asn Ala Gly Lys Ile Arg Pro Ser Phe Asn Ile Val 180 185 190 Pro Leu Thr Ser His Val Lys Pro Asp Pro Pro His Ile Lys Arg Leu 195 200 205 Phe Phe Gln Asn Gly Asn Leu Tyr Val Gln Trp Lys Asn Pro Gln Asn 210 215 220 Phe Tyr Ser Arg Cys Leu Ser Tyr Gln Val Glu Val Asn Asn Ser Gln 225 230 235 240 Thr Glu Thr His Asp Ile Phe Tyr Val Glu Glu Ala Lys Cys Gln Asn 245 250 255 Ser Glu Phe Glu Gly Asn Leu Glu Gly Thr Ile Cys Phe Met Val Pro 260 265 270 Gly Ile Leu Pro Asp Thr Leu Asn Thr Val Arg Ile Arg Val Arg Thr 275 280 285 Asn Lys Leu Cys Tyr Glu Asp Asp Arg Leu Trp Ser Asn Trp Ser Gln 290 295 300 Ala Met Ser Ile Gly Glu Asn Thr Asp Pro Thr 305 310 315 <210> 25 <211> 270 <212> PRT <213> Artificial sequence <220> <223> Domestic cat IL4R extracellular domain (without signal sequence) <400> 25 Ser Gly Ser Val Lys Val Leu Arg Ala Pro Thr Cys Phe Ser Asp Tyr 1 5 10 15 Phe Ser Thr Ser Val Cys Gln Trp Asn Met Asp Ala Pro Thr Asn Cys 20 25 30 Ser Ala Glu Leu Arg Leu Ser Tyr Gln Leu Asn Phe Met Gly Ser Glu 35 40 45 Asn Arg Thr Cys Val Pro Glu Asn Gly Glu Gly Ala Ala Cys Ala Cys 50 55 60 Ser Met Leu Met Asp Asp Phe Val Glu Ala Asp Val Tyr Gln Leu His 65 70 75 80 Leu Trp Ala Gly Thr Gln Leu Leu Trp Ser Gly Ser Phe Lys Pro Ser 85 90 95 Ser His Val Lys Pro Arg Ala Pro Gly Asn Leu Thr Val His Pro Asn 100 105 110 Val Ser His Thr Trp Leu Leu Arg Trp Ser Asn Pro Tyr Pro Pro Glu 115 120 125 Asn His Leu His Ala Glu Leu Thr Tyr Met Val Asn Ile Ser Ser Glu 130 135 140 Asp Asp Pro Thr Asp Val Ser Val Cys Ala Ser Gly Phe Leu Cys His 145 150 155 160 Leu Leu Gly Leu Arg Arg Val Glu Thr Gly Ala Pro Gly Ala Arg Leu 165 170 175 Pro Pro Trp Leu Cys Ala Pro Arg Pro Arg Arg Val Pro Gly Ser Gln 180 185 190 Cys Ala Val Ile Ser Cys Cys Arg Trp Val Leu Ile Ala Leu Thr Ser 195 200 205 Arg Gly Gly Arg Trp Arg Leu Thr Pro Gly Leu Arg Ser Gln Thr Arg 210 215 220 Tyr Val Ser Val Ala Glu Gly Leu Phe Gly Ala Thr Pro Arg Val Leu 225 230 235 240 Cys Pro Gly Thr Gln Ala Gly Leu Ala Ser Ala Ala Arg Glu Gln Met 245 250 255 Ser Pro Asp Pro Ser Ala Phe His Ser Ile Asp Tyr Glu Pro 260 265 270 <210> 26 <211> 315 <212> PRT <213> Artificial sequence <220> <223> Horse IL13R extracellular domain (without signal sequence) <400> 26 Thr Glu Ser Gln Pro Pro Val Thr Asn Leu Ser Val Ser Val Glu Asn 1 5 10 15 Leu Cys Thr Val Ile Trp Thr Trp Asn Pro Pro Glu Gly Val Ser Pro 20 25 30 Asn Cys Ser Leu Trp Tyr Phe Ser His Phe Gly Asn Lys Gln Asp Lys 35 40 45 Lys Ile Ala Pro Glu Thr His Arg Ser Lys Glu Val Pro Leu Asn Glu 50 55 60 Arg Ile Cys Leu Gln Val Gly Ser Gln Cys Ser Thr Asn Glu Ser Asp 65 70 75 80 Asn Pro Ser Ile Leu Val Glu Lys Cys Ile Ser Pro Pro Glu Gly Asp 85 90 95 Pro Glu Ser Ala Val Thr Glu Leu Gln Cys Val Trp His Asn Leu Ser 100 105 110 Tyr Met Lys Cys Thr Trp Leu Pro Gly Lys Asn Ala Ser Pro Asp Thr 115 120 125 Asn Tyr Thr Leu Tyr Tyr Trp His Ser Ser Leu Gly Lys Ile Leu Gln 130 135 140 Cys Glu Asp Ile Tyr Arg Glu Gly Gln His Ile Gly Cys Ser Phe Ala 145 150 155 160 Leu Thr Glu Val Lys Asp Ser Ile Phe Glu Gln His Ser Val Gln Ile 165 170 175 Met Val Lys Asp Asn Ala Gly Lys Ile Arg Pro Phe Phe Asn Ile Val 180 185 190 Pro Leu Thr Ser His Val Lys Pro Asp Pro Pro His Ile Lys Lys Leu 195 200 205 Phe Phe Gln Asn Gly Asp Leu Tyr Val Gln Trp Lys Asn Pro Gln Asn 210 215 220 Phe Tyr Ser Arg Cys Leu Ser Tyr Gln Val Glu Val Asn Asn Ser Gln 225 230 235 240 Thr Glu Thr Arg Asp Ile Phe Ser Val Glu Glu Ala Lys Cys Gln Asn 245 250 255 Pro Glu Phe Glu Gly Asp Leu Glu Gly Thr Ile Cys Phe Met Val Pro 260 265 270 Gly Val Leu Pro Asp Thr Val Asn Thr Val Arg Ile Arg Val Lys Thr 275 280 285 Asn Lys Leu Cys Tyr Glu Asp Asp Lys Leu Trp Ser Asn Trp Ser Gln 290 295 300 Ala Met Ser Ile Gly Lys Lys Ala Asp Pro Thr 305 310 315 <210> 27 <211> 205 <212> PRT <213> Artificial Sequence <220> <223> Equine IL4R extracellular domain (without signal sequence) <400> 27 Ser Gly Ser Val Lys Val Leu His Leu Thr Ala Cys Phe Ser Asp Tyr 1 5 10 15 Ile Ser Ala Ser Thr Cys Glu Trp Lys Met Asp Arg Pro Thr Asn Cys 20 25 30 Ser Ala Gln Leu Arg Leu Ser Tyr Gln Leu Asn Asp Glu Phe Ser Asp 35 40 45 Asn Leu Thr Cys Ile Pro Glu Asn Arg Glu Asp Glu Val Cys Val Cys 50 55 60 Arg Met Leu Met Asp Asn Ile Val Ser Glu Asp Val Tyr Glu Leu Asp 65 70 75 80 Leu Trp Ala Gly Asn Gln Leu Leu Trp Asn Ser Ser Phe Lys Pro Ser 85 90 95 Arg His Val Lys Pro Arg Ala Pro Gln Asn Leu Thr Val His Ala Ile 100 105 110 Ser His Thr Trp Leu Leu Thr Trp Ser Asn Pro Tyr Pro Leu Lys Asn 115 120 125 His Leu Trp Ser Glu Leu Thr Tyr Leu Val Asn Ile Ser Lys Glu Asp 130 135 140 Asp Pro Thr Asp Phe Lys Ile Tyr Asn Val Thr Tyr Met Asp Pro Thr 145 150 155 160 Leu Arg Val Thr Ala Ser Thr Leu Lys Ser Arg Ala Thr Tyr Ser Ala 165 170 175 Arg Val Lys Ala Arg Ala Gln Asn Tyr Asn Ser Thr Trp Ser Glu Trp 180 185 190 Ser Pro Ser Thr Thr Trp His Asn Tyr Tyr Glu Gln Pro 195 200 205 <210> 28 <211> 829 <212> PRT <213> Artificial Sequence <220> <223> Feline IL13R-IL4R-IgG2 (without signal sequence) <400> 28 Ser Gln Thr Gln Pro Pro Val Thr Asn Leu Ser Val Ser Val Glu Asn 1 5 10 15 Leu Cys Thr Val Ile Trp Thr Trp Asp Pro Pro Glu Gly Ala Ser Pro 20 25 30 Asn Cys Thr Leu Arg Tyr Phe Ser His Phe Asp Asn Lys Gln Asp Lys 35 40 45 Lys Ile Ala Pro Glu Thr His Arg Ser Lys Glu Val Pro Leu Asn Glu 50 55 60 Arg Ile Cys Leu Gln Val Gly Ser Gln Cys Ser Thr Asn Glu Ser Asp 65 70 75 80 Asn Pro Ser Ile Leu Val Glu Lys Cys Thr Pro Pro Pro Glu Gly Asp 85 90 95 Pro Glu Ser Ala Val Thr Glu Leu Gln Cys Val Trp His Asn Leu Ser 100 105 110 Tyr Met Lys Cys Thr Trp Leu Pro Gly Arg Asn Thr Ser Pro Asp Thr 115 120 125 Asn Tyr Thr Leu Tyr Tyr Trp His Ser Ser Leu Gly Lys Ile Leu Gln 130 135 140 Cys Glu Asn Ile Tyr Arg Glu Gly Gln His Ile Gly Cys Ser Phe Ala 145 150 155 160 Leu Thr Asn Leu Lys Asp Ser Ser Phe Glu Gln His Ser Val Gln Ile 165 170 175 Val Val Lys Asp Asn Ala Gly Lys Ile Arg Pro Ser Phe Asn Ile Val 180 185 190 Pro Leu Thr Ser His Val Lys Pro Asp Pro Pro His Ile Lys Arg Leu 195 200 205 Phe Phe Gln Asn Gly Asn Leu Tyr Val Gln Trp Lys Asn Pro Gln Asn 210 215 220 Phe Tyr Ser Arg Cys Leu Ser Tyr Gln Val Glu Val Asn Asn Ser Gln 225 230 235 240 Thr Glu Thr His Asp Ile Phe Tyr Val Glu Glu Ala Lys Cys Gln Asn 245 250 255 Ser Glu Phe Glu Gly Asn Leu Glu Gly Thr Ile Cys Phe Met Val Pro 260 265 270 Gly Ile Leu Pro Asp Thr Leu Asn Thr Val Arg Ile Arg Val Arg Thr 275 280 285 Asn Lys Leu Cys Tyr Glu Asp Asp Arg Leu Trp Ser Asn Trp Ser Gln 290 295 300 Ala Met Ser Ile Gly Glu Asn Thr Asp Pro Thr Gly Gly Gly Ser Gly 305 310 315 320 Ser Ser Gly Ser Val Lys Val Leu Arg Ala Pro Thr Cys Phe Ser Asp 325 330 335 Tyr Phe Ser Thr Ser Val Cys Gln Trp Asn Met Asp Ala Pro Thr Asn 340 345 350 Cys Ser Ala Glu Leu Arg Leu Ser Tyr Gln Leu Asn Phe Met Gly Ser 355 360 365 Glu Asn Arg Thr Cys Val Pro Glu Asn Gly Glu Gly Ala Ala Cys Ala 370 375 380 Cys Ser Met Leu Met Asp Asp Phe Val Glu Ala Asp Val Tyr Gln Leu 385 390 395 400 His Leu Trp Ala Gly Thr Gln Leu Leu Trp Ser Gly Ser Phe Lys Pro 405 410 415 Ser Ser His Val Lys Pro Arg Ala Pro Gly Asn Leu Thr Val His Pro 420 425 430 Asn Val Ser His Thr Trp Leu Leu Arg Trp Ser Asn Pro Tyr Pro Pro 435 440 445 Glu Asn His Leu His Ala Glu Leu Thr Tyr Met Val Asn Ile Ser Ser 450 455 460 Glu Asp Asp Pro Thr Asp Val Ser Val Cys Ala Ser Gly Phe Leu Cys 465 470 475 480 His Leu Leu Gly Leu Arg Arg Val Glu Thr Gly Ala Pro Gly Ala Arg 485 490 495 Leu Pro Pro Trp Leu Cys Ala Pro Arg Pro Arg Arg Val Pro Gly Ser 500 505 510 Gln Cys Ala Val Ile Ser Cys Cys Arg Trp Val Leu Ile Ala Leu Thr 515 520 525 Ser Arg Gly Gly Arg Trp Arg Leu Thr Pro Gly Leu Arg Ser Gln Thr 530 535 540 Arg Tyr Val Ser Val Ala Glu Gly Leu Phe Gly Ala Thr Pro Arg Val 545 550 555 560 Leu Cys Pro Gly Thr Gln Ala Gly Leu Ala Ser Ala Ala Arg Glu Gln 565 570 575 Met Ser Pro Asp Pro Ser Ala Phe His Ser Ile Asp Tyr Glu Pro Ser 580 585 590 Pro Lys Thr Ala Ser Thr Ile Glu Ser Lys Thr Gly Glu Cys Pro Lys 595 600 605 Cys Pro Val Pro Glu Ile Pro Gly Ala Pro Ser Val Phe Ile Phe Pro 610 615 620 Pro Lys Pro Lys Asp Thr Leu Ser Ile Ser Arg Thr Pro Glu Val Thr 625 630 635 640 Cys Leu Val Val Asp Leu Gly Pro Asp Asp Ser Asn Val Gln Ile Thr 645 650 655 Trp Phe Val Asp Asn Thr Glu Met His Thr Ala Lys Thr Arg Pro Arg 660 665 670 Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Ile 675 680 685 Leu His Gln Asp Trp Leu Lys Gly Lys Glu Phe Lys Cys Lys Val Asn 690 695 700 Ser Lys Ser Leu Pro Ser Ala Met Glu Arg Thr Ile Ser Lys Ala Lys 705 710 715 720 Gly Gln Pro His Glu Pro Gln Val Tyr Val Leu Pro Pro Thr Gln Glu 725 730 735 Glu Leu Ser Glu Asn Lys Val Ser Val Thr Cys Leu Ile Lys Gly Phe 740 745 750 His Pro Pro Asp Ile Ala Val Glu Trp Glu Ile Thr Gly Gln Pro Glu 755 760 765 Pro Glu Asn Asn Tyr Gln Thr Thr Pro Pro Gln Leu Asp Ser Asp Gly 770 775 780 Thr Tyr Phe Leu Tyr Ser Arg Leu Ser Val Asp Arg Ser His Trp Gln 785 790 795 800 Arg Gly Asn Thr Tyr Thr Cys Ser Val Ser His Glu Ala Leu His Ser 805 810 815 His His Thr Gln Lys Ser Leu Thr Gln Ser Pro Gly Lys 820 825 <210> 29 <211> 829 <212> PRT <213> Artificial Sequence <220> <223> Feline IL4R-IL13R-IgG2 (without signal sequence) <400> 29 Ser Gly Ser Val Lys Val Leu Arg Ala Pro Thr Cys Phe Ser Asp Tyr 1 5 10 15 Phe Ser Thr Ser Val Cys Gln Trp Asn Met Asp Ala Pro Thr Asn Cys 20 25 30 Ser Ala Glu Leu Arg Leu Ser Tyr Gln Leu Asn Phe Met Gly Ser Glu 35 40 45 Asn Arg Thr Cys Val Pro Glu Asn Gly Glu Gly Ala Ala Cys Ala Cys 50 55 60 Ser Met Leu Met Asp Asp Phe Val Glu Ala Asp Val Tyr Gln Leu His 65 70 75 80 Leu Trp Ala Gly Thr Gln Leu Leu Trp Ser Gly Ser Phe Lys Pro Ser 85 90 95 Ser His Val Lys Pro Arg Ala Pro Gly Asn Leu Thr Val His Pro Asn 100 105 110 Val Ser His Thr Trp Leu Leu Arg Trp Ser Asn Pro Tyr Pro Pro Glu 115 120 125 Asn His Leu His Ala Glu Leu Thr Tyr Met Val Asn Ile Ser Ser Glu 130 135 140 Asp Asp Pro Thr Asp Val Ser Val Cys Ala Ser Gly Phe Leu Cys His 145 150 155 160 Leu Leu Gly Leu Arg Arg Val Glu Thr Gly Ala Pro Gly Ala Arg Leu 165 170 175 Pro Pro Trp Leu Cys Ala Pro Arg Pro Arg Arg Val Pro Gly Ser Gln 180 185 190 Cys Ala Val Ile Ser Cys Cys Arg Trp Val Leu Ile Ala Leu Thr Ser 195 200 205 Arg Gly Gly Arg Trp Arg Leu Thr Pro Gly Leu Arg Ser Gln