ANTI-IL23 AND ANTI-TNFalpha ANTIBODIES: COMPOSITIONS AND Veterinary USES

Abstract

Various embodiments related to canine, feline, or horse-derived antibodies that bind to canine, feline, and / or horse IL23 and / or TNF [alpha], including bispecific antibodies that bind to both IL23 and TNF [alpha], are provided. Such antibodies may be used alone or in combination in methods of treating canine, cat and / or horse subjects suffering from inflammatory disorders, such as inflammatory disorders of dogs and cats, such as inflammatory bowel disease (IBD), osteoarthritis and gastroenteritis.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 387,777, filed December 16, 2022, each of which is incorporated herein by reference in its entirety. Technical Field

[0003] The present disclosure relates to caninized, feline, and equine anti-IL23 and anti-TNFα antibodies and their uses, e.g., for the treatment of canine, feline, or equine IL23-mediated and / or TNFα-mediated disorders, such as chronic bowel disease - inflammatory bowel disease (IBD), psoriasis, rheumatoid arthritis, osteoarthritis, sepsis, and multiple sclerosis (MS).

[0004] Sequence Listing Reference

[0005] An official copy of the sequence listing is filed with the USPTO Patent Center as an XML file in WIPO standard ST.26 format simultaneously with this specification, with the file name "20136-001WO1.xml", the creation date being December 13, 2023, and the size being 53,419 bytes. This sequence listing filed with the USPTO Patent Center is a part of this specification and is incorporated herein by reference in its entirety. Background Art

[0006] Interleukin-23 (IL23) is a heterodimeric cytokine composed of the IL-12B (IL-12p40) subunit (shared with IL-12) and the IL-23A (IL-23p19) subunit. IL23 is an inflammatory cytokine that is part of the IL-12 cytokine family. It has been shown to be a key cytokine for the maintenance and expansion of type 17 helper T cells (Th17 cells). IL23 is associated with a variety of chronic inflammatory conditions in humans, including diseases such as psoriasis, rheumatoid arthritis, psoriatic arthritis, osteoarthritis, multiple sclerosis, and inflammatory bowel disease (IBD).

[0007] Like humans, companion animals such as cats, dogs, and horses also suffer from chronic inflammatory conditions, however, to date there is no evidence that anti-IL23 can be used to treat such diseases in companion animals. In addition, proteins with significant human-derived amino acid sequence content may be immunogenic in non-human animals and may not bind to companion animal IL23 in a manner that provides an equivalent beneficial therapeutic effect in companion animals. See Mauldin et al., August 2010, 21(4):373-382.

[0008] Inflammatory bowel disease (IBD) is a general term used to describe conditions associated with chronic inflammation of the gastrointestinal tract. Although the exact cause of IBD in companion animals remains unclear, changes in the immune system's tolerance to dietary antigens, intestinal microbiota, and genetic susceptibility may all play a role. Canine, feline, and equine IBD may share similar causes to the conditions that occur in humans (also known as Crohn's disease and ulcerative colitis), but the clinical syndrome and histological changes may be slightly different. Canine IBD is characterized by persistent or recurring signs such as vomiting, diarrhea, abdominal pain, weight loss, and / or changes in appetite, as well as GI tract inflammation. (Ettinger and Feldman; Suchodolski JS et al. "The fecal microbiome in dogs with acute diarrhea and idiopathic inflammatory bowel disease." PLoS ONE. 2012; 7(12):e51907. doi:10.1371 / journal.pone.0051907; Suchodolski JS. "Companion-animals symposium: Microbes and gastrointestinal health of dogs and cats." JAnim Sci. 2010; 89(5):1520-1530. Feline IBD is characterized by vomiting, diarrhea, and weight loss, as well as inflammation of the GI tract, most commonly the small intestine (Jergens AE (2012). Feline idiopathic inflammatory bowel disease: what we know and what remains to be unraveled. Journal of feline medicine and surgery, 14(7), 445–458. https: / / doi.org / 10.1177 / 1098612X12451548).IBD in horses is characterized by lethargy, diarrhea, colic, and weight loss (Boshuizen, B., et al. (2018). Inflammatory bowel disease (IBD) in horses: a retrospective study exploring the value of different diagnostic approaches. BMC veterinary research, 14(1), 21. https: / / doi.org / 10.1186 / s12917-018-1343-1).

[0009] Accurate diagnosis of IBD in companion animals can be challenging, and the term "idiopathic" is often used when the exact pathogen cannot be identified. A complete history and physical examination, followed by laboratory tests, diagnostic imaging, and intestinal biopsy (to demonstrate the presence of inflammation) are generally recommended. Prognosis and response to traditional therapies vary in canine, feline, and equine IBD and can range from moderate to poor. However, studies evaluating cytokines in companion animals with IBD have demonstrated similar upregulation of cytokines such as IL-23 and TNFα as seen in humans (Jergens, AE, & Simpson, KW (2012). Inflammatory bowel disease in veterinary medicine. Frontiers in biopsy (elite ed.), 4(4), 1404–1419. https: / / doi.org / 10.2741 / e470 ; Jergens AE (2012). Feline idiopathic inflammatory bowel disease: what we know and whatremains to be unraveled. Journal of feline medicine and surgery, 14(7), 445–458. https: / / doi.org / 10.1177 / 1098612X12451548; Cerquetella, M., Spaterna, A., Laus, F., Tesei, B., Rossi, G., Antonelli, E., Villanacci, V., & Bassotti, G. (2010). Inflammatory bo wel disease in the dog: differences and similarities with humans. Wo rld journal of gastroenterology, 16(9), 1050–1056. https: / / doi.org / 10.3748 / wjg.v16.i9.1050 Olofsson, KM, Hjertner, B., Fossum, C., Press, C. M., & Lindberg, R. (2015). Expression of T helper type 17 (Th17)-associated cytokines and toll-like receptor 4 and their correlation with Foxp3-positive cells in rectal biopsies of horses with clinical signs of inflammatory bowel disease. Veterinary journal (London, England: 1997), 206(1), 97–104. https: / / doi.org / 10.1016 / j.tvjl.2015.07.003). However, many of the often-prescribed biologics successfully used to treat IBD in humans cannot be used in companion animals, resulting in limited therapeutic efficacy.

[0010] Gastroenteritis in humans and companion animals is associated with inflammation of the GI tract. Gastroenteritis in dogs and felines often presents with diarrhea and vomiting. Similar to IBD, studies evaluating gastroenteritis have found that cytokines involved in inflammation, such as IL-23 and TNFα, are also involved (Godinez, I., Keestra, A.M., Spees, A., & AJ(2011).The IL-23axis in Salmonella gastroenteritis.Cellular microbiology,13(11),1639-1647;Mu S., Gómez-García, A., Millán-Ibarra, J., Giono-Cerezo, S., & Yépez-Mulia, L. (2010). Giardia lamblia: interleukin 6 and tumornecrosis factor-alpha release from mast cells induced through an Ig-independent pathway. Experimental parasitology, 126(3), 298–303. ht tps: / / doi.org / 10.1016 / j.exppara.2010.06.013 ) suggests that targeting IL-23 and TNFα cytokines may be helpful in treating gastroenteritis (Zhou, P., Li, E., SHEA-DONOHUE, T., & Singer, SM (2007). Tumour necrosis factor α controls to protect against Giardia lamblia infection in mice. Parasit e immunology, 29(7), 367-374). Current treatments for gastroenteritis in companion animals are limited to electrolytes and antibiotics, and additional compounds that address GI tract inflammation may be beneficial.

[0011] In newborn foals, sepsis is an inflammatory response syndrome (SIRS) caused by a failure of passive transfer, and the foal presents with symptoms including pneumonia, meningoencephalitis, and / or arthritis (Taylor S. (2015). A review of equine sepsis. Equine veterinary education, 27(2), 99–109. https: / / doi.org / 10.1111 / eve.12290). Neonatal sepsis has a low survival rate, ranging from 45% to 60% (Taylor S. (2015). A review of equine sepsis. Equine veterinary education, 27(2), 99–109. http s: / / doi.org / 10.1111 / eve.12290). Proinflammatory cytokines such as TNFα and other cytokines downstream of IL-23 were found to be present at high levels in septic foals and were directly associated with the development of multiple organ dysfunction syndrome (Taylor S. (2015). A review of equ ine sepsis. Equine veterinary education, 27(2), 99–109. https: / / doi.org / 10.1111 / eve.12290 To date, there are no compounds for the treatment of sepsis, and compounds targeting IL-23 and TNFα cytokines may be beneficial for sepsis treatment.

[0012] Anti-TNFα and anti-IL-23 monoclonal antibodies have been investigated for the treatment of human rheumatoid arthritis (Smolen, J.S., Agarwal, S.K., Ilivanova, E., Xu, X.L., Miao, Y., Zhuang, Y., & Baker, D. (2017). A randomised phase II study evaluating the efficacy and safety of subcutaneously administered ustekinumab and guselkumab in patients with active rheumatoid arthritis despite treatment with methotrexate. Annals of the rheumatic diseases, 76(5), 831-839.; Chimenti, M.S., Talamonti, M., Novelli, L., Teoli, M., Galluzzo, M., Triggianese, P., & Perricone, R. (2015). Long-term ustekinumab therapy of psoriasis in patients with coexisting rheumatoid arthritis and syndrome.Report of two cases and review of literature.Journal ofdermatological case reports,9(3),71–75. https: / / doi.org / 10.3315 / jdcr.2015.1207; de Avila Machado, MA, Maciel, AA, de Lemos, LLP, Costa, JO, Kakehasi, AM, Andrade, EIG,... & de Assis Acurcio, F. (2013). Adalimu mab inrheumatoid arthritis treatment: a systematic review and meta-analysis ofrandomized clinical trials. Revista Brasileira de Reumatol ogia (English version), 53(5), 419-430. Schiff, MH, Burmester, GR, Kent, JD, Pangan, AL, Kupper, H., Fitzpatrick, SB, & Donova n, C. (2006). Safety analyzes of adalimumab (HUMIRA) inglobal clinical trials and US postmarketing surveillance of patients with rheumatoid arthritis. Annals of the rheumatic arthritis Diseases, 65(7), 889–894. https: / / doi.org / 10.1136 / ard.2005.043166). However, in a recent study of human osteoarthritis, it has been demonstrated that both TNFα and IL-23 are present in the synovial fluid of patients with OA (Scanzello, CR, & Goldring, SR (2012). The role of synovitis in osteoarthritis pathogenesis. Bone, 51(2), 249–257. https: / / doi.org / 10.1016 / j.bone.2012.02.012; Askari, A., Naghizadeh, MM, Homay ounfar, R., Shahi, A., Afsarian, MH, Paknahad, A., Kennedy, D., & Ataollahi, MR (2016). IncreasedSerum Levels of IL-17A and IL-23Are Associated with Decreased Vitamin D3 andIncreased Pai n in Osteoarthritis.PloS one,11(11),e0164757. https: / / doi.org / 10.1371 / journal.pone.0164757 Scanzello, CR, Umoh, E., Pessler, F., Diaz-Torne, C., Miles, T., Dicarlo, E., & Crow, MK (2009). Local cytokine profiles in kneeosteoarthritis: elevated synovial fluid interleukin-15 differentiates early from end-stage disease. Osteoarthritis and cartilage, 17(8), 1040-1048.). Since osteoarthritis is characterized by joint inflammation, the evidence for the presence of these proinflammatory cytokines in synovial fluid is not surprising. Similar to humans, companion animals will often develop OA with aging. As in humans, OA causes pain, joint swelling, slowness, stiffness, or refusal to move, to name just a few of the symptoms (Brown, DC (2017). What can we learn from osteoarthritis pain in companion animals. Clin. Exp. Rheumatol, 35(Suppl 107), 53-58). Treatments for OA in companion animals include anti-inflammatory drugs; however, these can be harmful to the kidneys of older animals. Compounds targeting the IL-23 and TNFα cytokines may be safer and more effective for the treatment of OA in companion animals.

