Il-4 / il-13 and il-4 / il-13 / il-31 receptor trap constructs

Abstract

Receptor traps for the neutralization of cytokines, optionally neutralization of Interleukin (IL)-4 and IL-13, also IL-31, for the treatment of inflammation.

Description

U.S. Provisional Patent Application Attorney Docket No.2920951-492977 IL-4 / IL-13 AND IL-4 / IL-13 / IL-31 RECEPTOR TRAP CONSTRUCTS REFERENCE TO RELATED PATENT APPLICATIONS

[0001] This International Patent Application claims priority to U.S. Provisional PatentApplication No.63 / 615,693, filed 28 December 2023; U.S. Provisional Patent Application No. 63 / 615,704, filed 28 December 2023; and U.S. Provisional Patent Application No.63 / 615,705, filed 28 December 2023, the disclosures of each of which are herein incorporated by reference in their entireties. FIELD OF THE INVENTION

[0002] The present disclosure relates to receptor traps for neutralization of Interleukin (IL)-4 andInterleukin-13 (IL-13), in addition to Interleukin-31 (IL-31), in for the treatment of <inflammation, including inflammation associated with atopic dermatitis and allergic dermatitis. REFERENCE TO SEQUENCE LISTING

[0003] Pursuant to 37 C.F.R. 1.821(c), a sequence listing is submitted herewith as an ASCIIcompliant text file named 2920951-492977-SEQ-LISTING, created on 20.12.2024 and having a size of 265 kilobytes. The content of the aforementioned file is hereby incorporated by reference in its entirety. BACKGROUND

[0004] Atopic dermatitis is a frequent allergic skin disorder that is characterized by aberrant andexcessive Th2 cell and ILC2 activation, with robust expression of type 2 cytokines, including interleukin (IL)-4, IL-5, IL-13 and IL-31, and variable activation of other cytokines, in particular IL-22 and IL-33, but also IL-17, IL-9 and IFN-γ (Moyle et al. Experimental Dermatology. 2019; 28:756–768; Renert-Yuval & Guttman-Yassky, Dermatol Clin.2019; 37:205–213). Atopic dermatitis is not only a frequent disorder in humans, but also in animals, in particular dogs. In fact, atopic dermatitis is the most common allergy in dogs and affects approximately 10% of the dog population, resulting in 15 million to 20 million dogs suffering from the disease in Europe and the United States alone (Griffin, et al., The ACVD task force on canine atopic dermatitis (XIV): clinical manifestations of canine atopic dermatitis. Veterinary immunology and immunopathology.2001; 81(3-4), 255-269). The itching or pruritus which is caused by thisU.S. Provisional Patent Application Attorney Docket No.2920951-492977 allergic skin disease is usually recurrent or chronic. It deeply impacts the quality of life for both the dogs and their owners.

[0005] There exists a need in the art for the treatment of atopic dermatitis and allergic dermatitisin canines. SUMMARY OF THE INVENTION

[0006] In an embodiment, a fusion protein can comprise:an IL-4-binding extracellular domain (IL-4-Rα1-ECD); and a fragment crystallizable region of an antibody (Fc).

[0007] In an embodiment, a fusion protein can comprise:an IL-13-binding extracellular domain (IL-13-Rα2-ECD); and a fragment crystallizable region of an antibody (Fc).

[0008] In an embodiment, a fusion protein can comprise:an IL-4-binding extracellular domain (IL-4-Rα1-ECD); an IL-13-binding extracellular domain (IL-13-Rα2-ECD); and a fragment crystallizable region of an antibody (Fc).

[0009] In an embodiment, a fusion protein can comprise:an IL-4-binding extracellular domain (IL-4-Rα1-ECD); an IL-13-binding extracellular domain (IL-13-Rα2-ECD); an cOSRM^-ligand binding domain (LBD); and a fragment crystallizable region of an antibody (Fc).

[0010] In an embodiment, a fusion protein can comprise:an IL-4-binding extracellular domain (IL-4-Rα1-ECD); an IL-13-binding extracellular domain (IL-13-Rα2-ECD); an OSRM^-ligand binding domain (LBD); IL-31RA-ligand binding domain (LBD); and a fragment crystallizable region of an antibody (Fc).

[0011] In an embodiment, the C-terminal of the IL-4-binding extracellular domain is linkedto the N-terminal of IL-13-binding extracellular domain.

[0012] In an embodiment, the C-terminal of the IL-13-binding extracellular domain is linkedto the N-terminal of the IL-4-binding extracellular domain.U.S. Provisional Patent Application Attorney Docket No.2920951-492977

[0013] In an embodiment, the Fc is linked to the C-terminal of the IL-4-binding extracellulardomain.

[0014] In an embodiment, the Fc is linked to the C-terminal of the IL-13-bindingextracellular domain.

[0015] In an embodiment, the Fc is linked to the C-terminal of the OSRM^-LBD.

[0016] In an embodiment, the Fc is linked to the C-terminal of the IL-31RA-ligand bindingdomain (LBD).

[0017] In an embodiment, the cIL-31RA-ligand binding domain (LBD) is linked to thecOSRM^-LBD.

[0018] In an embodiment, the C-terminus of the IL-4-binding extracellular domain is linkedto the N-terminus IL-13-binding extracellular domain which is linked to the N-terminus of the fragment crystallizable region of an antibody.

[0019] In an embodiment, the C-terminus of the IL-13-binding extracellular domain is linkedto the N-terminus of the IL-4-binding extracellular domain which is linked to the N-terminus of the fragment crystallizable region of an antibody.

[0020] In an embodiment, the C-terminus of the IL-4-binding extracellular domain (IL-4-Rα1-ECD) is linked to the N-terminus of the IL-13-binding extracellular domain (IL-13-RA2- ECD) which is linked to the N-terminus of the OSRM^-ligand binding domain (LBD) which is linked to the N-terminus of the fragment crystallizable region of an antibody (Fc).

[0021] In an embodiment, the C-terminus of the IL-4-binding extracellular domain (IL-4-Rα1-ECD) is linked to the N-terminus of the IL-13-binding extracellular domain (IL-13-RA2- ECD) which is linked to the N-terminus of the OSRM^-ligand binding domain (LBD) which is linked to the N-terminus of the IL-31RA-ligand binding domain (LBD) which is linked to the N- terminus of the fragment crystallizable region of an antibody (Fc).

[0022] In an embodiment, the IL-4-binding extracellular domain (IL-4-Rα1-ECD) and cFcare joined by a linker.

[0023] In an embodiment, the IL-13-binding extracellular domain (IL-13-RA2-ECD) andcFc are joined by a linker.

[0024] In an embodiment, the IL-4-binding extracellular domain (IL-4-Rα1-ECD) and theIL-13-binding extracellular domain (IL-13-RA2-ECD) are joined by a linker.U.S. Provisional Patent Application Attorney Docket No.2920951-492977

[0025] In an embodiment, the IL-13-binding extracellular domain (IL-13-RA2-ECD) andOSRM^-ligand binding domain (LBD) are joined by a linker.

[0026] In an embodiment, the OSRM^-ligand binding domain (LBD) and the IL-31RA-ligand binding domain (LBD) are joined by a linker.

[0027] In an embodiment, the IL-31RA-ligand binding domain (LBD) and fragmentcrystallizable region of an antibody (Fc) are joined by a linker. The linker is an oligopeptide can comprise about 8 to 20 amino acids.

[0028] In an embodiment, the IL-4-binding extracellular domain is a canine sequence. TheIL-4-binding extracellular domain can comprise an amino acid sequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 2. The IL-4-binding extracellular domain can comprise an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 2. The IL-4-binding extracellular domain can further comprise an ER import signal and a hexa-histidine tag. The IL-4-binding extracellular domain can comprise the amino acid sequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 3. The IL-4-binding extracellular domain can comprise the amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the SEQ ID NO: 3. The IL-4-binding extracellular domain can be encoded by a nucleic acid with at least 75% homology to the nucleic acid sequence of SEQ ID NO: 4. The IL-4-binding extracellular domain can be encoded by a nucleic acid with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the nucleic acid sequence of SEQ ID NO: 4.

[0029] In an embodiment, the Fc sequence is a canine sequence. The Fc sequence cancomprise an amino acid sequence with at least 75% homology to the amino acid sequence ofSEQ ID NO: 7. The Fc sequence can comprise an amino acid sequence with at least 80%, 85%,90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 7.

[0030] In an embodiment, the canine IL-4alpha1 receptor ECD-canine IgG Fc fusion proteincomprises an amino acid sequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 8. The fusion protein can comprise an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the aminoU.S. Provisional Patent Application Attorney Docket No.2920951-492977 acid sequence of SEQ ID NO: 8. The canine IL-4alpha1 receptor ECD-canine IgG Fc fusion protein can further comprise an ER import signal and a hexa-histidine tag. The canine IL- 4alpha1 receptor ECD-canine IgG Fc fusion protein can be encoded by a nucleic acid with at least 75% homology to the nucleic acid sequence of SEQ ID NO: 9. The canine IL-4alpha1 receptor ECD-canine IgG Fc fusion protein can be encoded by a nucleic acid with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the nucleic acid sequence of SEQ ID NO: 9.

[0031] In an embodiment, the IL-13-binding extracellular domain is a canine sequence. TheIL-13-binding extracellular domain can comprise an amino acid sequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 12. The IL-13-binding extracellular domain can comprise an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 12.

[0032] In an embodiment, the IL-13Ralpha2 ECD-canine IgG Fc fusion protein can comprisean amino acid sequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 13. The fusion protein can comprise an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 13. The IL-13Ralpha2 ECD-canine IgG Fc fusion protein can be encoded by a nucleic acid with at least 75% homology to the nucleic acid sequence of SEQ ID NO: 14. The IL-13Ralpha2 ECD-canine IgG Fc fusion protein can be encoded by a nucleic acid with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the nucleic acid sequence of SEQ ID NO: 14.

[0033] In an embodiment, the cIL-13Rα2-cIL-4Rα1-ECD-cFc fusion protein can comprisean amino acid sequence with at least about 75% sequence homology to SEQ ID NO: 16. The cIL-13Rα2-cIL-4Rα1-ECD-cFc fusion protein can comprise an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 16.

[0034] The cIL-13Rα2-cIL-4Rα1-ECD-cFc fusion protein can be encoded by a nucleic acidwith at least 75% homology to the nucleic acid sequence of SEQ ID NO: 17. The cIL-13Rα2- cIL-4Rα1-ECD-cFc fusion protein can be encoded by a nucleic acid with at least 80%, 85%,U.S. Provisional Patent Application Attorney Docket No.2920951-492977 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the nucleic acid sequence of SEQ ID NO: 17.

[0035] In an embodiment, the cIL-4Rα1-cIL-13α2-ECD-cFc fusion protein can comprise anamino acid sequence with at least about 75% sequence homology to SEQ ID NO: 19. The cIL- 4Rα1-cIL-13α2-ECD-cFc fusion protein can comprise an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 19.

[0036] The cIL-4Rα1-cIL-13α2-ECD-cFc fusion protein can be encoded by a nucleic acidwith at least 75% homology to the nucleic acid sequence of SEQ ID NO: 20. The cIL-4Rα1-cIL- 13α2-ECD-cFc fusion protein can be encoded by a nucleic acid with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the nucleic acid sequence of SEQ ID NO: 20.

[0037] In an embodiment, the IL-31RA sequence is a canine sequence. The IL-31RAsequence can comprise an amino acid sequence with at least 75% homology to the amino acidsequence of SEQ ID NO: 23. The IL-31RA sequence can comprise an amino acid sequence withat least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 23.

[0038] In an embodiment, the OSMR^ sequence is a canine sequence. The OSMR^sequence can comprise an amino acid sequence with at least 75% homology to the amino acidsequence of SEQ ID NO: 25. The OSMR^ sequence can comprise an amino acid sequence withat least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 25.

[0039] In an embodiment, the IL-4 / IL-13 / IL-31 fusion protein comprises an amino acidsequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 26. Thefusion protein can comprise an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 26. The IL-4 / IL-13 / IL-31 fusion protein can be encoded by a nucleic acid with at least 75% homology to the nucleic acid sequence of SEQ ID NO: 27. The IL-4 / IL-13 / IL-31 fusion protein can be encoded by a nucleic acid with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the nucleic acid sequence of SEQ ID NO: 27.U.S. Provisional Patent Application Attorney Docket No.2920951-492977

[0040] In an embodiment, a fusion protein can comprisean IL-4-binding extracellular domain (IL-4-R^-ECD); and a fragment crystallizable region of an antibody (Fc).

[0041] In an embodiment, a fusion protein can comprisean IL-13-binding extracellular domain (IL-13-Ralpha2-ECD); and a fragment crystallizable region of an antibody (Fc).

[0042] In an embodiment, a fusion protein can comprisean IL-4-binding extracellular domain (IL-4-R^-ECD); an IL-13-binding extracellular domain (IL-13-Ralpha2-ECD); and a fragment crystallizable region of an antibody (Fc).

[0043] In an embodiment, the IL-4-binding extracellular domain is a human sequence. TheIL-4-binding extracellular domain can comprise an amino acid sequence with at least 75%homology to the amino acid sequence of SEQ ID NO: 30. The IL-4-binding extracellulardomain can comprise an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 30.

[0044] In an embodiment, the IL-13-binding extracellular domain is a human sequence. TheIL-13-binding extracellular domain can comprise an amino acid sequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 32. The IL-13-binding extracellular domain can comprise an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 32.

[0045] In an embodiment, the Fc sequence is a human sequence. The Fc sequence cancomprise an amino acid sequence with at least 75% homology to the amino acid sequence ofSEQ ID NO: 33. The Fc sequence can comprise an amino acid sequence with at least 80%, 85%,90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 33.

[0046] In an embodiment, the fusion protein comprises a hIL-13Rα2-hIL-4Rα1-ECD-cFcprotein construct comprising an amino acid sequence with at least about 75% sequence homology to SEQ ID NO: 34. The fusion protein comprises a hIL-13Rα2-hIL-4Rα1-ECD-cFcU.S. Provisional Patent Application Attorney Docket No.2920951-492977 protein construct can comprise an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 34. The hIL-13Rα2-hIL-4Rα1-ECD-cFc fusion protein can be encoded by a nucleic acid with at least 75% homology to the nucleic acid sequence of SEQ ID NO: 35. The hIL-13Rα2- hIL-4Rα1-ECD-cFc fusion protein can be encoded by a nucleic acid with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the nucleic acid sequence of SEQ ID NO: 35.

[0047] In an embodiment, a fusion protein can comprise:an IL-4-binding extracellular domain (IL-4-Ralpha1-ECD); and a fragment crystallizable region of an antibody (Fc).

[0048] In an embodiment, a fusion protein can comprise:an IL-13-binding extracellular domain (IL-13-Ralpha2-ECD); and a fragment crystallizable region of an antibody (Fc).

[0049] In an embodiment, a fusion protein can comprise:an IL-4-binding extracellular domain (IL-4-Ralpha1-ECD); an IL-13-binding extracellular domain (IL-13-Ralpha2-ECD); and a fragment crystallizable region of an antibody (Fc).

[0050] In an embodiment, the IL-4-binding extracellular domain is a feline sequence. TheIL-4-binding extracellular domain can comprise an amino acid sequence with at least 75%homology to the amino acid sequence of SEQ ID NO: 39. The IL-4-binding extracellulardomain can comprise an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 39. The IL-4-binding extracellular domain can comprise an amino acid sequence with atleast 75% homology to the amino acid sequence of SEQ ID NO: 40. The IL-4-bindingextracellular domain can comprise an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 40.

[0051] In an embodiment, the IL-13-binding extracellular domain is a feline sequence. TheIL-13-binding extracellular domain can comprise an amino acid sequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 42. The IL-13-binding extracellularU.S. Provisional Patent Application Attorney Docket No.2920951-492977 domain can comprise an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 42.

[0052] In an embodiment, the IL-13-binding extracellular domain is a feline sequence. TheIL-13-binding extracellular domain can comprise an amino acid sequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 43. The IL-13-binding extracellular domain can comprise an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 43.

[0053] In an embodiment, the Fc sequence is a feline sequence. The Fc sequence cancomprise an amino acid sequence with at least 75% homology to the amino acid sequence ofSEQ ID NO: 45. The Fc sequence can comprise an amino acid sequence with at least 80%, 85%,90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 45.

[0054] In an embodiment, the Fc sequence is a feline sequence. The Fc sequence cancomprise an amino acid sequence with at least 75% homology to the amino acid sequence ofSEQ ID NO: 46. The Fc sequence can comprise an amino acid sequence with at least 80%, 85%,90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 46.

[0055] In an embodiment, the fusion protein comprises a fIL-13Rα2-fIL-4Rα1-ECD-cFcprotein construct comprising an amino acid sequence with at least about 75% sequence homology to SEQ ID NO: 47. The fusion protein comprises a fIL-13Rα2-fIL-4Rα1-ECD-cFc protein construct can comprise an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 47. The fIL-13Rα2-fIL-4Rα1-ECD-cFc fusion protein can be encoded by a nucleic acid with at least 75% homology to the nucleic acid sequence of SEQ ID NO: 50. The fIL-13Rα2-fIL- 4Rα1-ECD-cFc fusion protein can be encoded by a nucleic acid with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the nucleic acid sequence of SEQ ID NO: 50.U.S. Provisional Patent Application Attorney Docket No.2920951-492977

[0056] In an embodiment, the fusion protein comprises a fIL-4Rα1- fIL-13Rα2 -cFc proteinconstruct comprising an amino acid sequence with at least about 75% sequence homology to SEQ ID NO: 48. The fusion protein comprises a fIL-4Rα1- fIL-13Rα2--cFc protein construct can comprise an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 48.

[0057] SHARED EMBODIMENTS

[0058] In an embodiment, the linker can be an oligopeptide comprising about 8 to 20 aminoacids. The oligopeptide can be a polyglycine oligopeptide. The oligopeptide can comprise the amino acid sequence of SEQ ID NO: 81. The oligopeptide can be a polyglycine / serine oligopeptide. The polyglycine / serine oligopeptide can comprise the amino acid sequence of SEQ ID NO: 82, 83, or 84. The oligopeptide can comprise the amino acid sequence of SEQ ID NO: 85.

[0059] The fusion protein can further comprise an ER import signal sequence on the N-terminal. The ER import signal sequence can comprise the amino acid sequence of SEQ ID NO: 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, or 76.

[0060] The fusion protein further can comprise an affinity tag on the C-terminal. The affinitytag can be a hexahistidine-tag.

[0061] In an embodiment, a composition can comprise a fusion protein described herein. Thecomposition can be a pharmaceutical composition. The pharmaceutical composition can further comprise a pharmaceutical excipient, carrier, diluent, adjuvant, or a combination thereof. The composition can be formulated for intravenous, subcutaneous, infusion, oral, intrathecal, intraperitoneal, parenteral administration, or a combination thereof. The composition can further comprise an anti IL-31 antibody, anti IL-5 antibody, anti-IL-22 antibody, or a combination thereof.

[0062] In an embodiment, a nucleotide sequence encoding a fusion protein described herein.

[0063] In an embodiment, an expression vector can comprise a nucleotide sequence encodinga fusion protein described herein.

[0064] In an embodiment, a recombinant host cell can comprise a nucleotide sequenceencoding a fusion protein described herein.U.S. Provisional Patent Application Attorney Docket No.2920951-492977

[0065] In an embodiment, a recombinant host cell can comprise an expression vectorcomprising the nucleotide sequence encoding a fusion protein described herein.

[0066] In an embodiment, a method of treating or preventing inflammation comprisingadministering to a mammal in need thereof an effective amount of a fusion protein described herein.

[0067] In an embodiment, a method of treating or preventing inflammation comprisingadministering to a mammal in need thereof an effective amount of the composition comprising an effective amount of a fusion protein described herein.

[0068] In an embodiment, the dermatological condition comprises at least one skin disorderselected from psoriasis, atopic dermatitis, skin rash, skin irritation, skin sensitization, allergic reactions, pruritus, and combinations thereof.

[0069] In an embodiment, the inflammation is associated with atopic dermatitis.

[0070] In an embodiment, the inflammation is associated with allergic dermatitis.

[0071] In an embodiment, the mammal is a non-human mammal.

[0072] In an embodiment, the mammal is a canine.

[0073] In an embodiment, the mammal is a dog. The dog can be at least 9 months of age.The dog can be at least 12 months of age.

[0074] In an embodiment, the mammal is a cat.

[0075] In an embodiment, the administering is performed daily.

[0076] In an embodiment, the administering is performed twice daily.

[0077] In an embodiment, the administering is performed weekly.

[0078] In an embodiment, the administering is performed monthly.

[0079] In an embodiment, the effective amount is between about 0.1 and 10 mg / kg. Theeffective amount can be between about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 mg / kg. The effective amount can be between about 0.1 and 1 mg / kg, 0.5 and 2 mg / kg, 0.75 and 5 mg / kg, or 1 and 10 mg / kg.

[0080] Use of a fusion protein described herein for the manufacture of a medicament for thetreatment of inflammation in a mammal. The inflammation can be associated with atopic dermatitis or allergic dermatitis. BRIEF DESCRIPTION OF THE DRAWINGSU.S. Provisional Patent Application Attorney Docket No.2920951-492977

[0081] FIG. 1 depicts the sequence of a cIL13Ra2-cIL4RΑ1-cFc construct (SEQ ID NO: 16).

[0082] FIG. 2 depicts the sequence of a cIL4RΑ1-cIL13Ra2-cFc construct (SEQ ID NO: 19).

[0083] FIG. 3A depicts the general structure of a canine Fc-linked canine IL-4 / IL-13 / IL-31triple receptor trap protein

[0084] FIG. 3B depicts the sequence of a IL-4-IL-13-OSRM^ IL-31 LBD-IL-31RA LBD-cFcConstruct (SEQ ID NO: 26)

[0085] FIG. 4 depicts the DH82-pcDNA3.1-STAT6-SEAP cells stimulated with either 10 ng / mlcIL-4 or cIl-13 in the presence of cIL-4-R-ECD.

[0086] FIG. 5 depicts the Binding of receptor traps (can-IL4R-ECD-canFc and cIL-13RA2-cFc)to ELISA plate-immobilized cIL-4. OD405nm indicates the activity of the detection enzyme alkaline phosphatase. Standard deviations from triplicates are indicated.

[0087] FIG. 6 depicts DH82-pcDNA3.1-STAT6-SEAP cells stimulated with either 10 ng / mlcIL-4 in the presence of can-IL4R-ECD-canFc.