Thr Arg 210 215 220 Tyr Val Ser Val Ala Glu Gly Leu Phe Gly Ala Thr Pro Arg Val Leu 225 230 235 240 Cys Pro Gly Thr Gln Ala Gly Leu Ala Ser Ala Ala Arg Glu Gln Met 245 250 255 Ser Pro Asp Pro Ser Ala Phe His Ser Ile Asp Tyr Glu Pro Gly Gly 260 265 270 Gly Ser Gly Ser Ser Gln Thr Gln Pro Pro Val Thr Asn Leu Ser Val 275 280 285 Ser Val Glu Asn Leu Cys Thr Val Ile Trp Thr Trp Asp Pro Pro Glu 290 295 300 Gly Ala Ser Pro Asn Cys Thr Leu Arg Tyr Phe Ser His Phe Asp Asn 305 310 315 320 Lys Gln Asp Lys Lys Ile Ala Pro Glu Thr His Arg Ser Lys Glu Val 325 330 335 Pro Leu Asn Glu Arg Ile Cys Leu Gln Val Gly Ser Gln Cys Ser Thr 340 345 350 Asn Glu Ser Asp Asn Pro Ser Ile Leu Val Glu Lys Cys Thr Pro Pro 355 360 365 Pro Glu Gly Asp Pro Glu Ser Ala Val Thr Glu Leu Gln Cys Val Trp 370 375 380 His Asn Leu Ser Tyr Met Lys Cys Thr Trp Leu Pro Gly Arg Asn Thr 385 390 395 400 Ser Pro Asp Thr Asn Tyr Thr Leu Tyr Tyr Trp His Ser Ser Leu Gly 405 410 415 Lys Ile Leu Gln Cys Glu Asn Ile Tyr Arg Glu Gly Gln His Ile Gly 420 425 430 Cys Ser Phe Ala Leu Thr Asn Leu Lys Asp Ser Ser Phe Glu Gln His 435 440 445 Ser Val Gln Ile Val Val Lys Asp Asn Ala Gly Lys Ile Arg Pro Ser 450 455 460 Phe Asn Ile Val Pro Leu Thr Ser His Val Lys Pro Asp Pro Pro His 465 470 475 480 Ile Lys Arg Leu Phe Phe Gln Asn Gly Asn Leu Tyr Val Gln Trp Lys 485 490 495 Asn Pro Gln Asn Phe Tyr Ser Arg Cys Leu Ser Tyr Gln Val Glu Val 500 505 510 Asn Asn Ser Gln Thr Glu Thr His Asp Ile Phe Tyr Val Glu Glu Ala 515 520 525 Lys Cys Gln Asn Ser Glu Phe Glu Gly Asn Leu Glu Gly Thr Ile Cys 530 535 540 Phe Met Val Pro Gly Ile Leu Pro Asp Thr Leu Asn Thr Val Arg Ile 545 550 555 560 Arg Val Arg Thr Asn Lys Leu Cys Tyr Glu Asp Asp Arg Leu Trp Ser 565 570 575 Asn Trp Ser Gln Ala Met Ser Ile Gly Glu Asn Thr Asp Pro Thr Ser 580 585 590 Pro Lys Thr Ala Ser Thr Ile Glu Ser Lys Thr Gly Glu Cys Pro Lys 595 600 605 Cys Pro Val Pro Glu Ile Pro Gly Ala Pro Ser Val Phe Ile Phe Pro 610 615 620 Pro Lys Pro Lys Asp Thr Leu Ser Ile Ser Arg Thr Pro Glu Val Thr 625 630 635 640 Cys Leu Val Val Asp Leu Gly Pro Asp Asp Ser Asn Val Gln Ile Thr 645 650 655 Trp Phe Val Asp Asn Thr Glu Met His Thr Ala Lys Thr Arg Pro Arg 660 665 670 Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Ile 675 680 685 Leu His Gln Asp Trp Leu Lys Gly Lys Glu Phe Lys Cys Lys Val Asn 690 695 700 Ser Lys Ser Leu Pro Ser Ala Met Glu Arg Thr Ile Ser Lys Ala Lys 705 710 715 720 Gly Gln Pro His Glu Pro Gln Val Tyr Val Leu Pro Pro Thr Gln Glu 725 730 735 Glu Leu Ser Glu Asn Lys Val Ser Val Thr Cys Leu Ile Lys Gly Phe 740 745 750 His Pro Pro Asp Ile Ala Val Glu Trp Glu Ile Thr Gly Gln Pro Glu 755 760 765 Pro Glu Asn Asn Tyr Gln Thr Thr Pro Pro Gln Leu Asp Ser Asp Gly 770 775 780 Thr Tyr Phe Leu Tyr Ser Arg Leu Ser Val Asp Arg Ser His Trp Gln 785 790 795 800 Arg Gly Asn Thr Tyr Thr Cys Ser Val Ser His Glu Ala Leu His Ser 805 810 815 His His Thr Gln Lys Ser Leu Thr Gln Ser Pro Gly Lys 820 825 <210> 30 <211> 754 <212> PRT <213> Artificial Sequence <220> <223> Equine IL13R-IL4R-IgG2 (without signal sequence) <400> 30 Thr Glu Ser Gln Pro Pro Val Thr Asn Leu Ser Val Ser Val Glu Asn 1 5 10 15 Leu Cys Thr Val Ile Trp Thr Trp Asn Pro Pro Glu Gly Val Ser Pro 20 25 30 Asn Cys Ser Leu Trp Tyr Phe Ser His Phe Gly Asn Lys Gln Asp Lys 35 40 45 Lys Ile Ala Pro Glu Thr His Arg Ser Lys Glu Val Pro Leu Asn Glu 50 55 60 Arg Ile Cys Leu Gln Val Gly Ser Gln Cys Ser Thr Asn Glu Ser Asp 65 70 75 80 Asn Pro Ser Ile Leu Val Glu Lys Cys Ile Ser Pro Pro Glu Gly Asp 85 90 95 Pro Glu Ser Ala Val Thr Glu Leu Gln Cys Val Trp His Asn Leu Ser 100 105 110 Tyr Met Lys Cys Thr Trp Leu Pro Gly Lys Asn Ala Ser Pro Asp Thr 115 120 125 Asn Tyr Thr Leu Tyr Tyr Trp His Ser Ser Leu Gly Lys Ile Leu Gln 130 135 140 Cys Glu Asp Ile Tyr Arg Glu Gly Gln His Ile Gly Cys Ser Phe Ala 145 150 155 160 Leu Thr Glu Val Lys Asp Ser Ile Phe Glu Gln His Ser Val Gln Ile 165 170 175 Met Val Lys Asp Asn Ala Gly Lys Ile Arg Pro Phe Phe Asn Ile Val 180 185 190 Pro Leu Thr Ser His Val Lys Pro Asp Pro Pro His Ile Lys Lys Leu 195 200 205 Phe Phe Gln Asn Gly Asp Leu Tyr Val Gln Trp Lys Asn Pro Gln Asn 210 215 220 Phe Tyr Ser Arg Cys Leu Ser Tyr Gln Val Glu Val Asn Asn Ser Gln 225 230 235 240 Thr Glu Thr Arg Asp Ile Phe Ser Val Glu Glu Ala Lys Cys Gln Asn 245 250 255 Pro Glu Phe Glu Gly Asp Leu Glu Gly Thr Ile Cys Phe Met Val Pro 260 265 270 Gly Val Leu Pro Asp Thr Val Asn Thr Val Arg Ile Arg Val Lys Thr 275 280 285 Asn Lys Leu Cys Tyr Glu Asp Asp Lys Leu Trp Ser Asn Trp Ser Gln 290 295 300 Ala Met Ser Ile Gly Lys Lys Ala Asp Pro Thr Gly Gly Gly Ser Gly 305 310 315 320 Ser Ser Gly Ser Val Lys Val Leu His Leu Thr Ala Cys Phe Ser Asp 325 330 335 Tyr Ile Ser Ala Ser Thr Cys Glu Trp Lys Met Asp Arg Pro Thr Asn 340 345 350 Cys Ser Ala Gln Leu Arg Leu Ser Tyr Gln Leu Asn Asp Glu Phe Ser 355 360 365 Asp Asn Leu Thr Cys Ile Pro Glu Asn Arg Glu Asp Glu Val Cys Val 370 375 380 Cys Arg Met Leu Met Asp Asn Ile Val Ser Glu Asp Val Tyr Glu Leu 385 390 395 400 Asp Leu Trp Ala Gly Asn Gln Leu Leu Trp Asn Ser Ser Phe Lys Pro 405 410 415 Ser Arg His Val Lys Pro Arg Ala Pro Gln Asn Leu Thr Val His Ala 420 425 430 Ile Ser His Thr Trp Leu Leu Thr Trp Ser Asn Pro Tyr Pro Leu Lys 435 440 445 Asn His Leu Trp Ser Glu Leu Thr Tyr Leu Val Asn Ile Ser Lys Glu 450 455 460 Asp Asp Pro Thr Asp Phe Lys Ile Tyr Asn Val Thr Tyr Met Asp Pro 465 470 475 480 Thr Leu Arg Val Thr Ala Ser Thr Leu Lys Ser Arg Ala Thr Tyr Ser 485 490 495 Ala Arg Val Lys Ala Arg Ala Gln Asn Tyr Asn Ser Thr Trp Ser Glu 500 505 510 Trp Ser Pro Ser Thr Thr Trp His Asn Tyr Tyr Glu Gln Pro Asp Met 515 520 525 Ser Lys Cys Pro Lys Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser 530 535 540 Val Phe Ile Phe Pro Pro Asn Pro Lys Asp Thr Leu Met Ile Ser Arg 545 550 555 560 Thr Pro Val Val Thr Cys Val Val Val Asn Leu Ser Asp Gln Tyr Pro 565 570 575 Asp Val Gln Phe Ser Trp Tyr Val Asp Asn Thr Glu Val His Ser Ala 580 585 590 Ile Thr Lys Gln Arg Glu Ala Gln Phe Asn Ser Thr Tyr Arg Val Val 595 600 605 Ser Val Leu Pro Ile Gln His Gln Asp Trp Leu Ser Gly Lys Glu Phe 610 615 620 Lys Cys Ser Val Thr Asn Val Gly Val Pro Gln Pro Ile Ser Arg Ala 625 630 635 640 Ile Ser Arg Gly Lys Gly Pro Ser Arg Val Pro Gln Val Tyr Val Leu 645 650 655 Pro Pro His Pro Asp Glu Leu Ala Lys Ser Lys Val Ser Val Thr Cys 660 665 670 Leu Val Lys Asp Phe Tyr Pro Pro Asp Ile Ser Val Glu Trp Gln Ser 675 680 685 Asn Arg Trp Pro Glu Leu Glu Gly Lys Tyr Ser Thr Thr Pro Ala Gln 690 695 700 Leu Asp Gly Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Leu Glu 705 710 715 720 Thr Ser Arg Trp Gln Gln Val Glu Ser Phe Thr Cys Ala Val Met His 725 730 735 Glu Ala Leu His Asn His Tyr Thr Lys Thr Asp Ile Ser Glu Ser Leu 740 745 750 Gly Lys <210> 31 <211> 754 <212> PRT <213> Artificial Sequence <220> <223> Equine IL4R-IL13R-IgG2 (without signal sequence) <400> 31 Ser Gly Ser Val Lys Val Leu His Leu Thr Ala Cys Phe Ser Asp Tyr 1 5 10 15 Ile Ser Ala Ser Thr Cys Glu Trp Lys Met Asp Arg Pro Thr Asn Cys 20 25 30 Ser Ala Gln Leu Arg Leu Ser Tyr Gln Leu Asn Asp Glu Phe Ser Asp 35 40 45 Asn Leu Thr Cys Ile Pro Glu Asn Arg Glu Asp Glu Val Cys Val Cys 50 55 60 Arg Met Leu Met Asp Asn Ile Val Ser Glu Asp Val Tyr Glu Leu Asp 65 70 75 80 Leu Trp Ala Gly Asn Gln Leu Leu Trp Asn Ser Ser Phe Lys Pro Ser 85 90 95 Arg His Val Lys Pro Arg Ala Pro Gln Asn Leu Thr Val His Ala Ile 100 105 110 Ser His Thr Trp Leu Leu Thr Trp Ser Asn Pro Tyr Pro Leu Lys Asn 115 120 125 His Leu Trp Ser Glu Leu Thr Tyr Leu Val Asn Ile Ser Lys Glu Asp 130 135 140 Asp Pro Thr Asp Phe Lys Ile Tyr Asn Val Thr Tyr Met Asp Pro Thr 145 150 155 160 Leu Arg Val Thr Ala Ser Thr Leu Lys Ser Arg Ala Thr Tyr Ser Ala 165 170 175 Arg Val Lys Ala Arg Ala Gln Asn Tyr Asn Ser Thr Trp Ser Glu Trp 180 185 190 Ser Pro Ser Thr Thr Trp His Asn Tyr Tyr Glu Gln Pro Gly Gly Gly 195 200 205 Ser Gly Ser Thr Glu Ser Gln Pro Pro Val Thr Asn Leu Ser Val Ser 210 215 220 Val Glu Asn Leu Cys Thr Val Ile Trp Thr Trp Asn Pro Pro Glu Gly 225 230 235 240 Val Ser Pro Asn Cys Ser Leu Trp Tyr Phe Ser His Phe Gly Asn Lys 245 250 255 Gln Asp Lys Lys Ile Ala Pro Glu Thr His Arg Ser Lys Glu Val Pro 260 265 270 Leu Asn Glu Arg Ile Cys Leu Gln Val Gly Ser Gln Cys Ser Thr Asn 275 280 285 Glu Ser Asp Asn Pro Ser Ile Leu Val Glu Lys Cys Ile Ser Pro Pro 290 295 300 Glu Gly Asp Pro Glu Ser Ala Val Thr Glu Leu Gln Cys Val Trp His 305 310 315 320 Asn Leu Ser Tyr Met Lys Cys Thr Trp Leu Pro Gly Lys Asn Ala Ser 325 330 335 Pro Asp Thr Asn Tyr Thr Leu Tyr Tyr Trp His Ser Ser Leu Gly Lys 340 345 350 Ile Leu Gln Cys Glu Asp Ile Tyr Arg Glu Gly Gln His Ile Gly Cys 355 360 365 Ser Phe Ala Leu Thr Glu Val Lys Asp Ser Ile Phe Glu Gln His Ser 370 375 380 Val Gln Ile Met Val Lys Asp Asn Ala Gly Lys Ile Arg Pro Phe Phe 385 390 395 400 Asn Ile Val Pro Leu Thr Ser His Val Lys Pro Asp Pro Pro His Ile 405 410 415 Lys Lys Leu Phe Phe Gln Asn Gly Asp Leu Tyr Val Gln Trp Lys Asn 420 425 430 Pro Gln Asn Phe Tyr Ser Arg Cys Leu Ser Tyr Gln Val Glu Val Asn 435 440 445 Asn Ser Gln Thr Glu Thr Arg Asp Ile Phe Ser Val Glu Glu Ala Lys 450 455 460 Cys Gln Asn Pro Glu Phe Glu Gly Asp Leu Glu Gly Thr Ile Cys Phe 465 470 475 480 Met Val Pro Gly Val Leu Pro Asp Thr Val Asn Thr Val Arg Ile Arg 485 490 495 Val Lys Thr Asn Lys Leu Cys Tyr Glu Asp Asp Lys Leu Trp Ser Asn 500 505 510 Trp Ser Gln Ala Met Ser Ile Gly Lys Lys Ala Asp Pro Thr Asp Met 515 520 525 Ser Lys Cys Pro Lys Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser 530 535 540 Val Phe Ile Phe Pro Pro Asn Pro Lys Asp Thr Leu Met Ile Ser Arg 545 550 555 560 Thr Pro Val Val Thr Cys Val Val Val Asn Leu Ser Asp Gln Tyr Pro 565 570 575 Asp Val Gln Phe Ser Trp Tyr Val Asp Asn Thr Glu Val His Ser Ala 580 585 590 Ile Thr Lys Gln Arg Glu Ala Gln Phe Asn Ser Thr Tyr Arg Val Val 595 600 605 Ser Val Leu Pro Ile Gln His Gln Asp Trp Leu Ser Gly Lys Glu Phe 610 615 620 Lys Cys Ser Val Thr Asn Val Gly Val Pro Gln Pro Ile Ser Arg Ala 625 630 635 640 Ile Ser Arg Gly Lys Gly Pro Ser Arg Val Pro Gln Val Tyr Val Leu 645 650 655 Pro Pro His Pro Asp Glu Leu Ala Lys Ser Lys Val Ser Val Thr Cys 660 665 670 Leu Val Lys Asp Phe Tyr Pro Pro