[0013] Therefore, there remains a need for methods and compounds that can be used to bind to companion animal IL23 in companion animals to treat IL23-related disorders in companion animals. Ideally, such compounds would bind specifically to companion animal IL23 and have a half-life in plasma long enough to be viable for therapy, but would not be highly immunogenic in companion animals. Summary of the Invention

[0014] The present disclosure generally relates to caninized, feline, and equinized antibodies that specifically bind to IL-23 and / or TNFα, and the use of these antibodies in compositions and methods for treating inflammatory conditions in dogs, cats, and horses, such as inflammatory bowel disease (IBD), osteoarthritis, and gastroenteritis. This summary is intended to introduce the subject matter of the present disclosure and is not intended to encompass every embodiment, combination, or variation contemplated and described herein. Further embodiments are contemplated and described through the detailed description, drawings, and claims.

[0015] In at least one embodiment, the present disclosure provides a caninized anti-IL23 antibody, a caninized anti-TNFα antibody, a feline anti-IL23 antibody, or a feline anti-TNFα antibody. The present disclosure provides bispecific antibodies capable of binding to both canine IL23 and canine TNFα (or both feline IL23 and feline TNFα), as well as methods for using such bispecific antibodies for treating dogs or cats, for example, for treating IBD, sepsis, and gastroenteritis. The present disclosure provides methods and compositions for treating IBD in companion animals in combination with anti-TNFα antibodies, and thus relates to the fields of biology, molecular biology, and veterinary medicine.

[0016] Various embodiments provided by the present disclosure include, but are not limited to:

[0017] Embodiment 1 An anti-IL23 antibody that binds to canine, feline and / or equine IL23, comprising (i) a first light chain hypervariable region (HVR-L1), a second light chain hypervariable region (HVR-L2), and a third light chain hypervariable region (HVR-L3), and / or (ii) a first heavy chain hypervariable region (HVR-H1), a second heavy chain hypervariable region (HVR-H2), and a third heavy chain hypervariable region (HVR-H3), wherein: (a) the HVR-L1 region comprises the amino acid sequence of RASQGISSWLA (SEQ ID NO: 4), the HVR-L2 region comprises the amino acid sequence of YAASSLQS (SEQ ID NO: 5), and the HVR-L3 region comprises the amino acid sequence of QQYNIYPYT (SEQ ID NO: 6); and / or (b) the HVR-H1 region comprises the amino acid sequence of KGSGYSFTTYWLG (SEQ ID NO: 8), the HVR-H2 region comprises the amino acid sequence of IMSPVDSDIR (SEQ ID NO: 9); NO:9), and the HVR-H3 region comprises the amino acid sequence ARRRPGQGYFDF (SEQ ID NO:10).

[0018] Embodiment 2 The antibody of embodiment 1, wherein the antibody is caninized, felineized, or equineized.

[0019] Embodiment 3 The antibody of any one of embodiments 1 to 2, wherein the antibody comprises a light chain variable domain (VL) amino acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 3, 11, 13, and 15; and / or a heavy chain variable domain (VH) amino acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 7, 12, 14, and 16; optionally, wherein: (i) the light chain variable domain (VL) comprises a variant of SEQ ID NOs: 3, 11, 13, and 15, wherein 1 to 6 amino acids of the light chain variable domain (VL) are substituted by different amino acids; and / or (ii) the heavy chain variable domain (VH) comprises a variant of SEQ ID NOs: 7, 12, 14, and 16, wherein 1 to 6 amino acids of the heavy chain variable domain (VH) are substituted by different amino acids.

[0020] Embodiment 4 The antibody of any one of embodiments 1 to 3, wherein the antibody comprises a light chain variable domain (VL) and / or a heavy chain variable domain (VH), the light chain variable domain comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 3, 11, 13 and 15, and the heavy chain variable domain comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 7, 12, 14 and 16; optionally, wherein: (i) the antibody comprises a light chain variable domain (VL) and a heavy chain variable domain (VH), the light chain variable domain comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 3, and the heavy chain variable domain comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 7; (ii) the antibody comprises a light chain variable domain (VL) and a heavy chain variable domain (VH), the light chain variable domain comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 11, and the heavy chain variable domain comprising an amino acid sequence selected from the group consisting of SEQ ID NO: NO:12; (iii) the antibody comprises a light chain variable domain (VL) comprising an amino acid sequence selected from SEQ ID NO:13, and the heavy chain variable domain comprises an amino acid sequence selected from SEQ ID NO:14; or (iv) the antibody comprises a light chain variable domain (VL) and a heavy chain variable domain (VH), the light chain variable domain comprises an amino acid sequence selected from SEQ ID NO:15, and the heavy chain variable domain comprises an amino acid sequence selected from SEQ ID NO:16.

[0021] Embodiment 5 The antibody of any one of embodiments 1 to 4, wherein the antibody comprises: a light chain (LC) amino acid sequence that is at least 90% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 17, 19, 22, and 24, and / or a heavy chain (HC) amino acid sequence that is at least 90% identical to SEQ ID NOs: 18, 20, 21, 23, and 25; optionally, wherein the antibody comprises: (i) the LC amino acid sequence of SEQ ID NO: 17 and the HC amino acid sequence of SEQ ID NO: 18; (ii) the LC amino acid sequence of SEQ ID NO: 19 and the HC amino acid sequence of SEQ ID NO: 20; (iii) the LC amino acid sequence of SEQ ID NO: 19 and the HC amino acid sequence of SEQ ID NO: 21; (iv) the LC amino acid sequence of SEQ ID NO: 22 and the HC amino acid sequence of SEQ ID NO: 23; or (v) the LC amino acid sequence of SEQ ID NO: 24 and the HC amino acid sequence of SEQ ID NO: 25.

[0022] Embodiment 6 An anti-TNFα antibody that binds to canine, feline and / or equine TNFα comprises (i) a first light chain hypervariable region (HVR-L1), a second light chain hypervariable region (HVR-L2), and a third light chain hypervariable region (HVR-L3), and / or (ii) a first heavy chain hypervariable region (HVR-H1), a second heavy chain hypervariable region (HVR-H2), and a third heavy chain hypervariable region (HVR-H3), wherein: (a) the HVR-L1 region comprises the amino acid sequence of RASQGIRNYLA (SEQ ID NO: 27), the HVR-L2 region comprises the amino acid sequence of AASTLQ (SEQ ID NO: 28), and HVR-L3 comprises the amino acid sequence of QRYNRAPYT (SEQ ID NO: 29); and / or (a) the HVR-H1 region comprises the amino acid sequence of FTFDDYAMH (SEQ ID NO: 31), the HVR-H2 region comprises the amino acid sequence of AITWNSGHIDYADSVEGR (SEQ ID NO: 32); NO:32), and the HVR-H3 region comprises the amino acid sequence AKVSYLSTASSLDY (SEQ ID NO:33).

[0023] Embodiment 7 The antibody of embodiment 6, wherein the antibody is caninized, felineized, or equineized.

[0024] Embodiment 8 The antibody of any one of embodiments 6 to 7, wherein the antibody comprises a light chain variable domain (VL) amino acid sequence that is at least 90% identical to a sequence selected from SEQ ID NOs: 26, 34, and 36; and / or a heavy chain variable domain (VH) amino acid sequence that is at least 90% identical to a sequence selected from SEQ ID NOs: 30, 35, and 37; optionally, wherein: (i) the light chain variable domain (VL) comprises a variant of SEQ ID NOs: 26, 34, and 36, wherein 1 to 6 amino acids of the light chain variable domain (VL) are substituted by different amino acids; and / or (ii) the heavy chain variable domain (VH) comprises a variant of SEQ ID NOs: 30, 35, and 37, wherein 1 to 6 amino acids of the heavy chain variable domain (VH) are substituted by different amino acids.

[0025] Embodiment 9 The antibody of any one of embodiments 6 to 8, wherein the antibody comprises a light chain variable domain (VL) and / or a heavy chain variable domain (VH), the light chain variable domain comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 26, 34 and 36, and the heavy chain variable domain comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 30, 35 and 37; optionally, wherein the antibody comprises: (i) a light chain variable domain (VL) and a heavy chain variable domain (VH), the light chain variable domain comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 26, and the heavy chain variable domain comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 30; (ii) a light chain variable domain (VL) and a heavy chain variable domain (VH), the light chain variable domain comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 34, and the heavy chain variable domain comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 35; (iii) a light chain variable domain (VL) and a heavy chain variable domain (VH), the light chain variable domain comprising an amino acid sequence selected from the group consisting of SEQ ID NO: NO:36, and the heavy chain variable domain comprises an amino acid sequence selected from SEQ ID NO:37; (iv) a light chain variable domain (VL) and a heavy chain variable domain (VH), the light chain variable domain comprises an amino acid sequence selected from SEQ ID NO:36, and the heavy chain variable domain comprises an amino acid sequence selected from SEQ ID NO:37; (v) a light chain variable domain (VL) and a heavy chain variable domain (VH), the light chain variable domain comprises an amino acid sequence selected from SEQ ID NO:36, and the heavy chain variable domain comprises an amino acid sequence selected from SEQ ID NO:30; or (vi) a light chain variable domain (VL) and a heavy chain variable domain (VH), the light chain variable domain comprises an amino acid sequence selected from SEQ ID NO:26, and the heavy chain variable domain comprises an amino acid sequence selected from SEQ ID NO:37.