[0088] FIG. 7 depicts neutralization of 10 ng / ml canine IL-4 action in blood of dogs in thepresence and absence of 1000 ng / ml, 100 ng / ml and 10 ng / ml can-IL4R-ECD-His6 and can- IL4R-ECD-canFc. RNA preparations were analysed by qPCR for TARC (CCL-17) induction. Relative ΔΔCq values are given in relation to control blood (dosed with 10 ng / ml cIL-4, but not the receptor traps) set to 1.

[0089] FIG. 8 depicts (A) binding of receptor traps (can-IL4R-ECD-canFc and cIL-13RA2-cFc)to ELISA plate-immobilized cIL-4. OD405nm indicates the activity of the detection enzyme alkaline phosphatase. Standard deviations from triplicates are indicated. (B) PK assay for the double receptor traps: ELISA format.

[0090] FIG. 9 depicts DH82-pcDNA3.1-STAT6-SEAP cells stimulated with either 10 ng / mlcIL-4 or 10 ng / ml cIL-13 in the presence of cIL-13RA2-cFc.

[0091] FIG. 10 depicts DH82-pcDNA3.1-STAT6-SEAP cells stimulated with either 10 ng / mlcIL-4 (A) or cIL-13 (B) in the presence of cIL-4Rα1-cIL-13Rα2-ECD-cFc or cIL-13Rα2-ECD- cIL-4Rα1-cFc.

[0092] FIG. 11 depicts neutralization of 10 ng / ml canine IL-4 action in blood of dogs in thepresence and absence of 20 ng / ml, 200 ng / ml and 2000 ng / ml cIL-4Rα1-cIL-13Rα2-ECD-cFc. (D-F) Neutralization of 10 ng / ml canine IL-4 action in blood of dogs in the presence and absenceU.S. Provisional Patent Application Attorney Docket No.2920951-492977 of 20 ng / ml, 200 ng / ml and 2000 ng / ml cIL-13Rα2-cIL-4Rα1-ECD-cFc. RNA preparations were analysed by qPCR for TARC (CCL-17) induction. Relative ΔΔCq values are given in relation to control blood (either dosed with 10 ng / ml cIL-4, or untreated). (A,D): Linear ΔΔcq scale. (B,E): log10 ΔΔcq scale. (C,F): Calculation of % inhibition relative to the cIL-4 no inhibitor control.

[0093] FIG. 12 depicts the neutralization of 10 ng / ml canine IL-13 action in blood of dogs in thepresence and absence of 20 ng / ml, 200 ng / ml and 2000 ng / ml cIL-13Rα2-cIL-4Rα1-ECD-cFc or cIL-4Rα1-cIL-13Rα2-ECD-cFc. RNA preparations were analysed by qPCR for TARC (CCL-17) induction. Relative ΔΔCq values are given in relation to control blood (either dosed with 10 ng / ml cIL-4, or untreated). (A): Linear ΔΔcq scale. (B): log10 ΔΔcq scale. (C): Calculation of % inhibition relative to the cIL-4 no inhibitor control.

[0094] FIG. 13 depicts (Dog 4) the neutralization of 1 ng / ml cIL-4 (A,B) or 10 ng / ml canine IL-13 (C,D) action in blood of dogs injected with 1 mg / kg cIL-13Rα2-cIL-4Rα1-ECD-cFc. RNA preparations were analysed by qPCR for TARC (CCL-17) induction. (A,C): Relative ΔΔCq values (linear scale) are given in relation to control blood (untreated). (B,D): Calculation of % inhibition relative to the day -3 value set as 0% inhibition.

[0095] FIG. 14 depicts (Dog 5) the neutralization of 1 ng / ml cIL-4 (A,B) or 10 ng / ml canine IL-13 (C,D) action in blood of dogs injected with 1 mg / kg cIL-13Rα2-cIL-4Rα1-ECD-cFc. RNA preparations were analysed by qPCR for TARC (CCL-17) induction. (A,C): Relative ΔΔCq values (linear scale) are given in relation to control blood (untreated). (B,D): Calculation of % inhibition relative to the day -3 value set as 0% inhibition.

[0096] FIG. 15 depicts (Dog 6) neutralization of 1 ng / ml cIL-4 (A,B) or 10 ng / ml canine IL-13(C,D) action in blood of dogs injected with 1 mg / kg cIL-13Rα2-cIL-4Rα1-ECD-cFc. RNA preparations were analysed by qPCR for TARC (CCL-17) induction. (A,C): Relative ΔΔCq values (linear scale) are given in relation to control blood (untreated). (B,D): Calculation of % inhibition relative to the day –3 value set as 0% inhibition.

[0097] FIG. 16 DH82-pcDNA3.1-STAT6-SEAP cells stimulated with either 1 ng / ml cIL-4 (A)or 10 ng / ml cIL-13 (B) in the presence of variable concentrations of cIL4 / 13R-cOS31RA-cFc.

[0098] FIG. 17 depicts the neutralization of 1 ng / ml canine IL-4 (A) or IL-13 (B) action in bloodof dogs in the presence and absence of 20 ng / ml, 200 ng / ml and 2000 ng / ml cIL4 / 13R- cOS31RA-cFc. RNA preparations were analysed by qPCR for TARC (CCL-17) induction inU.S. Provisional Patent Application Attorney Docket No.2920951-492977 comparison to ^-actin as housekeeping mRNA. Relative ΔΔCq values are given in relation to control blood.

[0099] FIG. 18 depicts the binding of biotinylated cIL-31 to ELISA plates coated with either thetriple receptor trap cIL4 / 13R-cOS31RA-cFc, or the dual receptor traps IL13Ra2-cIL4RΑ1-cFc- and cIL4RΑ1-cIL13Ra2-cFc, or to non-coated plates.

[0100] FIG. 19 depicts HEKblue IL-4 / IL-13 cells stimulated with either 1 ng / ml human IL-4 (A)or human IL-13 (B) in the presence of variable concentrations of huIL13Ra2-huIL4Ra-huFc.

[0101] FIG. 20 depicts CRFK-pcDNA3.1-STAT6-SEAP cells stimulated with either feline IL-4(fel-IL-4) or feline IL-13 (fel-IL-13).

[0102] FIG. 21A-B depicts CRFK-pcDNA3.1-STAT6-SEAP cells stimulated with either 5ng / ml fIL-4 (A) or 10 ng / ml fIL-13 (B) in the presence of fIL-13Rα2-fIL-4Rα1-fFc. Definitions

[0103] Unless otherwise defined, all technical and scientific terms used herein have the samemeaning as commonly understood by one of ordinary skill in the art. In case of conflict, the present document, including definitions, will control. Preferred methods and materials are described below, although methods and materials similar or equivalent to those described herein can be used in practice or testing of the present invention. All publications, patent applications, patents and other references mentioned herein are incorporated by reference in their entirety. The materials, methods, and examples disclosed herein are illustrative only and not intended to be limiting.

[0104] “Non-human mammals,” as used herein, refers broadly to non-human mammaliananimals, including but not limited to dogs, cats, mice, rats, guinea pigs, rabbits, ferrets, cows, horses, sheep, goats, and pigs. Non-human mammals can be mammalian pets or companion animals, including but not limited to dogs and cats and also mice, rats, guinea pigs, ferrets, and rabbits. The non-human mammal can be a dog or a cat.

[0105] “Treatment” refers broadly to both therapeutic treatment and prophylactic or preventativemeasures. Those in need of treatment include those already with the disorder as well as those in which the disorder is to be prevented. As used herein, the term “treating,” refers broadly to treating a disease, arresting, or reducing the development of the disease or its clinical symptoms, and / or relieving the disease, causing regression of the disease or its clinical symptoms. TherapyU.S. Provisional Patent Application Attorney Docket No.2920951-492977 encompasses prophylaxis, treatment, remedy, reduction, alleviation, and / or providing relief from a disease, signs, and / or symptoms of a disease. Therapy encompasses an alleviation of signs and / or symptoms in patients with ongoing disease signs and / or symptoms. Therapy also encompasses “prophylaxis”. The term “reduced”, for purpose of therapy, refers broadly to the clinical significant reduction in signs and / or symptoms. Therapy includes treating relapses or recurrent signs and / or symptoms. Therapy encompasses but is not limited to precluding the appearance of signs and / or symptoms anytime as well as reducing existing signs and / or symptoms and eliminating existing signs and / or symptoms. Therapy includes treating chronic disease (“maintenance”) and acute disease. For example, treatment includes treating or preventing relapses or the recurrence of signs and / or symptoms.

[0106] “Host cell,” as used herein refers broadly to the particular subject cell transfected with anucleic acid molecule and the progeny or potential progeny of such a cell. Progeny may not be identical to the parent cell transfected with the nucleic acid molecule due to mutations or environmental influences that may occur in succeeding generations or integration of the nucleic acid molecule into the host cell genome.

[0107] “Effective amount,” as used herein, refers broadly to the amount of an agent, e.g., acompound, antibody, vector or cells that, when administered to a patient for treating a disease, is sufficient to effect such treatment for the disease. The effective amount can be an amount effective for prophylaxis, and / or an amount effective for prevention. The effective amount can be an amount effective to reduce, an amount effective to prevent the incidence of signs / symptoms, to reduce the severity of the incidence of signs / symptoms, to eliminate the incidence of signs / symptoms, to slow the development of the incidence of signs / symptoms, to prevent the development of the incidence of signs / symptoms, and / or effect prophylaxis of the incidence of signs / symptoms. The “effective amount” can vary depending on the disease and its severity and the age, weight, medical history, susceptibility, and pre-existing conditions, of the patient to be treated. The term “effective amount” is synonymous with “therapeutically effective amount” for purposes described herein.

[0108] “Mammal,” as used herein, refers broadly to any and all warm-blooded vertebrateanimals of the class Mammalia, characterized by a covering of hair on the skin and, in the female, milk-producing mammary glands for nourishing the young. Mammals include, but areU.S. Provisional Patent Application Attorney Docket No.2920951-492977 not limited to, humans, domestic and farm animals, and zoo, sports, or pet animals. Mammal also includes any and all those listed on the Mammal Species of the World maintained by the National Museum of Natural History, Smithsonian Institution in Washington D.C. Similarly, the term “subject” or “patient” includes both human and veterinary subjects and / or patients.

[0109] “Fusion protein,” a used herein, refers broadly to the in frame genetic linkage of at leasttwo heterologous polypeptides. Upon transcription / translation, a single protein is made. In this way, multiple proteins, or fragments thereof can be incorporated into a single polypeptide. “Operably linked” refers to the functional linkage between two or more elements. For example, an operable linkage between two polypeptides fuses both polypeptides together in frame to produce a single polypeptide fusion protein. In an aspect, the fusion protein can further comprise a third polypeptide which can comprise a linker sequence.

[0110] “Variant,” as used herein refers broadly to a polypeptide that possesses a similar oridentical function as the IL-4-binding extracellular domain or IL-13-binding extracellular domain, but does not necessarily comprise a similar or identical amino acid sequence of an the IL-4-binding extracellular domain or IL-13-binding extracellular domain, or possesses a similar or identical structure of an the IL-4-binding extracellular domain or IL-13-binding extracellular domain. A variant having a similar amino acid identity refers to a polypeptide that satisfies at least one of the following: (a) a polypeptide comprising, or alternatively consisting of, an amino acid sequence that is at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or at least 99% identical to the amino acid sequence of an the IL-4-binding extracellular domain or IL-13-binding extracellular domain; (b) a polypeptide encoded by a nucleotide sequence, the complementary sequence of which hybridizes under stringent conditions to a nucleotide sequence encoding an the IL-4-binding extracellular domain or IL-13- binding extracellular domain, of at least 5 amino acid residues, at least 10 amino acid residues, at least 15 amino acid residues, at least 20 amino acid residues, at least 25 amino acid residues, at least 30 amino acid residues, at least 40 amino acid residues, at least 50 amino acid residues, at least 60 amino residues, at least 70 amino acid residues, at least 80 amino acid residues, at least 90 amino acid residues, at least 100 amino acid residues, at least 125 amino acid residues, or at least 150 amino acid residues; and (c) a polypeptide encoded by a nucleotide sequence that is atU.S. Provisional Patent Application Attorney Docket No.2920951-492977 least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or at least 99%, identical to the nucleotide sequence encoding an the IL-4-binding extracellular domain or IL-13-binding extracellular domain. A polypeptide with similar structure to an IL-4- binding extracellular domain or IL-13-binding extracellular domain described herein refers to a polypeptide that has a similar secondary, tertiary or quaternary structure of an IL-4-binding extracellular domain or IL-13-binding extracellular domain as described herein. The structure of a polypeptide can be determined by methods known to those skilled in the art, including but not limited to, X-ray crystallography, nuclear magnetic resonance, and crystallographic electron microscopy. To determine the percent identity of two amino acid sequences or of two nucleic acid sequences, the sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced in the sequence of a first amino acid or nucleic acid sequence for optimal alignment with a second amino acid or nucleic acid sequence). The amino acid residues or nucleotides at corresponding amino acid positions or nucleotide positions are then compared. When a position in the first sequence is occupied by the same amino acid residue or nucleotide at the corresponding position in the second sequence, then the molecules are identical at that position. The percent identity between the two sequences is a function of the number of identical positionsshared by the sequences (e.g., % identity = number of identical overlapping positions / totalnumber of positions x 100%). In one embodiment, the two sequences are the same length.

[0111] The determination of percent identity between two sequences can be accomplished usinga mathematical algorithm known to those of skill in the art. An example of a mathematical algorithm for comparing two sequences is the algorithm of Karlin and Altschul Proc. Natl. Acad. Sci. USA 87:2264-2268 (1990), modified as in Karlin and Altschul Proc. Natl. Acad. Sci. USA 90:5873-5877 (1993). The BLASTn and BLASTx programs of Altschul, et al. J. Mol. Biol. 215:403-410(1990) have incorporated such an algorithm. BLAST nucleotide searches can be performed with the BLASTn program, score = 100, wordlength = 12 to obtain nucleotide sequences homologous to a nucleic acid molecules described herein. BLAST protein searches can be performed with the BLASTx program, score = 50, wordlength = 3 to obtain amino acid sequences homologous to a protein molecules described herein. To obtain gapped alignments for comparison purposes, Gapped BLAST can be utilized as described in Altschul et al. NucleicU.S. Provisional Patent Application Attorney Docket No.2920951-492977 Acids Res.25:3389-3402 (1997). Alternatively, PSI-BLAST can be used to perform an iterated search that detects distant relationships between molecules (Id.). When utilizing BLAST, Gapped BLAST, and PSI-BLAST programs, the default parameters of the respective programs (e.g., BLASTx and BLASTn) can be used.

[0112] Another example of a mathematical algorithm utilized for the comparison of sequences isthe algorithm of Myers and Miller, CABIOS (1989). The ALIGN program (version 2.0), which is part of the GCG sequence alignment software package, has incorporated such an algorithm. Other algorithms for sequence analysis known in the art include ADVANCE and ADAM asdescribed in Torellis and Robotti Comput. Appl. Biosci., 10 :3-5(1994); and FASTA described inPearson and Lipman Proc. Natl. Acad. Sci. 85:2444-8(1988). Within FASTA, ktup is a controloption that sets the sensitivity and speed of the search.

[0113] “Conservative” amino acid substitutions are those substitutions that do not substantiallyaffect or decrease the affinity of a protein, such as an the IL-4-binding extracellular domain or IL-13-binding extracellular domain against IL-4 or IL-13, respectively. For example, the IL-4- binding extracellular domain or IL-13-binding extracellular domain binds Il-14 or Il-13, respectively, can include at most about 1, at most about 2, at most about 5, at most about 10, or at most about 15 conservative substitutions and bind IL-4 or Il-13. The term “conservative variant” also includes the use of a substituted amino acid in place of an unsubstituted parent amino acid, provided that the IL-4-binding extracellular domain or IL-13-binding extracellular domain binds IL-4 or IL-13, respectively. Non-conservative substitutions are those that reduce binding to IL-4 or IL-13.

[0114] Conservative amino acid substitution tables providing functionally similar amino acidsare well known to one of ordinary skill in the art. The following six groups are examples of amino acids that are considered to be conservative substitutions for one another: 1) Alanine (A), Serine (S), Threonine (T); 2) Aspartic acid (D), Glutamic acid (E); 3) Asparagine (N), Glutamine (Q); 4) Arginine (R), Lysine (K); 5) Isoleucine (I), Leucine (L), Methionine (M), Valine (V); andU.S. Provisional Patent Application Attorney Docket No.2920951-492977 6) Phenylalanine (F), Tyrosine (Y), Tryptophan (W).

[0115] The term “derivative” as used herein, refers to a variant polypeptide described herein thatcomprises, or alternatively consists of, an amino acid sequence of an antibody described herein that binds to IL-4 or IL-13, which has been altered by the introduction of amino acid residue substitutions, deletions or additions. The term “derivative” as used herein also refers to an protein that binds IL-4 or IL-13 that has been modified, e.g., by the covalent attachment of any type of molecule to the polypeptide. For example, but not by way of limitation, an IL-4-binding extracellular domain or IL-13-binding extracellular domain can be modified, e.g., by glycosylation, acetylation, pegylation, phosphorylation, amidation, derivatization by known protecting / blocking groups, proteolytic cleavage, linkage to a cellular ligand or other protein, etc. A derivative of the IL-4-binding extracellular domain or IL-13-binding extracellular domain can be modified by chemical modifications using techniques known to those of skill in the art, including, but not limited to, specific chemical cleavage, acetylation, formylation, metabolic synthesis of tunicamycin, etc. Further, a derivative of an IL-4-binding extracellular domain or IL-13-binding extracellular domain can contain one or more non-classical amino acids. A polypeptide derivative possesses a similar or identical function as an IL-4-binding extracellular domain or IL-13-binding extracellular domain described herein.

[0116] “Host cell,” as used herein refers broadly to a cell transfected with a nucleic acidmolecule and the progeny or potential progeny of such a cell. Progeny may not be identical to the parent cell transfected with the nucleic acid molecule due to mutations or environmental influences that may occur in succeeding generations or integration of the nucleic acid molecule into the host cell genome. CANINE IL-4 / IL-13 DOUBLE RECEPTOR TRAPS

[0117] IL-4 and IL-13 are Th2 cytokines that are potent mediators of type 2-associatedinflammation, such as those found in atopic dermatitis and allergic dermatitis in canines. IL-13 and IL-4 are potent mediators of type 2–associated inflammation such as those found in atopic dermatitis. IL-4 shares overlapping biological functions with IL-13, a finding that is mainly explained by their ability to signal via the type 2 IL-4 receptor (R), which is composed of IL-4Rα in association with IL-13Rα1.U.S. Provisional Patent Application Attorney Docket No.2920951-492977

[0118] A receptor trap for neutralizing cytokines construct described herein can comprise, e.g.,the extracellular cytokine-binding receptor domain of a cytokine receptor complex fused to a fragment crystallizable region of an antibody (Fc), optionally a canine Fc. The receptor domain provides high ligand affinity, often in the picomolar range, while the Fc provides in vivo stability.

[0119] The IL-4 / IL-13 double receptor trap constructs described herein can comprise an IL-4extracellular domain (ECD) linked by a linker to an IL-13 extracellular domain (ECD) lined by a linker a fragment crystallizable region of an antibody (Fc). The IL-4 / IL-13 double receptor trap constructs described herein can comprise an IL-13 extracellular domain (ECD) linked by a linker to an IL-4 extracellular domain (ECD) lined by a linker a fragment crystallizable region of an antibody (Fc), optionally a canine Fc. The IL-4 and IL-13 can be linked by a linker comprising, for example, a polyglycine peptide (GGGGGGGG SID: 17) or a glycine-serine sequence (GGGGSGGGGSGGGGG SID: 16). For example, the IL-4 / IL-13 double receptor trap constructs described herein can comprise the following formulas: IL-4 ECD-linker-IL-13 ECD-linker-Fc IL-13 ECD-linker-IL-4 ECD-linker-Fc

[0120] The IL-4 / IL-13 double receptor trap constructs described herein can further comprise anartificial optimized ER import signal sequence, e.g. SEQ ID NOs: 66-67), at the N-terminus. The IL-4 / IL-13 double receptor trap constructs described herein can further comprise a polyhistidine tag, e.g., a stretch of 5-8 histidines. For example, the IL-4 / IL-13 double receptor trap constructs described herein can comprise the following formulas: ER import signal sequence-IL-4 ECD-linker-IL-13 ECD-linker-Fc IL-4 ECD-linker-IL-13 ECD-linker-Fc-polyhistidine tag ER import signal sequence-IL-4 ECD-linker-IL-13 ECD-linker-Fc-polyhistidine tag ER import signal sequence-IL-13 ECD-linker-IL-4 ECD-linker-Fc IL-13 ECD-linker-IL-4 ECD-linker-Fc-polyhistidine tag ER import signal sequence-IL-13 ECD-linker-IL-4 ECD-linker-Fc-polyhistidine tag

[0121] In an embodiment, the cIL-31RA-ligand binding domain (LBD) is linked to thecOSRM^-LBD.U.S. Provisional Patent Application Attorney Docket No.2920951-492977

[0122] In an embodiment, the C-terminus of the IL-4-binding extracellular domain is linkedto the N-terminus IL-13-binding extracellular domain which is linked to the N-terminus of the fragment crystallizable region of an antibody.

[0123] In an embodiment, the C-terminus of the IL-13-binding extracellular domain is linkedto the N-terminus of the IL-4-binding extracellular domain which is linked to the N-terminus of the fragment crystallizable region of an antibody.

[0124] In an embodiment, the C-terminus of the IL-4-binding extracellular domain (IL-4-Rα1-ECD) is linked to the N-terminus of the IL-13-binding extracellular domain (IL-13-RA2- ECD) which is linked to the N-terminus of the OSRM^-ligand binding domain (LBD) which is linked to the N-terminus of the fragment crystallizable region of an antibody (Fc).

[0125] In an embodiment, the C-terminus of the IL-4-binding extracellular domain (IL-4-Rα1-ECD) is linked to the N-terminus of the IL-13-binding extracellular domain (IL-13-RA2- ECD) which is linked to the N-terminus of the OSRM^-ligand binding domain (LBD) which is linked to the N-terminus of the IL-31RA-ligand binding domain (LBD) which is linked to the N- terminus of the fragment crystallizable region of an antibody (Fc).

[0126] In an embodiment, the IL-4-binding extracellular domain (IL-4-Rα1-ECD) and cFcare joined by a linker.