Asp Ile Ser Val Glu Trp Gln Ser 675 680 685 Asn Arg Trp Pro Glu Leu Glu Gly Lys Tyr Ser Thr Thr Pro Ala Gln 690 695 700 Leu Asp Gly Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Leu Glu 705 710 715 720 Thr Ser Arg Trp Gln Gln Val Glu Ser Phe Thr Cys Ala Val Met His 725 730 735 Glu Ala Leu His Asn His Tyr Thr Lys Thr Asp Ile Ser Glu Ser Leu 740 745 750 Gly Lys <210> 32 <211> 305 <212> PRT <213> Artificial Sequence <220> <223> Canine Mini-IL13R ECD <400> 32 Gln Pro Pro Val Thr Asn Leu Ser Val Ser Val Glu Asn Leu Cys Thr 1 5 10 15 Val Ile Trp Thr Trp Asp Pro Pro Glu Gly Ala Ser Pro Asn Cys Thr 20 25 30 Leu Arg Tyr Phe Ser His Phe Asp Asn Lys Gln Asp Lys Lys Ile Ala 35 40 45 Pro Glu Thr His Arg Ser Lys Glu Val Pro Leu Asn Glu Arg Ile Cys 50 55 60 Leu Gln Val Gly Ser Gln Cys Ser Thr Asn Glu Ser Asp Asn Pro Ser 65 70 75 80 Ile Leu Val Glu Lys Cys Thr Pro Pro Pro Glu Gly Asp Pro Glu Ser 85 90 95 Ala Val Thr Glu Leu Gln Cys Val Trp His Asn Leu Ser Tyr Met Lys 100 105 110 Cys Thr Trp Leu Pro Gly Arg Asn Thr Ser Pro Asp Thr Asn Tyr Thr 115 120 125 Leu Tyr Tyr Trp His Ser Ser Leu Gly Lys Ile Leu Gln Cys Glu Asp 130 135 140 Ile Tyr Arg Glu Gly Gln His Ile Gly Cys Ser Phe Ala Leu Thr Asn 145 150 155 160 Leu Lys Asp Ser Ser Phe Glu Gln His Ser Val Gln Ile Val Val Lys 165 170 175 Asp Asn Ala Gly Lys Ile Arg Pro Ser Phe Asn Ile Val Pro Leu Thr 180 185 190 Ser His Val Lys Pro Asp Pro Pro His Ile Lys Arg Leu Phe Phe Gln 195 200 205 Asn Gly Asn Leu Tyr Val Gln Trp Lys Asn Pro Gln Asn Phe Tyr Ser 210 215 220 Arg Cys Leu Ser Tyr Gln Val Glu Val Asn Asn Ser Gln Thr Glu Thr 225 230 235 240 Asn Asp Ile Phe Tyr Val Glu Glu Ala Lys Cys Gln Asn Ser Glu Phe 245 250 255 Glu Gly Asn Leu Glu Gly Thr Ile Cys Phe Met Val Pro Gly Val Leu 260 265 270 Pro Asp Thr Leu Asn Thr Val Arg Ile Arg Val Arg Thr Asn Lys Leu 275 280 285 Cys Tyr Glu Asp Asp Lys Leu Trp Ser Asn Trp Ser Gln Ala Met Ser 290 295 300 Ile 305 <210> 33 <211> 184 <212> PRT <213> Artificial Sequence <220> <223> Canine Mini-IL4R ECD <400> 33 Lys Val Leu His Glu Pro Ser Cys Phe Ser Asp Tyr Ile Ser Thr Ser 1 5 10 15 Val Cys Gln Trp Lys Met Asp His Pro Thr Asn Cys Ser Ala Glu Leu 20 25 30 Arg Leu Ser Tyr Gln Leu Asp Phe Met Gly Ser Glu Asn His Thr Cys 35 40 45 Val Pro Glu Asn Arg Glu Asp Ser Val Cys Val Cys Ser Met Pro Ile 50 55 60 Asp Asp Ala Val Glu Ala Asp Val Tyr Gln Leu Asp Leu Trp Ala Gly 65 70 75 80 Gln Gln Leu Leu Trp Ser Gly Ser Phe Gln Pro Ser Lys His Val Lys 85 90 95 Pro Arg Thr Pro Gly Asn Leu Thr Val His Pro Asn Ile Ser His Thr 100 105 110 Trp Leu Leu Met Trp Thr Asn Pro Tyr Pro Thr Glu Asn His Leu His 115 120 125 Ser Glu Leu Thr Tyr Met Val Asn Val Ser Asn Asp Asn Asp Pro Glu 130 135 140 Asp Phe Lys Val Tyr Asn Val Thr Tyr Met Gly Pro Thr Leu Arg Leu 145 150 155 160 Ala Ala Ser Thr Leu Lys Ser Gly Ala Ser Tyr Ser Ala Arg Val Arg 165 170 175 Ala Trp Ala Gln Thr Tyr Asn Ser 180 <210> 34 <211> 305 <212> PRT <213> Artificial Sequence <220> <223> Feline Mini-IL13R ECD <400> 34 Gln Pro Pro Val Thr Asn Leu Ser Val Ser Val Glu Asn Leu Cys Thr 1 5 10 15 Val Ile Trp Thr Trp Asp Pro Pro Glu Gly Ala Ser Pro Asn Cys Thr 20 25 30 Leu Arg Tyr Phe Ser His Phe Asp Asn Lys Gln Asp Lys Lys Ile Ala 35 40 45 Pro Glu Thr His Arg Ser Lys Glu Val Pro Leu Asn Glu Arg Ile Cys 50 55 60 Leu Gln Val Gly Ser Gln Cys Ser Thr Asn Glu Ser Asp Asn Pro Ser 65 70 75 80 Ile Leu Val Glu Lys Cys Thr Pro Pro Pro Glu Gly Asp Pro Glu Ser 85 90 95 Ala Val Thr Glu Leu Gln Cys Val Trp His Asn Leu Ser Tyr Met Lys 100 105 110 Cys Thr Trp Leu Pro Gly Arg Asn Thr Ser Pro Asp Thr Asn Tyr Thr 115 120 125 Leu Tyr Tyr Trp His Ser Ser Leu Gly Lys Ile Leu Gln Cys Glu Asn 130 135 140 Ile Tyr Arg Glu Gly Gln His Ile Gly Cys Ser Phe Ala Leu Thr Asn 145 150 155 160 Leu Lys Asp Ser Ser Phe Glu Gln His Ser Val Gln Ile Val Val Lys 165 170 175 Asp Asn Ala Gly Lys Ile Arg Pro Ser Phe Asn Ile Val Pro Leu Thr 180 185 190 Ser His Val Lys Pro Asp Pro Pro His Ile Lys Arg Leu Phe Phe Gln 195 200 205 Asn Gly Asn Leu Tyr Val Gln Trp Lys Asn Pro Gln Asn Phe Tyr Ser 210 215 220 Arg Cys Leu Ser Tyr Gln Val Glu Val Asn Asn Ser Gln Thr Glu Thr 225 230 235 240 His Asp Ile Phe Tyr Val Glu Glu Ala Lys Cys Gln Asn Ser Glu Phe 245 250 255 Glu Gly Asn Leu Glu Gly Thr Ile Cys Phe Met Val Pro Gly Ile Leu 260 265 270 Pro Asp Thr Leu Asn Thr Val Arg Ile Arg Val Arg Thr Asn Lys Leu 275 280 285 Cys Tyr Glu Asp Asp Arg Leu Trp Ser Asn Trp Ser Gln Ala Met Ser 290 295 300 Ile 305 <210> 35 <211> 266 <212> PRT <213> Artificial Sequence <220> <223> Feline Mini-IL4R ECD <400> 35 Lys Val Leu Arg Ala Pro Thr Cys Phe Ser Asp Tyr Phe Ser Thr Ser 1 5 10 15 Val Cys Gln Trp Asn Met Asp Ala Pro Thr Asn Cys Ser Ala Glu Leu 20 25 30 Arg Leu Ser Tyr Gln Leu Asn Phe Met Gly Ser Glu Asn Arg Thr Cys 35 40 45 Val Pro Glu Asn Gly Glu Gly Ala Ala Cys Ala Cys Ser Met Leu Met 50 55 60 Asp Asp Phe Val Glu Ala Asp Val Tyr Gln Leu His Leu Trp Ala Gly 65 70 75 80 Thr Gln Leu Leu Trp Ser Gly Ser Phe Lys Pro Ser Ser His Val Lys 85 90 95 Pro Arg Ala Pro Gly Asn Leu Thr Val His Pro Asn Val Ser His Thr 100 105 110 Trp Leu Leu Arg Trp Ser Asn Pro Tyr Pro Pro Glu Asn His Leu His 115 120 125 Ala Glu Leu Thr Tyr Met Val Asn Ile Ser Ser Glu Asp Asp Pro Thr 130 135 140 Asp Val Ser Val Cys Ala Ser Gly Phe Leu Cys His Leu Leu Gly Leu 145 150 155 160 Arg Arg Val Glu Thr Gly Ala Pro Gly Ala Arg Leu Pro Pro Trp Leu 165 170 175 Cys Ala Pro Arg Pro Arg Arg Val Pro Gly Ser Gln Cys Ala Val Ile 180 185 190 Ser Cys Cys Arg Trp Val Leu Ile Ala Leu Thr Ser Arg Gly Gly Arg 195 200 205 Trp Arg Leu Thr Pro Gly Leu Arg Ser Gln Thr Arg Tyr Val Ser Val 210 215 220 Ala Glu Gly Leu Phe Gly Ala Thr Pro Arg Val Leu Cys Pro Gly Thr 225 230 235 240 Gln Ala Gly Leu Ala Ser Ala Ala Arg Glu Gln Met Ser Pro Asp Pro 245 250 255 Ser Ala Phe His Ser Ile Asp Tyr Glu Pro 260 265 <210> 36 <211> 305 <212> PRT <213> Artificial Sequence <220> <223> Equine Mini-IL13R ECD <400> 36 Gln Pro Pro Val Thr Asn Leu Ser Val Ser Val Glu Asn Leu Cys Thr 1 5 10 15 Val Ile Trp Thr Trp Asn Pro Pro Glu Gly Val Ser Pro Asn Cys Ser 20 25 30 Leu Trp Tyr Phe Ser His Phe Gly Asn Lys Gln Asp Lys Lys Ile Ala 35 40 45 Pro Glu Thr His Arg Ser Lys Glu Val Pro Leu Asn Glu Arg Ile Cys 50 55 60 Leu Gln Val Gly Ser Gln Cys Ser Thr Asn Glu Ser Asp Asn Pro Ser 65 70 75 80 Ile Leu Val Glu Lys Cys Ile Ser Pro Pro Glu Gly Asp Pro Glu Ser 85 90 95 Ala Val Thr Glu Leu Gln Cys Val Trp His Asn Leu Ser Tyr Met Lys 100 105 110 Cys Thr Trp Leu Pro Gly Lys Asn Ala Ser Pro Asp Thr Asn Tyr Thr 115 120 125 Leu Tyr Tyr Trp His Ser Ser Leu Gly Lys Ile Leu Gln Cys Glu Asp 130 135 140 Ile Tyr Arg Glu Gly Gln His Ile Gly Cys Ser Phe Ala Leu Thr Glu 145 150 155 160 Val Lys Asp Ser Ile Phe Glu Gln His Ser Val Gln Ile Met Val Lys 165 170 175 Asp Asn Ala Gly Lys Ile Arg Pro Phe Phe Asn Ile Val Pro Leu Thr 180 185 190 Ser His Val Lys Pro Asp Pro Pro His Ile Lys Lys Leu Phe Phe Gln 195 200 205 Asn Gly Asp Leu Tyr Val Gln Trp Lys Asn Pro Gln Asn Phe Tyr Ser 210 215 220 Arg Cys Leu Ser Tyr Gln Val Glu Val Asn Asn Ser Gln Thr Glu Thr 225 230 235 240 Arg Asp Ile Phe Ser Val Glu Glu Ala Lys Cys Gln Asn Pro Glu Phe 245 250 255 Glu Gly Asp Leu Glu Gly Thr Ile Cys Phe Met Val Pro Gly Val Leu 260 265 270 Pro Asp Thr Val Asn Thr Val Arg Ile Arg Val Lys Thr Asn Lys Leu 275 280 285 Cys Tyr Glu Asp Asp Lys Leu Trp Ser Asn Trp Ser Gln Ala Met Ser 290 295 300 Ile 305 <210> 37 <211> 201 <212> PRT <213> Artificial Sequence <220> <223> Equine Mini-IL4R ECD <400> 37 Lys Val Leu His Leu Thr Ala Cys Phe Ser Asp Tyr Ile Ser Ala Ser 1 5 10 15 Thr Cys Glu Trp Lys Met Asp Arg Pro Thr Asn Cys Ser Ala Gln Leu 20 25 30 Arg Leu Ser Tyr Gln Leu Asn Asp Glu Phe Ser Asp Asn Leu Thr Cys 35 40 45 Ile Pro Glu Asn Arg Glu Asp Glu Val Cys Val Cys Arg Met Leu Met 50 55 60 Asp Asn Ile Val Ser Glu Asp Val Tyr Glu Leu Asp Leu Trp Ala Gly 65 70 75 80 Asn Gln Leu Leu Trp Asn Ser Ser Phe Lys Pro Ser Arg His Val Lys 85 90 95 Pro Arg Ala Pro Gln Asn Leu Thr Val His Ala Ile Ser His Thr Trp 100 105 110 Leu Leu Thr Trp Ser Asn Pro Tyr Pro Leu Lys Asn His Leu Trp Ser 115 120 125 Glu Leu Thr Tyr Leu Val Asn Ile Ser Lys Glu Asp Asp Pro Thr Asp 130 135 140 Phe Lys Ile Tyr Asn Val Thr Tyr Met Asp Pro Thr Leu Arg Val Thr 145 150 155 160 Ala Ser Thr Leu Lys Ser Arg Ala Thr Tyr Ser Ala Arg Val Lys Ala 165 170 175 Arg Ala Gln Asn Tyr Asn Ser Thr Trp Ser Glu Trp Ser Pro Ser Thr 180 185 190 Thr Trp His Asn Tyr Tyr Glu Gln Pro 195 200 <210> 38 <211> 4 <212> PRT <213> Artificial sequence <220> <223> GS connector <400> 38 Gly Ser Gly Ser 1 <210> 39 <211> 6 <212> PRT <213> Artificial sequence <220> <223> GS connector <400> 39 Gly Ser Gly Ser Gly Ser 1 5 <210> 40 <211> 6 <212> PRT <213> Artificial sequence <220> <223> GS connector <400> 40 Gly Gly Ser Gly Gly Ser 1 5 <210> 41 <211> 9 <212> PRT <213> Artificial sequence <220> <223> GS connector <400> 41 Gly Gly Ser Gly Gly Ser Gly Gly Ser 1 5 <210> 42 <211> 4 <212> PRT <213> Artificial sequence <220> <223> GS connector <400> 42 Gly Gly Gly Ser 1 <210> 43 <211> 8 <212> PRT <213> Artificial sequence <220> <223> GS connector <400> 43 Gly Gly Gly Ser Gly Gly Gly Ser 1 5 <210> 44 <211> 12 <212> PRT <213> Artificial sequence <220> <223> GS connector <400> 44 Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser 1 5 10 <210> 45 <211> 6 <212> PRT <213> Artificial sequence <220> <223> GS connector <400> 45 Gly Ser Ser Gly Ser Ser 1 5 <210> 46 <211> 9 <212> PRT <213> Artificial sequence <220> <223> GS connector <400> 46 Gly Ser Ser Gly Ser Ser Gly Ser Ser 1 5 <210> 47 <211> 4 <212> PRT <213> Artificial sequence <220> <223> GS connector <400> 47 Gly Gly Ser Ser 1 <210> 48 <211> 8 <212> PRT <213> Artificial sequence <220> <223> GS connector <400> 48 Gly Gly Ser Ser Gly Gly Ser Ser 1 5 <210> 49 <211> 12 <212> PRT <213> Artificial sequence <220> <223> GS connector <400> 49 Gly Gly Ser Ser Gly Gly Ser Ser Gly Gly Ser Ser 1 5 10 BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 It is a graph of the sequential binding of canine IL4R-IL13R-Fc to canine IL4 and IL13 or canine IL13 and IL4 using a concentration of 30 μg / mL of IL4 and IL13 in PBS.