[0026] Embodiment 10 The antibody of any one of embodiments 6 to 9, wherein the antibody comprises: a light chain (LC) amino acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 38, 40, and 41 and / or a heavy chain (HC) amino acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 39, 42, 43, and 44; optionally, wherein the antibody comprises: (i) the LC amino acid sequence of SEQ ID NO: 38 and the HC amino acid sequence of SEQ ID NO: 39; (ii) the LC amino acid sequence of SEQ ID NO: 40 and the HC amino acid sequence of SEQ ID NO: 42; (iii) the LC amino acid sequence of SEQ ID NO: 41 and the HC amino acid sequence of SEQ ID NO: 42; (iv) the LC amino acid sequence of SEQ ID NO: 40 and the HC amino acid sequence of SEQ ID NO: 43; (v) the LC amino acid sequence of SEQ ID NO: 41 and the HC amino acid sequence of SEQ ID NO: 43; (vi) the LC amino acid sequence of SEQ ID NO: 40 and the HC amino acid sequence of SEQ ID NO: 44; or (vii) the LC amino acid sequence of SEQ ID NO: 41 and the HC amino acid sequence of SEQ ID NO: 45. The LC amino acid sequence of SEQ ID NO:41 and the HC amino acid sequence of SEQ ID NO:44.

[0027] Embodiment 11 The antibody of any one of Embodiments 6 to 10, wherein the antibody is a scFv antibody; optionally, wherein the scFv antibody comprises an amino acid sequence having at least 90% sequence identity to a sequence selected from SEQ ID NOs: 45 and 46.

[0028] Embodiment 12 The antibody of any one of embodiments 1 to 11, wherein the antibody is an antibody fragment selected from Fv, scFv, Fab, Fab', F(ab')2 and Fab'-SH.

[0029] Embodiment 13 The antibody of any one of Embodiments 1 to 12, wherein the antibody comprises a canine heavy chain constant region selected from the group consisting of IgG-A, IgG-B, IgG-C, and IgG-D constant regions.

[0030] Embodiment 14 The antibody of any one of Embodiments 1 to 13, wherein the antibody comprises: (i) a canine light chain constant region and / or a canine heavy chain constant region; (ii) a feline light chain constant region and / or a feline heavy chain constant region; or (iii) an equine light chain constant region and / or an equine heavy chain constant region.

[0031] Embodiment 15 The antibody of any one of Embodiments 1 to 14, wherein the antibody comprises a heavy chain constant region having a "Y" mutation at position 252, EU numbering.

[0032] Embodiment 16 A bispecific antibody that binds to canine IL23 and canine TNFα, wherein the antibody comprises: a light chain (LC) comprising a VL domain, the VL domain having an HVR-L1 region comprising the amino acid sequence of RASQGISSWLA (SEQ ID NO: 4), an HVR-L2 region comprising the amino acid sequence of YAASSLQS (SEQ ID NO: 5), and an HVR-L3 region comprising the amino acid sequence of QQYNIYPYT (SEQ ID NO: 6); and a heavy chain (HC) comprising a VH domain, the VH domain having an HVR-H1 region comprising the amino acid sequence of KGSGYSFTTYWLG (SEQ ID NO: 8), an HVR-H2 region comprising the amino acid sequence of IMSPVDSDIR (SEQ ID NO: 9), and an HVR-H3 region comprising the amino acid sequence of ARRRPGQGYFDF (SEQ ID NO: 10). NO:10); and an scFv antibody fused to the HC, wherein the scFv antibody comprises a VL domain and a VH domain, the VL domain having an HVR-L1 region comprising the amino acid sequence of RASQGIRNYLA (SEQ ID NO:27), an HVR-L2 region comprising the amino acid sequence of AASTLQ (SEQ ID NO:28), and an HVR-L3 comprising the amino acid sequence of QRYNRAPYT (SEQ ID NO:29); and the VH domain having an HVR-H1 region comprising the amino acid sequence of FTFDDYAMH (SEQ ID NO:31), an HVR-H2 region comprising the amino acid sequence of AITWNSGHIDYADSVEGR (SEQ ID NO:32), and an HVR-H3 region comprising the amino acid sequence of AKVSYLSTASSLDY (SEQ ID NO:33).

[0033] Embodiment 17 The bispecific antibody of embodiment 16, wherein the antibody comprises: a light chain (LC) and a heavy chain (HC), wherein the light chain comprises a light chain variable domain (VL) amino acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 3, 11, 13, and 15, and the heavy chain comprises a heavy chain variable domain (VH) amino acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 7, 12, 14, and 16, wherein the HC is fused to a scFv antibody comprising a light chain variable domain (VL) amino acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 26, 34, and 36, and a heavy chain variable domain (VH) amino acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 30, 35, and 37.

[0034] Embodiment 18 The bispecific antibody of any one of Embodiments 16 to 17, wherein the light chain (LC) comprises an amino acid sequence having at least 90% identity to a sequence selected from SEQ ID NOs: 19 and 22; and the heavy chain (HC) comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NOs: 47 and 48; optionally, wherein the antibody comprises: (i) the LC amino acid sequence of SEQ ID NO: 19 and the HC amino acid sequence of SEQ ID NO: 47; (ii) the LC amino acid sequence of SEQ ID NO: 19 and the HC amino acid sequence of SEQ ID NO: 48; (iii) the LC amino acid sequence of SEQ ID NO: 22 and the HC amino acid sequence of SEQ ID NO: 47; or (iv) the LC amino acid sequence of SEQ ID NO: 22 and the HC amino acid sequence of SEQ ID NO: 48.

[0035] Embodiment 19 A bispecific antibody that binds to canine IL23 and canine TNFα, wherein the antibody comprises: a light chain (LC) comprising a VL domain and a heavy chain (HC) comprising a VH domain, wherein the VL domain has an HVR-L1 region comprising the amino acid sequence of RASQGIRNYLA (SEQ ID NO: 27), an HVR-L2 region comprising the amino acid sequence of AASTLQ (SEQ ID NO: 28), and an HVR-L3 comprising the amino acid sequence of QRYNRAPYT (SEQ ID NO: 29); and the VH domain has an HVR-H1 region comprising the amino acid sequence of FTFDDYAMH (SEQ ID NO: 31), an HVR-H2 region comprising the amino acid sequence of AITWNSGHIDYADSVEGR (SEQ ID NO: 32), and an HVR-H3 region comprising the amino acid sequence of AKVSYLSTASSLDY (SEQ ID NO: 33). NO:33); and a scFv antibody fused to the HC, wherein the scFv antibody comprises a VL domain and a VH domain, the VL domain having an HVR-L1 region comprising the amino acid sequence of RASQGISSWLA (SEQ ID NO:4), an HVR-L2 region comprising the amino acid sequence of YAASSLQS (SEQ ID NO:5), and an HVR-L3 region comprising the amino acid sequence of QQYNIYPYT (SEQ ID NO:6); and the VH domain having an HVR-H1 region comprising the amino acid sequence of KGSGYSFTTYWLG (SEQ ID NO:8), an HVR-H2 region comprising the amino acid sequence of IMSPVDSDIR (SEQ ID NO:9), and an HVR-H3 region comprising the amino acid sequence of ARRRPGQGYFDF (SEQ ID NO:10).

[0036] Embodiment 20 The bispecific antibody of embodiment 18, wherein the antibody comprises: a light chain (LC) comprising a light chain variable domain (VL) amino acid sequence at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 26, 34, and 36, and a heavy chain variable domain (VH) amino acid sequence at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 30, 35, and 37, wherein the HC is fused to a scFv antibody comprising a light chain variable domain (VL) amino acid sequence at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 3, 11, 13, and 15, and a heavy chain variable domain (VH) amino acid sequence at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 7, 12, 14, and 16.

[0037] Embodiment 21 A bispecific antibody that binds to canine, feline, and / or equine IL23 and canine, feline, and / or equine TNFα, wherein the antibody comprises:

[0038] (i) an anti-IL23 light chain (LC) comprising a VL domain having an HVR-L1 region comprising the amino acid sequence of RASQGISSWLA (SEQ ID NO:4), an HVR-L2 region comprising the amino acid sequence of YAASSLQS (SEQ ID NO:5), and an HVR-L3 region comprising the amino acid sequence of QQYNIYPYT (SEQ ID NO:6);

[0039] (ii) an anti-IL23 heavy chain (HC) comprising a VH domain having an HVR-H1 region comprising the amino acid sequence of KGSGYSFTTYWLG (SEQ ID NO: 8), an HVR-H2 region comprising the amino acid sequence of IMSPVDSDIR (SEQ ID NO: 9), and an HVR-H3 region comprising the amino acid sequence of ARRRPGQGYFDF (SEQ ID NO: 10);

[0040] (iii) an anti-TNFα light chain (LC) comprising a VL domain having an HVR-L1 region comprising the amino acid sequence of RASQGIRNYLA (SEQ ID NO:27), an HVR-L2 region comprising the amino acid sequence of AASTLQ (SEQ ID NO:28), and an HVR-L3 comprising the amino acid sequence of QRYNRAPYT (SEQ ID NO:29); and

[0041] (iv) an anti-TNFα heavy chain (HC) comprising a VH domain having an HVR-H1 region comprising the amino acid sequence of FTFDDYAMH (SEQ ID NO: 31), an HVR-H2 region comprising the amino acid sequence of AITWNSGHIDYADSVEGR (SEQ ID NO: 32), and an HVR-H3 region comprising the amino acid sequence of AKVSYLSTASSLDY (SEQ ID NO: 33).

[0042] Embodiment 22: The bispecific antibody of embodiment 21, wherein:

[0043] (i) the anti-IL23 light chain (LC) comprises a light chain variable domain (VL) amino acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 3, 11, 13, and 15;

[0044] (ii) the anti-IL23 heavy chain (HC) comprises a heavy chain variable domain (VH) amino acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 7, 12, 14, and 16;

[0045] (iii) the anti-TNFα light chain (LC) comprises a light chain variable domain (VL) amino acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 26, 34, and 36; and

[0046] (iv) the anti-TNFα heavy chain (HC) comprises a heavy chain variable domain (VH) amino acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 30, 35, and 37.