[0127] In an embodiment, the IL-13-binding extracellular domain (IL-13-RA2-ECD) andcFc are joined by a linker.

[0128] In an embodiment, the IL-4-binding extracellular domain (IL-4-Rα1-ECD) and theIL-13-binding extracellular domain (IL-13-RA2-ECD) are joined by a linker.

[0129] In an embodiment, the IL-13-binding extracellular domain (IL-13-RA2-ECD) andOSRM^-ligand binding domain (LBD) are joined by a linker.

[0130] In an embodiment, the OSRM^-ligand binding domain (LBD) and the IL-31RA-ligand binding domain (LBD) are joined by a linker.

[0131] In an embodiment, the IL-31RA-ligand binding domain (LBD) and fragmentcrystallizable region of an antibody (Fc) are joined by a linker. The linker is an oligopeptide can comprise about 8 to 20 amino acids. The oligopeptide can be a polyglycine oligopeptide, optionally comprising the amino acid sequence of SEQ ID NO: 81. The oligopeptide can be aU.S. Provisional Patent Application Attorney Docket No.2920951-492977 polyglycine / serine oligopeptide, optionally comprising the amino acid sequence of SEQ ID NO: 82, 83, or 84. The oligopeptide can comprise the amino acid sequence of SEQ ID NO: 85.

[0132] The IL-4α1-binding extracellular domain is a canine sequence. The IL-4α1-bindingextracellular domain can comprise an amino acid sequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 2. The IL-4α1-binding extracellular domain can comprise an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 2. The IL-4α1- binding extracellular domain can comprise an amino acid sequence of SEQ ID NO: 2.

[0133] The IL-4α1-binding extracellular domain can further comprise an ER import signaland a hexa-histidtine tag. The IL-4α1-binding extracellular domain can comprise the amino acid sequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 3. The IL-4α1- binding extracellular domain can comprise the amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the SEQ ID NO: 3. The IL-4α1-binding extracellular domain can comprise the amino acid sequence of SEQ ID NO: 3.

[0134] The IL-4α1-binding extracellular domain can be encoded by a nucleic acid with atleast 75% homology to the nucleic acid sequence of SEQ ID NO: 4. The IL-4α1-binding extracellular domain can be encoded by a nucleic acid with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the nucleic acid sequence of SEQ ID NO: 4. The IL-4α1-binding extracellular domain can be encoded by a nucleic acid of SEQ ID NO: 4.

[0135] The Fc sequence can be a canine sequence. The Fc sequence can comprise an aminoacid sequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 7. The Fcsequence can comprise an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 7. The Fc sequence can comprise an amino acid sequence of SEQ ID NO: 7.

[0136] The canine IL-4alpha1 receptor ECD-canine IgG Fc fusion protein comprises anamino acid sequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 8. The fusion protein can comprise an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence ofU.S. Provisional Patent Application Attorney Docket No.2920951-492977 SEQ ID NO: 8. The canine IL-4alpha1 receptor ECD-canine IgG Fc fusion protein can further comprise an ER import signal and a hexa-histidine tag. The canine IL-4alpha1 receptor ECD- canine IgG Fc fusion protein comprises an amino acid sequence of SEQ ID NO: 8.

[0137] The canine IL-4alpha1 receptor ECD-canine IgG Fc fusion protein can be encoded bya nucleic acid with at least 75% homology to the nucleic acid sequence of SEQ ID NO: 9. The canine IL-4alpha1 receptor ECD-canine IgG Fc fusion protein can be encoded by a nucleic acid with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the nucleic acid sequence of SEQ ID NO: 9. The canine IL-4alpha1 receptor ECD- canine IgG Fc fusion protein can be encoded by a nucleic acid of SEQ ID NO: 9.

[0138] The IL-13-binding extracellular domain can be a canine sequence. The IL-13-bindingextracellular domain can comprise an amino acid sequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 12. The IL-13-binding extracellular domain can comprise an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 12. The IL-13- binding extracellular domain can comprise an amino acid sequence of SEQ ID NO: 12.

[0139] The IL-13Ralpha2 ECD-canine IgG Fc fusion protein can comprise an amino acidsequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 13. The fusion protein can comprise an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 13. The IL-13Ralpha2 ECD-canine IgG Fc fusion protein can comprise an amino acid sequence of SEQ ID NO: 13.

[0140] The IL-13Ralpha2 ECD-canine IgG Fc fusion protein can be encoded by a nucleicacid with at least 75% homology to the nucleic acid sequence of SEQ ID NO: 14. The IL- 13Ralpha2 ECD-canine IgG Fc fusion protein can be encoded by a nucleic acid with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the nucleic acid sequence of SEQ ID NO: 14. The IL-13Ralpha2 ECD-canine IgG Fc fusion protein can be encoded by a nucleic acid of SEQ ID NO: 14.

[0141] The cIL-13Rα2-cIL-4Rα1-ECD-cFc fusion protein can comprise an amino acidsequence with at least about 75% sequence homology to SEQ ID NO: 16. The cIL-13Rα2-cIL- 4Rα1-ECD-cFc fusion protein can comprise an amino acid sequence with at least 80%, 85%,U.S. Provisional Patent Application Attorney Docket No.2920951-492977 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 16. The cIL-13Rα2-cIL-4Rα1-ECD-cFc fusion protein can comprise an amino acid sequence of SEQ ID NO: 16.

[0142] The cIL-13Rα2-cIL-4Rα1-ECD-cFc fusion protein can be encoded by a nucleic acidwith at least 75% homology to the nucleic acid sequence of SEQ ID NO: 17. The cIL-13Rα2- cIL-4Rα1-ECD-cFc fusion protein can be encoded by a nucleic acid with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the nucleic acid sequence of SEQ ID NO: 17. The cIL-13Rα2-cIL-4Rα1-ECD-cFc fusion protein can be encoded by a nucleic acid comprising the nucleotide sequence of SEQ ID NO: 17.

[0143] The cIL-13Rα2-cIL-4Rα1-ECD-cFc fusion protein can comprise an amino acidsequence with at least about 75% sequence homology to SEQ ID NO: 19. The cIL-13Rα2-cIL- 4Rα1-ECD-cFc fusion protein can comprise an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 19. The cIL-13Rα2-cIL-4Rα1-ECD-cFc fusion protein can comprise an amino acid sequence of SEQ ID NO: 19.

[0144] The cIL-13Rα2-cIL-4Rα1-ECD-cFc fusion protein can be encoded by a nucleic acidwith at least 75% homology to the nucleic acid sequence of SEQ ID NO: 20. The cIL-13Rα2- cIL-4Rα1-ECD-cFc fusion protein can be encoded by a nucleic acid with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the nucleic acid sequence of SEQ ID NO: 20. The cIL-13Rα2-cIL-4Rα1-ECD-cFc fusion protein can be encoded by a nucleic acid comprising the nucleotide sequence of SEQ ID NO: 20.

[0145] The IL-31RA sequence can be a canine sequence. The IL-31RA sequence cancomprise an amino acid sequence with at least 75% homology to the amino acid sequence ofSEQ ID NO: 23. The IL-31RA sequence can comprise an amino acid sequence with at least80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 23. The IL-31RA sequence can comprise an amino acid sequence of SEQ ID NO: 23.

[0146] The canine fusion protein can comprise polygycline, polyglycine / serine linker, or a14-mer linker.

[0147] CANINE IL-4 / IL-13 / IL-31 TRIPLE RECEPTOR TRAPSU.S. Provisional Patent Application Attorney Docket No.2920951-492977

[0148] A receptor trap for neutralizing cytokines construct described herein can comprise, e.g.,the extracellular cytokine-binding receptor domains of cytokine receptor complexes fused to a fragment crystallizable region of an antibody (Fc), optionally a canine Fc. The receptor domain provides high ligand affinity, often in the picomolar range, while the Fc provides in vivo stability.

[0149] The canine Fc-linked canine IL-4 / IL-13 / IL-31 triple receptor trap construct describedherein can comprise, from the N-terminus: IL-4α1-extracellular domain (ECD) linked by a linkerto IL-13Rα2-ECD linked by a linker to Oncostatin-M-specific receptor subunit beta (OSRM) ^- ligand binding domain (LBD) to IL-31RA-LBD linked by a linker to a fragment crystallizable region of an antibody (Fc). The canine Fc-linked canine IL-4 / IL-13 / IL-31 triple receptor trap construct described herein can further comprise ER import signal sequence at the N-terminus and / or a polyhistidine tag on the C-terminus.

[0150] The canine Fc-linked canine IL-4 / IL-13 / IL-31 triple receptor trap construct describedherein can comprise, from the N-terminus: a signal sequence directing ER import, the maturecIL-4Rα1 extracellular domain (ECD) sequence protein sequence, a glycine / serine (G / S)- containing 15 amino acid linker (GGGGSGGGGSGGGGG) (SEQ ID NO: 82), the mature cIL- 13Rα2 ECD, a 15 amino acid linker (GGGGSGGGGSGGGGG) (SEQ ID NO: 82), the cOSRM^ cIL-31 ligand binding domain (LBD) followed by an 20 amino acid linker (GGGGSGGGGSGGGGG) (SEQ ID NO: 83), the IL-31RA LBD, an octa-glycine linker, and Fc part from the canine IgG A gene. The C-terminus is formed by a hexa-histidine tag for protein purification purposes. For example, the IL-4 / IL-13 / IL-31 triple receptor trap constructs described herein can comprise the following general structure shown in FIG.3. ER signal-IL-4 ECD-linker-IL-13 ECD-linker-cOSRM^-LBD-cIL-31RA-LBD-Fc-hexaHis

[0151] The OSMR^ sequence can be a canine sequence. The OSMR^ sequence cancomprise an amino acid sequence with at least 75% homology to the amino acid sequence ofSEQ ID NO: 25. The OSMR^ sequence can comprise an amino acid sequence with at least80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 25. The OSMR^ sequence can comprise the amino acid sequence of SEQ ID NO: 25.U.S. Provisional Patent Application Attorney Docket No.2920951-492977

[0152] The IL-4 / IL-13 / IL-31 fusion protein can comprise an amino acid sequence with atleast 75% homology to the amino acid sequence of SEQ ID NO: 26. The fusion protein cancomprise an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 26. The IL-4 / IL-13 / IL-31 fusion protein can be encoded by a nucleic acid with at least 75% homology to the nucleic acid sequence of SEQ ID NO: 27. The IL-4 / IL-13 / IL-31 fusion protein can be encoded by a nucleic acid with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the nucleic acid sequence of SEQ ID NO: 27. The IL-4 / IL- 13 / IL-31 fusion protein can comprise an amino acid sequence of SEQ ID NO : 26.

[0153] HUMAN IL-4 / IL-13 DOUBLE RECEPTOR TRAPS

[0154] IL-4 and IL-13 are Th2 cytokines that are potent mediators of type 2-associatedinflammation, such as those found in atopic dermatitis and allergic dermatitis. IL-13 and IL-4 are potent mediators of type 2–associated inflammation such as those found in atopic dermatitis. IL- 4 shares overlapping biological functions with IL-13, a finding that is mainly explained by their ability to signal via the type 2 IL-4 receptor (R), which is composed of IL-4Rα in association with IL-13Rα1.

[0155] A receptor trap for neutralizing cytokines construct described herein can comprise, e.g.,the extracellular cytokine-binding receptor domain of a cytokine receptor complex fused to a fragment crystallizable region of an antibody (Fc), optionally a human Fc. The receptor domain provides high ligand affinity, often in the picomolar range, while the Fc provides in vivo stability.

[0156] The IL-4 / IL-13 double receptor trap constructs described herein can comprise an IL-4receptor extracellular domain (ECD) linked by a linker to an IL-13 receptor extracellular domain (ECD) lined by a linker a fragment crystallizable region of an antibody (Fc). The IL-4 / IL-13 double receptor trap constructs described herein can comprise an IL-13 extracellular domain (ECD) linked by a linker to an IL-4 extracellular domain (ECD) linked by a linker to a fragment crystallizable region of an antibody (Fc) , optionally a human Fc. The IL-4 and IL-13 receptor extracellular domains and Fc portion can be linked by a linker comprising, for example, a polyglycine peptide (SEQ ID NO: 81), a glycine-serine sequence (SEQ ID NO: 82, 83), or a 14-U.S. Provisional Patent Application Attorney Docket No.2920951-492977 mer linker (SEQ ID NO: 84). For example, the IL-4 / IL-13 double receptor trap constructs described herein can comprise the following formulas: IL-4R ECD-linker-IL-13R ECD-linker-Fc IL-13R ECD-linker-IL-4R ECD-linker-Fc

[0157] The IL-4 / IL-13 double receptor trap constructs described herein can further comprise anartificial optimized ER import signal sequence, e.g., (SEQ ID NOs: 67-76), at the N-terminus. The IL-4 / IL-13 double receptor trap constructs described herein can further comprise a polyhistidine tag, e.g., a stretch of 5-8 histidines, e.g., SEQ ID NO: 87. For example, the IL-4 / IL- 13 double receptor trap constructs described herein can comprise the following formulas: ER import signal sequence-IL-4R ECD-linker-IL-13R ECD-linker-Fc IL-4R ECD-linker-IL-13R ECD-linker-Fc-polyhistidine tag ER import signal sequence-IL-4R ECD-linker-IL-13R ECD-linker-Fc-polyhistidine tag ER import signal sequence-IL-13R ECD-linker-IL-4R ECD-linker-Fc IL-13R ECD-linker-IL-4R ECD-linker-Fc-polyhistidine tag ER import signal sequence-IL-13R ECD-linker-IL-4R ECD-linker-Fc-polyhistidine tag

[0158] The IL-4-binding extracellular domain can be a human sequence. The IL-4-bindingextracellular domain can comprise an amino acid sequence with at least 75% homology to theamino acid sequence of SEQ ID NO: 30. The IL-4-binding extracellular domain can comprisean amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 30. The IL-4-binding extracellular domain can comprise the amino acid sequence of SEQ ID NO: 30.

[0159] The IL-13-binding extracellular domain can be a human sequence. The IL-13-bindingextracellular domain can comprise an amino acid sequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 32. The IL-13-binding extracellular domain can comprise an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 32. The IL-13- binding extracellular domain can comprise an amino acid sequence of SEQ ID NO: 32.

[0160] In an embodiment, the Fc sequence can be a human sequence. The Fc sequence cancomprise an amino acid sequence with at least 75% homology to the amino acid sequence ofU.S. Provisional Patent Application Attorney Docket No.2920951-492977SEQ ID NO: 33. The Fc sequence can comprise an amino acid sequence with at least 80%, 85%,90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 33. The Fc sequence can comprise the amino acid sequence of SEQ ID NO: 33.

[0161] The fusion protein can comprise a hIL-13Rα2-hIL-4Rα1-ECD-cFc protein constructcomprising an amino acid sequence with at least about 75% sequence homology to SEQ ID NO: 34. The fusion protein comprises a hIL-13Rα2-hIL-4Rα1-ECD-cFc protein construct can comprise an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 34. The hIL-13Rα2-hIL-4Rα1-ECD-cFc fusion protein can be encoded by a nucleic acid with at least 75% homology to the nucleic acid sequence of SEQ ID NO: 35. The hIL-13Rα2-hIL-4Rα1- ECD-cFc fusion protein can be encoded by a nucleic acid with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the nucleic acid sequence of SEQ ID NO: 35. The fusion protein can comprise a hIL-13Rα2-hIL-4Rα1-ECD-cFc protein construct comprising an amino acid sequence of SEQ ID NO: 34.

[0162] FELINE IL-4 / IL-13 DOUBLE RECEPTOR TRAPS

[0163] IL-4 and IL-13 are Th2 cytokines that are potent mediators of type 2-associatedinflammation, such as those found in atopic dermatitis and allergic dermatitis in felines. IL-13 and IL-4 are potent mediators of type 2–associated inflammation such as those found in atopic dermatitis. IL-4 shares overlapping biological functions with IL-13, a finding that is mainly explained by their ability to signal via the type 2 IL-4 receptor (R), which is composed of IL- 4Rα1 in association with IL-13Rα1.

[0164] A receptor trap for neutralizing cytokines construct described herein can comprise, e.g.,the extracellular cytokine-binding receptor domain of a cytokine receptor complex fused to a fragment crystallizable region of an antibody (Fc), optionally a feline Fc. The receptor domain provides high ligand affinity, often in the picomolar range, while the Fc provides in vivo stability.

[0165] The IL-4 / IL-13 double receptor trap constructs described herein can comprise an IL-4extracellular domain (ECD) linked by a linker to an IL-13 extracellular domain (ECD) lined by a linker a fragment crystallizable region of an antibody (Fc). The IL-4 / IL-13 double receptor trapU.S. Provisional Patent Application Attorney Docket No.2920951-492977 constructs described herein can comprise an IL-13 extracellular domain (ECD) linked by a linker to an IL-4 extracellular domain (ECD) lined by a linker a fragment crystallizable region of an antibody (Fc), optionally a feline Fc. The IL-4 and IL-13 can be linked by a linker comprising, for example, a polyglycine peptide (GGGGGGGG) (SEQ ID NO: 81) or a glycine-serine sequence of SEQ ID NO: 82, 83). For example, the IL-4 / IL-13 double receptor trap constructs described herein can comprise the following formulas: IL-4 ECD-linker-IL-13 ECD-linker-Fc IL-13 ECD-linker-IL-4 ECD-linker-Fc

[0166] The IL-4 / IL-13 double receptor trap constructs described herein can further comprise anartificial optimized ER import signal sequence, e.g., MGWSCIILFLVATATGVHS (SEQ ID NO: 70), at the N-terminus. The IL-4 / IL-13 double receptor trap constructs described herein can further comprise a polyhistidine tag, e.g., a stretch of 5-8 histidines. For example, the IL-4 / IL-13 double receptor trap constructs described herein can comprise the following formulas: ER import signal sequence-IL-4 ECD-linker-IL-13 ECD-linker-Fc IL-4 ECD-linker-IL-13 ECD-linker-Fc-polyhistidine tag ER import signal sequence-IL-4 ECD-linker-IL-13 ECD-linker-Fc-polyhistidine tag ER import signal sequence-IL-13 ECD-linker-IL-4 ECD-linker-Fc IL-13 ECD-linker-IL-4 ECD-linker-Fc-polyhistidine tag ER import signal sequence-IL-13 ECD-linker-IL-4 ECD-linker-Fc-polyhistidine tag

[0167] The IL-4-binding extracellular domain can be a feline sequence. The IL-4-bindingextracellular domain can comprise an amino acid sequence with at least 75% homology to theamino acid sequence of SEQ ID NO: 39. The IL-4-binding extracellular domain can comprisean amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 39. The IL-4-binding extracellular domain can comprise an amino acid sequence with at least 75% homology to theamino acid sequence of SEQ ID NO: 40. The IL-4-binding extracellular domain can comprisean amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 40. The IL-4-binding extracellular domain can comprise an amino acid sequence of SEQ ID NO: 39.U.S. Provisional Patent Application Attorney Docket No.2920951-492977

[0168] The IL-13-binding extracellular domain can be a feline sequence. The IL-13-bindingextracellular domain can comprise an amino acid sequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 42. The IL-13-binding extracellular domain can comprise an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 42. The IL-13- binding extracellular domain can comprise an amino acid sequence of SEQ ID NO: 42.

[0169] The IL-13-binding extracellular domain can be a feline sequence. The IL-13-bindingextracellular domain can comprise an amino acid sequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 43. The IL-13-binding extracellular domain can comprise an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 43. The IL-13- binding extracellular domain can comprise an amino acid sequence of SEQ ID NO: 43.

[0170] The IL-13-binding extracellular domain can be a feline sequence. The IL-13-bindingextracellular domain can comprise an amino acid sequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 44. The IL-13-binding extracellular domain can comprise an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 44. The IL-13- binding extracellular domain can comprise an amino acid sequence of SEQ ID NO: 44.

[0171] The Fc sequence can be a feline sequence. The Fc sequence can comprise an aminoacid sequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 45. TheFc sequence can comprise an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 45. The Fc sequence can comprise an amino acid sequence of SEQ ID NO: 45.

[0172] The Fc sequence can be a feline sequence. The Fc sequence can comprise an aminoacid sequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 46. TheFc sequence can comprise an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 46. The Fc sequence can comprise an amino acid sequence of SEQ ID NO: 46.

[0173] The Fc sequence can be a feline sequence. The Fc sequence can comprise an aminoacid sequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 47. TheU.S. Provisional Patent Application Attorney Docket No.2920951-492977 Fc sequence can comprise an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 47. The Fc sequence can comprise an amino acid sequence of SEQ ID NO: 47.

[0174] The fIL-13Rα2-fIL-4Rα1-ECD-cFc fusion protein can comprise an amino acidsequence with at least about 75% sequence homology to SEQ ID NO: 48. The fusion protein comprises a fIL-13Rα2-fIL-4Rα1-ECD-cFc protein construct can comprise an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 48. The fIL-13Rα2-fIL-4Rα1- ECD-cFc fusion protein can comprise an amino acid sequence of SEQ ID NO: 48.

[0175] The fIL-13Rα2-fIL-4Rα1-ECD-cFc fusion protein can comprise an amino acidsequence with at least about 75% sequence homology to SEQ ID NO: 49. The fusion protein comprises a fIL-13Rα2-fIL-4Rα1-ECD-cFc protein construct can comprise an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 49. The fIL-13Rα2-fIL-4Rα1- ECD-cFc fusion protein can comprise an amino acid sequence of SEQ ID NO: 49.

[0176] The fIL-13Rα2-fIL-4Rα1-ECD-cFc fusion protein can be encoded by a nucleic acidwith at least 75% homology to the nucleic acid sequence of SEQ ID NO: 50. The fIL-13Rα2-fIL- 4Rα1-ECD-cFc fusion protein can be encoded by a nucleic acid with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the nucleic acid sequence of SEQ ID NO: 50. The fIL-13Rα2-fIL-4Rα1-ECD-cFc fusion protein can be encoded by a nucleic acid of SEQ ID NO: 50.