[0032] Figure 2 It is a graph showing the sequential binding of canine IL13R-IL4R-Fc to canine IL4 and IL13 or canine IL13 and IL4 using a concentration of 30 μg / mL of IL4 and IL13 in PBS.

[0033] Figure 3 Graph showing neutralization of canine IL4 activity by canine IL4R-IL13R-Fc in a TF1 cell proliferation assay. Canine IL4 (50 ng / mL or 3.85 nM) was used in the assay.

[0034] Sequence Description

[0035] Table 1 provides a listing of certain sequences mentioned herein.

[0036]

[0037]

[0038]

[0039]

[0040]

[0041]

[0042]

[0043]

[0044]

[0045]

[0046]

[0047]

[0048]

[0049]

[0050]

[0051]

[0052]

[0053]

[0054]

[0055]

[0056]

[0057]

[0058] Description of the implementation plan

[0059] A continuous polypeptide that binds canine IL13 and / or IL4, feline IL13 and / or IL4, and / or equine IL13 and / or IL4 is provided. In some embodiments, the continuous polypeptide comprises an extracellular domain of an IL13R polypeptide and an extracellular domain of an IL4R polypeptide. A method for producing or purifying a continuous polypeptide is also provided. A method for treating using a continuous polypeptide to bind IL13 and / or IL4 and inhibit IL13- and / or IL-4-mediated signaling is provided. Such methods include, but are not limited to, methods for treating IL13- and / or IL4-induced conditions in companion animal species. A method for detecting IL13 and / or IL4 in a sample from a companion animal species is also provided.

[0060] For the convenience of the reader, the following definitions of terms used herein are provided.

[0061] As used herein, numerical terms such as Kd are calculated based on scientific measurements and therefore have measurement errors. In some cases, numerical terms may include values ​​rounded to the nearest significant figure.

[0062] As used herein, "a" or "an" means "at least one" or "one or more" unless otherwise specified. As used herein, the term "or" means "and / or" unless otherwise specified. In the context of multiple dependent claims, the use of "or" when it refers to prior other claims refers only to those claims in the alternative.

[0063] IL13R / IL4R continuous peptide

[0064] Novel IL13R / IL4R contiguous polypeptides are provided, for example, contiguous polypeptides that bind canine IL13 and / or IL4, feline IL13 and / or IL4, and / or equine IL13 and / or IL4.

[0065] "Amino acid sequence" means the sequence of amino acid residues in a peptide or protein. The terms "polypeptide" and "protein" are used interchangeably to refer to polymers of amino acid residues, without limiting the minimum length. Such polymers of amino acid residues may contain natural or non-natural amino acid residues, and include, but are not limited to, peptides, oligopeptides, dimers, trimers and polymers of amino acid residues. The definition includes both full-length proteins and fragments thereof. The term also includes post-expression modifications of polypeptides, such as glycosylation, sialylation, acetylation, phosphorylation, etc. In addition, for the purposes of this disclosure, "polypeptide" refers to proteins including modifications to the native sequence, such as deletions, additions and substitutions (usually conservative substitutions in nature), as long as the protein maintains the desired activity. These modifications may be intentional, such as by site-directed mutagenesis, or may be accidental, such as by mutations of the host producing the protein or errors caused by PCR amplification.

[0066] The term "continuous polypeptide" herein is used to refer to an uninterrupted sequence of amino acids. A continuous polypeptide is usually translated from a single continuous DNA sequence. It can be made by genetic engineering, for example, by removing the stop codon from the DNA sequence of a first protein and then appending the DNA sequence of a second protein in frame so that the DNA sequence is expressed as a single protein. Typically, this is accomplished by cloning a cDNA into a frame with an existing gene in an expression vector.

[0067] As used herein, "IL4R" is a polypeptide comprising all or a fragment of the IL4 receptor subunit alpha that binds IL-4.

[0068] For example, unless otherwise indicated, "IL4R" refers to an IL4R polypeptide from any vertebrate source, including mammals such as primates (e.g., humans and cynomolgus monkeys), rodents (e.g., mice and rats), and companion animals (e.g., dogs, cats, and horses). In some embodiments, IL4R is an extracellular domain fragment that binds IL4. In some such embodiments, IL4R may be referred to as an IL4R extracellular domain (ECD). In some embodiments, IL4R comprises the amino acid sequence of SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 33, SEQ ID NO: 35, or SEQ ID NO: 37.

[0069] As used herein, "IL13R" is a polypeptide comprising all or a portion of the IL13 receptor subunit alpha-1 that binds IL-13.

[0070] For example, unless otherwise indicated, "IL13R" refers to an IL13R polypeptide from any vertebrate source, including mammals such as primates (e.g., humans and cynomolgus monkeys), rodents (e.g., mice and rats), and companion animals (e.g., dogs, cats, and horses). In some embodiments, IL13R is an extracellular domain fragment that binds IL13. In some such embodiments, IL13R may be referred to as an IL13R extracellular domain (ECD). In some embodiments, the IL13R polypeptide comprises the amino acid sequence of SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 32, SEQ ID NO: 34, or SEQ ID NO: 36.

[0071] The term "companion animal species" refers to animals suitable for the companionship of humans. In some embodiments, companion animal species are small mammals, such as canines, felines, dogs, cats, horses, rabbits, ferrets, guinea pigs, rodents, etc. In some embodiments, companion animal species are farm animals, such as horses, cows, pigs, etc.

[0072] An "extracellular domain" ("ECD") is a portion of a polypeptide that extends beyond the transmembrane domain into the extracellular space. As used herein, the term "extracellular domain" may include a complete extracellular domain or may include a truncated extracellular domain that binds its ligand lacking one or more amino acids. The composition of the extracellular domain may depend on the algorithm used to determine which amino acids are within the membrane. For a given protein, different algorithms may predict, and different systems may express, different extracellular domains.

[0073] The extracellular domain of the IL4R polypeptide may comprise the entire extracellular domain of IL4R or a truncated extracellular domain that binds IL4. As used herein, the terms "extracellular domain of an IL4R polypeptide," "IL4R ECD," and similar terms refer to an IL4R polypeptide that does not comprise a transmembrane domain or a cytoplasmic domain, even if the term is followed by an open transition word (such as "comprising," "comprises," etc.). In some embodiments, the extracellular domain of an IL4R polypeptide is an extracellular domain of an IL4R polypeptide derived from an animal of a companion species. For example, in some embodiments, the extracellular domain of an IL4R polypeptide is derived from a canine IL4R, a feline IL4R, or an equine IL4R. In some embodiments, the extracellular domain of an IL4R polypeptide comprises the amino acid sequence of SEQ ID NO: 23, SEQ ID NO: 25, or SEQ ID NO: 27, or any fragment thereof. In some embodiments, the extracellular domain of an IL4R polypeptide comprises the amino acid sequence of SEQ ID NO: 33, SEQ ID NO: 35, or SEQ ID NO: 37, or any fragment thereof.

[0074] The extracellular domain of the IL13R polypeptide may comprise the complete extracellular domain of IL13R or a truncated extracellular domain that binds IL13. As used herein, the term "extracellular domain of an IL13R polypeptide," "IL13R ECD," and similar terms refer to an IL13R polypeptide that does not comprise a transmembrane domain or a cytoplasmic domain, even if the term is followed by an open transition word such as "comprising," "comprises," etc. In some embodiments, the extracellular domain of an IL13R polypeptide is an extracellular domain of an IL13R polypeptide derived from an animal of a companion species. For example, in some embodiments, the extracellular domain of an IL13R polypeptide is derived from a canine IL13R, a feline IL13R, or an equine IL13R. In some embodiments, the extracellular domain of an IL13R polypeptide comprises the amino acid sequence of SEQ ID NO: 22, SEQ ID NO: 24, or SEQ ID NO: 26, or any fragment thereof. In some embodiments, the extracellular domain of the IL13R polypeptide comprises the amino acid sequence of SEQ ID NO: 32, SEQ ID NO: 34, or SEQ ID NO: 36, or any fragment thereof.

[0075] The terms "IL13R / IL4R consecutive polypeptide" and "IL4R / IL13R consecutive polypeptide" are used interchangeably to refer to a consecutive polypeptide comprising an IL13R polypeptide and an IL4R polypeptide, wherein the terms do not indicate the order in which the IL13R and IL4R polypeptides appear in the consecutive polypeptide unless the order indicates otherwise. For example, an IL13R / IL4R consecutive polypeptide or an IL4R / IL13R consecutive polypeptide may refer to an IL4R polypeptide that sequentially precedes or follows an IL13R polypeptide. In addition, an IL13R / IL4R consecutive polypeptide or an IL4R / IL13R consecutive polypeptide may refer to an IL13R polypeptide that sequentially precedes or follows an IL4R polypeptide.

[0076] In some embodiments, the IL13R / IL4R contiguous polypeptide comprises an IL13R polypeptide bound to an IL4R polypeptide at the C-terminus of the IL13R polypeptide or at the N-terminus of the IL13R polypeptide. In some embodiments, the IL13R / IL4R contiguous polypeptide comprises an IL4R polypeptide bound to an IL13R polypeptide at the C-terminus of the IL4R polypeptide or at the N-terminus of the IL4R polypeptide.