[0047] Embodiment 23 The bispecific antibody of any one of Embodiments 21 to 22, wherein:

[0048] (i) the anti-IL23 light chain (LC) comprises an amino acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 19 and 22;

[0049] (ii) the anti-IL23 heavy chain (HC) comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NOs: 20, 21 and 23;

[0050] (iii) the anti-TNFα light chain (LC) comprises an amino acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 40 and 41; and

[0051] (iv) the anti-TNFα heavy chain (HC) comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NOs: 42, 43 and 44.

[0052] Embodiment 24. One or more isolated nucleic acids encoding the antibody of any one of embodiments 1 to 23.

[0053] Embodiment 25. A host cell comprising one or more nucleic acids according to embodiment 24.

[0054] Embodiment 26 A method of producing an antibody, comprising culturing the host cell of embodiment 25 and isolating the antibody.

[0055] Embodiment 27 A pharmaceutical composition comprising the antibody of any one of Embodiments 1 to 23 and a pharmaceutically acceptable carrier.

[0056] Embodiment 28. A method of treating a dog, cat, or horse having a disorder associated with IL23, the method comprising administering to the dog, cat, or horse a therapeutically effective amount of the antibody of any one of Embodiments 1 to 23 or the pharmaceutical composition of Embodiment 27.

[0057] Embodiment 29. A method of maintaining remission of an IL23-associated disorder in a dog, cat, or horse, comprising administering to the dog, cat, or horse a therapeutically effective amount of the antibody of any one of embodiments 1 to 23 or the pharmaceutical composition of embodiment 27.

[0058] Embodiment 30 The method of any one of embodiments 28 to 29, wherein the IL23-related disorder is an inflammatory disease.

[0059] Embodiment 31 The method of any one of embodiments 28 to 30, wherein the IL23-related disorder is a gastrointestinal inflammatory disease.

[0060] Embodiment 32 The method of any one of embodiments 28 to 31, wherein the disorder associated with IL23 is inflammatory bowel disease.

[0061] Embodiment 33 The method of any one of embodiments 28 to 32, wherein the disorder associated with IL23 is ankylosing spondylitis, asthma, cancer, Crohn's disease, idiopathic arthritis, psoriasis, plaque psoriasis, psoriatic arthritis, rheumatoid arthritis, osteoarthritis, or ulcerative colitis.

[0062] Embodiment 34. A method of treating a dog, cat, or horse having a disorder associated with IL23 and TNFα, comprising administering to the dog, cat, or horse a therapeutically effective amount of an IL23 antibody according to embodiments 1 to 5 or 12 to 23, and an anti-TNFα antibody according to any one of embodiments 6 to 23, or a pharmaceutical composition according to embodiment 24.

[0063] Embodiment 35 The method of embodiment 34, wherein the anti-IL23 antibody is administered in combination with an anti-TNFα antibody or in the form of a bispecific IL23 / TNFα antibody, wherein the condition associated with IL23 or IL23 / TNFα is a gastrointestinal inflammatory disease.

[0064] Embodiment 36 The method of any one of embodiments 34 to 35, wherein the condition associated with IL23 or IL23 / TNFα is ankylosing spondylitis, asthma, cancer, Crohn's disease, idiopathic arthritis, psoriasis, plaque psoriasis, psoriatic arthritis, rheumatoid arthritis, osteoarthritis, or ulcerative colitis.

[0065] Embodiment 37 The method of any one of Embodiments 28 to 36, wherein the antibody or the pharmaceutical composition is administered parenterally.

[0066] Embodiment 38 The method of any one of Embodiments 28 to 37, wherein the antibody or the pharmaceutical composition is administered by an intramuscular route, an intraperitoneal route, a cerebrospinal fluid route, a subcutaneous route, an intraarterial route, an intrasynovial route, an intrathecal route, or an inhalation route.

[0067] Embodiment 39 The method of any one of embodiments 28 to 38, wherein the method further comprises administering an IL17 antibody, an IL-5 antibody, an IL-31 antibody, an IL4 antibody, an IL13 antibody, an IL23 antibody, an IgE antibody, a CD11α antibody, an IL6R antibody, an α4-integrin antibody, a β9-integrin, an IL12 antibody, an IL1β antibody, or an anti-BlyS antibody.

[0068] Embodiment 40 The method of any one of Embodiments 28 to 39, wherein the antibody is administered in an amount ranging from 0.01 mg / kg to 100 mg / kg body weight per dose.

[0069] Embodiment 41 A method of reducing canine, feline or equine IL23 and / or TNFα signaling function in a cell, the method comprising exposing the cell to the antibody of any one of Embodiments 1 to 23 under conditions permissive for binding to IL23 and / or TNFα, thereby reducing binding of the cell to IL23 and / or TNFα signaling function.

[0070] Embodiment 42 The method of embodiment 41, wherein the cell is exposed to the antibody or the pharmaceutical composition ex vivo.

[0071] Embodiment 43 The method of embodiment 41, wherein the cell is exposed to the antibody or the pharmaceutical composition in vivo.

[0072] Embodiment 44 The method of any one of Embodiments 41 to 43, wherein the cell is a canine cell, a feline cell, or a horse cell.

[0073] Embodiment 45 A method for detecting IL23 and / or TNFα in a sample from a companion animal species, the method comprising contacting the sample with the antibody of any one of Embodiments 1 to 23 under conditions permissive for binding of the antibody to IL23 and / or TNFα, and detecting whether a complex is formed between the antibody and IL23 and / or TNFα in the sample.

[0074] Embodiment 46 The method of embodiment 45, wherein the sample is a biological sample obtained from a dog, a cat, or a horse. DETAILED DESCRIPTION

[0075] The present disclosure provides specific embodiments (including examples) of antibodies that specifically bind to canine IL23 and / or feline IL23, antibodies that specifically bind to canine TNFα and / or feline TNFα, and bispecific antibodies that bind to both canine IL23 and canine TNFα (or feline IL23 and feline TNF). The present disclosure provides various exemplary forms of these antibodies, including full-length antibodies and scFv antibodies, as well as uses of these antibodies, including methods for treating various diseases and conditions mediated by or associated with the binding activity of IL23 and / or TNFα. Also provided are methods for designing, producing, or purifying bispecific antibodies against canine IL23 and canine TNFα. Methods for detecting IL23 and / or TNFα in samples from companion animals are provided.

[0076] Unless the context clearly dictates otherwise, for purposes of the description herein and the appended claims, the singular forms "a," "an," and "an" include plural referents. Thus, for example, reference to "a protein" includes more than one protein, and reference to "a compound" refers to more than one compound. It is further noted that claims may be drafted to exclude any optional element. Therefore, this description is intended to serve as a precondition for the use of such exclusive terminology as "only," "only," and the like in connection with the recitation of claim elements, or for the use of a "negative" limitation. The use of "comprise," "comprises," "comprising," "include," "includes," and "including" are interchangeable and are not intended to be limiting. It should also be further understood that while the description of various embodiments uses the term "comprising," those skilled in the art will understand that, in some specific instances, the embodiments may also be described using the language "consisting essentially of" or "consisting of."

[0077] When providing a numerical range, unless the context clearly stipulates otherwise, it should be understood that each intermediate integer of the value, and each intermediate integer of the value, unless the context clearly stipulates otherwise, otherwise any other specified value or intermediate value between the upper and lower limits of the range and within the specified range is encompassed within the present invention. The upper and lower limits of these smaller ranges can be independently included in the smaller range and are also encompassed within the present invention, subject to the constraints of any specific limits excluded within the range. When the range includes one or two of these limits, the scope of (i) one or (ii) two of the limits included is also encompassed within the present invention. For example, "1 to 50" includes "2 to 25", "5 to 20", "25 to 50", "1 to 10" etc.

[0078] Generally, the nomenclature used herein and the techniques and procedures described herein include those well understood and commonly employed by those skilled in the art, such as those described in, for example, Green and Sambrook, Molecular Cloning: A Laboratory Manual (4th ed.), Vols. 1-3, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, 2012 (hereinafter referred to as “Sambrook”); and Current Protocols in Molecular Biology, F.M. Ausubel et al., eds., originally published as a book in 1987 by Greene Publishing Associates, Inc. and John Wiley & Sons, Inc., with regular supplements in 2011, and now available online as a journal Current Protocols Molecular Biology, Vol. 00-130 , (1987-2020), Common Techniques and Methods published by Wiley & Sons, Inc. in the Wiley Online Library (hereinafter referred to as "Ausubel").

[0079] All publications, patents, patent applications, and other documents cited in this disclosure are hereby incorporated by reference in their entirety for all purposes to the same extent as if each individual publication, patent, patent application, or other document were individually indicated to be incorporated by reference herein for all purposes.

[0080] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which the present invention belongs. It should be understood that the terms used herein are only used to describe specific embodiments and are not intended to be limiting. For the purpose of explaining this disclosure, the following terminology will apply, and where appropriate, terms used in the singular will also include the plural form, and vice versa.

[0081] Unless otherwise indicated, as used herein, "IL23" or "IL-23" refers to the cytokine protein interleukin 23 (or IL-23), and encompasses IL23 proteins from any vertebrate source, including mammals, such as primates (e.g., humans and cynomolgus monkeys) and rodents (e.g., mice and rats) and companion animals (e.g., dogs, cats, and horses). The term also includes naturally occurring variants of IL23, e.g., splice variants or allelic variants. The amino acid sequences of exemplary recombinant forms of canine IL23 and feline IL23 proteins are provided in Table 1 below and in the accompanying sequence listing.

[0082] Unless otherwise indicated, as used herein, "TNF" or "TNFa" or "TNFα" refers to the cytokine protein, tumor necrosis factor, TNFα protein from any vertebrate source, including mammals, such as primates (e.g., humans and cynomolgus monkeys) and rodents (e.g., mice and rats) and companion animals (e.g., dogs, cats and horses). The term also includes naturally occurring variants of TNFα, e.g., splice variants or allelic variants.