[0177] Polypeptide Variants

[0178] The polypeptides described herein can have sequence identity polypeptides of at least65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% of to the sequences. In further embodiments, the polypeptides have at least 60%, 65%, 70%,71%, 72%, 73%, 74%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% of identity in their amino acid sequences with other polypeptides to which they are compared. Nucleic Acids, Vectors, and Host CellsU.S. Provisional Patent Application Attorney Docket No.2920951-492977

[0179] Described herein are isolated nucleic acid molecules encoding the IL-4 / IL-13 doublereceptor trap constructs and / or IL-4 / IL-13 / IL-31 triple receptor trap proteins described herein (including molecules comprising, or alternatively consisting of, fragments or variants thereof). The nucleic acids can be present in whole cells, in a cell lysate, or in a partially purified or substantially pure form. A nucleic acid can be isolated by purification away from other cellular components or other contaminants (e.g., other cellular nucleic acids or proteins) by standard techniques, including alkaline / SDS treatment, CsCI banding, column chromatography, agarose gel electrophoresis and others well known in the art (Ausubel, et al. (2011) Current Protocols in Molecular Biology John Wiley & Sons, Inc). A nucleic acid described herein can be, for example, DNA or RNA and can or cannot contain intronic sequences. The nucleic acid can be a cDNA molecule. Nucleic acids described herein can be obtained using standard molecular biology techniques. Specifically, degenerate codon substitutions can be achieved by generating, e.g., sequences in which the third position of one or more selected codons is substituted with mixed-base and / or deoxyinosine residues (Batzer et al., Nucleic Acid Res., 19:5081, 1991; Ohtsuka et al., J. Biol. Chem., 260: 2605-8, 1985; Rossolini et al., Mol. Cell. Probes, 8:91-8, 1994).

[0180] Described herein are methods for recombinantly producing IL-4 / IL-13 double receptortrap constructs and / or IL-4 / IL-13 / IL-31 triple receptor trap proteins described herein. Methods of producing the recombinant proteins are well known to those of ordinary skill in the art. The IL- 4 / IL-13 double receptor trap constructs and / or IL-4 / IL-13 / IL-31 triple receptor trap proteins described herein can also be produced by constructing, using conventional techniques well known to those of ordinary skill in the art, an expression vector containing an operon and a DNA sequence encoding the IL-4 / IL-13 double receptor trap constructs and / or IL-4 / IL-13 / IL-31 triple receptor trap proteins described herein (e.g., vectors, especially plasmids, cosmids, viruses, bacteriophages and other vectors common in genetic engineering, which contain the above- mentioned nucleic acid molecules). The nucleic acid molecules contained in the vectors can be linked to regulatory elements that ensure the transcription in prokaryotic and eukaryotic cells.

[0181] Vectors contain elements that facilitate manipulation for the expression of a foreignprotein within the target host cell. Manipulation of sequences and production of DNA for transformation can be, for example, first performed in a bacterial host (e.g., E. coli) and usuallyU.S. Provisional Patent Application Attorney Docket No.2920951-492977 vectors include sequences to facilitate such manipulations, including a bacterial origin of replication and appropriate bacterial selection marker. Selection markers encode proteins necessary for the survival or growth of transformed host cells grown in a selective culture medium. Host cells not transformed with the vector containing the selection gene do not survive in the culture medium. Typical selection genes encode proteins that confer resistance to antibiotics or other toxins, complement auxotrophic deficiencies, or supply critical nutrients not available from complex media. Exemplary vectors and methods for transformation of yeast are described in the art (Burke et al., Methods in Yeast Genetics Cold Spring Harbor Laboratory Press, 2000).

[0182] The polynucleotide coding for the IL-4 / IL-13 double receptor trap constructs and / or IL-4 / IL-13 / IL-31 triple receptor trap proteins described herein can be operably linked to transcriptional and translational regulatory sequences that provide for expression of thepolypeptide in yeast cells. These vector components can include, but are not limited to, one ormore of the following: an enhancer element, a promoter, and a transcription termination sequence. Sequences for the secretion of the polypeptide can also be included (e.g., a signal sequence).

[0183] Nucleic acids are “operably linked” when placed into a functional relationship withanother nucleic acid sequence. For example, DNA for a signal sequence is operably linked to DNA for a polypeptide if it is expressed as a preprotein that participates in the secretion of the polypeptide; a promoter or enhancer is operably linked to a coding sequence if it affects the transcription of the sequence. Generally, “operably linked” refers broadly to contiguous linked DNA sequences, and, in the case of a secretory leader, contiguous and in reading frame. However, enhancers do not have to be contiguous.

[0184] Promoters are untranslated sequences located upstream (5’) to the start codon of astructural gene (generally within about 100 to 1000 bp) that control the transcription and translation of particular nucleic acid sequences to which they are operably linked. Such promoters fall into several classes: inducible, constitutive, and repressible promoters (e.g., that increase levels of transcription in response to absence of a repressor). Inducible promoters can initiate increased levels of transcription from DNA under their control in response to someU.S. Provisional Patent Application Attorney Docket No.2920951-492977 change in culture conditions (e.g., the presence or absence of a nutrient or a change in temperature).

[0185] The expression vectors are transfected into a host cell by convention techniques wellknown to those of ordinary skill in the art to produce a transfected host cell, said transfected host cell cultured by conventional techniques well known to those of ordinary skill in the art to produce said IL-4 / IL-13 double receptor trap constructs and / or IL-4 / IL-13 / IL-31 triple receptor trap proteins described herein.

[0186] The host cells used to express the IL-4 / IL-13 double receptor trap constructs and / or IL-4 / IL-13 / IL-31 triple receptor trap proteins described herein can be either a bacterial cell such as E.coli, yeast (e.g., S. cerevisiae), or a eukaryotic cell (e.g., a mammalian cell line). A mammalian cell of a well-defined type for this purpose, such as a myeloma cell, 3T3, HeLa, C6A2780, Vero, MOCK II, a Chinese hamster ovary (CHO), Sf9, Sf21, COS, NS0, or HEK293 cell line can be used.

[0187] The general methods by which the vectors can be constructed, transfection methodsrequired to produce the host cell and culturing methods required to produce the antibodies, and fragments thereof, from said host cells all include conventional techniques. Although preferably the cell line used to produce the IL-4 / IL-13 double receptor trap constructs and / or IL-4 / IL-13 / IL- 31 triple receptor trap proteins described herein is a mammalian cell line, any other suitable cell line, such as a bacterial cell line such as an E. coli-derived bacterial strain, or a yeast cell line, can be used.

[0188] Similarly, once produced IL-4 / IL-13 double receptor trap constructs and / or IL-4 / IL-13 / IL-31 triple receptor trap proteins described herein can be purified according to standard procedures in the art, such as for example cross-flow filtration, ammonium sulphate precipitation, and affinity column chromatography. Compositions

[0189] The IL-4 / IL-13 double receptor trap constructs and / or IL-4 / IL-13 / IL-31 triple receptortrap proteins described herein can be administered in combination with additional therapeutics. In an embodiment, the IL-4 / IL-13 double receptor trap constructs and / or IL-4 / IL-13 / IL-31 triple receptor trap proteins described herein can be administered in combination with one or more steroids. For example, prednisone, prednisolone, triamcinolone, betamethasone, dexamethasone,U.S. Provisional Patent Application Attorney Docket No.2920951-492977 flumethasone, fludrocortisone, hydrocortisone, methylprednisolone, and combinations thereof. In another embodiment, the present receptor traps can be administered in combination with one or more antihistamines. For example, hydroxyzine, chlorpheniramine, chlorphenamine, dimetindene, diphenhydramine, loratadine, cetirizine, clemastine, and combinations thereof. In yet another embodiment, the present receptor traps can be administered in combination one or more with calcineurin inhibitors. For example, cyclosporine, tacrolimus, and a combination thereof. In yet another embodiment, the present receptor traps can be administered in combination with one or more JAK inhibitors. For example, oclacitinib. In another embodiment, the IL-4 / IL-13 double receptor trap constructs and / or IL-4 / IL-13 / IL-31 triple receptor trap proteins described herein can be administered in combination with one or more neutralizing monoclonal antibodies. For example, anti-IL-4 antibodies, anti-IL-5 antibodies, anti-IL-13 antibodies, anti-IL-17A antibodies, anti-IL-17C antibodies, anti-IL-22 antibodies, anti-IL-31 antibodies, anti-IL-33 antibodies, and combinations thereof. In another embodiment, the IL-4 / IL- 13 double receptor trap constructs and / or IL-4 / IL-13 / IL-31 triple receptor trap proteins described herein can be administered in combination with one or more nonsteroidal anti-inflammatory drugs (NSAID). For example, meloxicam. The IL-4 / IL-13 double receptor trap constructs and / or IL-4 / IL-13 / IL-31 triple receptor trap proteins described herein can be administered concurrently or sequentially with the additional therapeutics using the same or a different route of administration. For example, the IL-4 / IL-13 double receptor trap constructs and / or IL-4 / IL- 13 / IL-31 triple receptor trap proteins described herein can be administered concurrently or sequentially with anti IL-31 antibodies, anti IL-5 antibodies, anti-IL-22 antibodies, or a combination thereof. Effective Amount

[0190] The effective amount of the IL-4 / IL-13 double receptor trap constructs and / or IL-4 / IL-13 / IL-31 triple receptor trap proteins described herein in a composition in an amount sufficient to treat a dermatological condition. The dermatological condition comprises at least one skin disorder selected from psoriasis, atopic dermatitis, skin rash, skin irritation, skin sensitization, allergic reactions, pruritus, and combinations thereof.U.S. Provisional Patent Application Attorney Docket No.2920951-492977

[0191] The effective amount of the IL-4 / IL-13 double receptor trap constructs and / or IL-4 / IL-13 / IL-31 triple receptor trap proteins described herein can range from about 1 nanogram (ng) to 1 gram (g).

[0192] The effective amount of the IL-4 / IL-13 double receptor trap constructs and / or IL-4 / IL-13 / IL-31 triple receptor trap proteins described herein can be about 1 ng to 1,000 ng.

[0193] The effective amount may be between about 1 ng and 100 ng, 10 ng and 500 ng, 200 ngand 800 ng, or 250 ng and 750 ng.

[0194] The effective amount of the IL-4 / IL-13 double receptor trap constructs and / or IL-4 / IL-13 / IL-31 triple receptor trap proteins described herein can be about 1 ng, 2 ng, 3 ng, 4 ng, 5 ng, 6 ng, 7 ng, 8 ng, 9 ng, 10 ng, 11 ng, 12 ng, 13 ng, 14 ng, 15 ng, 16 ng, 17 ng, 18 ng, 19 ng, 20 ng, 21 ng, 22 ng, 23 ng, 24 ng, 25 ng, 26 ng, 27 ng, 28 ng, 29 ng, 30 ng, 31 ng, 32 ng, 33 ng, 34 ng, 35 ng, 36 ng, 37 ng, 38 ng, 39 ng, 40 ng, 41 ng, 42 ng, 43 ng, 44 ng, 45 ng, 46 ng, 47 ng, 48 ng, 49 ng, 50 ng, 51 ng, 52 ng, 53 ng, 54 ng, 55 ng, 56 ng, 57 ng, 58 ng, 59 ng, 60 ng, 61 ng, 62 ng, 63 ng, 64 ng, 65 ng, 66 ng, 67 ng, 68 ng, 69 ng, 70 ng, 71 ng, 72 ng, 73 ng, 74 ng, 75 ng, 76 ng, 77 ng, 78 ng, 79 ng, 80 ng, 81 ng, 82 ng, 83 ng, 84 ng, 85 ng, 86 ng, 87 ng, 88 ng, 89 ng, 90 ng, 91 ng, 92 ng, 93 ng, 94 ng, 95 ng, 96 ng, 97 ng, 98 ng, 99 ng, 100 ng, 110 ng, 120 ng, 130 ng, 140 ng, 150 ng, 160 ng, 170 ng, 180 ng, 190 ng, 200 ng, 210 ng, 220 ng, 230 ng, 240 ng, 250 ng, 260 ng, 270 ng, 280 ng, 290 ng, 300 ng, 310 ng, 320 ng, 330 ng, 340 ng, 350 ng, 360 ng, 370 ng, 380 ng, 390 ng, 400 ng, 410 ng, 420 ng, 430 ng, 440 ng, 450 ng, 460 ng, 470 ng, 480 ng, 490 ng, 500 ng, 525 ng, 550 ng, 575 ng, 600 ng, 625 ng, 650 ng, 675 ng, 700 ng, 725 ng, 750 ng, 775 ng, 800 ng, 825 ng, 850 ng, 875 ng, 900 ng, 950 ng, or 975 ng.

[0195] The effective amount of the IL-4 / IL-13 double receptor trap constructs and / or IL-4 / IL-13 / IL-31 triple receptor trap proteins described herein can be about 1 µg to 1,000 µg.

[0196] The effective amount can be between about 1 µg and 100 µg, 10 µg and 500 µg, 200 µgand 800 µg, or 250 µg and 750 µg.

[0197] The effective amount of the IL-4 / IL-13 double receptor trap constructs and / or IL-4 / IL-13 / IL-31 triple receptor trap proteins described herein can be about 1 µg, 2 µg, 3 µg, 4 µg, 5 µg, 6 µg, 7 µg, 8 µg, 9 µg, 10 µg, 11 µg, 12 µg, 13 µg, 14 µg, 15 µg, 16 µg, 17 µg, 18 µg, 19 µg, 20 µg, 21 µg, 22 µg, 23 µg, 24 µg, 25 µg, 26 µg, 27 µg, 28 µg, 29 µg, 30 µg, 31 µg, 32 µg, 33 µg, 34 µg, 35 µg, 36 µg, 37 µg, 38 µg, 39 µg, 40 µg, 41 µg, 42 µg, 43 µg, 44 µg, 45 µg, 46 µg, 47U.S. Provisional Patent Application Attorney Docket No.2920951-492977 µg, 48 µg, 49 µg, 50 µg, 51 µg, 52 µg, 53 µg, 54 µg, 55 µg, 56 µg, 57 µg, 58 µg, 59 µg, 60 µg, 61 µg, 62 µg, 63 µg, 64 µg, 65 µg, 66 µg, 67 µg, 68 µg, 69 µg, 70 µg, 71 µg, 72 µg, 73 µg, 74 µg, 75 µg, 76 µg, 77 µg, 78 µg, 79 µg, 80 µg, 81 µg, 82 µg, 83 µg, 84 µg, 85 µg, 86 µg, 87 µg, 88 µg, 89 µg, 90 µg, 91 µg, 92 µg, 93 µg, 94 µg, 95 µg, 96 µg, 97 µg, 98 µg, 99 µg, 100 µg, 110 µg, 1203µg, 130 µg, 140 µg, 150 µg, 160 µg, 170 µg, 180 µg, 190 µg, 200 µg, 210 µg, 220 µg, 230 µg, 240 µg, 250 µg, 260 µg, 270 µg, 280 µg, 290 µg, 300 µg, 310 µg, 320 µg, 330 µg, 340 µg, 350 µg, 360 µg, 370 µg, 380 µg, 390 µg, 400 µg, 410 µg, 420 µg, 430 µg, 440 µg, 450 µg, 460 µg, 470 µg, 480 µg, 490 µg, 500 µg, 525 µg, 550 µg, 575 µg, 600 µg, 625 µg, 650 µg, 675 µg, 700 µg, 725 µg, 750 µg, 775 µg, 800 µg, 825 µg, 850 µg, 875 µg, 900 µg, 950 µg, or 975 µg.

[0198] The effective amount of the IL-4 / IL-13 double receptor trap constructs and / or IL-4 / IL-13 / IL-31 triple receptor trap proteins described herein can be about 1 mg to 1,000 mg.

[0199] The effective amount can be between about 1 mg and 100 mg, 10 mg and 500 mg, 200mg and 800 mg, or 250 mg and 750 mg.

[0200] The effective amount of the IL-4 / IL-13 double receptor trap constructs and / or IL-4 / IL-13 / IL-31 triple receptor trap proteins described herein can be about 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, 10 mg, 11 mg, 12 mg, 13 mg, 14 mg, 15 mg, 16 mg, 17 mg, 18 mg, 19 mg, 20 mg, 21 mg, 22 mg, 23 mg, 24 mg, 25 mg, 26 mg, 27 mg, 28 mg, 29 mg, 30 mg, 31 mg, 32 mg, 33 mg, 34 mg, 35 mg, 36 mg, 37 mg, 38 mg, 39 mg, 40 mg, 41 mg, 42 mg, 43 mg, 44 mg, 45 mg, 46 mg, 47 mg, 48 mg, 49 mg, 50 mg, 51 mg, 52 mg, 53 mg, 54 mg, 55 mg, 56 mg, 57 mg, 58 mg, 59 mg, 60 mg, 61 mg, 62 mg, 63 mg, 64 mg, 65 mg, 66 mg, 67 mg, 68 mg, 69 mg, 70 mg, 71 mg, 72 mg, 73 mg, 74 mg, 75 mg, 76 mg, 77 mg, 78 mg, 79 mg, 80 mg, 81 mg, 82 mg, 83 mg, 84 mg, 85 mg, 86 mg, 87 mg, 88 mg, 89 mg, 90 mg, 91 mg, 92 mg, 93 mg, 94 mg, 95 mg, 96 mg, 97 mg, 98 mg, 99 mg, 100 mg, 110 mg, 1203mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, 350 mg, 360 mg, 370 mg, 380 mg, 390 mg, 400 mg, 410 mg, 420 mg, 430 mg, 440 mg, 450 mg, 460 mg, 470 mg, 480 mg, 490 mg, 500 mg, 525 mg, 550 mg, 575 mg, 600 mg, 625 mg, 650 mg, 675 mg, 700 mg, 725 mg, 750 mg, 775 mg, 800 mg, 825 mg, 850 mg, 875 mg, 900 mg, 950 mg, or 975 mg.

[0201] The effective amount of the IL-4 / IL-13 double receptor trap constructs and / or IL-4 / IL-13 / IL-31 triple receptor trap proteins described herein can be about 0.1 mg / kg to 10 mg / kg.U.S. Provisional Patent Application Attorney Docket No.2920951-492977

[0202] The effective amount can be between about 0.1 mg / kg and 5 mg / kg, 1 mg / kg and 5mg / kg, 0.2 mg / kg and 8 mg / kg, or 0.25 mg / kg and 10 mg / kg.

[0203] The effective amount of the IL-4 / IL-13 double receptor trap constructs and / or IL-4 / IL-13 / IL-31 triple receptor trap proteins described herein can be about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 mg / kg. The effective amount can be between about 0.1 and 1 mg / kg, 0.5 and 2 mg / kg, 0.75 and 5 mg / kg, or 1 and 10 mg / kg.

[0204] Therapeutic Methods

[0205] The IL-4 / IL-13 double receptor trap constructs and / or IL-4 / IL-13 / IL-31 triple receptortrap proteins described herein and / or composition comprising an effective amount thereof, can used to treat a dermatological condition. The dermatological condition can comprise at least one skin disorder selected from psoriasis, atopic dermatitis, skin rash, skin irritation, skin sensitization, allergic reactions, pruritus, and combinations thereof.

[0206] The route of administration of the IL-4 / IL-13 double receptor trap constructs and / or IL-4 / IL-13 / IL-31 triple receptor trap proteins described herein and compositions comprising the same can be by known routes, e.g. injection or infusion by intravenous, intraperitoneal, intracerebral, subcutaneous, intramuscular, intraocular, inhaled, optionally intransal, intrapulmonary, intraarterial, intracerebrospinal, or intralesional routes, or by sustained release systems. Preferably The IL-4 / IL-13 double receptor trap constructs and / or IL-4 / IL-13 / IL-31 triple receptor trap proteins described herein are given systemically.

[0207] Therapeutic and pharmaceutical compositions comprising the IL-4 / IL-13 double receptortrap constructs and / or IL-4 / IL-13 / IL-31 triple receptor trap proteins described herein can be used to treat dermatological conditions. The dermatological condition can comprise at least one skin disorder selected from psoriasis, atopic dermatitis, skin rash, skin irritation, skin sensitization, allergic reactions, pruritus, and combinations thereof.

[0208] As a general proposition, the initial pharmaceutically effective amount of the IL-4 / IL-13double receptor trap constructs and / or IL-4 / IL-13 / IL-31 triple receptor trap proteins described herein administered parenterally will be in the range of about 0.1 to 50 mg / kg of patient body weight per day, with the typical initial range of the IL-4 / IL-13 double receptor trap constructs and / or IL-4 / IL-13 / IL-31 triple receptor trap proteins described herein used being 0.3 to 20 mg / kg / day, more preferably 0.3 to 15 mg / kg / day. The desired dosage can be delivered by aU.S. Provisional Patent Application Attorney Docket No.2920951-492977 single bolus administration, by multiple bolus administrations, or by continuous infusion administration of the IL-4 / IL-13 double receptor trap constructs and / or IL-4 / IL-13 / IL-31 triple receptor trap proteins described herein, depending on the pattern of pharmacokinetic decay that the practitioner wishes to achieve.

[0209] Further, the effective amount of the IL-4 / IL-13 double receptor trap constructs and / or IL-4 / IL-13 / IL-31 triple receptor trap proteins described herein administered (optionally parenterally) can be in the range of about 0.1 to 50 mg / kg of patient body weight. The IL-4 / IL-13 double receptor trap constructs and / or IL-4 / IL-13 / IL-31 triple receptor trap proteins described herein can be administered every 1, 2, 3, or 4 weeks. The IL-4 / IL-13 double receptor trap constructs and / or IL-4 / IL-13 / IL-31 triple receptor trap proteins described herein can be administered every 1, 2, 3, 4, 5, or 6 months. For example the IL-4 / IL-13 double receptor trap constructs and / or IL-4 / IL-13 / IL-31 triple receptor trap proteins described herein can be administered every 2-3 months. The IL-4 / IL-13 double receptor trap constructs and / or IL-4 / IL- 13 / IL-31 triple receptor trap proteins described herein can be administered every 2-3 weeks.

[0210] Kits

[0211] A pharmaceutical pack or kit may comprise one or more containers filled with one ormore of the ingredients of the pharmaceutical compositions comprising the IL-4 / IL-13 double receptor trap constructs and / or IL-4 / IL-13 / IL-31 triple receptor trap proteins described herein. Optionally associated with such container(s) can be a notice in the form prescribed by a governmental agency regulating the manufacture, use or sale of pharmaceuticals or biological products, which notice reflects approval by the agency of manufacture, use or sale for veterinary uses.

[0212] Kits that can be used in the methods described herein. A kit may comprise the IL-4 / IL-13double receptor trap constructs and / or IL-4 / IL-13 / IL-31 triple receptor trap proteins described herein or compositions comprising the same, in one or more containers.