[0077] The IL13R / IL4R contiguous polypeptides of the invention may comprise an extracellular domain of an IL13R polypeptide and / or an extracellular domain of an IL4R polypeptide, wherein the polypeptides are derived from a companion animal species. For example, the contiguous polypeptide may comprise an extracellular domain of an IL4R polypeptide from a dog, a cat or a horse and / or may comprise an extracellular domain of an IL13R polypeptide from a dog, a cat or a horse.

[0078] "Wild-type" refers to a non-mutated form of a polypeptide or a fragment thereof that occurs in nature. A wild-type polypeptide can be produced recombinantly. "Wild-type IL13R ECD" or "wild-type IL4R ECD" refers to a protein having an amino acid sequence identical to the same portion of the extracellular domain of IL13R or IL4R that occurs in nature.

[0079] A "variant" is a nucleic acid molecule or polypeptide that differs from a reference nucleic acid molecule or polypeptide by single or multiple amino acid substitutions, deletions, and / or additions, and substantially retains at least one biological activity of the reference nucleic acid molecule or polypeptide.

[0080] "Biologically active" entities or entities with "biological activity" are entities with any function related or associated with a metabolic or physiological process and / or with a structural function, regulatory function or biochemical function of a naturally occurring molecule. Biologically active polynucleotide fragments are those polynucleotide fragments that exhibit an activity similar to, but not necessarily identical to, the activity of the polynucleotides of the present invention. Biologically active polypeptides or fragments thereof include polypeptides or fragments thereof that can participate in biological reactions (including, but not limited to, ligand-receptor interactions or antigen-antibody binding). Biological activity may include improved desired activity, or reduced undesired activity. When an entity participates in a molecular interaction (such as hybridization) with another molecule, when an entity has therapeutic value for alleviating a disease condition, when an entity has preventive value for inducing an immune response, when an entity has diagnostic and / or prognostic value in determining the presence of a molecule (such as a biologically active fragment of a polynucleotide, which can be detected as being unique to the polynucleotide molecule), and when an entity can be used as a primer in a polymerase chain reaction (PCR), the entity can be shown to have biological activity.

[0081] As used herein, "percent (%) amino acid sequence identity" and "homology" with respect to nucleic acid molecules or polypeptide sequences are defined as the percentage of nucleotides or amino acid residues in a reference sequence that are identical to the nucleotides or amino acid residues in a particular nucleic acid molecule or polypeptide sequence, after aligning the sequences and introducing gaps (if necessary) to obtain the maximum percentage of sequence identity, and without considering any conservative substitutions as part of the sequence identity. Alignment for the purpose of determining percentage sequence identity can be achieved in a variety of ways within the capabilities of those skilled in the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN or MEGALINETM (DNASTAR) software. Those skilled in the art can determine appropriate parameters for measuring alignment, including any algorithm required to achieve maximum alignment over the full length of the sequence being compared.

[0082] In some embodiments, after aligning the sequences and introducing gaps (if necessary) to achieve maximum percentage sequence identity, and without considering any conservative substitutions as part of the sequence identity, the variant has at least about 50% sequence identity with the reference nucleic acid molecule or polypeptide. Such variants include, for example, polypeptides in which one or more amino acid residues are added or deleted at the N-terminus or C-terminus of the polypeptide. In some embodiments, the variant has at least about 50% sequence identity, at least about 60% sequence identity, at least about 65% sequence identity, at least about 70% sequence identity, at least about 75% sequence identity, at least about 80% sequence identity, at least about 85% sequence identity, at least about 90% sequence identity, at least about 95% sequence identity, at least about 98% sequence identity with the sequence of the reference nucleic acid or polypeptide.

[0083] In some embodiments, the IL13R / IL4R contiguous polypeptide comprises an extracellular domain of an IL13R polypeptide having at least 85%, at least 90%, at least 95%, at least 98% sequence identity to the amino acid sequence of SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 32, SEQ ID NO: 34, or SEQ ID NO: 36. In some embodiments, the IL13R / IL4R contiguous polypeptide comprises an extracellular domain of an IL4R polypeptide having at least 85%, at least 90%, at least 95%, at least 98% sequence identity to the amino acid sequence of SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 33, SEQ ID NO: 35, or SEQ ID NO: 37.

[0084] In some embodiments, the IL13R / IL4R contiguous polypeptide comprises an extracellular domain of an IL13R polypeptide comprising a cysteine ​​at a position corresponding to position 18 of SEQ ID NO: 22, at a position corresponding to position 18 of SEQ ID NO: 24, or at a position corresponding to position 18 of SEQ ID NO: 26. In some embodiments, the IL13R / IL4R contiguous polypeptide comprises an extracellular domain of an IL13R polypeptide comprising a cysteine ​​at position 18 of SEQ ID NO: 22, at position 18 of SEQ ID NO: 24, at position 18 of SEQ ID NO: 26, at position 15 of SEQ ID NO: 32, at position 15 of SEQ ID NO: 34, or at position 15 of SEQ ID NO: 36.

[0085] A "point mutation" is a mutation involving a single nucleotide or amino acid residue. The mutation may be a loss of a nucleotide or amino acid, a substitution of a nucleotide or amino acid residue for another nucleotide or amino acid, or an insertion of another nucleotide or amino acid residue.

[0086] Amino acid substitutions may include, but are not limited to, replacing an amino acid in a polypeptide with another amino acid. Exemplary substitutions are shown in Table 2. Amino acid substitutions may be introduced into a molecule of interest, and the product may be screened for a desired activity (e.g., retained / improved antigen binding, reduced immunogenicity, or improved ADCC or CDC, or enhanced pharmacokinetics).

[0087] Table 2

[0088]

[0089]

[0090] Amino acids can be grouped according to common side chain properties:

[0091] (1) Hydrophobicity: norleucine, Met, Ala, Val, Leu, Ile;

[0092] (2) Neutral hydrophilicity: Cys, Ser, Thr, Asn, Gln;

[0093] (3) Acidic: Asp, Glu;

[0094] (4) Basic: His, Lys, Arg;

[0095] (5) Residues that affect chain direction: Gly, Pro;

[0096] (6) Aromaticity: Trp, Tyr, Phe.

[0097] Non-conservative substitutions will involve exchanging a member of one of these classes for another class.

[0098] As used herein, "fusion partner" refers to an additional component of the IL13R / IL4R contiguous polypeptide, such as an additional polypeptide, such as albumin, an albumin binding fragment, or a fragment of an immunoglobulin molecule. The fusion partner may comprise an oligomerization domain, such as the Fc domain of a heavy chain immunoglobulin.

[0099] A "fragment crystallizable (Fc) polypeptide" is a portion of an antibody molecule that interacts with effector molecules and cells. It comprises the C-terminal portion of an immunoglobulin heavy chain. As used herein, an Fc polypeptide comprises a fragment of an Fc domain that has one or more biological activities of an intact Fc polypeptide. An "effector function" of an Fc polypeptide is any action or activity that an antibody performs in whole or in part in response to a stimulus, and may include complement fixation or ADCC (antibody-dependent cellular cytotoxicity) induction.

[0100] The term "IgX" or "IgX Fc" means that the Fc region is derived from a specific antibody isotype (e.g., IgG, IgA, IgD, IgE, IgM, etc.), where "X" represents the antibody isotype. Thus, "IgG" or "IgGFc" represents the Fc region of the γ chain, "IgA" or "IgA Fc" represents the Fc region of the α chain, "IgD" or "IgD Fc" represents the Fc region of the δ chain, "IgE" or "IgE Fc" represents the Fc region of the ε chain, "IgM" or "IgM Fc" represents the Fc region of the μ chain, etc. In some embodiments, the IgG Fc region comprises CH1, hinge, CH2, CH3 and CL1. "IgXN" or "IgXN Fc" indicates that the Fc region is derived from an antibody isotype of a particular subclass (such as canine IgG subclass A, B, C or D; feline IgG subclass 1, 2a or 2b; or equine IgG subclass IgG1, IgG2, IgG3, IgG4, IgG5, IgG6 or IgG7, etc.), where "N" indicates the subclass.

[0101] In some embodiments, IgX or IgXN district is derived from companion animal, such as dog, cat or horse. In some embodiments, IgG district is separated from dog γ heavy chain (such as IgGA, IgGB, IgGC or IgGD). In some cases, IgG Fc district is separated from cat γ heavy chain (such as IgG1, IgG2a or IgG2b). In other cases, IgG district is separated from horse γ heavy chain (such as IgG1, IgG2, IgG3, IgG4, IgG5, IgG6 or IgG7). The polypeptide comprising the Fc district of IgGA, IgGB, IgGC or IgGD can provide higher expression level in the recombinant production system.

[0102] "Signal sequence" refers to an amino acid residue sequence, or a polynucleotide encoding such an amino acid residue sequence, that promotes the secretion of a polypeptide of interest and is usually cleaved after the polypeptide is exported to the outside of the cell surface membrane.

[0103] "Linker" refers to one or more amino acid residues that connect a first polypeptide to a second polypeptide.

[0104] In some embodiments, the linker is a flexible, non-structural linker that is glycine-rich, serine-rich, or GS-rich. In some embodiments, the linker comprises the amino acids G (Gly) and / or S (Ser). For example, the linker can comprise G or repeats of G (e.g., GG, GGG, etc.); GS or repeats of GS (e.g., GSGS (SEQ ID NO:38), GSGSGS (SEQ ID NO:39), etc.); GGS or repeats of GGS (e.g., GGSGGS (SEQ ID NO:40), GGSGGSGGS (SEQ ID NO:41), etc.); GGGS (SEQ ID NO:42) or repeats of GGGS (SEQ ID NO:42) (e.g., GGGSGGGS (SEQ ID NO:43), GGGSGGGSGGGS (SEQ ID NO:44), etc.); GSS or repeats of GSS (e.g., GSSGSS (SEQ ID NO:45), GSSGSSGSS (SEQ ID NO:46), etc.); or GGSS (SEQ ID NO:47) or repeats of GGSS (SEQ ID NO:47) (e.g., GGSSGGSS (SEQ ID NO:48), GGSSGGSSGGSS (SEQ ID NO:49), etc.). NO:49) etc.).

[0105] In some embodiments, the IL13R / IL4R continuous polypeptide comprises at least one fusion partner and / or at least one linker. In some embodiments, the IL13R / IL4R continuous polypeptide comprises an optional signal sequence, an optional fusion partner, and at least one optional linker. In some embodiments, the IL13R / IL4R continuous polypeptide does not comprise a signal sequence, a fusion partner, or a linker. In some embodiments, the IL13 / IL4 continuous polypeptide is translated to have a signal sequence, but the signal sequence is cleaved from the IL13R / IL4R continuous polypeptide.

[0106] In some embodiments, the IL13R / IL4R contiguous polypeptide comprises:

[0107] Formula (I): IL13R-L1-IL4R-L2-FP or

[0108] Formula (II): IL4R-L1-IL13R-L2-FP,

[0109] wherein IL13R is an IL13R extracellular domain (ECD) polypeptide derived from a companion animal species, IL4R is an IL4R ECD polypeptide derived from a companion animal species, L1 is a first optional linker, L2 is a second optional linker, and FP is an optional fusion partner.

[0110] In some embodiments, the IL13R / IL4R contiguous polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31.

[0111] IL13R / IL4R continuous polypeptide expression and production

[0112] Provided are polynucleotide sequences encoding all or part of an IL13R / IL4R contiguous polypeptide (e.g., an extracellular domain) with or without a signal sequence. If a homologous signal sequence (i.e., a signal sequence of a native IL-4R or IL13R) is not used in the construction of the nucleic acid molecule, another signal sequence may be used, for example, any of the signal sequences described in PCT US06 / 02951.

[0113] Typically, a nucleotide sequence encoding a polypeptide of interest (such as an IL13R / IL4R contiguous polypeptide) is inserted into an expression vector suitable for expression in a selected host cell.

[0114] A "vector" is a plasmid that can be used to transfer a DNA sequence from one organism to another or to express a gene of interest. A vector typically includes a replication origin and regulatory sequences that regulate the expression of the gene of interest, and may or may not carry a selectable marker gene, such as an antibiotic resistance gene. The vector is suitable for use in a host cell in which the vector is to be expressed. When a gene of interest is present in a vector, the vector may be referred to as a "recombinant vector."

[0115] "Host cell" refers to a cell that can be or has been a recipient of a vector or isolated polynucleotide. Host cells can be prokaryotic or eukaryotic. Exemplary eukaryotic cells include mammalian cells, such as primate or non-primate cells; fungal cells, such as yeast; plant cells; and insect cells. Non-limiting exemplary mammalian cells include, but are not limited to, NS0 cells, Host cells include cells (Crucell), 293 cells and CHO cells, and derivatives thereof, such as 293-6E, DG44, CHO-S and CHO-K cells. Host cells include progeny of a single host cell, and due to natural, accidental or deliberate mutations, the progeny are not necessarily identical to the original parent cell (in morphology or in genomic DNA complement). Host cells include cells transfected in vivo with one or more polynucleotides encoding one or more amino acid sequences provided herein.

[0116] The term "isolated" as used herein refers to a molecule that has been separated from at least some of the components commonly found or produced in nature. For example, a polypeptide is said to be "isolated" when it is separated from at least some of the components of the cell that produced it. In the case where the polypeptide is secreted by a cell after expression, the supernatant containing the polypeptide is physically separated from the cell that produced it, and is considered to be "isolating" the polypeptide. Similarly, a polynucleotide is said to be "isolated" when it is not part of a larger polynucleotide commonly found in nature (such as, for example, genomic DNA or mitochondrial DNA in the case of a DNA polynucleotide), or when it is separated from at least some of the components of the cell that produced it (for example, in the case of an RNA polynucleotide). Therefore, a DNA polynucleotide contained in a vector within a host cell can be referred to as "isolated".

[0117] In some embodiments, the IL13R / IL4R contiguous polypeptides are separated using chromatography, such as size exclusion chromatography, ion exchange chromatography, protein A column chromatography, hydrophobic interaction chromatography, and CHT chromatography.

[0118] The terms "label" and "detectable label" mean a moiety attached to an IL13R / IL4R contiguous polypeptide to render it detectable. In some embodiments, the label is a detectable marker that produces a signal that can be detected visually or instrumentally, for example, a polypeptide incorporating a radiolabeled amino acid or attached to a biotin moiety that can be detected by labeled avidin (e.g., streptavidin containing a fluorescent marker or enzymatic activity that can be detected by optical or colorimetric methods). Examples of labels for polypeptides include, but are not limited to, the following: a radioisotope or radionuclide (e.g., 3 H. 14 C. 35 S. 90 Y. 99 Tc, 111 In, 125 I. 131 I. 177 Lu, 166 Ho or 153Sm); chromogens, fluorescent labels (e.g., FITC, rhodamine, lanthanide phosphors), enzyme labels (e.g., horseradish peroxidase, luciferase, alkaline phosphatase); chemiluminescent labels; biotinyl groups; predicted polypeptide epitopes recognized by secondary reporter molecules (e.g., leucine zipper pair sequences, binding sites of secondary antibodies, metal binding domains, epitope tags); and magnetic agents, such as gadolinium chelates. Representative examples of labels commonly used in immunoassays include light-generating moieties (e.g., acridine compounds) and fluorescence-generating moieties, such as fluorescein. In this regard, the moiety itself may not be detectably labeled, but may become detectable after reacting with another moiety.