[0083] As used herein, "IL23-mediated conditions" or "IL23-mediated diseases" encompass any disease or condition associated with the specific binding of IL23 to an IL23 receptor or other specific IL23 binding target, and may include diseases associated with, caused by, or characterized by elevated levels or altered gradients of IL23 concentrations. For example, specific binding of IL23 stimulates the production of Th17 cells that participate in an immune response. Thus, IL23-mediated diseases may include, but are not limited to, any disease or condition mediated by and / or responsive to an antagonist or inhibitor of IL23 binding to an IL23 receptor or other IL23 target. Particular exemplary diseases or conditions include, but are not limited to, Crohn's disease, inflammatory bowel disease (IBD), psoriasis (including plaque psoriasis), psoriatic arthritis, rheumatoid arthritis, ulcerative colitis, osteoarthritis, multiple sclerosis, and other chronic inflammatory disorders.

[0084] As used herein, "TNFα-mediated condition" or "TNFα-mediated disease" encompasses any disease or condition associated with the specific binding of TNFα to a TNFα receptor or other specific TNFα binding target, and may include diseases associated with, caused by, or characterized by elevated levels or altered gradients of TNFα concentrations. Thus, a TNFα-mediated disease may include, but is not limited to, any disease or condition mediated by and / or responsive to an antagonist or inhibitor of the binding of TNFα to a TNFα receptor or other specific TNFα binding target. Specific exemplary diseases are provided elsewhere herein.

[0085] As used herein, "antibody" refers to a molecule comprising one or more polypeptide chains that specifically binds to or immunoreacts with a specific antigen. Exemplary antibodies of the present disclosure include monoclonal antibodies, polyclonal antibodies, chimeric antibodies, caninized antibodies, felinized antibodies, multispecific (or heteroconjugate) antibodies (e.g., bispecific antibodies), monovalent antibodies (e.g., one-armed antibodies), multivalent antibodies, antigen-binding fragments (e.g., Fab', F(ab')2, Fab, Fv, rIgG, and scFv fragments), antibody fusions, and synthetic antibodies (or antibody mimetics).

[0086] "Anti-IL23 antibody" and "antibody that binds to IL23" refer to an antibody that binds to IL23 with sufficient affinity such that the antibody can be used as a diagnostic and / or therapeutic agent targeting IL23. Anti-IL23 antibodies may further specifically refer to antibodies that bind to a specific type of IL23, such as canine IL23 (e.g., "anti-canine IL23" or "anti-IL23 that binds to canine IL23"). In some embodiments, the extent of binding of an anti-IL23-specific antibody to an unrelated, non-IL23 antigen is less than about 20%, less than about 15%, less than about 10%, or less than about 5% of the binding of the antibody to IL23, as measured, for example, by radioimmunoassay (RIA) or surface plasmon resonance (SPR). In some embodiments, the dissociation constant (K) of the antibody that binds to IL23 is less than about 20%, less than about 15%, less than about 10%, or less than about 5% of the binding of the antibody to IL23. D ) is <1 μM, <100 nM, <10 nM, <1 nM, <0.1 nM, <0.01 nM or <1 pM (e.g., 10 -8 M or lower, e.g., 10 -8 M to 10 -13 M, for example, 10 -9 M to 10 -13 M).

[0087] "Anti-TNFα antibody" and "antibody that binds to TNFα" refer to an antibody that binds to TNFα with sufficient affinity such that the antibody is useful as a diagnostic and / or therapeutic agent targeting TNFα. Anti-TNFα antibodies may further specifically refer to antibodies that bind to a specific type of TNFα, such as canine TNFα (e.g., "anti-canine TNFα" or "anti-TNFα that binds canine TNFα"). In some embodiments, the extent of binding of an anti-TNFα-specific antibody to an unrelated, non-TNFα antigen is less than about 20%, less than about 15%, less than about 10%, or less than about 5% of the binding of the antibody to TNFα, as measured, for example, by radioimmunoassay (RIA) or surface plasmon resonance (SPR). In some embodiments, the dissociation constant (K) of the antibody that binds to TNFα is less than about 20%, less than about 15%, less than about 10%, or less than about 5% of the binding of the antibody to TNFα. D ) is <1 μM, <100 nM, <10 nM, <1 nM, <0.1 nM, <0.01 nM or <1 pM (e.g., 10 -8 M or lower, e.g., 10 -8 M to 10 -13 M, for example 10 -9 M to 10 -13 M).

[0088] "Full length antibody," "intact antibody," or "whole antibody" are used interchangeably herein to refer to an antibody that has a structure substantially similar to a native antibody structure or has heavy chains that contain an Fc region as defined herein.

[0089] The "class" of an antibody refers to the type of constant domain or constant region possessed by its heavy chain. There are five major classes of human antibodies: IgA, IgD, IgE, IgG and IgM, and several of these are further divided into subclasses (isotypes), for example, IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2. The heavy chain constant domains corresponding to different classes of immunoglobulins are referred to as α, δ, ε, γ and μ, respectively. Dogs, cats and horses have different classes of antibodies, which are shared by many other mammalian species. For example, dog species have antibody classes IgGA, IgGB, IgGC, IgGD, and cat species have antibody classes IgGA1, IgGA2, IgGB.

[0090] "Variable region" or "variable domain" refers to the domain of an antibody heavy chain or light chain that is involved in binding the antibody to the antigen. The variable domains of the heavy and light chains of a natural antibody (V H and V L ) generally have a similar structure, wherein each domain comprises four conserved framework regions (FR) and three hypervariable regions (HVR) (see, e.g., Kindt et al., Kuby Immunology, 6th ed., WH Freeman and Co., p. 91). A single V H or V L The V domain may be sufficient to confer antigen binding specificity. In addition, the V domain from an antibody that binds a specific antigen is used. H domain or V L domains to screen for complementary V L domain or V H Domain libraries can be used to isolate antibodies that bind the antigen (see, eg, Portolano et al., J. Immunol. 150:880-887 (1993); Clarkson et al., Nature 352:624-628 (1991)).

[0091] As used herein, "hypervariable region" or "HVR" refers to each region of an antibody variable domain that is hypervariable in sequence and / or forms structurally defined loops ("hypervariable loops"). Generally, a native antibody comprises four chains and six HVRs; three in the heavy chain variable domain, V H (HVR-H1, HVR-H2, HVR-H3), and three in the light chain variable domain, V L(HVR-L1, HVR-L2, HVR-L3). HVRs generally comprise amino acid residues from hypervariable loops and / or from "complementarity determining regions" (CDRs). A variety of hypervariable region divisions are in use and are encompassed herein. The Kabat complementarity determining regions (CDRs) are based on sequence variability and are the most commonly used (Kabat et al., Sequences of Proteins of Immunological Interest, 5th ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991)). Chothia refers to the position of structural loops (Chothia and Lesk J. Mol. Biol. 196:901-917 (1987)). The AbM hypervariable regions represent a compromise between the Kabat CDRs and the Chothia structural loops and are used by Oxford Molecular's AbM antibody modeling software. The "contact" hypervariable regions are based on analysis of available complex crystal structures. The following table indicates the residues from each of these hypervariable regions.

[0092]

[0093] Unless otherwise indicated, HVR residues and other residues in the variable domain (eg, FR residues) are numbered herein according to Kabat et al., supra.

[0094] As used herein, a hypervariable region may include an extension or alternative hypervariable region as follows: L 24-36 or 24-34 (L1), 46-56 or 50-56 (L2) and 89-97 or 89-96 (L3) in the domain, and V H 26-35 or 30-35 (H1), 50-61, 50-65 or 49-65 (H2), and 93-102, 94-102 or 95-102 (H3) in the domain. For each of these definitions, the variable domain residues are numbered according to Kabat et al., supra.

[0095] As used herein, "complementarity determining region" or "CDR" refers to the region within the variable domain HVR with the highest sequence variability and / or involved in antigen recognition. Generally, natural antibodies contain four chains and six CDRs; three in the heavy chain variable domain, V H (H1, H2, H3), and three of the light chain variable domains, V L(L1, L2, L3). Exemplary CDRs (CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2, and CDR-H3) occur at amino acid residues 24-34 of L1, amino acid residues 50-56 of L2, amino acid residues 89-97 of L3, amino acid residues 31-35 of H1, amino acid residues 50-61 of H2, and amino acid residues 95-102 of H3. (Numbering is according to Kabat et al., supra).

[0096] "Framework" or "FR" refers to the variable domain residues other than the hypervariable region (HVR) residues. The FR of a variable domain is generally composed of four FR domains: FR1, FR2, FR3, and FR4. Thus, the HVR and FR sequences are arranged in a V H (or V L ) usually appear in the following sequence: FR1-H1(L1)-FR2-H2(L2)-FR3-H3(L3)-FR4.

[0097] "Native antibodies" refer to naturally occurring immunoglobulin molecules. For example, natural IgG antibodies are heterotetrameric glycoproteins of approximately 150,000 daltons composed of two identical light chains and two identical heavy chains linked by disulfide bonds. From N-terminus to C-terminus, each heavy chain has a variable region (V H ) (also called a variable heavy chain domain or a heavy chain variable domain), followed by three constant domains (CH1, CH2, and CH3). Similarly, from N-terminus to C-terminus, each light chain has a variable region (V L The light chains of antibodies are primarily composed of two amino acid sequences: a variable light domain (κ) and a variable light domain (λ). The light chains of antibodies are primarily composed of two amino acid sequences: a variable light domain (κ) and a variable light domain (λ). The variable light domains are followed by a constant light (CL) domain. The light chains of antibodies are primarily composed of two amino acid sequences: a variable light domain (κ) and a variable light domain (λ).

[0098] As used herein, " monoclonal antibody " refers to the antibody obtained from a substantially homogeneous antibody population, i.e., the single antibody comprising the population is identical and / or binds to the same epi-position, except for possible variant antibodies (e.g., variant antibodies contain naturally occurring mutations or mutations that occur during the production of monoclonal antibodies, and generally exist in less amounts). In contrast to polyclonal antibody preparations that typically include different antibodies for different determinants (epitopes), each monoclonal antibody of a monoclonal antibody preparation is directed to a single determinant on the antigen. Therefore, the term " monoclonal " represents the characteristic of the antibody obtained from a substantially homogeneous antibody population and should not be construed as needing to produce the antibody by any ad hoc method. For example, monoclonal antibodies can be prepared by various techniques, including but not limited to hybridoma methods, recombinant DNA methods, phage display methods, and methods utilizing transgenic animals containing all or part of human immunoglobulin loci, and such methods and other exemplary methods for preparing monoclonal antibodies are described herein.