[0213] Routes of administration

[0214] The IL-4 / IL-13 double receptor trap constructs and / or IL-4 / IL-13 / IL-31 triple receptortrap proteins described herein can be administered parentally by injection or by gradual infusion over time. Although the tissue to be treated can typically be accessed in the body by systemic administration and therefore most often treated by intravenous administrationU.S. Provisional Patent Application Attorney Docket No.2920951-492977 of therapeutic compositions, other tissues and delivery means are contemplated where there is a likelihood that the tissue targeted contains the target molecule. Thus, the IL-4 / IL-13 double receptor trap constructs and / or IL-4 / IL-13 / IL-31 triple receptor trap proteins described herein may be administered intravenously, intraperitoneally, intramuscularly, subcutaneously, intracaviatary, intravesically, transdermally, topically, intraocually, orally, intranasally, or a combination thereof.

[0215] Further details described herein can be found in the following example, which furtherdefines the scope described herein. All references cited throughout the specification, and the references cited therein, are hereby expressly incorporated by reference in their entirety. EXAMPLES EXAMPLE 1 CANINE IL-4 / IL-13 DUAL RECEPTOR TRAP PROTEINS Design of canine IL-4 receptor trap—canine IL-4-Ra1-ECD-His6

[0216] The canine interleukin 4 receptor α1 subunit gene was examined and the deduced proteinsequence was extracted (SEQ ID NO: 1). Based on a SignalP-5.0 analysis, the predicted ER import signal sequence of IL-4Rα1 spans amino acids (AA) 1-24. However, the peak predicting the cleavage site is broad and could also encompass AA1-12, as predicted for the human sequence. Therefore the N-terminus of mature canine IL-4Rα1 for initial expression experiments was defined as ‘GSVK….’ (SEQ ID NO: 88).

[0217] With respect to the prediction of the transmembrane helix of canine IL-4Ra1, TMHMM2.0 analysis gave no conclusive result, while the DAS transmembrane prediction tool resulted in a sufficiently extended (19 AA) and likely region between AA235-253. This prediction overlaps with the known transmembrane (TM) helix in human of IL-4Rα1. Given that the sequence identity in the alignment with the human sequence is high overall, and specifically in this region, transmembrane helix was defined as “LPLGVSISCLVILAICLSCYFSII” (SEQ ID NO: 78).

[0218] Based on the mature canine IL-4Rα1-ECD protein sequence, an expression constructwith a N-terminally added artificial ER import signal sequence and a C-terminally added hexa- histidine (H6) was designed (SEQ ID NO: 3). This protein sequence was backtranslated into a DNA sequence taking account for the human codon usage bias for expression in HEK293 cells. Furthermore, 5’ upstream of the ATG start codon, a Kozak consensus sequence (GCCACC)U.S. Provisional Patent Application Attorney Docket No.2920951-492977 (SEQ ID NO: 86) was added. In addition, at the 5’ end of the synthetic gene, a unique EcoRI, and at the 3’ end a unique Hind III restriction enzyme site were added for subcloning into the mammalian expression vector pcDNA3.4 (SEQ ID NO: 4). The subcloning of this IL-4Rα1- ECD-His6 DNA resulted in the expression vector pcDNA3.4-canIL4R-ECD (SEQ ID NO: 5).

[0219] HEK293 cells were transiently transfected with recombinant plasmid at the ~ 100 mlculture scale by the CRO (Genscript), by employing standard molecular biology techniques. Culture supernatant was subjected to HisTrap FF crude (Ni2+NTA) column chromatography purification and the recombinant canIL-4-R-ECD (15.40 mg) was transferred into PBS, pH 7.2. The identity and purity of the recombinant product was confirmed by reducing SDS- PAGE / Coomassie Blue staining, where a broad band was observed between 29 kDa and 44 kDa apparent molecular mass. Nonreducing conditions did not lead to a size increase, suggesting that canIL-4-R-ECD does not form disulfide-linked dimers. Analysis of the mature sequence by Protparam predicted a molecular mass of 24670.31 Da. The bands of canIL-4-R-ECD in SDS- PAGE migrate at a considerably higher mass range. NetNGlyc1.0 analysis predicted 7 N- glycosylation sites, six of which have the potential to be occupied. The variable occupancy and a variable degree of sialylation of canIL-4-R-ECD may account for the observed heterogeneity and increased apparent molecular mass in SDS-PAGE analysis.

[0220] To investigate whether cIL-4-R-ECD is exhibiting biological activity, i.e., binding to andneutralizing canine IL-4, a canine DH82 monocyte IL-4 / IL-13 reporter cell line, DH82- pcDNA3.1-STAT6-SEAP, was stimulated with 10 ng / ml canine IL-4 (cIL-4, R&D Systems, 754-CL-025 / CF), or 10 ng / ml canine IL-13 (cIL-13, own E. coli expression at a CRO, Genscript) (Fig.10). CanIL-4-R-ECD was added to these cytokines in a 1:2 dilution titration starting at 10 mg / ml, before application to the DH82-pcDNA3.1-STAT6-SEAP cells. In the case of cIL-13, not expected to interact with canIL-4-R-ECD, no decrease in reporter gene (SEAP) signal was observed (Fig. 10). By contrast, when cIL-4 was used as stimulating cytokine, canIL-4-R-ECD led to a progressive decrease in reporter gene signal, starting already at ~ 20 ng / ml and reaching complete inhibition at ~ 1000 ng / ml (Fig.4). This suggests that canIL-4-R-ECD may be a potent and specific tool to inhibit canine IL-4. EXAMPLE 2 CANINE IL-4R^1-ECD-cFcU.S. Provisional Patent Application Attorney Docket No.2920951-492977

[0221] To generate a more stable and dimeric version of canine IL-4Rα1-ECD, a fusion proteinwith an immunoglobulin G (IgG) fragment crystallizable region (Fc region) was designed. To this end, the Canis lupus (Canis lupus familiaris) IgG heavy chain A protein was chosen (SEQ ID NO: 8).

[0222] The N-terminus of the Fc region of the canine IgG A (SEQ ID NO: 6) was fused to theC-terminus of the IL-4Rα1-ECD (including the ER import signal sequence) via an octa-glycine linker sequence. A His6 sequence was appended to the Fc C-terminus resulting in IL-4Rα1-ECD- cFc-His6(SEQ ID NO: 8). This protein sequence was backtranslated into a DNA sequence taking account for the human codon usage bias for expression in HEK293 cells. Furthermore, 5’ upstream of the ATG start codon, a Kozak consensus sequence (GCCACC) (SEQ ID NO: 86) was added. In addition, at the 5’ end of the synthetic gene, a unique EcoRI, and at the 3’ end a unique Hind III restriction enzyme site were added for subcloning into the mammalian expression vector pcDNA3.4 (SEQ ID NO: 9). The subcloning of this IL-4Rα1-ECD-canFc- His6 DNA resulted in the expression vector pcDNA3.4-can-IL4R-ECD-cFc (SEQ ID NO: 10).

[0223] HEK293 cells were transiently transfected with recombinant plasmid at the ~ 100 mlculture scale by the CRO (Genscript), by employing standard molecular biology techniques. Culture supernatant was subjected to HisTrap FF crude (Ni2+NTA) column chromatography purification and the recombinant canIL-4-R-ECD-canFc (4.04 mg) was transferred into PBS, pH 7.2. The identity and purity of the recombinant product was confirmed by reducing SDS- PAGE / Coomassie Blue staining, where a broad band was observed between 60 kDa and 70 kDa apparent molecular mass. Nonreducing conditions did lead to protein ~ 140 kDa in size, suggesting that can-IL4R-ECD-canFc does form disulfide-linked dimers, probably mediated by the canine Fc domain. Analysis of the mature sequence by Protparam predicted a molecular mass of 51904.03 Da for the reduced polypeptide. The band of reduced can-IL4R-ECD-canFc in SDS- PAGE migrates at a considerably higher mass range. NetNGlyc1.0 analysis predicted 8 N- glycosylation sites, 7 of which have the potential to be occupied. The variable occupancy and a variable degree of sialylation of canIL-4-R-ECD may account for the observed heterogeneity and increased apparent molecular mass in SDS-PAGE analysis.

[0224] In next step, it was investigated whether can-IL4R-ECD-canFc was able to bind toELISA plate immobilized cIL-4 (see procedure below).U.S. Provisional Patent Application Attorney Docket No.2920951-492977 ELISA procedure for detection of receptor trap binding to cytokines:

[0225] In the ELISA format using plate-immobilized cIL-4, dose-dependent binding of can-IL4R-ECD-canFc was detected starting around 50 ng / ml (Fig. 5). The specificity of the assay was ensured by the application of a receptor trap expected to be specific for cIL-13 (cIL-13RA2- cFc). This protein did not generate any specific signal in the cIl-4 binding assay (Fig.5).

[0226] To investigate whether can-IL4R-ECD-canFc is exhibiting biological activity, i.e.,binding to and neutralizing canine IL-4, the canine DH82 monocyte IL-4 / IL-13 reporter cell line, DH82-pcDNA3.1-STAT6-SEAP was stimulated with 10 ng / ml canine IL-4 in the presenceU.S. Provisional Patent Application Attorney Docket No.2920951-492977 of 1:2 dilutions of can-IL4R-ECD-canFc, starting at 10 ^g / ml (Fig. 6). The presence of can- IL4R-ECD-canFc led to a progressive decrease in reporter gene signal, starting already at ~ 50 ng / ml and reaching complete inhibition at ~ 10000 ng / ml (Fig. 6).

[0227] IL-4 induces expression of thymus- and activation-regulated chemokine (TARC or CCL17)at the mRNA and protein level, at least in human and mouse blood cells. In dogs, upregulation of TARC in atopic dermatitis has been documented, although the direct connection to IL-4 exposure has not been shown.

[0228] TARC is a positive marker (strong mRNA upregulation) of canine blood exposure to IL-4.The inhibition of IL-4-induced TARC mRNA upregulation by can-IL4R-ECD-His6 and canine- IL4R-ECD-canFc in ex vivo dog blood samples was assessed to investigate the potential of these proteins as in vivo cIL-4 blockers: -EDTA-stabilized blood was taken from naive dogs.- 500 ^l blood was supplemented with canine IL-4 (R&D systems, 754-CL-025 / CF) to 10ng / ml, in the presence and absence of 1000 ng / ml, 100 ng / ml and 10 ng / ml can-IL4R-ECD- His6 and can-IL4R-ECD-canFc, and the blood was then incubated for 6 h at 35°C, 5% (v / v) CO2and 96% relative humidity -Blood lysis and RNA stabilization was done with RNAprotect Animal Blood Tubes 500μl(Qiagen 76554) and an incubation for 2 h at room temperature -RNA isolation was then done with the RNeasy Protect Animal Blood Kit (Qiagen 73224)- Isolated RNA was analysed and quantified using an Implen NanoPhotometer®, type NP80.- Quantitative RT- PCR (qPCR) was performed with the TaqMan® Assay Cf02622128_m1(Thermo) with probe and primers for canine CCL-17 (TARC) and the QuantiNova Probe RT-PCR Kit (Qiagen 208354), with the primers, reaction mixtures and conditions outlined by the manufacturers. Canine ^-actin TaqMan probe and PCR primers were used as a housekeeping gene control. Typically 25 ng total RNA was used as template. -The qPCR was performed using a CFX96 Real-Time System (BioRad)

[0229] Concomitant addition of the IL-4 receptor traps can-IL4R-ECD-His6 or can-IL4R-ECD-canFc to canine blood inhibits 10 ng / ml IL-4-induced CCL017 / TARC mRNA production and / or accumulation in a dose-dependent manner. Already at 10 ng / ml of the receptor traps inhibit 60- 80 % of the qPCR signal, with 1000 ng / ml attaining 98-100 % inhibition of the signal (Fig. 7).

[0230] Collectively the available data support that, canIL-4-R-ECD-His6 as well as can-IL4R-ECD-canFc are potent and specific tools to inhibit canine IL-4 in in vitro in ELISA and canine cell line assays, and likely also in vivo, as witnessed by dog blood ex vivo assays.U.S. Provisional Patent Application Attorney Docket No.2920951-492977 EXAMPLE 3 DESIGN OF A CANINE IL-13 RECEPTOR TRAP

[0231] The deduced protein sequence was extracted from the canine interleukin-13 receptorsubunit alpha-2 (IL-13-Rα2) gene (SEQ ID NO: 11).

[0232] Based on a SignalP-5.0 analysis, the predicted ER import signal sequence of IL-13Rα2spans amino acids (AA) 1-27. Therefore the N-terminus of mature canine IL-13Rα2 for initial expression experiments was defined as ‘EIKV….’ (SEQ ID NO: 89).

[0233] With respect to the prediction of the transmembrane helix of canine IL-13Rα2, theTMHMM 2.0 transmembrane prediction tool resulted in a sufficiently extended (23 AA) and likely region between AA340-362. These data define the mature canine IL-13Rα2-ECD as a polypeptide of 312 AA starting with ‘EIKV…’ (SEQ ID NO: 89) and ending with ‘…KETL’ (SEQ ID NO: 90).

[0234] Based on the cIL-13Rα2-ECD protein sequence, an expression construct with theendogeneous ER import signal sequence, and a C-terminally added canine Fc part from the canine IgG A gene followed by a hexa-histidine (His6) sequence serving as an affinity tag for Ni2+NTA column purification. (SEQ ID NO: 8) This protein sequence was backtranslated into a DNA sequence taking account for the human codon usage bias for expression in HEK293 cells. Furthermore, 5’ upstream of the ATG start codon, a Kozak consensus sequence (GCCACC) (SEQ ID NO: 86) was added. In addition, at the 5’ end of the synthetic gene, a unique EcoRI, and at the 3’ end a unique Hind III restriction enzyme site were added for subcloning into the mammalian expression vector pcDNA3.4 (SEQ ID NO: 9). The subcloning of this IL-13Rα2- cFc DNA resulted in the expression vector pcDNA3.4-cIL13RA2-cFc (SEQ ID NO: 10).

[0235] HEK293 cells were transiently transfected with recombinant plasmid at the ~ 100 mlculture scale by the CRO (Genscript), by employing standard molecular biology techniques. Culture supernatant was subjected to His TrapFF crude (Ni2+NTA) column chromatography purification and the recombinant cIL13RA2-ECD-cFc (17.28 mg) was transferred into PBS, pH 7.2. The identity and purity of the recombinant product was confirmed by reducing SDS- PAGE / Coomassie Blue staining, where a broad band was observed around ~ 70-80 kDa apparent molecular mass. Nonreducing conditions did lead to protein ~ 140 - 160 kDa in size, suggestingU.S. Provisional Patent Application Attorney Docket No.2920951-492977 that cIL13RA2-ECD-cFc does form disulfide-linked dimers, probably mediated by the canine Fc domain. Analysis of the mature sequence by Protparam predicted a molecular mass of 64268.34 Da for the reduced polypeptide. The band of reduced can-IL4R-ECD-canFc in SDS-PAGE migrates at a slightly higher mass range. NetNGlyc1.0 analysis predicted 4 N-glycosylation sites, all of which have the potential to be occupied. The variable occupancy and a variable degree of sialylation of canIL-4-R-ECD may account for the increased apparent molecular mass in SDS- PAGE analysis.

[0236] In next step, it was investigated whether cIL-13RA2-ECD-cFc is able to bind to ELISAplate immobilized cIL-13 (see procedure below). ELISA procedure for detection of receptor trap binding to cytokines: 1) Coating of polystyrene ELISA plates (384 well: Thermo Maxisorp, CatNo. 464718)with10 µl / well of 5µg / ml dissolved recombinant protein of immune products cIL13 and cIL4 in coating buffer PBS (ThermoFisher Phosphate-Buffered Saline - pH 7.2, CatNo. 20012-019, or equivalent) at 40C with closed lid. 2) Removal of coating solution, washings 3 x with PBS (Thermo Fisher Phosphate-BufferedSaline pH 7.2, Cat.No.20012-019 or equivalent), 0.05% (v / v) Tween 20, 35 µl / well. 3) Blocking of nonspecific binding sites with 35 µl / well PBS, 0.05% (v / v) Tween 20, 5%(w / v) milk powder (= blocking solution). Incubation at room temperature (RT) for > 1 hour (h). 4) Removal of blocking solution (discard), addition of 20 µl / well of sampledilution / preparation. Start at 10 µg / ml a 1:2 dilution series from cIL13RA2-cFc (Genscript, U1355FG060), canIL4-ECD-canFc (Genscript, U368JGD210) in blocking solution on a nonadsorptive ELISA plate. Incubation at room temperature (RT) for > 1 hour (h). 5) Removal of solutions, washings 3 x with PBS 0.05% (v / v) Tween 20, 35 µl / well.6) Addition of 20 µl / well dilutions of 1:2000 in blocking solution diluted Rabbit IgG anti-Dog IgG (Fc)-Alk. Phos., MinX none (Jackson Immuno Research: 304-055-008, Lot: 145922-0.6 mg / ml) (Generally aim at: 100-400 ng / ml conjugate) Incubate at room temperature (RT) for > 1 h. 7) Removal of conjugate solution, washing 2 x with PBS 0.05% (v / v) Tween 20, 35 µl / well.8) Washing 1 x with AP buffer (50 mM NaHCO3 / Na2CO3, 2 mM MgCl2, pH 9.6), 50 µl / well.9) Develop ELISA by adding 90 µl / well of 5 mM 4-nitrophenyl phosphate disodium salthexahydrate (pNPP, Applichem, A1442,0050) in 50 mM NaHCO3 / Na2CO3, 2 mM MgCl2, pH 9.6 buffer. Kinetic monitoring of the optical density (OD) at 405 nm in an ELISA reader (ΔOD / min) at RT. Determination of the curve slope in a linear range. Slopes > 200 mOD / min are considered problematic.U.S. Provisional Patent Application Attorney Docket No.2920951-492977 10) One-point measurement after 1 and 2 hour.

[0237] In the ELISA format using plate-immobilized cIL-13, dose-dependent binding of cIL-13RA2-cFc was detected starting around 500 ng / ml (Fig.8). The specificity of the assay was ensured by the application of a receptor trap expected to be specific for cIL-4 (can-IL4R-ECD- canFc). This protein did not generate any specific signal in the cIL-13 binding assay (Fig.8).

[0238] To investigate whether cIL-13RA2-cFc is exhibiting biological activity, i. e. binding toand neutralizing canine IL-13, the canine DH82 monocyte IL-4 / IL-13 reporter cell line, DH82- pcDNA3.1-STAT6-SEAP was stimulated with 10 ng / ml canine IL-13 in the presence of 1:2 dilutions of cIL-13RA2-cFc, starting at 10 mg / ml (Fig.9). The presence of cIL-13RA2-cFc led to a progressive decrease in reporter gene signal, starting already at ~ 5 ng / ml and reaching complete inhibition at ~ 40 ng / ml (Fig. 9) . Stimulation of DH82-pcDNA3.1-STAT6-SEAP with 10 ng / ml cIL-4 was not sensitive to the presence of cIL-13RA2-cFc (Fig.9). EXAMPLE 4 DESIGN OF CANINE IL4 / IL-13 DUAL RECEPTOR TRAPS IL-13-Ra2-IL-4Ra1-ECD-cFc

[0239] Based on the cIL-13Rα2-ECD protein sequence followed by a glycine / serine (G / S)-containing 15 AA linker (GGGGSGGGGSGGGGG) (SEQ ID NO: 82), followed by the mature cIL-4Rα1-ECD sequence , followed by an octa-glycine linker (GGGGGGGG) (SEQ ID NO: 81) and Fc part from the canine IgG A gene (SEQ ID NO: 7). Upon submission of this sequence to the CRO (Genscript) for gene synthesis and transient expression / protein purification. In this process the CRO applied their own signal sequence prediction program, which predicted a cleavage site N-terminally of ‘SMLS…’ (SEQ ID NO: 91), some 6 amino acids upstream of that identified by SignalP 5.0. This alternative site was also seen in Signal P5.0, but as a minor peak in the prediction algorithm. Therefore, the cIL-13Rα2-ECD component of the IL-13-Rα2-IL- 4Rα1-ECD-cFc construct starts with ‘SMLS…’ (SEQ ID NO: 91) (confirmed by SignalP5.0) instead of ‘EIKV…’ (SEQ ID NO: 89) as in the IL-13-Rα2-ECD-cFc construct.

[0240] This protein sequence (SEQ ID NO: 16) was backtranslated into a DNA sequence takingaccount for the human codon usage bias for expression in HEK293 cells. (SEQ ID NO: 17) Furthermore, 5’ upstream of the ATG start codon, a Kozak consensus sequence (GCCACC)U.S. Provisional Patent Application Attorney Docket No.2920951-492977 (SEQ ID NO: 86) was added. In addition, at the 5’ end of the synthetic gene, a unique EcoRI, and at the 3’ end a unique Hind III restriction enzyme site were added for subcloning into the mammalian expression vector pcDNA3.4 (SEQ ID NO: 17). The subcloning of this IL-13Rα2- cIL-4Rα1-cFc DNA resulted in the expression vector pcDNA3.4-cIL13RA2-cIL-4Rα1-cFc (SEQ ID NO: 18).

[0241] HEK293 cells were transiently transfected with recombinant plasmid at the ~ 100 mlculture scale by the CRO (Genscript), by employing standard molecular biology techniques. Culture supernatant was subjected to His TrapFF crude (Ni2+NTA) column chromatography purification and the recombinant cIL-13Rα2-cIL-4Rα1-ECD-cFc DNA (14.9 mg) was transferred into PBS, pH 7.2. The identity and purity of the recombinant product was confirmed by reducing SDS-PAGE / Coomassie Blue staining, where a broad band was observed at ~ 120 kDa apparent molecular mass. Nonreducing conditions did lead to protein ~ 250 kDa in size, suggesting that cIL-13Rα2-cIL-4Rα1-ECD-cFc does form disulfide-linked dimers, probably mediated by the canine Fc domain. Analysis of the mature sequence (SEQ ID NO: 16) by Protparam predicted a molecular mass of 90049.74 Da for the reduced polypeptide. The band of reduced cIL-13Rα2-cIL-4Rα1-ECD-cFc in SDS-PAGE migrates at a considerably higher mass range. NetNGlyc1.0 analysis predicted 11 N-glycosylation sites, 10 of which have the potential to be occupied. The variable occupancy and a variable degree of sialylation of cIL-13Rα2-cIL- 4Rα1-ECD-cFc may account for the observed heterogeneity and increased apparent molecular mass in SDS-PAGE analysis.