[0119] IL13R / IL4R contiguous peptides as decoy receptor traps

[0120] The IL13R / IL4R contiguous polypeptides of the present invention can act as decoy receptors for capturing IL13 or IL4 and inhibiting their interaction with IL13R and IL4R on the cell surface. Decoy receptors, such as the decoy receptors of the present invention, recognize their ligands with high affinity and specificity, but are structurally incapable of signal transduction. They compete with wild-type receptors for ligand binding and participate in ligand / receptor interactions, thereby regulating the activity or number of functional receptors and / or cellular activity downstream of the receptors. Decoy receptors can act as molecular traps for agonistic ligands and thereby inhibit ligand-induced receptor activation.

[0121] As used herein, "IL13" refers to any natural IL13 produced from the expression and processing of IL13 in cells. Unless otherwise indicated, the term includes IL13 from any vertebrate, including mammals, such as primates (e.g., humans and cynomolgus monkeys) and rodents (e.g., mice and rats) and companion animals (e.g., dogs, cats and horses). The term also includes naturally occurring variants of IL13, e.g., splice variants or allelic variants.

[0122] In some embodiments, canine IL13 comprises the amino acid sequence of SEQ ID NO: 4. In some embodiments, feline IL13 comprises the amino acid sequence of SEQ ID NO: 5. In some embodiments, equine IL13 comprises the amino acid sequence of SEQ ID NO: 6.

[0123] As used herein, "IL4" refers to any natural IL4 produced from the expression and processing of IL14 in cells. Unless otherwise indicated, the term includes IL4 from any vertebrate, including mammals, such as primates (e.g., humans and cynomolgus monkeys) and rodents (e.g., mice and rats) and companion animals (e.g., dogs, cats and horses). The term also includes naturally occurring variants of IL4, e.g., splice variants or allelic variants.

[0124] In some embodiments, canine IL4 comprises the amino acid sequence of SEQ ID NO: 1. In some embodiments, feline IL4 comprises the amino acid sequence of SEQ ID NO: 2. In some embodiments, equine IL4 comprises the amino acid sequence of SEQ ID NO: 3.

[0125] The present invention provides IL13R / IL4R contiguous polypeptides as therapeutic agents. The IL13R / IL4R contiguous polypeptides of the present invention bind IL13 and / or IL4, which, as described in more detail herein, have been shown to be associated with allergic diseases. In various embodiments, the IL13R / IL4R contiguous polypeptides can bind IL13 and IL4 with very high affinity. In various embodiments, the IL13R / IL4R contiguous polypeptides can interfere with IL13 and IL4 signaling.

[0126] The term "affinity" refers to the sum of the strengths of non-covalent interactions between a single binding site of a molecule (e.g., a receptor) and its binding partner (e.g., a ligand). The affinity of a molecule X for its partner Y can generally be expressed by the dissociation constant (K D ) Affinity can be measured by common methods known in the art, such as, for example, immunoblotting, ELISA KD, KinEx A, biolayer interference assay (BLI), or surface plasmon resonance instrumentation.

[0127] The term "K D ”, “K d "," Kd "or "Kd value" refers to the equilibrium dissociation constant of a receptor fusion-ligand interaction. In some embodiments, the K of a fusion molecule for its ligand is d The biofilm interference assay was performed using a biosensor (such as The binding rate constant (k) was determined using the ForteBio system (Pall ForteBio LLC, Fremont, CA). Briefly, biotinylated antigen was bound to the sensor tip and the binding of the fusion molecule was monitored for 90 seconds and the dissociation was monitored for 600 seconds. The buffer used for the dilution and binding steps was 20 mM phosphate, 150 mM NaCl, pH 7.2. A blank curve of buffer only was subtracted to correct for any drift. The data were fitted to a 2:1 binding model using ForteBio data analysis software to determine the binding rate constant (k on ), dissociation rate constant (k off ) and Kd. Equilibrium dissociation constant (K d ) is calculated as k off / k on The term “k on ” refers to the rate constant for the binding of molecule X to its partner Y, and the term “k off ” refers to the rate constant for the dissociation of molecule X or partner Y from the molecule X / partner Y complex.

[0128] The term "binding" to a substance is a well-known term in the art, and methods for determining such binding are also well-known in the art. A molecule is said to exhibit "binding" if it reacts, binds, or has affinity for a specific cell or substance, and the reaction, binding, or affinity can be detected by one or more methods known in the art (such as, for example, immunoblotting, ELISA KD, KinEx A, biofilm layer interference assay (BLI), surface plasmon resonance equipment, or the like).

[0129] "Surface plasmon resonance" refers to an optical phenomenon that can be used to analyze real-time biospecific interactions by detecting changes in protein concentration in a biosensor matrix, such as using BIAcore TM System (BIAcore International AB, GE Healthcare, Uppsala, Sweden and Piscataway, NJ). For further description, see Jonsson et al. (1993) Ann. Biol. Clin. 51: 19-26.

[0130] "Biofilm interferometry" refers to an optical analysis technique that analyzes the interference pattern of light reflected from an immobilized protein layer on a biosensor tip and an internal reference layer. Changes in the number of molecules bound to the biosensor tip result in a shift in the interference pattern that can be measured in real time. A non-limiting exemplary device for biofilm interferometry is System (Pall ForteBio LLC). See, e.g., Abdiche et al., 2008, Anal. Biochem. 377: 209-277.

[0131] In some embodiments, the IL13R / IL4R contiguous polypeptide binds canine IL13 and / or IL4, feline IL13 and / or IL4, or equine IL13 and / or IL4 with a dissociation constant (Kd) measured by a biofilm interference assay of less than 5 x 10 -6 M, less than 1x 10 -6 M, less than 5x 10 -7 M, less than 1x 10 -7 M, less than 5x 10 -8 M, less than 1x 10 -8 M, less than 5x 10 -9 M, less than 1x 10 -9 M, less than 5x10 -10 M, less than 1x 10 -10 M, less than 5x 10 -11 M, less than 1x 10 -11 M, less than 5x 10 -12 M, or less than 1x 10 -12 In some embodiments, the IL13R / IL4R contiguous polypeptide binds canine IL13 and / or IL4, feline IL13 and / or IL4, or equine IL13 and / or IL4 with a dissociation constant (Kd) measured by a biofilm interference assay of between 5 x 10 - 6 M and 1x 10 -6 Between M, between 5x10 -6 M and 5x 10 -7 M, between 5x 10 -6 M and 1x 10 -7 M, between 5x 10 - 6 M and 5x 10 -8 Between M, 5x 10 -6 M and 1x 10 -8 M, between 5x 10 -6 M and 5x 10 -9 M, between 5x 10 -6 M and 1x10 -9 M, between 5x 10 -6 M and 5x 10 -10 M, between 5x 10 -6 M and 1x 10 -10 M, between 5x 10-6 M and 5x10 -11 M, between 5x 10 -6 M and 1x 10 -11 M, between 5x 10 -6 M and 5x 10 -12 M, between 5x 10 -6 M and 1x 10 -12 M, between 1x 10 -6 M and 5x 10 -7 M, between 1x 10 -6 M and 1x 10 -7 M, between 1x 10 -6 M and 5x 10 -8 Between M, 1x 10 -6 M and 1x 10 -8 M, between 1x 10 -6 M and 5x 10 -9 M, between 1x 10 -6 M and 1x10 -9 M, between 1x 10 -6 M and 5x 10 -10 M, between 1x 10 -6 M and 1x 10 -10 M, between 1x 10 -6 M and 5x10 -11 M, between 1x 10 -6 M and 1x 10 -11 M, between 1x 10 -6 M and 5x 10 -12 M, between 1x 10 -6 M and 1x 10 -12 M, between 5x 10 -7 M and 1x 10 -7 M, between 5x 10 -7 M and 5x 10 -8 Between M, 5x 10 -7 M and 1x10 -8 M, between 5x 10 -7 M and 5x 10 -9 M, between 5x 10 -7 M and 1x 10 -9 M, between 5x 10 -7 M and 5x10 -10 M, between 5x 10 -7 M and 1x 10 -10 M, between 5x 10-7 M and 5x 10 -11 M, between 5x 10 -7 M and 1x 10 -11 M, between 5x 10 -7 M and 5x 10 -12 M, between 5x 10 -7 M and 1x 10 -12 M, between 1x 10 -7 M and 5x 10 -8 Between M, 1x 10 -7 M and 1x 10 -8 M, between 1x 10 -7 M and 5x 10 -9 M, between 1x 10 -7 M and 1x10 -9 M, between 1x 10 -7 M and 5x 10 -10 M, between 1x 10 -7 M and 1x 10 -10 M, between 1x 10 -7 M and 5x10 -11 M, between 1x 10 -7 M and 1x 10 -11 M, between 1x 10 -7 M and 5x 10 -12 M, between 1x 10 -7 M and 1x 10 -12 M, between 5x 10 -8 M and 1x 10 -8 M, between 5x 10 -8 M and 5x 10 -9 M, between 5x 10 -8 M and 1x 10 -9 M, between 5x 10 -8 M and 5x 10 -10 M, between 5x 10 -8 M and 1x 10 -10 M, between 5x 10 - 8 M and 5x 10 -11 M, between 5x 10 -8 M and 1x 10 -11 M, between 5x 10 -8 M and 5x 10 -12 Between M, between 5x10 -8 M and 1x 10-12 Between M, 1x 10 -8 M and 5x 10 -9 M, between 1x 10 -8 M and 1x 10 -9 M, between 1x 10 - 8 M and 5x 10 -10 M, between 1x 10 -8 M and 1x 10 -10 M, between 1x 10 -8 M and 5x 10 -11 Between M, between 1x10 -8 M and 1x 10 -11 M, between 1x 10 -8 M and 5x 10 -12 M, between 1x 10 -8 M and 1x 10 -12 M, between 5x 10 -9 M and 1x 10 -9 Between M, between 5x10 -9 M and 5x 10 -10 M, between 5x 10 -9 M and 1x 10 -10 M, between 5x 10 -9 M and 5x 10 -11 M, between 5x 10 -9 M and 1x 10 -11 M, between 5x 10 -9 M and 5x 10 -12 M, between 5x 10 -9 M and 1x 10 -12 M, between 1x 10 -9 M and 5x10 -10 M, between 1x 10 -9 M and 1x 10 -10 M, between 1x 10 -9 M and 5x 10 -11 M, between 1x 10 -9 M and 1x 10 -11 M, between 1x 10 -9 M and 5x 10 -12 M, between 1x 10 -9 M and 1x 10 -12 M, between 5x 10 -10 M and 1x 10 -10 M, between 5x 10-10 M and 5x 10 - 11 M, between 1x 10 -10 M and 5x 10 -11 Between M, 1x 10 -10 M and 1x 10 -11 M, between 1x 10 -10 M and 5x 10 -12 Between M, between 1x10 -10 M and 1x 10 -12 M, between 5x 10 -11 M and 1x 10 -12 M, between 5x 10 -11 M and 5x10 -12 M, between 5x 10 -11 M and 1x 10 -12 M, between 1x 10 -11 M and 5x 10 -12 M or between 1x 10 -11 M and 1x 10 -12 In some embodiments, the IL13R / IL4R contiguous polypeptide binds canine IL13 and / or IL4, feline IL13 and / or IL4, and / or equine IL13 and / or IL4.

[0132] "Reduce" or "inhibit" means to reduce, decrease or prevent activity, function or quantity compared to a reference. In some embodiments, "reduce" or "inhibit" refers to the ability to cause an overall reduction of 20% or more. In some embodiments, "reduce" or "inhibit" refers to the ability to cause an overall reduction of 50% or more. In some embodiments, "reduce" or "inhibit" refers to the ability to cause an overall reduction of 75%, 85%, 90%, 95% or more. In some embodiments, the above amount is suppressed or reduced over a period of time relative to a control dose (such as a placebo) in the same time period. "Reference" as used herein refers to any sample, standard or level for comparison purposes. Reference can be obtained from a healthy or non-diseased sample. In some instances, the reference is obtained from a non-diseased or untreated sample of a companion animal. In some instances, the reference is obtained from one or more healthy animals of a particular species, and the healthy animal is not the animal being tested or treated.

[0133] As used herein, the term "substantially reduce" represents a sufficiently high degree of reduction between a numerical value and a reference value so that a person skilled in the art will consider the difference between the two values ​​to be statistically significant within the range of the biological property measured by the value. In some embodiments, the substantially reduced numerical value is reduced by greater than about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, or any one of 100% compared to a reference value.

[0134] In some embodiments, the IL13R / IL4R contiguous polypeptides can reduce IL13 and / or IL4 signaling in a companion animal species by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or 100% compared to IL13 and / or IL4 signaling in the absence of the fusion molecule. In some embodiments, signaling is measured by a reduction in IL4-dependent TF-1 cell proliferation. In some embodiments, the reduction in IL13 and / or IL4 signaling or the reduction in proliferation is between 10% and 15%, between 10% and 20%, between 10% and 25%, between 10% and 30%, between 10% and 35%, between 10% and 40%, between 10% and 45%, between 10% and 50%, between 10% and 60%, between 10% and 70%, between 10% and 80%, between 10% and 90%, between 10% and 100%, between 15% and 20% between 15% and 25%, between 15% and 30%, between 15% and 35%, between 15% and 40%, between 15% and 45%, between 15% and 50%, between 15% and 60%, between 15% and 70%, between 15% and 80%, between 15% and 90%, between 15% and 100%, between 20% and 25%, between 20% and 30%, between 20% and 35%, between 20% and 40%, between 20% and 45%, between 2 Between 0% and 50%, Between 20% and 60%, Between 20% and 70%, Between 20% and 80%, Between 20% and 90%, Between 20% and 100%, Between 25% and 30%, Between 25% and 35%, Between 25% and 40%, Between 25% and 45%, Between 25% and 50%, Between 25% and 60%, Between 25% and 70%, Between 25% and 80%, Between 25% and 90%, Between 25% and 100%, Between 30 ...35%, Between 25% and 35%, Between 25% and 35%, Between 25% and 35%, Between 25% and 35%, Between 25% and 35%, Between 25% and 35%, Between 25% and 35%, Between 25% and 35%, Between 25% and 35%, Between 25% and 35%, Between 25% and 35%, Between 25% and 35%, Between 25% and 3 5%, between 30% and 40%, between 30% and 45%, between 30% and 50%, between 30% and 60%, between 30% and 70%, between 30% and 80%, between 30% and 90%, between 30% and 100%, between 35% and 40%, between 35% and 45%, between 35% and 50%, between 35% and 60%, between 35% and 70%, between 35% and 80%, between 35% and 90%, between 35% and 100%,Between 40% and 45%, Between 40% and 50%, Between 40% and 60%, Between 40% and 70%, Between 40% and 80%, Between 40% and 90%, Between 40% and 100%, Between 45% and 50%, Between 45% and 60%, Between 45% and 70%, Between 45% and 80%, Between 45% and 90%, Between 45% and 100%, Between 50% and 60%, Between between 50% and 70%, between 50% and 80%, between 50% and 90%, between 50% and 100%, between 60% and 70%, between 60% and 80%, between 60% and 90%, between 60% and 100%, between 70% and 80%, between 70% and 90%, between 70% and 100%, between 80% and 90%, between 80% and 100% or between 90% and 100%.