[0099] A "chimeric antibody" refers to an antibody in which a portion of the heavy and / or light chains are derived from a particular source or species, while the remaining heavy and / or light chains are derived from a different source or species. A "canine chimeric antibody" refers to a chimeric antibody having at least a portion of the heavy chain or a portion of the light chain derived from a dog. In some embodiments, a canine chimeric antibody may comprise mouse VH and / or VL sequences and canine heavy and light chain constant domains. In some embodiments, the antibody is a chimeric antibody comprising a murine heavy chain variable domain (VH) framework region or a murine light chain variable domain (VL) framework region.

[0100] A "caninized antibody" refers to a chimeric antibody comprising an amino acid sequence from a non-canine HVR and an amino acid sequence from a canine FR. In certain embodiments, a caninized antibody will comprise substantially all of at least one and typically two variable domains, wherein all or substantially all of the HVRs correspond to those of a non-canine antibody, and all or substantially all of the FRs correspond to those of a canine antibody. A caninized antibody optionally may comprise at least a portion of an antibody constant region derived from a canine antibody. A "caninized form" of an antibody (e.g., a non-canine antibody) refers to an antibody that has undergone caninization.

[0101] A "felinized antibody" refers to a chimeric antibody comprising an amino acid sequence from a non-feline HVR and an amino acid sequence from a feline FR. In certain embodiments, a felinized antibody will comprise substantially all of at least one and typically two variable domains, wherein all or substantially all of the HVRs correspond to those of a non-feline antibody, and all or substantially all of the FRs correspond to those of a feline antibody. A felinized antibody optionally may comprise at least a portion of an antibody constant region derived from a feline antibody. A "felinized form" of an antibody (e.g., a non-feline antibody) refers to an antibody that has undergone felinization.

[0102] "Canine antibody" refers to an antibody having an amino acid sequence corresponding to an antibody produced by a dog or canine cell, or an amino acid sequence derived from a non-canine source utilizing a canine antibody library or other canine antibody encoding sequence. This definition of a canine antibody specifically excludes caninized antibodies that contain non-canine antigen-binding residues.

[0103] The consensus framework is the L or V H The framework of the most commonly occurring amino acid residues in the framework sequence. L or V H The sequence is selected from a subset of variable domain sequences. Generally, the subset of sequences is as described in Kabat et al., Sequences of Proteins of Immunological Interest, Fifth Edition, NIH Press 91-3242, Bethesda MD (1991), Volumes 1-3. In some embodiments, for V L , subgroup is subgroup κI as in Kabat et al., supra. In some embodiments, for V H , subgroup is subgroup III as in Kabat et al., supra.

[0104] As used herein, an "acceptor framework" is a construct comprising a light chain variable domain (V L ) framework or heavy chain variable domain (V H ) framework. An acceptor framework "derived from" an immunoglobulin framework or a consensus framework may comprise the same amino acid sequence thereof, or it may contain amino acid sequence changes. In some embodiments, the number of amino acid changes is 10 or less, 9 or less, 8 or less, 7 or less, 6 or less, 5 or less, 4 or less, 3 or less, or 2 or less. In some embodiments, V L Receptor framework sequence and V L The immunoglobulin framework sequences or consensus framework sequences are identical.

[0105] "Fc region" refers to a dimeric complex comprising the C-terminal polypeptide sequence of an immunoglobulin heavy chain, wherein the C-terminal polypeptide sequence is a sequence obtainable by papain digestion of an intact antibody. The Fc region may comprise a native or variant Fc sequence. The Fc sequence of an immunoglobulin generally comprises two constant domains, a CH2 domain and a CH3 domain, and optionally comprises a CH4 domain. The boundaries of the Fc sequence of an immunoglobulin heavy chain may vary depending on the immunoglobulin class and species.

[0106] The term "IgX Fc" means that the Fc region is derived from a specific antibody isotype (e.g., IgG, IgA, IgD, IgE, IgM, etc.), where "X" represents the antibody isotype. Thus, "IgG Fc" represents the Fc region of the γ chain, "IgA Fc" represents the Fc region of the α chain, "IgDFc" represents the Fc region of the δ chain, "IgE Fc" represents the Fc region of the ε chain, "IgM Fc" represents the Fc region of the μ chain, and the like. In some embodiments, the IgG Fc region comprises CH1, hinge, CH2, CH3, and CL1. "IgX-N-Fc" represents that the Fc region is derived from a specific subclass of an antibody isotype (such as canine IgG subclass A, B, C, or D; or feline IgG subclass 1, 2a, or 2b), where "N" represents the subclass. In some embodiments, the IgX Fc or IgX-N-Fc region is derived from a companion animal, such as a dog. In some embodiments, the IgG Fc region is isolated from a canine gamma heavy chain, such as IgG-A, IgG-B, IgG-C, or IgG-D. Antibodies comprising an Fc region of IgG-A, IgG-B, IgG-C, or IgG-D can provide higher expression levels in recombinant production systems. "IgX Fc" and "IgX Fc polypeptide" are intended to include wild-type IgX Fc polypeptides and variant IgX Fc polypeptides.

[0107] "Effector functions" refer to those biological activities caused by the Fc region of an antibody, which vary with the antibody isotype. Examples of antibody effector functions include: Clq binding and complement-dependent cytotoxicity (CDC); Fc receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis and downregulation of cell surface receptors (e.g., B cell receptors); and B cell activation.

[0108] An "immunoconjugate" refers to an antibody conjugated to one or more heterologous molecules, including but not limited to a cytotoxic agent.

[0109] A "multispecific antibody" is an antibody that has at least two distinct binding sites, each with a different binding specificity. A multispecific antibody can be a full-length antibody or an antibody fragment, and each of the binding sites can bind to a different antigen, or the different binding sites can bind to two different epitopes on the same antigen.

[0110] "Fv fragment" refers to an antibody fragment that contains a complete antigen recognition and binding site. This region consists of a dimer of one heavy chain variable domain and one light chain variable domain in tight association, which can be covalent in nature, such as in scFv. It is this configuration in which the three HVRs of each variable domain interact that defines the V domain. H -V LThe antigen-binding site on the surface of the dimer. Collectively, the six HVRs or a subset thereof confer antigen-binding specificity to the antibody. However, even a single variable domain (or half of an Fv comprising only three HVRs specific for an antigen) has the ability to recognize and bind antigen, although generally at a lower affinity than the entire binding site.

[0111] A "Fab fragment" refers to an antibody fragment containing the variable and constant domains of the light chain and the variable and first constant domains (CH1) of the heavy chain. A "F(ab')2 fragment" comprises a pair of Fab fragments, which are typically covalently linked near their carboxyl termini by hinge cysteines. Other chemical couplings of antibody fragments are also known in the art.

[0112] As used herein, "antigen binding arm" refers to a component of an antibody that has the ability to specifically bind to a target molecule of interest. Typically, an antigen binding arm is a complex of immunoglobulin polypeptide sequences, such as HVR and / or variable domain sequences of immunoglobulin light and heavy chains.

[0113] "Single-chain Fv" or "scFv" refers to a V-chain fragment comprising an antibody H domain and V L In general, scFv antibodies comprise a single polypeptide chain having a V domain and a V domain. H Domain and V L The polypeptide linker between the polypeptides of the structural domain sequence. H domain and V L The domains allow the hypervariable regions of the scFv to form the desired antigen-binding structure.

[0114] As used herein, "polypeptide linker" or "polypeptide linker" refers to a chain of two or more amino acids, each end of which is covalently attached to a different polypeptide molecule, thereby conjugating or fusing different polypeptides.

[0115] "Diabodies" are small antibody fragments with two antigen-binding sites, said fragments being comprised of the same polypeptide chain (V H and V L ) and the light chain variable domain (V L ) linked to the heavy chain variable domain (V H ). By using a linker that is too short to allow pairing between the two domains on the same chain, the domains are forced to pair with the complementary domains of another chain and create two antigen-binding sites.

[0116] "Linear antibodies" refer to the antibodies described in Zapata et al., Protein Eng., 8(10): 1057-1062 (1995). Briefly, these antibodies comprise a pair of tandem Fd segments (VH-CH1-VH-CH1) that, together with complementary light chain polypeptides, form a pair of antigen-binding regions. Linear antibodies can be bispecific or monospecific.

[0117] A "naked antibody" refers to an antibody that is not conjugated to a heterologous moiety (eg, a cytotoxic moiety) or radiolabel.

[0118] "Affinity" refers to the strength of the sum of all non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). "Binding affinity" refers to the intrinsic binding affinity that reflects a 1:1 interaction between members of a binding pair (e.g., an antibody and an antigen). The affinity of a molecule X for its partner Y can generally be expressed in terms of the equilibrium dissociation constant (K D ). Affinity can be measured by common methods known in the art, including those described herein. Specific illustrative and exemplary embodiments for measuring binding affinity are described below.

[0119] "Binding specificity" or "specific binding" refers to an antibody binding affinity to an antigen with a value of no more than about 1 x 10 - 7 In some embodiments, an antibody may have a secondary affinity for an antigen other than the antigen to which it specifically binds, wherein "secondary affinity" will generally refer to an antibody having a binding affinity value for the secondary antigen greater than about 10 nM, as described elsewhere herein. Although an antibody may have a secondary affinity for a secondary antigen, such an antibody will nevertheless specifically bind to the primary antigen.

[0120] An "isolated antibody" is one that has been separated from components of its natural environment. In some embodiments, the antibody is purified to a purity greater than 95% or 99% as determined by, for example, electrophoresis (e.g., SDS-PAGE, isoelectric focusing (IEF), capillary electrophoresis) or chromatographic methods (e.g., ion exchange or reversed-phase HPLC). For a review of methods for evaluating antibody purity, see, e.g., Flatman et al., J. Chromatogr. B 848:79-87.

[0121] As used herein, "percent (%) amino acid sequence identity" and "homology" with respect to polypeptide or antibody sequences are defined as the percentage of amino acid residues in a candidate sequence that are identical with the amino acid residues in a specified peptide or polypeptide sequence, after aligning the sequences and introducing gaps (if necessary) to achieve the maximum percent sequence identity, and not considering any conservative substitutions as part of the sequence identity. Alignment for the purpose of determining percent amino acid sequence identity can be achieved in various ways that are within the skill in the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN, or MEGALINE TM (DNASTAR) software. Those skilled in the art can determine appropriate parameters for measuring alignment, including any algorithms needed to achieve maximal alignment over the full length of the sequences being compared.