[0242] IL-4R-α1-ECD-IL-13-Ra2-cFc

[0243] Based on the mature cIL-4Rα1-ECD sequence followed by a glycine / serine (G / S)-containing 15 AA linker (GGGGSGGGGSGGGGG) (SEQ ID NO: 82), followed by the mature cIL-13Rα2-ECD protein sequence, followed by an octa-glycine linker (GGGGGGGG) (SEQ ID NO: 81) and Fc part from the canine IgG A gene was joined to the protein construct IL-4R-α1- ECD-IL-13-Rα2-cFc (SEQ ID NO: 19).

[0244] This IL-4R- α1-ECD-IL-13-R α2-cFc protein sequence (SEQ ID NO: 19) wasbacktranslated into a DNA sequence taking account for the human codon usage bias for expression in HEK293 cells. Furthermore, 5’ upstream of the ATG start codon, a Kozak consensus sequence (GCCACC) (SEQ ID NO: 86) was added. In addition, at the 5’ end of theU.S. Provisional Patent Application Attorney Docket No.2920951-492977 synthetic gene, a unique EcoRI, and at the 3’ end a unique Hind III restriction enzyme site were added for subcloning into the mammalian expression vector pcDNA3.4 (SEQ ID NO: 20). The subcloning of this IL-13Rα2-cFc DNA resulted in the expression vector pcDNA3.4-cIL13RA2- cFc (SEQ ID NO: 21).

[0245] HEK293 cells were transiently transfected with recombinant plasmid at the ~ 100 mlculture scale by the CRO (Genscript), by employing standard molecular biology techniques. Culture supernatant was subjected to His TrapFF crude (Ni2+NTA) chromatography purification and the recombinant cIL-4Rα1-cIL-13Rα2-ECD-cFc DNA (12.46 mg) was transferred into PBS, pH 7.2. The identity and purity of the recombinant product was confirmed by reducing SDS- PAGE / Coomassie Blue staining, where a broad band was observed at ~ 120 kDa apparent molecular mass (Fig. 40). Nonreducing conditions did lead to protein ~ 250 kDa in size, suggesting that cIL-4Rα1-cIL-13Rα2-ECD-cFc does form disulfide-linked dimers, probably mediated by the canine Fc domain. Analysis of the mature sequence (SEQ ID NO: 19) by Protparam predicted a molecular mass of 89446.05 Da for the reduced polypeptide. The band of reduced cIL-4Rα1-cIL-13Rα2-ECD-cFc in SDS-PAGE migrates at a considerably higher mass range. NetNGlyc1.0 analysis predicted 11 N-glycosylation sites, 9 of which have the potential to be occupied. The variable occupancy and a variable degree of sialylation of cIL-4Rα1-cIL- 13Rα2-ECD-cFc may account for the observed heterogeneity and increased apparent molecular mass in SDS-PAGE analysis. EXAMPLE 5 BIOLOGICAL EFFICACY OF IL4 / IL-13 DUAL RECEPTOR TRAPS: CELL CULTURE Inhibition of cIL-4 and cIL-13 action on a dog monocyte reporter cell line, DH82-pcDNA3.1- STAT6-SEAP

[0246] To investigate whether the dual receptor traps are exhibiting the desired biologicalactivity, i.e., binding to and neutralizing canine IL-4 as well as canine IL-13, a canine DH82 monocyte IL-4 / IL-13 reporter cell line, DH82-pcDNA3.1-STAT6-SEAP (separate report), was stimulated with 10 ng / ml canine IL-4 (cIL-4, R&D Systems, 754-CL-025 / CF), or 10 ng / ml canine IL-13 (cIL-13, own E. coli expression at a CRO, Genscript) (Fig.10). Either cIL-4Rα1- cIL-13Rα2-ECD-cFc or cIL-13Rα2-ECD-cIL-4Rα1-cFc was added to these cytokines in a 1:2U.S. Provisional Patent Application Attorney Docket No.2920951-492977 dilution titration starting at 2.5 ^g / ml, before application to the DH82-pcDNA3.1-STAT6-SEAP cells. In the case of cIL-4, both versions of the receptor trap, cIL-4Rα1-cIL-13Rα2-ECD-cFc or cIL-13Rα2-ECD-cIL-4Rα1-cFc, were able to inhibit the reporter enzyme readout completely, with estimated EC50s of ~ 25 ng / ml in both cases, and complete inhibition reached around 500 ng / ml (Fig.10A). When cIL-13 was used as stimulating cytokine, again both versions of receptor trap led to a progressive steep decrease in reporter gene signal, starting already at ~ 10 ng / ml and reaching complete inhibition at ~ 50 ng / ml (Fig.10B). This suggests that both receptor trap versions, cIL-4Rα1-cIL-13Rα2-ECD-cFc and cIL-13Rα2-ECD-cIL-4Rα1-cFc are highly potent dual action cIL-4 and cIL-13 inhibitors. EXAMPLE 6 BIOLOGICAL EFFICACY OF IL4 / IL-13 DUAL RECEPTOR TRAPS Study using naïve dog blood

[0247] Both IL-4 and IL-13 are induce expression of thymus- and activation-regulated chemokine(TARC or CCL17) at the mRNA and protein level, at least in human and mouse blood cells. In dogs, upregulation of TARC in atopic dermatitis has been documented, although the direct connection to IL-4 exposure has not been shown. Previous studies by the inventor established TARC as a positive marker (strong mRNA upregulation) of canine blood exposure to IL-4 and IL- 13. The inhibition of IL-4- and IL-13-induced TARC mRNA upregulation by cIL-4Rα1-cIL- 13Rα2-ECD-cFc and cIL-13Rα2-ECD-cIL-4Rα1-cFc in ex vivo dog blood samples was assessed to investigate the potential of these proteins as in vivo cIL-4 / cIL-13 blockers: -EDTA-stabilized blood was taken from naive dogs.- 500 ml blood was supplemented with canine IL-4 (R&D systems, 754-CL-025 / CF) orcanine IL-13 (cIL-13, own E. coli expression at a CRO, Genscript) to 10 ng / ml, in thepresence and absence of 2000 ng / ml, 200 ng / ml and 20 ng / ml cIL-4Rα1-cIL-13Rα2-ECD- cFc and cIL-13Rα2-ECD-cIL-4Rα1-cFc, and the blood was then incubated for 6 h at 35°C, 5% (v / v) CO2and 96% relative humidity -Blood lysis and RNA stabilization was done with RNAprotect Animal Blood Tubes 500μl(Qiagen 76554) and an incubation for 2 h at room temperature -RNA isolation was then done with the RNeasy Protect Animal Blood Kit (Qiagen 73224)- Isolated RNA was analysed and quantified using an Implen NanoPhotometer®, type NP80.- Quantitative RT- PCR (qPCR) was performed with the TaqMan® Assay Cf02622128_m1(Thermo) with probe and primers for canine CCL-17 (TARC) and the QuantiNova ProbeU.S. Provisional Patent Application Attorney Docket No.2920951-492977 RT-PCR Kit (Qiagen 208354), with the primers, reaction mixtures and conditions outlined by the manufacturers. Canine ^-actin TaqMan probe and PCR primers were used as a housekeeping gene control. Typically 25-100 ng total RNA was used as template. -The qPCR was performed using a CFX96 Real-Time System (BioRad)

[0248] Concomitant addition of the IL-4 / IL-13 dual receptor traps (cIL-13Rα2-ECD-cIL-4Rα1-cFc and cIL-4Rα1-cIL-13Rα2-ECD-cFc) to canine blood inhibits 10 ng / ml IL-4-induced CCL017 / TARC mRNA production and / or accumulation in a dose-dependent manner. Already at 20 ng / ml of the cIL-13Rα2-ECD-cIL-4Rα1-cFc inhibited ~ 95% of the qPCR signal, with 200 ng / ml and 2000 ng / ml attaining ~ 98% inhibition of the signal (Fig.11 A,B,C). In the case of cIL-4Rα1-cIL-13Rα2-ECD-cFc 20 ng / ml inhibited ~ 80% of the qPCR signal, while 200 ng / ml and 2000 ng / ml achieved 95% and 98% inhibition, respectively (Fig. 11 D,E,F)

[0249] In the case of blood stimulation with canine IL-13, both IL-4 / IL-13 dual receptor traps(cIL-13Rα2-ECD-cIL-4Rα1-cFc and cIL-4Rα1-cIL-13Rα2-ECD-cFc) inhibited the induced CCL017 / TARC mRNA production and / or accumulation in a dose-dependent manner. At 20 ng / ml of the cIL-13Rα2-ECD-cIL-4Rα1-cFc inhibited ~ 85% of the qPCR signal, with 200 ng / ml and 2000 ng / ml attaining ~ 98% % inhibition of the signal (Fig.12 A,B,C). In the case of cIL-4Rα1-cIL-13Rα2-ECD-cFc 20 ng / ml inhibited ~ 65% of the qPCR signal, while 200 ng / ml and 2000 ng / ml achieved again 98% inhibition, (Fig. 12 A,B,C).

[0250] in vivo study using blood from dual receptor trap-i. v. injected dogs

[0251] Three dogs were injected intravenously (i.v.) with 1 mg / kg of the dual receptor trap cIL-13Rα2-cIL-4Rα1-ECD-cFc. EDTA stabilized blood samples were taken three days before injection (day -3), one day (24 h) after injection, three days (72 h) after injection, and 7 days (168 h after injection). cIL-4 (1 ng / ml) or cIL-13 (10 ng / ml were added for 6 hours, and then the blood samples were lysed, total RNA was prepared, and qRT-PCR was performed for the cIL- 4 / cIL-13-induced mRNA for canine CCL-17 (TARC) (see procedure below). -EDTA-stabilized blood was taken from the dual receptor trap cIL-13Rα2-cIL-4Rα1-ECD-cFc i. v. dosed dogs at different time points post injection. -500 µl blood was supplemented with canine IL-4 (Genscript, HEK293 cell-expressed at aCRO, Genscript) or canine IL-13 (cIL-13, own E. coli expression at a CRO, Genscript) to1 ng / ml and 10 ng / ml, respectively, and the blood was then incubated for 6 h at 35°C, 5% (v / v) CO2and 96% relative humidityU.S. Provisional Patent Application Attorney Docket No.2920951-492977 -Blood lysis and RNA stabilization was done with RNAprotect Animal Blood Tubes 500μl(Qiagen 76554) and an incubation for 2 h at room temperature -RNA isolation was then done with the RNeasy Protect Animal Blood Kit (Qiagen 73224)- Isolated RNA was analysed and quantified using an Implen NanoPhotometer®, type NP80.- Quantitative RT- PCR (qPCR) was performed with the TaqMan® Assay Cf02622128_m1(Thermo) with probe and primers for canine CCL-17 (TARC) and the QuantiNova Probe RT-PCR Kit (Qiagen 208354), with the primers, reaction mixtures and conditions outlined by the manufacturers. Homemade canine ^-actin TaqMan probe and PCR primers were used as a housekeeping gene control. Typically 25-100 ng total RNA was used as template. -The qPCR was performed using a CFX96 Real-Time System (BioRad)

[0252] All cIL-13Rα2-cIL-4Rα1-ECD-cFc dual receptor trap treated dogs showed between ~75% and 95% inhibition of cIL4-induced CCL017 (TARC) induction in their blood compared to pre-dosing levels (Figs.13, 14, 15 A, B). This partial suppression of cIL-4 action persisted to day 3 in the blood of all three animals, ranging from 40% to > 90%, and in 2 out of 3 animals even to day 7, where > 70% inhibition was seen in dog 6 and > 20% in dog 5, while no inhibition was seen at this time point in dog 4.

[0253] In the case of cIL-13 inhibition was > 90-100 % in all three dogs at day 1. At the thirdday, 2 out of 3 dogs showed inhibition ranging from 65% (dog 5) to > 90% (dog 6), which was still > 30% (dog 5) or 85% (dog 6) at day 7, while in dog 4 no inhibition of cIL-13 action was seen anymore. EXAMPLE 7 CANINE IL-4 / IL-13 / IL-31 TRIPLE RECEPTOR TRAP PROTEIN CONSTRUCTS

[0254] Canine IL-31 receptor A (IL-31RA) and canine and OSMR^ receptor subunit proteinsequences

[0255] Canine IL-31 receptor A (IL-31RA)

[0256] The Canis lupus (Canis lupus familiaris) interleukin 31 receptor A (IL31RA), mRNAtranscript variant X4, was translated (SEQ ID NO: 22).

[0257] SignalP5.0 analysis of the protein sequence suggests the presence of an endoplasmicreticulum (ER) import signal sequence with a cleavage site after amino acid (AA) 19 (Fig.2). This information, together with the knowledge from the human IL-31RA sequence about the IL-U.S. Provisional Patent Application Attorney Docket No.2920951-492977 31 binding fibronectin-like domains 1 and 2 was used to define the canine IL-31 binding domain in the translated XM_038658914.1 DNA sequence. The fibronectin type-III domains 1 and 2 (AA 21 – AA 225) from the canine IL-31RA protein sequence (SEQ ID NO: 23) were identified.

[0258] canine OSMR^ receptor

[0259] The Canis lupus (Canis lupus familiaris) oncostatin-M-specific receptor subunit beta(OSMR^) was identified in the NCBI database (SEQ ID NO: 24). SignalP5.0 analysis of the protein sequence suggests the presence of an endoplasmic reticulum (ER) import signal sequence with a cleavage site after amino acid 23. Amino acids 24-424 of the canine protein sequence were identified as the domain important for cIL-31 ligand binding. The cIL-31 binding fibronectin type-III domains 1 and 2 of the canine OSMR^ is shown in SEQ ID NO: 25.

[0260] Design and production of a canine IL-4 / IL-13 / IL-31 triple receptor trap protein:cIL4 / 13R-cOS31RA-cFc.

[0261] Using the information on the ligand binding regions on canine IL-31RA and the canineOSMR^ protein sequences, together with the information from EXAMPLE 1, a protein construct was designed striving to combine cIL-4, cIL-13 and cIL-31 ligand binding properties, a ‘cIL- 4 / cIL-13 / cIL-31 receptor trap (Fig. 3A).

[0262] The following protein sequences were added to each other starting from the N-terminus:a signal sequence directing ER import, the mature cIL-4Rα1 extracellular domain (ECD) sequence protein sequence, a glycine / serine (G / S)-containing 15 AA linker (GGGGSGGGGSGGGGG) (SEQ ID NO: 82), the mature cIL-13Rα2 ECD, a 15 AA linker (GGGGSGGGGSGGGGG) (SEQ ID NO: 82), the cOSRM^ cIL-31 ligand binding domain (LBD) followed by an 20 AA linker (GGGGSGGGGSGGGGGGGGGS) (SEQ ID NO: 83), the IL-31RA LBD, an octa-glycine linker, and Fc part from the canine IgG A gene. The C-terminus is formed by a hexa-histidine tag for protein purification purposes (SEQ ID NO: 87).

[0263] This protein cIL4 / 13R-cOS31RA-cFc sequence (SEQ ID NO: 26) was backtranslatedinto a DNA sequence taking account for the human codon usage bias for expression in HEK293 cells. Furthermore, 5’ upstream of the ATG start codon, a Kozak consensus sequence (GCCACC) was added. In addition, at the 5’ end of the synthetic gene, a unique EcoRI, and at the 3’ end a unique Hind III restriction enzyme site were added for subcloning into theU.S. Provisional Patent Application Attorney Docket No.2920951-492977 mammalian expression vector pcDNA3.4 (SEQ ID NO: 27). The subcloning of this IL-13Rα2- cIL4R^1-cOS31RA-cFc DNA resulted in the expression vector pcDNA3.4- cIL4 / 13R- cOS31RA-cFc (SEQ ID NO: 28).

[0264] HEK293 cells were transiently transfected with recombinant plasmid at the ~ 100 mlculture scale by the CRO (Genscript), by employing standard molecular biology techniques. Culture supernatants from two different transfections were subjected to His Trap FF crude (Ni2+-NTA) column chromatography, or protein A (MabSelect SuRe LX ) purification, and the recombinant cIL4 / 13R-cOS31RA-cFc (420 ^g and 945 µg, respectively) were transferred into PBS, pH 7.2. The identity and purity of the recombinant product was confirmed for both expression products by reducing SDS-PAGE / Coomassie Blue staining, where a broad band was observed at ~ 200 kDa apparent molecular mass. Nonreducing conditions did lead to protein considerably larger in size, suggesting that cIL4 / 13R-cOS31RA-cFc does form disulfide-linked dimers, probably mediated by the canine Fc domain. Analysis of the mature sequence (SEQ ID NO: 26) by Protparam predicted a molecular mass of 161905.50 Da for the reduced polypeptide. The band of reduced cIL4 / 13R-cOS31RA-cFc in SDS-PAGE migrates at a considerably higher mass range. NetNGlyc1.0 analysis predicted 28 N-glycosylation sites, 26 of which have the potential to be occupied. The variable occupancy and a variable degree of sialylation of cIL4 / 13R-cOS31RA-cFc may account for the observed heterogeneity and increased apparent molecular mass in SDS-PAGE analysis. EXAMPLE 8 FUNCTIONAL CHARACTERIZATION OF cIL4 / 13R-cOS31RA-cFc

[0265] Inhibition of cIL-4 and cIL-13 action by cIL4 / 13R-cOS31RA-cFc on a dog monocytereporter cell line, DH82-pcDNA3.1-STAT6-SEAP

[0266] To investigate whether the triple receptor traps are exhibiting biological activity, withrespect to the binding and neutralization of canine IL-4 as well as canine IL-13, a canine DH82 monocyte IL-4 / IL-13 reporter cell line, DH82-pcDNA3.1-STAT6-SEAP (separate report), was stimulated with 1 ng / ml canine IL-4 (cIL-4, R&D Systems, 754-CL-025 / CF), or 10 ng / ml canine IL-13 (cIL-13, own E. coli expression at a CRO, Genscript) (Fig. 16). cIL4 / 13R-cOS31RA-cFc was added to these cytokines in a 1:2 dilution titration starting at 10 ^g / ml, before application toU.S. Provisional Patent Application Attorney Docket No.2920951-492977 the DH82-pcDNA3.1-STAT6-SEAP cells. In the case of cIL-4 cIL4 / 13R-cOS31RA-cFc was able to inhibit the reporter enzyme readout completely, with estimated EC50s of ~ 200 ng / ml in both cases, and complete inhibition reached around 2000 ng / ml (Fig. 16A). When cIL-13 was used as stimulating cytokine, again both versions of receptor trap led to a progressive steep decrease in reporter gene signal, starting already at ~ 100 ng / ml and reaching complete inhibition at ~ 1000 ng / ml (Fig. 16B). This suggests that cIL4 / 13R-cOS31RA-cFc is a highly potent dual action cIL-4 and cIL-13 inhibitor.

[0267] Inhibition of cIL-4 and cIL-13 action by cIL4 / 13R-cOS31RA-cFc in ex vivo cytokinestimulation studies using naïve dog blood

[0268] Both IL-4 and IL-13 induce expression of thymus- and activation-regulated chemokine(TARC or CCL17) at the mRNA and protein level, at least in human and mouse blood cells. In dogs, upregulation of TARC in atopic dermatitis has been documented, although the direct connection to IL-4 exposure has not been shown. Through previous experiments, the inventors established TARC as a positive marker (strong mRNA upregulation) of canine blood exposure to IL-4 and IL-13. The inhibition of IL-4- and IL-13-induced TARC mRNA upregulation by cIL4 / 13R-cOS31RA-cFc in ex vivo dog blood samples was assessed to investigate the potential of these proteins as in vivo cIL-4 / cIL-13 blockers: -EDTA-stabilized blood was taken from naive dogs.- 500 ml blood was supplemented with canine IL-4 (R&D systems, 754-CL-025 / CF) orcanine IL-13 (cIL-13, own E. coli expression at a CRO, Genscript) to 10 ng / ml, in thepresence and absence of 2000 ng / ml, 200 ng / ml and 20 ng / ml cIL4 / 13R-cOS31RA-cFc, and the blood was then incubated for 6 h at 35°C, 5% (v / v) CO2 and 96% relative humidity -Blood lysis and RNA stabilization was done with RNAprotect Animal Blood Tubes 500μl(Qiagen 76554) and an incubation for 2 h at room temperature -RNA isolation was then done with the RNeasy Protect Animal Blood Kit (Qiagen 73224)- Isolated RNA was analysed and quantified using an Implen NanoPhotometer®, type NP80.- Quantitative RT- PCR (qPCR) was performed with the TaqMan® Assay Cf02622128_m1(Thermo) with probe and primers for canine CCL-17 (TARC) and the QuantiNova Probe RT-PCR Kit (Qiagen 208354), with the primers, reaction mixtures and conditions outlined by the manufacturers. canine b-actin TaqMan probe and PCR primers were used as a housekeeping gene control. Typically 25-100 ng total RNA was used as template. -The qPCR was performed using a CFX96 Real-Time System (BioRad)U.S. Provisional Patent Application Attorney Docket No.2920951-492977

[0269] Concomitant addition of the IL-4 / IL-13 / Il-31 triple receptor trap (cIL4 / 13R-cOS31RA-cFc) to canine blood inhibits 1 ng / ml IL-4-induced CCL017 / TARC mRNA production and / or accumulation in a dose-dependent manner. Already at 20 ng / ml and 200 ng / ml of cIL4 / 13R- cOS31RA-cFc inhibited ~ 60-70 % of the qPCR signal, and 2000 ng / ml attaining ~ 98% % inhibition of the IL-4-induced CCL017 qPCR signal (Fig.17A). In the case of IL-13 stimulation, 20 ng / ml cIL4 / 13R-cOS31RA-cFc inhibited ~ 60% of the qPCR signal, while 200 ng / ml and 2000 ng / ml achieved >95% inhibition (Fig. 17B).