[0135] Pharmaceutical composition

[0136] The terms "pharmaceutical formulation" and "pharmaceutical composition" refer to a preparation that is in such form as to permit the biological activity of one or more active ingredients to be effective and that contains no additional components that are unacceptably toxic to a subject to which the formulation would be administered.

[0137] "Pharmaceutically acceptable carrier" refers to a conventional non-toxic solid, semi-solid or liquid filler, diluent, encapsulating material, formulation auxiliary material or carrier in the art for use with a therapeutic agent and together forming a "pharmaceutical composition" for administration to a subject. A pharmaceutically acceptable carrier is non-toxic to the recipient at the dosage and concentration employed and is compatible with the other ingredients of the formulation. A pharmaceutically acceptable carrier is suitable for the formulation employed. Examples of pharmaceutically acceptable carriers include alumina; aluminum stearate; lecithin; serum proteins such as human serum albumin, canine or other animal albumin; buffers such as phosphate buffer, citrate buffer, tromethamine buffer or HEPES buffer; glycine; sorbic acid; potassium sorbate; partial glyceride mixtures of saturated vegetable fatty acids; water; salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silicon dioxide or magnesium trisilicate; polyvinyl pyrrolidone, cellulose-based materials; polyethylene glycol; sucrose; mannitol; or amino acids, including, but not limited to, arginine.

[0138] The pharmaceutical composition can be stored in a lyophilized form. Therefore, in some embodiments, the preparation process includes a lyophilization step. Then, before being applied to a dog, cat or horse, the lyophilized composition can be reformulated as an aqueous composition generally suitable for parenteral administration. In other embodiments, in particular, in the case where the fusion molecule is highly stable to heat and oxidative denaturation, the pharmaceutical composition can be stored in a liquid, that is, as an aqueous composition, it can be directly or in a suitable dilution to a dog, cat or horse. The lyophilized composition can be reconstituted with sterile water for injection (WFI). An antibacterial agent, such as benzyl alcohol, may be included. Therefore, the present invention provides a pharmaceutical composition in solid or liquid form.

[0139] When administered, the pH of the pharmaceutical composition can range from about pH 5 to about pH 8. If the compositions of the present invention are to be used for therapeutic purposes, they are sterile. Sterility can be achieved by any of several methods known in the art, including filtration through a sterile filtration membrane (e.g., a 0.2 micron membrane). Sterility can be maintained with or without an antibacterial agent.

[0140] Uses of IL13R / IL4R continuous polypeptides and pharmaceutical compositions

[0141] The IL13R / IL4R continuous polypeptide or a pharmaceutical composition comprising the IL13R / IL4R continuous polypeptide of the present invention may be useful for treating IL13- and / or IL4-induced conditions. As used herein, "IL13 or IL4-induced conditions" means a disease associated with, caused by, or characterized by elevated levels or altered distribution of IL13 or IL4. Such IL13 and / or IL4-induced conditions include, but are not limited to, pruritus or allergic diseases. In some embodiments, the IL13 and / or IL4-induced condition is atopic dermatitis, pruritus, asthma, psoriasis, scleroderma, or eczema. IL13 or IL4-induced conditions may be manifested in companion animals (including, but not limited to dogs, cats, or horses).

[0142] As used herein, "treatment" is a method for obtaining a beneficial or desired clinical outcome. As used herein, "treatment" encompasses any administration or application of a therapeutic agent for a disease in a mammal (including a companion animal). For the purposes of this disclosure, a beneficial or desired clinical outcome includes, but is not limited to, any one or more of the following: alleviation of one or more symptoms, reduction in the extent of the disease, prevention or delay of the spread of the disease, prevention or delay of the recurrence of the disease, delay or slowing of the progression of the disease, improvement of the disease state, inhibition of the progression of the disease or the disease, inhibition or slowing of the disease or its progression, prevention of its development, and alleviation (whether in part or in whole). "Treatment" also includes reducing the pathological results of proliferative diseases. Any one or more of these aspects of the method provided herein are contemplated for treatment. Consistent with the above, the term treatment does not require one hundred percent removal of all aspects of the disease.

[0143] In some embodiments, IL13R / IL4R contiguous polypeptides or pharmaceutical compositions comprising IL13R / IL4R contiguous polypeptides can be used according to the methods herein to treat IL13 or IL4 induced conditions. In some embodiments, IL13R / IL4R contiguous polypeptides or pharmaceutical compositions are administered to companion animals (such as dogs, cats, or horses) to treat IL13 and IL4 induced conditions.

[0144] A "therapeutically effective amount" of a substance / molecule, agonist or antagonist may vary according to factors such as the type of disease to be treated, the disease state, the severity and course of the disease, the type of therapeutic objective, any previous treatment, clinical history, response to previous treatment, the discretion of the attending veterinarian, the age, sex and weight of the animal, and the ability of the substance / molecule, agonist or antagonist to elicit a desired response in the animal. A therapeutically effective amount is also an amount in which the therapeutically beneficial effects of the substance / molecule, agonist or antagonist exceed any toxic or deleterious effects. A therapeutically effective amount may be delivered in one or more administrations. A therapeutically effective amount refers to an amount that effectively achieves the desired therapeutic or preventive outcome at the necessary dosage and within the necessary time period.

[0145] In some embodiments, the IL13R / IL4R continuous polypeptide or a pharmaceutical composition comprising the IL13R / IL4R continuous polypeptide is administered parenterally, by subcutaneous administration, intravenous infusion or intramuscular injection. In some embodiments, the IL13R / IL4R continuous polypeptide or a pharmaceutical composition comprising the IL13R / IL4R continuous polypeptide is administered as a bolus or continuous infusion over a period of time. In some embodiments, the IL13R / IL4R continuous polypeptide or a pharmaceutical composition comprising the IL13R / IL4R continuous polypeptide is administered by intramuscular, intraperitoneal, intracerebrospinal, subcutaneous, intraarterial, intrasynovial, intrathecal or inhalation route.

[0146] The IL13R / IL4R contiguous polypeptides described herein can be administered in an amount ranging from 0.1 mg / kg to 100 mg / kg body weight per dose. In some embodiments, the anti-IL13 / IL4 fusion can be administered in an amount ranging from 0.5 mg / kg to 50 mg / kg body weight per dose. In some embodiments, the anti-IL13 / IL4 fusion can be administered in an amount ranging from 1 mg / kg to 10 mg / kg body weight per dose. In some embodiments, the fusion molecule can be administered in an amount ranging from 0.5 mg / kg body weight to 100 mg / kg body weight, 1 mg / kg body weight to 100 mg / kg body weight, 5 mg / kg body weight to 100 mg / kg body weight, 10 mg / kg body weight to 100 mg / kg body weight, 20 mg / kg body weight to 100 mg / kg body weight, 50 mg / kg body weight to 100 mg / kg body weight, 1 mg / kg body weight to 10 mg / kg body weight, 5 mg / kg body weight to 10 mg / kg body weight, 0.5 mg / kg body weight to 10 mg / kg body weight, or 5 mg / kg body weight to 50 mg / kg body weight.

[0147] The IL13R / IL4R continuous polypeptides or pharmaceutical compositions comprising the IL13R / IL4R continuous polypeptides can be administered to companion animals once or over a series of treatments. For example, the IL13R / IL4R continuous polypeptides or pharmaceutical compositions comprising IL13R and IL4R can be administered at least once, more than once, at least twice, at least three times, at least four times, or at least five times.

[0148] In some embodiments, the dose is administered once a week for at least two or three consecutive weeks, and in some embodiments, this treatment cycle is repeated two or more times, optionally distributed in one or more weeks of no treatment. In other embodiments, the therapeutically effective dose is administered once a day for two to five consecutive days, and in some embodiments, this treatment cycle is repeated two or more times, optionally distributed in one or more days or weeks of no treatment.

[0149] Administration "in combination" with one or more additional therapeutic agents includes simultaneous (concurrent) and continuous or sequential administration in any order. The term "concurrently" is used herein to refer to the administration of two or more therapeutic agents, where at least part of the administration overlaps in time or relative to the administration of another therapeutic agent, and the administration of one therapeutic agent falls within a short period of time. For example, the two or more therapeutic agents are administered at a time interval not exceeding a specified number of minutes. The term "sequentially" is used herein to refer to the administration of two or more therapeutic agents, wherein one or more medicaments are continued to be administered after the administration of one or more other medicaments is stopped, or wherein one or more medicaments are administered before the administration of one or more other medicaments. For example, the administration of the two or more therapeutic agents is administered at a time interval of more than the specified number of minutes. As used herein, "in conjunction with" refers to the administration of a therapeutic form other than another therapeutic form. Similarly, "in conjunction with" refers to the administration of a therapeutic form before, during or after the administration of another therapeutic form to an animal.

[0150] In some embodiments, the method comprises administering a IL13R / IL4R continuous polypeptide or a pharmaceutical composition comprising a IL13R / IL4R continuous polypeptide in combination with a Jak inhibitor, a PI3K inhibitor, an AKT inhibitor, or a MAPK inhibitor. In some embodiments, the method comprises administering a IL13R / IL4R continuous polypeptide or a pharmaceutical composition comprising a IL13R / IL4R continuous polypeptide in combination with an anti-IL17 antibody, an anti-TNFα antibody, an anti-CD20 antibody, an anti-CD19 antibody, an anti-CD25 antibody, an anti-IL31 antibody, an anti-IL23 antibody, an anti-IgE antibody, an anti-CD11α antibody, an anti-IL6R antibody, an anti-α4-integrin antibody, an anti-IL12 antibody, an anti-IL1β antibody, or an anti-BlyS antibody.

[0151] Provided herein are methods for exposing cells to IL13R / IL4R continuous polypeptides or pharmaceutical compositions comprising IL13R / IL4R continuous polypeptides under conditions that permit binding to IL13 or IL4. In some embodiments, the cells are exposed to IL13R / IL4R continuous polypeptides or pharmaceutical compositions in vitro. In some embodiments, the cells are exposed to IL13R / IL4R continuous polypeptides or pharmaceutical compositions in vivo. In some embodiments, cells are exposed to IL13R / IL4R continuous polypeptides. In some embodiments, cells are exposed to IL13R / IL4R continuous polypeptides or pharmaceutical compositions under conditions that permit binding of fusion molecules to extracellular IL13 or IL4. In some embodiments, cells can be exposed to IL13R / IL4R continuous polypeptides or pharmaceutical compositions in vivo by any one or more of the administration methods described herein (including but not limited to intraperitoneal, intramuscular, and intravenous injection into a subject). In some embodiments, the cells can be exposed to IL13R / IL4R continuous polypeptides or pharmaceutical compositions in vitro by exposing the cells to a culture medium containing a fusion molecule or pharmaceutical composition. In some embodiments, the permeability of the cell membrane is affected prior to exposing the cells to culture medium containing the fusion molecule or pharmaceutical composition using any number of methods understood by one of skill in the art, such as electroporating the cells or exposing the cells to a solution containing calcium chloride.

[0152] In some embodiments, the exposure results in a decrease in IL13 or IL4 signaling function of the cell. In some embodiments, the IL13R / IL4R contiguous polypeptide can reduce IL13 or IL4 signaling in the cell by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or 100% compared to IL13 or IL4 signaling function in the absence of the IL13R / IL4R contiguous polypeptide. In some embodiments, the reduction in IL13 or IL4 signaling or the reduction in TF-1 proliferation is between 10% and 15%, between 10% and 20%, between 10% and 25%, between 10% and 30%, between 10% and 35%, between 10% and 40%, between 10% and 45%, between 10% and 50%, between 10% and 60%, between 10% and 70%, between 10% and 80%, between 10% and 90%, between 10% and 100%, between 15% and 20%. %, between 15% and 25%, between 15% and 30%, between 15% and 35%, between 15% and 40%, between 15% and 45%, between 15% and 50%, between 15% and 60%, between 15% and 70%, between 15% and 80%, between 15% and 90%, between 15% and 100%, between 20% and 25%, between 20% and 30%, between 20% and 35%, between 20% and 40%, between 20% and 45%, between Between 20% and 50%, Between 20% and 60%, Between 20% and 70%, Between 20% and 80%, Between 20% and 90%, Between 20% and 100%, Between 25% and 30%, Between 25% and 35%, Between 25% and 40%, Between 25% and 45%, Between 25% and 50%, Between 25% and 60%, Between 25% and 70%, Between 25% and 80%, Between 25% and 90%, Between 25% and 100%, Between 30 ...35%, Between 25% and 35%, Between 25% and 35%, Between 25% and 35%, Between 25% and 35%, Between 25% and 35%, Between 25% and 35%, Between 25% and 35%, Between 25% and 35%, Between 25% and 35%, Between 25% and 35%, Between 25% and 35%, Between 25% and 35%, Between 25% and 5%, between 30% and 40%, between 30% and 45%, between 30% and 50%, between 30% and 60%, between 30% and 70%, between 30% and 80%, between 30% and 90%, between 30% and 100%, between 35% and 40%, between 35% and 45%, between 35% and 50%, between 35% and 60%, between 35% and 70%, between 35% and 80%, between 35% and 90%, between 35% and 100%,Between 40% and 45%, Between 40% and 50%, Between 40% and 60%, Between 40% and 70%, Between 40% and 80%, Between 40% and 90%, Between 40% and 100%, Between 45% and 50%, Between 45% and 60%, Between 45% and 70%, Between 45% and 80%, Between 45% and 90%, Between 45% and 100%, Between 50% and 60%, Between between 50% and 70%, between 50% and 80%, between 50% and 90%, between 50% and 100%, between 60% and 70%, between 60% and 80%, between 60% and 90%, between 60% and 100%, between 70% and 80%, between 70% and 90%, between 70% and 100%, between 80% and 90%, between 80% and 100%, or between 90% and 100%.