[0122] "Amino acid substitution" refers to the replacement of an amino acid in a polypeptide by another amino acid. In some embodiments, the amino acid substitution is a conservative substitution. Amino acid substitutions can be introduced into a molecule of interest, and the product can be screened for desired activity, for example, retained / improved antigen binding, reduced immunogenicity, or improved ADCC or CDC, or enhanced pharmacokinetics.

[0123] The term "vector" is used to describe a polynucleotide that can be engineered to contain one or more cloned polynucleotides that can be propagated in a host cell. A vector may contain one or more of the following elements: an origin of replication, one or more regulatory sequences that regulate expression of a polypeptide of interest (such as, for example, a promoter or enhancer), or one or more selectable marker genes (such as, for example, antibiotic resistance genes and genes that can be used for colorimetric assays, e.g., β-galactosidase). The term "expression vector" refers to a vector that is used to express a polypeptide of interest in a host cell.

[0124] "Host cell" refers to a cell that can be or has been a recipient of a vector or isolated polynucleotide. Host cells can be prokaryotic or eukaryotic. Exemplary eukaryotic cells include mammalian cells, such as primate cells or non-primate cells; fungal cells, such as yeast; plant cells; and insect cells. Non-limiting exemplary mammalian cells include, but are not limited to, NSO cells, PER. Host cells include Crucell, 293 cells and CHO cells and their derivatives, such as 293-6E cells, DG44 cells, CHO-S cells and CHO-K cells. Host cells include progeny of a single host cell, and the progeny may not be completely identical (in morphology or in genomic DNA composition (complement)) to the original parent cell due to natural, accidental or deliberate mutations. Host cells include cells transfected in vivo with polynucleotides encoding the amino acid sequences provided herein.

[0125] The term "companion animal species" refers to animals suitable as companions for humans. In some embodiments, the companion animal species is a small mammal, such as a dog, a feline, a dog, a cat, a horse, a rabbit, a ferret, a guinea pig, a rodent, etc. In some embodiments, the companion animal species is a large animal, such as a camel or a farm animal, such as a horse, a cow, a pig, etc.

[0126] "Reduce" or "inhibit" means to reduce, decrease or prevent a certain activity, function or amount compared to a reference value. In some embodiments, "reduce" or "inhibit" means the ability to cause an overall reduction of 20% or more. In some embodiments, "reduce" or "inhibit" means the ability to cause an overall reduction of 50% or more. In some embodiments, "reduce" or "inhibit" means the ability to cause an overall reduction of 75%, 85%, 90%, 95% or more. In some embodiments, the above amount is suppressed or reduced over the same time period relative to a control dose (such as a placebo). As used herein, "reference" refers to any sample, standard or level used for comparison purposes. A reference can be obtained from a healthy or non-diseased sample. In some instances, a reference value is obtained from a sample of a companion animal that has never been sick or treated. In some instances, a reference value is obtained from one or more healthy animals of a particular species that are not the animals being tested or treated.

[0127] As used herein, "substantially similar" or "substantially identical" refers to a sufficiently high degree of similarity between two values ​​(e.g., one associated with a test antibody and the other associated with a reference antibody) such that one skilled in the art would consider the difference between the two values ​​to be in the biological characteristic measured by the values ​​(e.g., K D values) have little or no biological and / or statistical significance.

[0128] As used herein, "substantially different" refers to a sufficiently high difference between two values ​​(generally one associated with a molecule and the other associated with a reference molecule) that one of skill in the art would consider the difference between the two values ​​to be significant in the biological characteristic measured by the values ​​(e.g., K D values) is statistically significant.

[0129] "Treatment," "treat," or "treating" refers to an intervention that attempts to alter the natural course of the condition being treated, and may be performed for prevention or during the course of clinical pathology. Desired therapeutic outcomes include, but are not limited to, preventing the onset or recurrence of the condition, alleviating symptoms, eliminating any direct or indirect pathological consequences of the condition, preventing metastasis, reducing the rate of progression, ameliorating or palliating the disease state, and alleviating or improving prognosis. For example, treatment may comprise administering to a subject a therapeutically effective amount of a pharmaceutical formulation comprising an anti-IL23 antibody to delay the development of or slow the progression of a disease or condition mediated by IL23 or in which IL23 may play a role in the pathogenesis and / or progression. Treatment does not require 100 percent ablation of all aspects of the condition.

[0130] A "pharmaceutical formulation" refers to a formulation that is in a form that permits the biological activity of the active ingredient to be effective and that does not contain additional components that are toxic to the subject to which the formulation is administered. A pharmaceutical formulation may include one or more active agents. For example, a pharmaceutical formulation may include an anti-IL23 antibody as the sole active agent of the formulation, or may include an anti-IL23 antibody and one or more additional active agents.

[0131] "Pharmaceutically acceptable carrier" refers to ingredients other than the active ingredient in a pharmaceutical formulation that are non-toxic to the subject to which it is administered. Pharmaceutically acceptable carriers include, but are not limited to, buffers, excipients, stabilizers, or preservatives.

[0132] A "therapeutically effective amount" refers to an amount of an active ingredient or agent (e.g., a pharmaceutical formulation) that achieves a desired therapeutic or preventive result (e.g., treating or preventing a disease, condition, or disorder in a subject). For IL23-mediated diseases or disorders, a therapeutically effective amount of a therapeutic agent is an amount that reduces, prevents, inhibits, and / or alleviates to some extent one or more symptoms associated with the disease, condition, or disorder.

[0133] "Individual" or "subject" refers to mammals, including but not limited to domestic or companion animals (e.g., cows, sheep, cats, dogs, and horses), primates (e.g., humans and non-human primates such as monkeys), rabbits, and rodents (e.g., mice and rats).

[0134] Anti-IL23 and anti-TNFα antibodies

[0135] Table 1 below provides a summary description of the sequences cited in this disclosure, including canine IL23 protein, feline IL23 protein, and various anti-IL23 and anti-TNFα antibodies of the disclosure, and their sequence identifiers. The sequences are also included in the accompanying sequence listing.

[0136] Table 1: Description of exemplary antibody sequences

[0137]

[0138]

[0139]

[0140]

[0141]

[0142]

[0143]

[0144]

[0145] Provided herein are novel antibodies against canine, feline, or equine IL23 and / or canine, feline, or equine TNFα. Anti-IL23 and / or anti-TNFα antibodies provided herein include, but are not limited to, monoclonal antibodies, chimeric antibodies, caninized antibodies, feline antibodies, or equinized antibodies, scFv antibodies, and bispecific antibodies that bind to both IL23 and TNFα. Also provided herein are amino acid sequences of monoclonal antibodies. For example, light and heavy chain hypervariable regions (HVR-L1, HVR-L2, HVR-L3, HVR-H1, HVR-H2, HVR-H3), heavy chain variable domains (VH), light chain variable domains (VL), variable region heavy chain framework sequences, and variable region light chain framework sequences of the monoclonal antibodies described herein are provided. In addition, amino acid sequences, framework sequences, light chain variable domain (VL) sequences, and heavy chain variable domain (VH) sequences of HVRs of different caninized and feline light and heavy chains are also provided.

[0146] The novel anti-IL23 antibodies provided herein are monoclonal antibodies designated "Clone 340" or "C340." As shown in Table 1, the amino acid sequence of the C340 light chain variable domain (VL) is provided as SEQ ID NO: 3, and the amino acid sequence of the C340 heavy chain variable domain (VH) is provided as SEQ ID NO: 7. The corresponding hypervariable regions (HVR-L1, HVR-L2, and HVR-L3) of the C340 VL domain are provided as SEQ ID NOs: 4, 5, and 6, respectively. The corresponding hypervariable regions (HVR-H1, HVR-H2, and HVR-H3) of the C340 VH domain are provided as SEQ ID NOs: 8, 9, and 10, respectively.

[0147] In at least one embodiment, the present disclosure provides chimeric anti-IL23 antibodies derived from C340. In one embodiment of the chimeric anti-IL23 antibody, the antibody comprises the VL and VH domains of SEQ ID NOs: 3 and 7, respectively. The chimeric VL domain is linked to a canine kappa light chain constant region, and the chimeric VH domain is linked to a canine IgG-B constant region. The light and heavy chain amino acid sequences of these anti-IL23 chimeric antibodies are provided as SEQ ID NOs: 17 and 18, respectively. The tight binding affinity of the anti-IL23 chimeric antibodies to canine IL23 was measured as described in the Examples.

[0148] In at least one embodiment, the present disclosure provides caninized versions of the C340 anti-IL23 antibody. Two different caninized versions of the VL domain are provided as SEQ ID NOs: 11 and 13. Two different caninized versions of the VH domain are provided as SEQ ID NOs: 12 and 14. The caninized versions of the VL domain comprise the sequences of the C340 VL domain, HVR-L1, HVR-L2, HVR-L3, and SEQ ID NOs: 4, 5, and 6, respectively. The caninized versions of the VH domain comprise the sequences of the C340 VH domain, HVR-H1, HVR-H2, HVR-H3, and SEQ ID NOs: 8, 9, and 10, respectively.

[0149] Exemplary caninized anti-IL23 antibody light chain (LC) and heavy chain (HC) sequences comprising caninized VL domains of SEQ ID NOs: 11 and 13 and caninized VH domains of SEQ ID NOs: 12 and 14 are provided in Table 1. Exemplary caninized anti-IL23 light chains comprising canine kappa light chain constant regions are SEQ ID NOs: 19 and 22. Exemplary caninized anti-IL23 heavy chains comprising canine Fc IgG-B constant regions are provided as SEQ ID NOs: 20, 21, and 23. It is contemplated that a range of exemplary caninized anti-IL23 antibodies can be prepared using any combination of the LC sequences of SEQ ID NOs: 19 and 22 and the HC sequences of SEQ ID NOs: 20, 21, and 23.

[0150] In at least one embodiment, the present disclosure also provides a felinized version of the C340 anti-IL23 antibody. An exemplary felinized version of the VL domain is provided as SEQ ID NO: 15, and a felinized version of the VH domain is provided as SEQ ID NO: 16. The felinized version of the VL domain of SEQ ID NO: 15 comprises the C340 HVR-L1, HVR-L2, and HVR-L3 sequences of SEQ ID NOs: 4, 5, and 6, respectively. Similarly, the felinized version of the VH domain of SEQ ID NO: 16 comprises the C340 HVR-H1, HVR-H2, and HVR-H3 sequences of SEQ ID NOs: 8, 9, and 10, respectively.