[0270] Binding of canine IL-31 to cIL4 / 13R-cOS31RA-cFc

[0271] To investigate the capacity of the triple receptor trap cIL4 / 13R-cOS31RA-cFc to bindcanine IL-31 (cIL-31), an ELISA format was devised where cIL4 / 13R-cOS31RA-cFc was immobilized to the plate to serve as a potential binding partner to biotinylated cIL-31. Biotinylation of cIL31 1) Equilibration of a Zeba 0.5 ml Spin desalting columns 40K with 50mM NaHCO3150mM NaCl pH 8.5 2) 10-fold concentration of cIL31 (Genscript U4939FI210-30.82mg / ml) via a 10 kDa spinning membrane device (Sartorius, Vivaspin) 3) Concentrated purified cIL31 (100 ^l) is passed through the equilibrated column (buffer exchange) two times 4) Addition of 20µl 10mM EZ-Link™ Sulfo-NHS-LC-Biotin in PBS (Thermo Scientific 21338) 5) Incubation for 4 h at 37°C 6) Addition of 1µl 1M Tris pH 8.0 7) Equilibration of a Zeba 0.5 ml Spin desalting columns 40K with PBS pH 7.2 8) Passing of the reaction mixture through the PBS equilibrated column. 9) Filling up of the cIL-31-Biotin solution to 200µl with PBS. Assumed concentration: 4.1mg / ml cIL31-Biotin Indirect ELISA for binding of biotinylated cIL-31 to cOSMRIL31RA coated plates 1) Coating of polystyrene ELISA plates (384 well: Thermo Maxisorp, CatNo. 464718) with10 µl / well of 1µg / ml cOSMRIL31RA (GenScript U0737FG130-4), cOS31RA-cFc (GenScript U6448GH270-4), cIL4 / 13R-cOS31RA-cFc (GenScript U862RGH270-4) and cIL13Ra2-cIL4RΑ1-cFc (GenScript U3071GF150-4) cIL4RΑ1-cIL13Ra2-cFc (GenScriptU.S. Provisional Patent Application Attorney Docket No.2920951-492977 U713XGF150-4) dissolved in coating buffer PBS (ThermoFisher Phosphate-Buffered Saline - pH 7.2, CatNo.20012-019, or equivalent). Incubation overnight (O / N) at 40C with closed lid. 2) Removal of coating solution, washings 3 x with PBS (Thermo Fisher Phosphate-Buffered Saline pH 7.2, Cat.No.20012-019 or equivalent), 0.05% (v / v) Tween 20, 35 µl / well. 3) Blocking of nonspecific binding sites with 35 µl / well PBS, 0.05% (v / v) Tween 20, 5% (w / v) non-fat dry milk powder (MP) (Roth T145.2) (= blocking solution). Incubation at room temperature (RT) for > 1 (h) with closed lid. 4) Removal of blocking solution (discard), addition of 20 µl / well of sample dilution / preparation. start a 1:1,5 dilution series from 5µg / ml of cIL31-Biotin in blocking solution (MP) on a nonadsorptive ELISA plate. Incubation at room temperature (RT) for > 1 hour (h). 5) Removal of solutions, washings 4 x with PBS 0.05% (v / v) Tween 20, 35 µl / well. 6) Addition of 20 µl / well dilutions of 1:17,000 Extravidin-AP (Sigma E2636) in: PBS, 0.05% (v / v) Tween 20, 5% (v / v) gelatin (from cold water fish skin, 40-50% in H2O, Sigma C 7765). (Generally, aim at: 100-400 ng / ml conjugate) Incubate at room temperature (RT) for > 1 h. 7) Removal of conjugate solution, washing 2 x with PBS 0.05% (v / v) Tween 20, 35 µl / well. 8) Washing 1 x with AP buffer (50 mM NaHCO3 / Na2CO3, 2 mM MgCl2, pH 9.6), 50 µl / well. 9) Develop ELISA by adding 90 µl / well of 5 mM 4-nitrophenyl phosphate disodium salt hexahydrate (pNPP, Applichem, A1442,0050) in 50 mM NaHCO3 / Na2CO3, 2 mM MgCl2, pH 9.6 buffer. Kinetic monitoring of the optical density (OD) at 405 nm in an ELISA reader (mOD / min) at RT. Determination of the curve slope in a linear range. Vmax star: Vmax measurements over points 2-5 Vmax variable: Vmax use of the 4 measuring points with maximal slope within the points 1-7 One point measuring at 405 nm after 22min, 60min, 180 min and 240min

[0272] Control were coatings with IL13Ra2-cIL4RΑ1-cFc and cIL4RΑ1-cIL13Ra2-cFc as well asabsence of coating. Biotinylated cIL-31 was titrated from 5000 ng / ml in 1:1.5 dilution steps down to ~ 20 ng / ml. While noncoated plates as well as IL13Ra2-cIL4RΑ1-cFc- and cIL4RΑ1-cIL13Ra2- cFc-coated plates did not show a signal below ~ 2000 ng / ml, while the triple receptor trap constructU.S. Provisional Patent Application Attorney Docket No.2920951-492977 cIL4 / 13R-cOS31RA-cFc specifically and progressively immobilized biotinylated cIL-31, starting already at 100 ng / ml cytokine (Fig.18).

[0273] The triple receptor trap cIL4 / 13R-cOS31RA-cFc comprising four different caninecytokine receptor extracellular domains linked to a canine Fc was designed and successfully expressed. In vitro cellular stimulation data demonstrate that the triple receptor trap is able to bind to and neutralize both cIL-4 and cIL-13. Ex vivo dog blood stimulation and qPCR studies demonstrate that the triple receptor trap is able the neutralize both cIL-4 and cIL-13 also in a in vivo-like dog blood context. An canine IL-31 binding assay demonstrates that the triple receptor trap is able to specifically bind to this cytokine, demonstrating functionality also for this third function. EXAMPLE 9 Human IL-4 / IL-13 dual receptor trap protein, huIL13Ra2-huIL4Ra-huFc

[0274] The mature extracellular domains of the human Interleukin-13 receptor subunit alpha-2(huIL13RA2) and the human Interleukin-4 receptor subunit alpha (huIL4Ra) were identified. Using an artificial ER import signal sequence, a tandem protein of the two mature polypeptides huIL13Ralpha2 (SEQ ID NO: 31) and huIL4Ralpha (SEQ ID NO: 29) separated by the 15 AA linker (GGGGS)3 (SEQ ID NO: 84) was constructed. This construct was followed by a 14mer linker (GGGGGGGGIEGRMD) (SEQ ID NO: 85) and a human IgG1 heavy chain constant region (SEQ ID NO: 33). The entire dual receptor trap construct huIL13RA2-huIL4Ra-huFc encompasses 801 amino acids (SEQ ID NO: 34).

[0275] The huIL13RA2-huIL4Ra-huFc protein sequence was backtranslated into a DNAsequence taking account the human codon usage bias for expression in HEK293 cells. Furthermore, 5’ upstream of the ATG start codon, a Kozak consensus sequence (GCCACC) (SEQ ID NO: 86) was added. In addition, at the 5’ end of the synthetic gene, a unique EcoRI, and at the 3’ end a unique Hind III restriction enzyme site were added for subcloning into the mammalian expression vector pcDNA3.4 (SEQ ID NO: 35). The subcloning of this huIL13RA2- huIL4Ra-huFc-encoding DNA resulted in the expression vector pcDNA3.4-huIL13RA2- huIL4Ra-huFc (SEQ ID NO: 36).U.S. Provisional Patent Application Attorney Docket No.2920951-492977

[0276] HEK293 cells were transiently transfected with recombinant plasmid at the ~ 100 mlculture scale by the CRO (Genscript), by employing standard molecular biology techniques. Culture supernatant was subjected to HiTrap MabSelect Sure LX (protein A) column chromatography purification and the recombinant huIL-13Rα2-huIL-4Rα1-ECD-huFc DNA (0.5 mg) was transferred into PBS, pH 7.2. The identity and purity of the recombinant product was confirmed by reducing SDS-PAGE / Coomassie Blue staining, where a broad band was observed at ~ 120 kDa apparent molecular mass. Nonreducing conditions did lead to a fuzzy protein band at ~ 250 kDa in size, suggesting that huIL13Rα2-huIL4Rα1-huFc does form disulfide-linked dimers, probably mediated by the human Fc domain. In addition, aggregated material was visible. Analysis of the mature sequence by Protparam predicted a molecular mass of 88635.81 Da for the reduced polypeptide. The band of reduced huIL13Ra2-huIL4Ra-huFc in SDS-PAGE migrates at a considerably higher mass range. NetNGlyc1.0 analysis predicted 11 N- glycosylation sites, all of which have the potential to be occupied. The variable occupancy and a variable degree of sialylation of huIL13Ra2-huIL4Ra-huFc may account for the observed heterogeneity and increased apparent molecular mass in SDS-PAGE analysis. EXAMPLE 11 Inhibition of human IL-4 and human IL-13 action on a fully human HEKblue IL-4 / IL-13 reporter cell line by huIL13Ra2-huIL4Ra-huFc

[0277] To investigate whether the dual receptor trap huIL13Ra2-huIL4Ra-huFc is exhibiting thebiological activity, i.e., binding to and neutralizing human IL-4 as well as human IL-13 receptor engagement, a fully human HEKblue IL-4 / IL-13 reporter cell line (Invivogen) was stimulated with 1 ng / ml human IL-4 or human IL-13 (Fig.19). The dual receptor trap huIL13Ra2- huIL4Ra-huFc was added to these cytokines in a 1:2 dilution titration starting at 5 ^g / ml, before application to the HEKblue IL-4 / IL-13 cells. In the case of human IL-4, the dual receptor trap huIL13Ra2-huIL4Ra-huFc was able to inhibit the reporter enzyme readout completely, with estimated EC50s of ~ 600 ng / ml (Fig. 1A). When IL-13 was used as stimulating cytokine, the dual receptor trap led to a progressive steep decrease in reporter gene signal, starting already at ~U.S. Provisional Patent Application Attorney Docket No.2920951-492977 100 ng / ml and reaching complete inhibition at ~ 500 ng / ml (Fig.1B). This suggests that huIL13Ra2-huIL4Ra-huFc is a highly potent dual action human IL-4 and human IL-13 inhibitor. EXAMPLE 12 DESIGN AND EXPRESSION OF A FELINE IL-4-RΑ1-ECD / IL-13-RΑ2-ECD DUAL RECEPTOR TRAP

[0278] For the design of a feline IL-4-Rα1-ECD / IL-13-Rα2-ECD dual receptor trap protein(SEQ ID NO: 49), an endoplasmic reticulum (ER) import signal sequence of 19 amino acids was followed by the mature feline IL-13-Rα2-ECD and then, separated by a 15 amino acid Glycine / Serine (G / S) sequence, the mature feline IL-4-Rα1-ECD. To achieve a more stable and dimeric version of the planned IL-4-Rα1-ECD / IL-13-Rα2-ECD dual receptor trap, a fusion protnein with an immunoglobulin G (IgG) fragment crystallizable region (Fc region) was designed. To identify such a feline Fc region, a Felis catus partial mRNA for IgG1 heavy chain was identified, and the translation product aligned to the protein sequence of IgG A from Canis lupus familiaris. A large degree of sequence identity was noted, and a region equivalent to the canine Fc of IgG-A was identified starting with the sequence DKTV... (SEQ ID NO: 92) and ending with …QSPG (SEQ ID NO: 93). This sequence was appended to the feline IL-4-Rα1- ECD / IL-13-Rα2-ECD domain after an octa-glycine spacer (GGGGGGGG)(SEQ ID NO: 81). The C-terminus of the entire construct was then formed by a hexa-histidine sequence (HHHHHH) (SEQ ID NO: 87).

[0279] A fIL13Ra2-fIL4Ra1-fFc-His6 (SEQ ID NO: 48) protein sequence was developed andback-translated into a DNA sequence taking account for the human codon usage bias for expression in HEK293 cells. Furthermore, 5’ upstream of the ATG start codon, a Kozak consensus sequence (GCCACC) (SEQ ID NO: 86) was added. In addition, at the 5’ end of the synthetic gene, a unique EcoRI, and at the 3’ end a unique Hind III restriction enzyme site were added for subcloning into the mammalian expression vector pcDNA3.4 (SEQ ID NO: 50). The subcloning of this fIL13Ra2-fIL4Ra1-fFc-His6 DNA resulted in the expression vector pcDNA3.4-fIL13Ra2-fIL4Ra1-fFc (SEQ ID NO: 51).

[0280] HEK293 cells were transiently transfected with recombinant plasmid at the about 100 mLculture scale by the CRO (Genscript), by employing standard molecular biology techniques.U.S. Provisional Patent Application Attorney Docket No.2920951-492977 Culture supernatant was subjected to Ni2+NTA column chromatography purification and the recombinant fIL13Ra2-fIL4Ra1-fFc (8.64 mg) was transferred into PBS, pH 7.2. The identity and purity of the recombinant product was confirmed by reducing SDS-PAGE / Coomassie Blue staining, where a broad band was observed at about 110 kDa apparent molecular mass. Nonreducing conditions did lead to protein about 220 kDa in size, suggesting that fIL13Ra2- fIL4Ra1-fFc does form disulfide-linked dimers, probably mediated by the canine Fc domain. Analysis of the mature sequence (SEQ ID NO: 47) by Protparam predicted a molecular mass of 89256.87 Da for the reduced polypeptide. The band of reduced fIL13Ra2-fIL4Ra1-fFc in SDS- PAGE migrates at a considerably higher mass range. NetNGlyc1.0 analysis predicted 11 N- glycosylation sites, all of which have the potential to be occupied. The variable occupancy and a variable degree of sialylation of fIL13Ra2-fIL4Ra1-fFc may account for the observed heterogeneity and increased apparent molecular mass in SDS-PAGE analysis. EXAMPLE 13 FUNCTIONAL EXPRESSION OF FELINE IL-4

[0281] The feline interleukin 4 (IL-4) gene was viewed in Genbank, and the deduced proteinsequence was extracted (SEQ ID NO: 52).

[0282] Based on a SignalP-5.0 analysis, the predicted ER import signal sequence of feline IL-4spans amino acids (AA) 1-24, predicting as the N-terminus of the mature sequence ‘QNFN….’ (SEQ ID NO: 94).

[0283] Based on the mature feline IL-4 protein sequence, an expression construct with a N-terminally added artificial ER import signal sequence and a C-terminally added hexa-histidine (H6) was designed (SEQ ID NO: 53). This protein sequence was backtranslated into a DNA sequence taking account for the human codon usage bias for expression in HEK293 cells. Furthermore, 5’ upstream of the ATG start codon, a Kozak consensus sequence (GCCACC)U.S. Provisional Patent Application Attorney Docket No.2920951-492977 (SEQ ID NO: 84) was added. In addition, at the 5’ end of the synthetic gene, a unique EcoRI, and at the 3’ end a unique Hind III restriction enzyme site were added for subcloning into the mammalian expression vector pcDNA3.4 (SEQ ID NO: 54). The subcloning of this IL-4Rα1- ECD-His6 DNA resulted in the expression vector pcDNA3.4-fel-IL-4 (SEQ ID NO: 55). HEK293 cells were transiently transfected with recombinant plasmid at the about 100 ml culture scale by the CRO (Genscript), by employing standard molecular biology techniques. Culture supernatant was subjected to Ni2+NTA column chromatography purification and the recombinant fel-IL-4 (1.1 mg) was transferred into PBS, pH 7.2. The identity and purity of the recombinant product was confirmed by reducing SDS-PAGE / Coomassie Blue staining, where multiple bands and a smear was observed between 15 kDa and ~30 kDa apparent molecular mass. Nonreducing conditions did not change this banding pattern, suggesting that fel-IL-4 is monomeric. Analysis of the mature sequence (SEQ ID NO: 53) by Protparam predicted a molecular mass of 13255.3 Da for the reduced polypeptide. The band of reduced fel-IL4 in SDS-PAGE migrates at a considerably higher mass range. NetNGlyc1.0 analysis predicted 6 N-glycosylation sites, 5 of which have the potential to be occupied. The variable occupancy and a variable degree of sialylation of fel-IL-4 may account for the observed heterogeneity and increased apparent molecular mass in SDS-PAGE analysis. EXAMPLE 14 FUNCTIONAL EXPRESSION OF FELINE IL-13

[0284] Inspection of feline IL-13 sequences in the Genbank database revealed 6 distinct mRNAswith the corresponding translation products. SignalP5.0 analysis of these feline IL-13 translation products X1-X6 suggested the absence of a functional ER import signal sequence. Multiple sequence alignment of these sequences revealed two groups, X1-X4 and X5-X6, which exhibited quite dissimilar, non-homologous N-termini. Multiple sequence alignment with a canine IL-13 versions that does possess a functional ER import signal sequence suggested that in the case of feline IL13 X1-X4 mRNAs, the N-terminus assignment in the Genbank database is not correct, and that the true N-terminus is 53 amino acids later than indicated in the database. Indeed, if the ‘truncated’ feline Il-13 X1-X4 versions are subjected to a SignalP5.0 analysis, a signal sequence is apparent, with a cleavage point equivalent to that seen in the canine sequence. Therefore, catU.S. Provisional Patent Application Attorney Docket No.2920951-492977 IL-13 splice variant X2 with newly assigned N-terminus was used for further studies (SEQ ID NO: 56).

[0285] Further IL-13 isoforms include but are not limited to IL-13 Isoform X1 (SEQ ID NO:60), IL-13 Isoform X2 (SEQ ID NO: 61), IL-13 Isoform X3 (SEQ ID NO: 63), IL-13 Isoform X4 (SEQ ID NO: 63), IL-13 Isoform X5 (SEQ ID NO: 64), and IL-13 Isoform X6 (SEQ ID NO: 65).

[0286] For E. coli expression, the sequence without predicted signal sequence wascomplemented with an N-terminal start methionine and a C-terminal hexa-histidine tag (SEQ ID NO: 57). The complete sequence from SEQ ID NO: 58 was subcloned into the pET30(+) E. coli expression vector via the restriction sites NdeI and HindIII (underlined) (SEQ ID NO: 59): E. coli strain BL21(DE3) was transformed with recombinant plasmid.

[0287] Purification and Analysis:

[0288] E. coli strain BL21 Star™ (DE3) was transformed with recombinant plasmid. A singlecolony was inoculated into LB medium containing related antibiotic; culture was incubated in 37°C at 200 rpm and then induced with IPTG. SDS-PAGE was used to monitor the expression.

[0289] Scale up Expression: Recombinant BL21(DE3) stored in glycerol was inoculated into TBmedium containing related antibiotic and cultured at 37℃. When the OD600 reached about 1.2, cell culture was induced with IPTG at 15 / 16hours. Bacteria were harvested by centrifugation.

[0290] Purification and Analysis: bacterial pellets were resuspended with lysis buffer followedby sonication. The precipitate after centrifugation was dissolved using denaturing agent. Target protein was obtained by one-step purification using Ni column. Target protein was sterilized by 0.22 μm filter before stored in aliquots. The concentration was determined by Bradford protein assay with BSA as standard. The protein purity and molecular weight were determined by standard SDS-PAGE.

[0291] SDS-PAGE Analysis: Buffer: Reducing Loading buffer: 300 mM Tris-HCl, 10% SDS,30% Glycerol, 0.5% bromophenol blue, 250 mM DTT, pH 6.8. Non-Reducing Loading buffer: 300 mM Tris-HCl,10% SDS, 30% Glycerol, 0.5% bromophenol blue, pH 6.8. Reducing and non- reducing loading buffer were added to protein sample respectively and the final concentration of protein was close to 0.5 mg / ml. After mixing the sample, heating at 100oC for 5-10 minutes was performed (only for reducing condition). The protein samples were centrifuged at 10000rpm forU.S. Provisional Patent Application Attorney Docket No.2920951-492977 1 minute, and then loaded onto a precast gel (Genscript, Cat.No. M42012) in a gel chamber and the appropriate running buffer. Electrophoresis was performed at 140V for approximately 60 min.

[0292] By the above procedure, approximately 10.57 mg of soluble (in phosphate-bufferedsaline, PBS) fel-IL-13-His6 were obtained from bacterial pellet of 1 L culture. The size of the dominant band in lane R of the SDS-PAGE (Fig. 28) is in good agreement with the prediction expected from the protein sequence (13592.58 Da, calculated from the mature sequence using Protparam program. EXAMPLE 15 Stimulation of a feline STAT6 reporter cell line, CRFK-pcDNA3.1-STAT6-SEAP, by feline IL-4 and feline IL-13

[0293] To investigate whether HEK293-expressed feline IL-4 as well as E. coli-expressed felineIL-13 (described herein) are biologically active, a homemade feline CRFK kidney IL-4 / IL-13 reporter cell line, CRFK-pcDNA3.1-STAT6-SEAP, was stimulated with 1:2 dilutions over 15 steps starting from 200 ng / ml concentration. Feline IL-4 already induced production and secretion of the reporter enzyme SEAP at ~ 20 pg / ml and exhibited an EC50of ~ 200-500 pg / ml (Fig.20). Feline IL-13 required at least 1 ng / ml to induce production and secretion of the reporter enzyme SEAP and exhibited an EC50 of ~ 10 ng / ml (Fig.20). EXAMPLE 16 Inhibition of feline IL-4 and feline IL-13 action on a cat kidney reporter cell line, CRFK- pcDNA3.1-STAT6-SEAP, by the dual receptor trap fIL13Ra2-fIL4Ra1-fFc

[0294] To investigate whether the fully feline dual receptor trap is exhibiting the expectedbiological activity, e.g.,. binding to and neutralizing feline IL-4 as well as feline IL-13, the feline CRFK kidney IL-4 / IL-13 reporter cell line, CRFK-pcDNA3.1-STAT6-SEAP, was stimulated with either 5 ng / ml feline IL-4 (fIL-4, own HEK293 cell expression, Genscript U790JGK290-4), or 10 ng / ml feline IL-13 (fIL-13, own E. coli expression, Genscript U2081GL220-1) (Fig.10). fIL-13Rα2-ECD-fIL-4Rα1-fFc was added to these cytokines in a 1:2 dilution titration starting at 1 ^g / ml, before application to the CRFK-pcDNA3.1-STAT6-SEAP cells. In the case of fIL-4,U.S. Provisional Patent Application Attorney Docket No.2920951-492977 fIL-13Rα2-ECD-fIL-4Rα1-fFc, was able to inhibit the reporter enzyme readout completely, with estimated EC50s of ~ 80 ng / ml, and complete inhibition reached around 500 ng / ml (Fig.21A). When fIL-13 was used as stimulating cytokine, the fully feline dual receptor trap led to a progressive steep decrease in reporter gene signal, starting already at less than 10 ng / ml and reaching complete inhibition below 50 ng / ml (Fig. 21B). This suggests that the fully feline dual IL-4 / IL-13 receptor trap fIL-13Rα2-fIL-4Rα1-fFc is a highly potent dual action fIL-4 and fIL-13 inhibitor.