[0153] Provided herein are methods for detecting, diagnosing, and monitoring IL13 or IL4-induced conditions using one or more IL13R / IL4R continuous polypeptides and polynucleotides. Provided herein are methods for determining whether a companion animal will respond to IL13R / IL4R continuous polypeptide therapy. In some embodiments, the method includes using IL13R / IL4R continuous polypeptides to detect whether the animal has cells expressing IL13 or IL4. In some embodiments, the method of detection includes contacting a sample with an antibody, polypeptide, or polynucleotide, and determining whether the level of binding is different from a reference or comparison sample (such as a control). In some embodiments, the method may be useful for determining whether the IL13R / IL4R continuous polypeptide described herein is an appropriate treatment for a subject animal.

[0154] In some embodiments, the sample is a biological sample. The term "biological sample" means a certain amount of material from a living thing or a previously living thing. In some embodiments, the biological sample is a cell or a cell / tissue lysate. In some embodiments, the biological sample includes, but is not limited to, blood (e.g., whole blood), plasma, serum, urine, synovial fluid, and epithelial cells.

[0155] In some embodiments, the cell or cell / tissue lysate is contacted with an IL13R / IL4R continuous polypeptide and the binding between the IL13R / IL4R continuous polypeptide and the cell is determined. When the test cell shows binding activity compared to a reference cell of the same tissue type, this may indicate that the subject will benefit from treatment with the IL13R / IL4R continuous polypeptide. In some embodiments, the test cell is from a companion animal tissue.

[0156] A variety of methods known in the art for detecting specific antibody-antigen binding can be used. Exemplary immunoassays that can be performed include fluorescence polarization immunoassay (FPIA), fluorescence immunoassay (FIA), enzyme immunoassay (EIA), turbidity inhibition immunoassay (NIA), enzyme-linked immunosorbent assay (ELISA), and radioimmunoassay (RIA). Indicator moieties or label groups can be attached to the subject antibody and are selected to meet the needs of various uses of the method, which are generally indicated by the availability of assay equipment and compatible immunoassay procedures. Suitable labels include, but are not limited to, radionuclides (e.g., 125 I. 131 I. 35 S. 3 H or 32 P), enzymes (e.g., alkaline phosphatase, horseradish peroxidase, luciferase, or β-galactosidase), fluorescent moieties or proteins (e.g., fluorescein, rhodamine, phycoerythrin, GFP, or BFP), or luminescent moieties (e.g., Qdot™ nanoparticles supplied by Quantum Dot Corporation, Palo Alto, Calif.). General techniques for performing the various immunoassays described above are known to those of ordinary skill in the art.

[0157] For diagnostic purposes, the IL13R / IL4R continuous polypeptides can be labeled with a detectable moiety, including but not limited to radioisotopes, fluorescent labels, and various enzyme substrate labels known in the art. Methods for conjugating labels to polypeptides are known in the art. In some embodiments, the IL13R / IL4R continuous polypeptide does not need to be labeled, and its presence can be detected, for example, using an antibody that binds to the IL13R / IL4R continuous polypeptide. In some embodiments, the IL13R / IL4R continuous polypeptide can be used in any known assay method (such as competitive binding assays, direct and indirect sandwich assays, and immunoprecipitation assays). Zola, Monoclonal Antibodies: A Manual of Techniques, pp. 147-158 (CRC Press, Inc. 1987). Anti-IL13 and IL4 antibodies and polypeptides can also be used in in vivo diagnostic assays, such as in vivo imaging. Typically, the antibody or polypeptide is labeled with a radionuclide (such as 111 In, 99 Tc, 14 C. 131 I. 125 I. 3H or any other radionuclide label, including those outlined herein), so that cells or tissues of interest can be located using immunoscintigraphy. The IL13R / IL4R contiguous polypeptides can also be used as staining agents in pathology using techniques well known in the art.

[0158] In some embodiments, the IL13R / IL4R contiguous polypeptide is used as a diagnostic agent and the IL13R / IL4R contiguous polypeptide is used as a therapeutic agent. In some embodiments, the first and second IL13R / IL4R contiguous polypeptides are different.

[0159] The following examples illustrate specific aspects of the present disclosure and are not intended to limit the present disclosure in any way. Example

[0160] Example 1

[0161] Expression and purification of canine IL4 and IL13

[0162] The nucleotide sequence encoding the canine IL13 protein (SEQ ID NO: 4) with a poly-His tag on the C-terminus was synthesized and cloned into a mammalian expression vector and transfected into 293 cells or CHOS. The same method was used to clone and express the nucleotide sequence encoding the canine IL4 protein (SEQ ID NO: 1) with a poly-His tag on the C-terminus.

[0163] The supernatant containing canine IL13 protein was collected and filtered. Protein A affinity resin (Repligen) was used to purify canine IL13. The same method was used to purify canine IL4.

[0164] Example 2

[0165] Extracellular domains of IL13R and IL4R

[0166] The extracellular domains of canine, feline and equine IL4R responsible for binding to canine, feline and equine IL4 and / or IL13 were identified and delimited. The full-length extracellular domains of canine IL4R, feline IL4R and equine IL4R were identified as SEQ ID NO: 23, SEQ ID NO: 25 and SEQ ID NO: 27, respectively. The extracellular domain fragments of canine IL4R, feline IL4R and equine IL4R that are claimed to retain biological activity were identified as SEQ ID NO: 33, SEQ ID NO: 35 and SEQ ID NO: 37, respectively.

[0167] The extracellular domains of canine, feline and equine IL13R responsible for binding to canine, feline and equine IL4 and / or IL13 were identified and delimited. The full-length extracellular domains of canine IL13R, feline IL13R and equine IL13R were identified as SEQ ID NO: 22, SEQ ID NO: 24 and SEQ ID NO: 26, respectively. The extracellular domain fragments of canine IL13R, feline IL13R and equine IL13R that are claimed to retain biological activity were identified as SEQ ID NO: 32, SEQ ID NO: 34 and SEQ ID NO: 36, respectively.

[0168] The unpaired cysteine ​​(Cys) in canine IL13R (position 18 of SEQ ID NO: 22), feline IL13R (position 18 of SEQ ID NO: 24), and equine IL13R (position 18 of SEQ ID NO: 26) were identified informatively and determined to be embedded (unexposed) based on 3-D modeling. Unpaired cysteines are unlikely to form disulfide bonds and have a low probability of aggregation. Therefore, site-directed mutagenesis of this Cys residue was not introduced.

[0169] Example 3

[0170] Expression and purification of canine IL13R / IL4R contiguous polypeptides from CHO cells

[0171] A nucleotide sequence with a signal sequence encoding a canine IL13R / IL4R contiguous polypeptide linked to Fc IgGB was designed. For the contiguous polypeptide "IL13R-IL4R-IgGB" (SEQ ID NO: 20), the extracellular domain of IL13R (SEQ ID NO: 22) precedes the extracellular domain of IL4R (SEQ ID NO: 23). For the contiguous polypeptide IL4R-IL13R-IgGB (SEQ ID NO: 21), the extracellular domain of IL4R precedes the extracellular domain of IL13R.

[0172] These nucleotide sequences are chemically synthesized and inserted into expression vectors suitable for transfection into CHO host cells. After transfection into CHO cells, the fusion protein is secreted from the cells. For example, the fusion protein is purified by one-step protein A column chromatography.

[0173] Each of IL13R-IL4R-IgGB and IL4R-IL13R-IgGB can be expressed and purified in a single step using a protein A column or other chromatography methods such as ion exchange column chromatography, hydrophobic interaction column chromatography, mixed mode column chromatography (such as CHT), or multi-modal column chromatography (such as CaptoMMC). Low pH or other viral inactivation and viral removal steps can be applied. The purified protein can be mixed with an excipient and sterilized by filtration to prepare a pharmaceutical composition of the invention. The pharmaceutical composition can be administered to dogs with atopic dermatitis or asthma in an amount sufficient to bind and / or inhibit IL13 and / or IL4.

[0174] The vector is then used to create the image using FreestyleMax TM Transfection was performed on a pilot scale in CHO-S cells using the TRANSFERR® transfection reagent (Life Technologies). The supernatant was harvested by clarifying the conditioned medium. The protein was purified using a single pass protein A chromatography step and used for further studies.

[0175] Example 4

[0176] Demonstration of IL13 and IL4 binding activity

[0177] This example demonstrates that both IL13R-IL4R-IgGB (SEQ ID NO:20) and IL4R-IL13R-IgGB (SEQ ID NO:21) bind canine IL4 and IL13 with the kinetics necessary for therapeutic activity.

[0178] Binding analysis was performed using the biosensor Octet as follows. Briefly, canine IL4 (produced using 293 cells) was biotinylated. Free unreacted biotin was removed from the biotinylated IL4 by extensive dialysis. Biotinylated canine IL4 was captured on the tip of the streptavidin sensor. Monitoring of IL4 bound to IL13R-IL4R-IgGB (SEQ ID NO: 20) at multiple concentrations (12, 16, and 44 nM) lasted for ninety seconds. Monitoring dissociation lasted for 600 seconds. Subtracting a blank curve of only buffer to correct for any drift. Using ForteBioTM data analysis software, the data was fitted to a 1:1 binding model to determine k on , k off The buffer used for dilution and all binding steps was: 20 mM phosphate, 150 mM NaCl, pH 7.2. The Kd of IL13R-IL4R-IgGB and ligand IL4 was 8x10 -11 .

[0179] Canine IL4 binding to IL4R-IL13R-IgGB (SEQ ID NO: 21) at multiple concentrations (40.7 and 140 nM) was monitored for ninety seconds. Dissociation was monitored for 600 seconds. A buffer-only blank curve was subtracted to correct for any drift. TM The data analysis software fits the data to a 1:1 binding model to determine k on , k off The buffer used for dilution and all binding steps was: 20 mM phosphate, 150 mM NaCl, pH 7.2. The Kd of IL4R-IL13R-IgGB and ligand IL4 was 1.1x10 -11 .

[0180] Canine IL4 and canine IL13 with C-terminal poly-His tags were expressed and purified from 293 cells. EZ-Link NHS-LC-Biotin was obtained from Thermo Scientific (Cat. #21336), and streptavidin biosensor was obtained from ForteBio (Cat. #18-509).

[0181] Sequential binding experiments of IL4 and IL13 to IL13R-IL4R-IgGB (SEQ ID NO: 20) were performed. Biotinylated canine IL13R-IL4R-IgGB was captured on a streptavidin sensor tip. Canine IL13R-IL4R-IgGB was exposed to (1) canine IL4 followed by IL13 or (2) canine IL13 followed by IL14 using IL4 and IL13 at a concentration of 30 μg / mL in PBS. Figure 1 ). This experiment demonstrated that once IL13R-IL4R-IgGB binds IL13, it cannot bind IL4, and once bound to IL4, its ability to bind IL13 is reduced.

[0182] IL4 and IL13 sequential binding experiments with IL4R-IL13R-IgGB (SEQ ID NO: 21) were performed. Biotinylated canine IL4R-IL13R-IgGB was captured on a streptavidin sensor tip. Canine IL4R-IL13R-IgGB was exposed to (1) canine IL4 followed by IL13 or (2) canine IL13 followed by IL14 using concentrations of IL4 and IL13 at 30 μg / mL in PBS. Figure 2 ). These experiments demonstrated that once IL4R-IL13R-IgGB binds IL13, it can no longer bind IL4, and once bound to IL4, its ability to bind IL13 is reduced.

[0183] The tight binding of IL13R-IL4R-IgGB and IL4R-IL13R-IgGB to IL4 or IL13 is thought to be due to the simultaneous binding contribution from both IL4R and IL13R.

[0184] Example 5

[0185] Cellular Functional Activity of Canine IL4R-IL13R-Fc(SINK)

[0186] TF1 cells (ATCC cat #CRL-2003), which is a human erythroleukemia cell line expressing endogenous interleukin 4 receptors on the cell surface, are used for proliferation assays. Cells grown in the logarithmic growth phase in RPMI1640 (Gibco, registration number 11875) supplemented with heat-inactivated 10% fetal bovine serum (Sigma, Cat #2868) and 2nM / ml human GM-CSF (R&D system, registration number 215-GM-010) are used for the assay. The cells are washed twice with PBS and resuspended in the above culture medium without GM-CSF. 20,000 cells per well are plated in a 96-well plate (Corning, registration number 3610). Canine IL4R-IL13R-Fc (SINK) is added at a series of dilutions, followed by canine IL4 (Sino Biological Inc, registration number 70021-DNAE-5) at 50ng / ml. The cells were incubated for 48 h at 37 °C, 5% CO2 in a total volume of 100 μl. At the end of the incubation, the cells were cooled at room temperature and washed with The luminescent cell viability assay (Promega, Cat# G7570) measures cellular ATP content to determine proliferation / variability.

[0187] In this assay, 100 μl premixed reagent A and B are added to each hole. After 2 minutes of shaking on an orbital shaker, the cells are lysed. In the presence of Mg2+ and ATP in the cell, the monooxygenation reaction of luciferin is catalyzed by luciferase, which results in the generation of a luminescent signal proportional to the ATP amount in the cell. The amount of ATP is directly proportional to the number of cells present in the culture. Plates were incubated at room temperature for 10 minutes to stabilize the luminescent signal, and Synergy HT microplate reader (Biotek, Winooski, VT) was used to detect luminescence.

[0188] The data were analyzed using a 4-parameter logistic fit and the IC50 was 2.0 nM. Figure 3 .

Claims

1. A polypeptide comprising an extracellular domain of an IL13R polypeptide and an extracellular domain of an IL4R polypeptide, wherein the IL13R and IL4R polypeptides are derived from dogs, The polypeptide consists of an amino acid sequence selected from the group consisting of: SEQ ID NO: 20 or SEQ ID NO: 21; and The polypeptide has a concentration of less than 1 x 10 -10 The dissociation constant (Kd) of M binding to canine IL4 was measured by the biofilm interference assay.

2. The polypeptide of claim 1, wherein the polypeptide is present in an amount less than 1 x 10 -10 The dissociation constant (Kd) of M binds to canine IL13 as measured by a biofilm interference assay.

3. The polypeptide of claim 1, wherein the polypeptide reduces IL13 and / or IL4 signaling in a canine compared to IL13 and / or IL4 signaling in the absence of the polypeptide.

4. An isolated nucleic acid encoding the polypeptide of claim 1 or 2.

5. An isolated or non-human host cell comprising the nucleic acid of claim 4.

6. A method for producing the polypeptide of claim 1, the method comprising culturing the host cell of claim 5 and isolating the polypeptide.

7. A pharmaceutical composition comprising the polypeptide of claim 1 and a pharmaceutically acceptable carrier.

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