[0295] Summary:

[0296] The fully feline dual receptor trap fIL-13Rα2-fIL-4Rα1-fFc consisting of two differentfeline cytokine receptor extracellular domains linked to a feline Fc was designed and successfully expressed. Feline IL-4 and feline IL-13 as well as a feline STAT6 reporter cell line (CRFK-STAT6-SEAP) were generated as tools for the qualification of fIL-13Rα2-fIL-4Rα1-fFc. In vitro cellular stimulation data using CRFK-STAT6-SEAP demonstrated that the dual receptor trap is able to bind to and neutralize both fIL-4 and fIL-13.

Claims

U.S. Provisional Patent Application Attorney Docket No.2920951-492977 What is claimed: CANINE CONSTRUCTS1. A fusion protein comprising:an IL-4-binding extracellular domain (IL-4-Ralpha1-ECD); and a fragment crystallizable region of an antibody (Fc).

2. A fusion protein comprising:an IL-13-binding extracellular domain (IL-13-Ralpha2-ECD); and a fragment crystallizable region of an antibody (Fc).

3. A fusion protein comprising:an IL-4-binding extracellular domain (IL-4-Ralpha1-ECD); an IL-13-binding extracellular domain (IL-13-Ralpha2-ECD); and a fragment crystallizable region of an antibody (Fc).

4. A fusion protein comprising:an IL-4-binding extracellular domain (IL-4-Ralpha1-ECD); an IL-13-binding extracellular domain (IL-13-Ralpha2-ECD); an cOSRM^-ligand binding domain (LBD); and a fragment crystallizable region of an antibody (Fc).

5. A fusion protein comprising:an IL-4-binding extracellular domain (IL-4-Ralpha1-ECD); an IL-13-binding extracellular domain (IL-13-Ralpha2-ECD); an OSRM^-ligand binding domain (LBD); IL-31RA-ligand binding domain (LBD); and a fragment crystallizable region of an antibody (Fc).

6. The fusion protein of any one of claims 1-5, wherein the C-terminal of the IL-4-bindingextracellular domain is linked to the N-terminal of IL-13-binding extracellular domain.

7. The fusion protein of any one of claims 1-6, wherein the C-terminal of the IL-13-bindingextracellular domain is linked to the N-terminal of the IL-4-binding extracellular domain.

8. The fusion protein of any one of claims 1-7, wherein the Fc is linked to the C-terminal ofthe IL-4-binding extracellular domain.U.S. Provisional Patent Application Attorney Docket No.2920951-4929779. The fusion protein of any one of claims 1-8, wherein the Fc is linked to the C-terminal ofthe IL-13-binding extracellular domain.

10. The fusion protein of any one of claims 1-9, wherein the Fc is linked to the C-terminal ofthe OSRM^-LBD.

11. The fusion protein of any one of claims 1-10, wherein the Fc is linked to the C-terminalof the IL-31RA-ligand binding domain (LBD).

12. The fusion protein of any one of claims 1-11, wherein the cIL-31-ligand binding domain(LBD) is linked to the cOSRM^-LBD.

13. The fusion protein of any one of claims 1-12, wherein the C-terminus of the IL-4-bindingextracellular domain is linked to the N-terminus IL-13-binding extracellular domain which is linked to the N-terminus of the fragment crystallizable region of an antibody.

14. The fusion protein of any one of claims 1-13, wherein the C-terminus of the IL-13-binding extracellular domain is linked to the N-terminus of the IL-4-binding extracellular domain which is linked to the N-terminus of the fragment crystallizable region of an antibody.

15. The fusion protein of any one of claims 1-14, wherein the C-terminus of the IL-4-bindingextracellular domain (IL-4-RΑ1-ECD) is linked to the N-terminus of the IL-13-binding extracellular domain (IL-13-RA2-ECD) which is linked to the N-terminus of the OSRM^-ligand binding domain (LBD) which is linked to the N-terminus of the fragment crystallizable region of an antibody (Fc).

16. The fusion protein of any one of claims 1-15, wherein the C-terminus of the IL-4-bindingextracellular domain (IL-4-RΑ1-ECD) is linked to the N-terminus of the IL-13-binding extracellular domain (IL-13-RA2-ECD) which is linked to the N-terminus of the OSRM^-ligand binding domain (LBD) which is linked to the N-terminus of the IL- 31RA-ligand binding domain (LBD) which is linked to the N-terminus of the fragment crystallizable region of an antibody (Fc).

17. The fusion protein of any one of claims 1-16, wherein the IL-4-binding extracellulardomain (IL-4-RΑ1-ECD) and cFc are joined by a linker.

18. The fusion protein of any one of claims 1-17, wherein the IL-13-binding extracellulardomain (IL-13-RA2-ECD) and cFc are joined by a linker.U.S. Provisional Patent Application Attorney Docket No.2920951-49297719. The fusion protein of any one of claims 1-18, wherein the IL-4-binding extracellulardomain (IL-4-Ralpha1-ECD) and the IL-13-binding extracellular domain (IL-13- Ralpha2-ECD) are joined by a linker.

20. The fusion protein of any one of claims 1-19, wherein the IL-13-binding extracellulardomain (IL-13-Ralpha2-ECD) and OSRM^-ligand binding domain (LBD) are joined by a linker.

21. The fusion protein of any one of claims 1-20, wherein the OSRM^-ligand bindingdomain (LBD) and the IL-31RA-ligand binding domain (LBD) are joined by a linker.

22. The fusion protein of any one of claims 1-21, wherein the IL-31RA-ligand bindingdomain (LBD) and fragment crystallizable region of an antibody (Fc) are joined by a linker. CANINE SEQUENCES23. The fusion protein of any one of claims 1-22, wherein the IL-4-binding extracellulardomain is a canine sequence.

24. The fusion protein of any one of claims 1-23, wherein the IL-4-binding extracellulardomain comprises an amino acid sequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 2.

25. The fusion protein of any one of claims 1-24, wherein the IL-4-binding extracellulardomain comprises an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 2.

26. The fusion protein of any one of claims 1-25, wherein the IL-13-binding extracellulardomain is a canine sequence.

27. The fusion protein of any one of claims 1-26, wherein the IL-13-binding extracellulardomain comprises an amino acid sequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 12.

28. The fusion protein of any one of claims 1-27, wherein the IL-13-binding extracellulardomain comprises an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 12.U.S. Provisional Patent Application Attorney Docket No.2920951-49297729. The fusion protein of any one of claims 1-28, wherein the Fc sequence is a caninesequence.

30. The fusion protein of any one of claims 1-29, wherein the Fc sequence comprises anamino acid sequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 7.

31. The fusion protein of any one of claims 1-30, wherein the Fc sequence comprises anamino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 7.

32. The fusion protein of claim 1, wherein the canine IL-4alpha1 receptor ECD-canine IgGFc fusion protein comprises an amino acid sequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 8.

33. The fusion protein of claim 32, wherein the canine IL-4alpha1 receptor ECD-canine IgGFc fusion protein comprises an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 8.

34. The fusion protein of claim 2 wherein the IL-13Ralpha2 ECD-canine IgG Fc fusionprotein comprises an amino acid sequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 9.

35. The fusion protein of claim 34, wherein the IL-13Ralpha2 ECD-canine IgG Fc fusionprotein comprises an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 9.

36. The fusion protein of claim 3, wherein the fusion protein comprises a cIL-13Rα2-cIL-4Rα1-ECD-cFc protein construct comprising an amino acid sequence with at least about 75% sequence homology to SEQ ID NO: 16.

37. The fusion protein of claim 36, wherein the fusion protein comprises a cIL-13Rα2-cIL-4Rα1-ECD-cFc protein construct comprising an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 16.U.S. Provisional Patent Application Attorney Docket No.2920951-49297738. The fusion protein of any one of claims 1-39, wherein the fusion protein comprises a cIL-4Rα1-ECD-cIL-13Rα2-cFc protein construct comprising an amino acid sequence with at least about 75% sequence homology to SEQ ID NO: 19.

39. The fusion protein of any one of claims 1-40, wherein the fusion protein comprises a cIL-4Rα1-ECD-cIL-13Rα2-cFc protein construct comprising an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 19.

40. The fusion protein of any one of claims 1-39, wherein the IL-31RA sequence is a caninesequence.

41. The fusion protein of claim 40, wherein the IL-31RA sequence comprises an amino acidsequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 23.

42. The fusion protein of claim 41, wherein the IL-31RA sequence comprises an amino acidsequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 23.

43. The fusion protein of any one of claims 1-42, wherein the OSMR^ sequence is a caninesequence.

44. The fusion protein of claim 43, wherein the OSMR^ sequence comprises an amino acidsequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 25.

45. The fusion protein of claim 44, wherein the OSMR^ sequence comprises an amino acidsequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 25.

46. The fusion protein of claim 5, wherein the IL-4 / IL-13 / IL-31 fusion protein comprises anamino acid sequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 26.

47. The fusion protein of claim 46, wherein the IL-4 / IL-13 / IL-31 fusion protein comprises anamino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO:

26. HUMAN CONSTRUCTS48. A fusion protein comprising:an IL-4-binding extracellular domain (IL-4-R^-ECD); andU.S. Provisional Patent Application Attorney Docket No.2920951-492977 a fragment crystallizable region of an antibody (Fc).

49. A fusion protein comprising:an IL-13-binding extracellular domain (IL-13-Ralpha2-ECD); and a fragment crystallizable region of an antibody (Fc).

50. A fusion protein comprising:an IL-4-binding extracellular domain (IL-4-R^-ECD); an IL-13-binding extracellular domain (IL-13-Ralpha2-ECD); and a fragment crystallizable region of an antibody (Fc).

51. The fusion protein of any one of claims 48-50, wherein the C-terminal of the IL-4-binding extracellular domain is linked to the N-terminal of IL-13-binding extracellular domain.

52. The fusion protein of any one of claims 48-51, wherein the C-terminal of the IL-13-binding extracellular domain is linked to the N-terminal of the IL-4-binding extracellular domain.

53. The fusion protein of any one of claims 48-52, wherein the Fc is linked to the C-terminalof the IL-4-binding extracellular domain.

54. The fusion protein of any one of claims 48-53, wherein the Fc is linked to the C-terminalof the IL-13-binding extracellular domain.

55. The fusion protein of any one of claims 48-54, wherein the C-terminus of the IL-4-binding extracellular domain is linked to the N-terminus IL-13-binding extracellular domain which is linked to the N-terminus of the fragment crystallizable region of an antibody.

56. The fusion protein of any one of claims 48-55, wherein the C-terminus of the IL-13-binding extracellular domain is linked to the N-terminus of the IL-4-binding extracellular domain which is linked to the N-terminus of the fragment crystallizable region of an antibody.

57. The fusion protein of claim 48 or 50, wherein the IL-4-binding extracellular domain (IL-4-RΑ1-ECD) and cFc are joined by a linker.

58. The fusion protein of claim 49 or 53, wherein the IL-13-binding extracellular domain (IL-13-RA2-ECD) and cFc are joined by a linker.U.S. Provisional Patent Application Attorney Docket No.2920951-49297759. The fusion protein of any one of claims 48-58, wherein the IL-4-binding extracellulardomain (IL-4-RΑ1-ECD) and the IL-13-binding extracellular domain (IL-13-RA2-ECD) are joined by a linker. HUMAN SEQUENCES60. The fusion protein of any one of claims 48-59, wherein the IL-4-binding extracellulardomain is a human sequence.

61. The fusion protein of claim 60, wherein the IL-4-binding extracellular domain comprisesan amino acid sequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 30.

62. The fusion protein of claim 61, wherein the IL-4-binding extracellular domain comprisesan amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 30.

63. The fusion protein of any one of claims 48-62, wherein the IL-13-binding extracellulardomain is a human sequence.

64. The fusion protein of claim 63, wherein the IL-13-binding extracellular domaincomprises an amino acid sequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 32.

65. The fusion protein of claim 64, wherein the IL-13-binding extracellular domaincomprises an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 32.

66. The fusion protein of any one of claims 48-65, wherein the Fc sequence is a humansequence.

67. The fusion protein of claim 66, wherein the Fc sequence comprises an amino acidsequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 33.

68. The fusion protein of claim 67, wherein the Fc sequence comprises an amino acidsequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 33.U.S. Provisional Patent Application Attorney Docket No.2920951-49297769. The fusion protein of any one of claims 48-68, wherein the fusion protein comprises ahIL-4Rα1-hIL-13Rα2-ECD-cFc protein construct comprising an amino acid sequence with at least about 75% sequence homology to SEQ ID NO: 34.

70. The fusion protein of any one of claims 48-69, wherein the fusion protein comprises ahIL-13Rα2-hIL-4Rα1-ECD-cFc protein construct comprising an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO:

34. FELINE CONSTRUCTS71. A fusion protein comprising:an IL-4-binding extracellular domain (IL-4-RΑ1-ECD); and a fragment crystallizable region of an antibody (Fc).

72. A fusion protein comprising:an IL-13-binding extracellular domain (IL-13-Ra2-ECD); and a fragment crystallizable region of an antibody (Fc).

73. A fusion protein comprising:an IL-4-binding extracellular domain (IL-4-RΑ1-ECD); an IL-13-binding extracellular domain (IL-13-Ra2-ECD); and a fragment crystallizable region of an antibody (Fc).

74. The fusion protein of any one of claims 71-73, wherein the C-terminal of the IL-4-binding extracellular domain is linked to the N-terminal of IL-13-binding extracellular domain.

75. The fusion protein of any one of claims 71-74, wherein the C-terminal of the IL-13-binding extracellular domain is linked to the N-terminal of the IL-4-binding extracellular domain.

76. The fusion protein of any one of claims 71-75, wherein the Fc is linked to the C-terminalof the IL-4-binding extracellular domain.

77. The fusion protein of any one of claims 71-76, wherein the Fc is linked to the C-terminalof the IL-13-binding extracellular domain.

78. The fusion protein of any one of claims 71-77, wherein the C-terminus of the IL-4-binding extracellular domain is linked to the N-terminus IL-13-binding extracellularU.S. Provisional Patent Application Attorney Docket No.2920951-492977 domain which is linked to the N-terminus of the fragment crystallizable region of an antibody.

79. The fusion protein of any one of claims 71-78, wherein the C-terminus of the IL-13-binding extracellular domain is linked to the N-terminus of the IL-4-binding extracellular domain which is linked to the N-terminus of the fragment crystallizable region of an antibody.

80. The fusion protein of claim 71 or 73, wherein the IL-4-binding extracellular domain (IL-4-RΑ1-ECD) and cFc are joined by a linker.

81. The fusion protein of claim 72 or 74, wherein the IL-13-binding extracellular domain (IL-13-RA2-ECD) and cFc are joined by a linker.

82. The fusion protein of any one of claims 71-81, wherein the IL-4-binding extracellulardomain (IL-4-RΑ1-ECD) and the IL-13-binding extracellular domain (IL-13-RA2-ECD) are joined by a linker. FELINE SEQUENCES83. The fusion protein of any one of claims 71-82, wherein the IL-4-binding extracellulardomain is a feline sequence.

84. The fusion protein of any one of claims 71-83, wherein the IL-4-binding extracellulardomain comprises an amino acid sequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 39.

85. The fusion protein of any one of claims 71-84, wherein the IL-4-binding extracellulardomain comprises an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 39.

86. The fusion protein of any one of claims 71-85, wherein the IL-4-binding extracellulardomain comprises an amino acid sequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 40.

87. The fusion protein of any one of claims 71-86, wherein the IL-4-binding extracellulardomain comprises an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 40.U.S. Provisional Patent Application Attorney Docket No.2920951-49297788. The fusion protein of any one of claims 71-87, wherein the IL-13-binding extracellulardomain is a feline sequence.

89. The fusion protein of any one of claims 71-88, wherein the IL-13-binding extracellulardomain comprises an amino acid sequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 43.

90. The fusion protein of any one of claims 71-89, wherein the IL-13-binding extracellulardomain comprises an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 43.

91. The fusion protein of any one of claims 71-90, wherein the IL-13-binding extracellulardomain comprises an amino acid sequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 44.

92. The fusion protein of any one of claims 71-91, wherein the IL-13-binding extracellulardomain comprises an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 44.

93. The fusion protein of any one of claims 71-92, wherein the Fc sequence is a felinesequence.

94. The fusion protein of any one of claims 71-93, wherein the Fc sequence comprises anamino acid sequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 46.

95. The fusion protein of any one of claims 71-94, wherein the Fc sequence comprises anamino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 46.

96. The fusion protein of any one of claims 71-93, wherein the Fc sequence comprises anamino acid sequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 47.

97. The fusion protein of any one of claims 71-96, wherein the Fc sequence comprises anamino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 47.U.S. Provisional Patent Application Attorney Docket No.2920951-49297798. The fusion protein of any one of claims 71-97, wherein the fIL-13Rα2-fIL-4Rα1-ECD-cFc fusion protein comprises an amino acid sequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 48.

99. The fusion protein of any one of claims 71-98, wherein the fIL-13Rα2-fIL-4Rα1-ECD-cFc fusion protein comprises an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO: 48.

100. The fusion protein of any one of claims 71-99, wherein the fIL-4Rα1- fIL-13Rα2-ECD-cFc fusion protein comprises an amino acid sequence with at least 75% homology to the amino acid sequence of SEQ ID NO: 49.

101. The fusion protein of any one of claims 71-100, wherein the fIL-4Rα1- fIL-13Rα2-ECD-cFc fusion protein comprises an amino acid sequence with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology to the amino acid sequence of SEQ ID NO:

49. SHARED FEATURES102. The fusion protein of any one of claims 1-101, wherein the linker is an oligopeptidecomprising about 8 to 20 amino acids.

103. The fusion protein of claim 102, wherein the oligopeptide is a polyglycine oligopeptide.

104. The fusion protein of claim 106, wherein the oligopeptide comprises the amino acidsequence of SEQ ID NO: 81.

105. The fusion protein of claim 102, wherein the oligopeptide is a polyglycine / serineoligopeptide.

106. The fusion protein of claim 105, wherein the oligopeptide comprises the amino acidsequence of SEQ ID NO: 82, 83, or 84.

107. The fusion protein of claim 102, wherein the oligopeptide comprises the amino acidsequence of SEQ ID NO: 85.

108. The fusion protein of any one of claims 1-107, wherein the fusion protein furthercomprises an ER import signal sequence on the N-terminal.

109. The fusion protein of any one of claims 1-109, wherein the fusion protein furthercomprises an affinity tag on the C-terminal.U.S. Provisional Patent Application Attorney Docket No.2920951-492977110. The fusion protein of claim 109, wherein the affinity tag is a hexahistidine-tag.

111. A composition comprising the fusion protein of any one of claims 1-110.

112. The composition of claim 111, wherein the composition is a pharmaceutical composition.

113. The pharmaceutical composition of claim 112, wherein the pharmaceutical compositionfurther comprises a pharmaceutical excipient, carrier, diluent, adjuvant, or a combination thereof.

114. The composition of any one of claims 111-113, wherein the composition is formulatedfor intravenous, subcutaneous, infusion, oral, intrathecal, intraperitoneal, parenteral administration, or a combination thereof.

115. The composition of any one of claims 111-114, wherein the composition furthercomprises an anti IL-31 antibody, anti IL-5 antibody, anti-IL-22 antibody, or a combination thereof.

116. A nucleotide sequence encoding the fusion protein of any one of claims 1-110.

117. An expression vector comprising the nucleotide sequence of claim 116.

118. A recombinant host cell comprising the nucleotide sequence of claim 117.

119. A recombinant host cell comprising the expression vector of claim 117.

120. A method of treating or preventing inflammation comprising administering to a mammalin need thereof an effective amount of the fusion protein of any one of claims 1-110.

121. A method of treating or preventing inflammation comprising administering to a mammalin need thereof an effective amount of the composition any one of claims 111-115.

122. The method of claim 120 or 121, wherein the dermatological condition comprises at leastone skin disorder selected from psoriasis, atopic dermatitis, skin rash, skin irritation, skin sensitization, allergic reactions, pruritus, and combinations thereof.

123. The method of any one of claims 120-122, wherein the inflammation is associated withatopic dermatitis.

124. The method of any one of claims 120-123, wherein the inflammation is associated withallergic dermatitis.

125. The method of any one of claims 120-124, wherein the mammal is a human.

126. The method of any one of claims 120-125, wherein the mammal is a non-humanmammal.U.S. Provisional Patent Application Attorney Docket No.2920951-492977127. The method of any one of claims 120-126, wherein the mammal is a canine.

128. The method of any one of claims 120-127, wherein the mammal is a dog.

129. The method of claim 128, wherein the dog is at least 9 months of age.

130. The method of claim 129, wherein the dog is at least 12 months of age.

131. The method of any one of claims 120-124 wherein the mammal is a cat.

132. The method of any one of claims 120-131, wherein the administering is performed daily.

133. The method of any one of claims 120-132, wherein the administering is performed twicedaily.

134. The method of any one of claims 120-133, wherein the administering is performedweekly.

135. The method of any one of claims 120-134, wherein the administering is performedmonthly.

136. The method of any one of claims 120-135, wherein the effective amount is betweenabout 0.1 and 10 mg / kg.

137. The method of any one of claims 120-136, wherein the effective amount is betweenabout 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 mg / kg.

138. The method of any one of claims 120-137, wherein the effective amount is betweenabout 0.1 and 1 mg / kg, 0.5 and 2 mg / kg, 0.75 and 5 mg / kg, or 1 and 10 mg / kg.

139. A composition for comprising an effective amount of the fusion protein of any one ofclaims 1-110.

140. The composition of claim 139, wherein the composition is a pharmaceutical compositionfurther comprising a pharmaceutically acceptable excipient, carrier, diluent, vehicle, adjuvant, or a combination thereof.

141. The composition of claim 139 or 140, wherein the effective amount is between about 0.1and 10 mg / kg.

142. The composition of any one of claims 139-141, wherein the effective amount is betweenabout 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 mg / kg.

143. The composition of any one of claims 139-142, wherein the effective amount is betweenabout 0.1 and 1 mg / kg, 0.5 and 2 mg / kg, 0.75 and 5 mg / kg, or 1 and 10 mg / kg.U.S. Provisional Patent Application Attorney Docket No.2920951-492977144. The composition of any one of claims 139-143, wherein the inflammation is associatedwith atopic dermatitis.

145. The composition of any one of claims 139-144, wherein the inflammation is associatedwith allergic dermatitis.

146. Use of the fusion protein of any one of claims 1-110 for the manufacture of a medicamentfor the treatment of inflammation in a mammal.

147. The use of claim 146, wherein the inflammation is associated with atopic dermatitis orallergic dermatitis.

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