[0151] The present disclosure provides a novel anti-TNFα antibody, designated "D2E7," that specifically binds to canine, feline, and / or equine TNFα. As shown in Table 1, the amino acid sequence of the light chain variable domain (VL) of the D2E7 anti-TNFα antibody is provided as SEQ ID NO: 26, and the amino acid sequence of the heavy chain variable domain (VH) of C340 is provided as SEQ ID NO: 30. The corresponding hypervariable regions (HVR-L1, HVR-L2, and HVR-L3) of the D2E7 VL domain are provided as SEQ ID NOs: 27, 28, and 29, respectively. The corresponding hypervariable regions (HVR-H1, HVR-H2, and HVR-H3) of the D2E7 VH domain are provided as SEQ ID NOs: 31, 32, and 33, respectively.

[0152] In at least one embodiment, the present disclosure provides a chimeric anti-TNFα antibody derived from the exemplary anti-TNFα monoclonal antibody D2E7. In at least one embodiment, the chimeric anti-TNFα D2E7 antibody comprises the VL and VH domains of SEQ ID NOs: 26 and 30, respectively, linked to a canine kappa light chain constant region and a canine IgG-B constant region. Exemplary light chain (LC) and heavy chain (HC) amino acid sequences of the chimeric anti-TNFα D2E7 antibody are provided in Table 1 as SEQ ID NOs: 38 and 39, respectively.

[0153] In at least one embodiment, the present disclosure provides caninized versions of the VL and VH domains of the anti-TNFα D2E7 antibody, as SEQ ID NOs: 34 and 35, respectively. Exemplary caninized anti-TNFα antibody light chain (LC) and heavy chain (HC) sequences comprising the caninized VL domain of SEQ ID NO: 34 and the caninized VH domain of SEQ ID NO: 35 are provided in Table 1. Exemplary caninized anti-TNFα light chains comprising a canine kappa light chain constant region are provided as SEQ ID NOs: 40 and 41. Exemplary caninized anti-TNFα heavy chains comprising a canine Fc IgG-B constant region are provided as SEQ ID NOs: 42, 43, and 44. It is contemplated that a range of exemplary caninized anti-TNFα antibodies can be prepared using any combination of the LC sequences of SEQ ID NOs: 40 and 41 with the HC sequences of SEQ ID NOs: 42, 43, and 44.

[0154] In at least one embodiment, the present disclosure also provides a felinized version of the D2E7 anti-TNFα antibody. An exemplary felinized version of the VL domain is provided as SEQ ID NO: 36, and an exemplary felinized version of the VH domain is provided as SEQ ID NO: 37. The felinized version of the VL domain of SEQ ID NO: 36 comprises the D2E7 HVR-L1, HVR-L2, and HVR-L3 sequences of SEQ ID NOs: 27, 28, and 29, respectively. Similarly, the felinized version of the VH domain of SEQ ID NO: 37 comprises the D2E7 HVR-H1, HVR-H2, and HVR-H3 sequences of SEQ ID NOs: 31, 32, and 33, respectively.

[0155] In at least one embodiment, the present disclosure provides an anti-TNFα scFv antibody having a VL domain comprising the D2E7 HVR-L1, HVR-L2, HVR-L3 sequences of SEQ ID NOs: 27, 28, and 29, respectively, fused via a polypeptide linker to a VH domain comprising the D2E7 HVR-H1, HVR-H2, HVR-H3 sequences of SEQ ID NOs: 31, 32, and 33, respectively. In at least one embodiment, the scFv antibody VL domain comprises the amino acid sequence of SEQ ID NOs: 26, 34, or 40, and the scFv antibody VL domain comprises the amino acid sequence of SEQ ID NOs: 30, 35, or 41. In at least one embodiment, the polypeptide linker fusing the scFv antibody VL and VH domains comprises the amino acid sequence GGGGGSGGGGSGGGGGS (SEQ ID NO: 49). In at least one embodiment, the scFv antibody of the present disclosure may comprise an amino acid sequence having at least 90% sequence identity to a sequence selected from SEQ ID NOs: 45 and 46.

[0156] Although the polypeptide of SEQ ID NO: 49 is exemplified as an scFv antibody in Table 1, one of ordinary skill in the art will appreciate that a variety of polypeptide linkers are known in the art and can be used in the scFv antibodies and other polypeptide fusion compositions of the present disclosure. In general, polypeptides comprising polypeptide chains of 5 to 30 amino acids can be used to fuse the polypeptide components of the scFv antibodies and bispecific antibody structures of the present disclosure.

[0157] The present disclosure also provides bispecific antibodies capable of specifically binding to canine, feline, and / or equine IL23 and TNFα. In at least one embodiment, the bispecific antibody comprises six HVR sequences of the C340 anti-IL23 antibody and six HVR sequences of the D2E7 anti-TNFα antibody in a single fusion construct. Thus, in at least one embodiment, the present disclosure provides a "four-chain" anti-IL23 LC / HC + anti-TNFα LC / HC bispecific antibody structure that binds to canine, feline, and / or equine IL23 and canine, feline, and / or equine TNFα, wherein the antibody comprises:

[0158] (i) an anti-IL23 light chain (LC) comprising a VL domain having an HVR-L1 region comprising the amino acid sequence of RASQGISSWLA (SEQ ID NO:4), an HVR-L2 region comprising the amino acid sequence of YAASSLQS (SEQ ID NO:5), and an HVR-L3 region comprising the amino acid sequence of QQYNIYPYT (SEQ ID NO:6);

[0159] (ii) an anti-IL23 heavy chain (HC) comprising a VH domain having an HVR-H1 region comprising the amino acid sequence of KGSGYSFTTYWLG (SEQ ID NO: 8), an HVR-H2 region comprising the amino acid sequence of IMSPVDSDIR (SEQ ID NO: 9), and an HVR-H3 region comprising the amino acid sequence of ARRRPGQGYFDF (SEQ ID NO: 10);

[0160] (iii) an anti-TNFα light chain (LC) comprising a VL domain having an HVR-L1 region comprising the amino acid sequence of RASQGIRNYLA (SEQ ID NO:27), an HVR-L2 region comprising the amino acid sequence of AASTLQ (SEQ ID NO:28), and an HVR-L3 comprising the amino acid sequence of QRYNRAPYT (SEQ ID NO:29); and

[0161] (iv) an anti-TNFα heavy chain (HC) comprising a VH domain having an HVR-H1 region comprising the amino acid sequence of FTFDDYAMH (SEQ ID NO: 31), an HVR-H2 region comprising the amino acid sequence of AITWNSGHIDYADSVEGR (SEQ ID NO: 32), and an HVR-H3 region comprising the amino acid sequence of AKVSYLSTASSLDY (SEQ ID NO: 33).

[0162] In some embodiments, the bispecific antibody binds to canine IL23 and canine TNFα and comprises:

[0163] (i) the anti-IL23 light chain (LC) comprises a light chain variable domain (VL) amino acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 3, 11, 13, and 15;

[0164] (ii) the anti-IL23 heavy chain (HC) comprises a heavy chain variable domain (VH) amino acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 7, 12, 14, and 16;

[0165] (iii) the anti-TNFα light chain (LC) comprises a light chain variable domain (VL) amino acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 26, 34, and 36; and

[0166] (iv) the anti-TNFα heavy chain (HC) comprises a heavy chain variable domain (VH) amino acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 30, 35, and 37.

[0167] In some embodiments, the present disclosure provides a bispecific antibody that binds to canine IL23 and canine TNFα and comprises:

[0168] (i) the anti-IL23 light chain (LC) comprises an amino acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 19 and 22;

[0169] (ii) the anti-IL23 heavy chain (HC) comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NOs: 20, 21 and 23;

[0170] (iii) the anti-TNFα light chain (LC) comprises an amino acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 40 and 41; and

[0171] (iv) the anti-TNFα heavy chain (HC) comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NOs: 42, 43 and 44.

[0172] In addition to the anti-IL23 LC / HC + anti-TNFα LC / HC bispecific antibody structure, the present disclosure also provides a bispecific antibody comprising an anti-IL23 LC / HC antibody fused to an anti-TNFα scFv antibody structure, or an anti-TNFα LC / HC antibody fused to an anti-IL23 scFv antibody structure. Thus, in at least one embodiment, the present disclosure provides a bispecific antibody that binds to canine, feline, and / or equine IL23 and canine, feline, and / or equine TNFα, wherein the antibody comprises:

[0173] (i) an anti-IL23 light chain (LC) comprising a VL domain having an HVR-L1 region comprising the amino acid sequence of RASQGISSWLA (SEQ ID NO:4), an HVR-L2 region comprising the amino acid sequence of YAASSLQS (SEQ ID NO:5), and an HVR-L3 region comprising the amino acid sequence of QQYNIYPYT (SEQ ID NO:6);

[0174] (ii) an anti-IL23 heavy chain (HC) comprising a VH domain having an HVR-H1 region comprising the amino acid sequence of KGSGYSFTTYWLG (SEQ ID NO: 8), an HVR-H2 region comprising the amino acid sequence of IMSPVDSDIR (SEQ ID NO: 9), and an HVR-H3 region comprising the amino acid sequence of ARRRPGQGYFDF (SEQ ID NO: 10); and

[0175] (iii) an anti-TNFα scFv antibody fused to the HC, wherein the anti-TNFα scFv antibody comprises a VL domain and a VH domain, the VL domain having an HVR-L1 region comprising the amino acid sequence of RASQGIRNYLA (SEQ ID NO: 27), an HVR-L2 region comprising the amino acid sequence of AASTLQ (SEQ ID NO: 28), and an HVR-L3 comprising the amino acid sequence of QRYNRAPYT (SEQ ID NO: 29); and the VH domain having an HVR-H1 region comprising the amino acid sequence of FTFDDYAMH (SEQ ID NO: 31), an HVR-H2 region comprising the amino acid sequence of AITWNSGHIDYADSVEGR (SEQ ID NO: 32), and an HVR-H3 region comprising the amino acid sequence of AKVSYLSTASSLDY (SEQ ID NO: 33).

[0176] In another embodiment, the present disclosure also provides a bispecific antibody that binds to canine, feline and / or equine IL23 and canine, feline and / or equine TNFα, wherein the antibody comprises:

[0177] (i) an anti-TNFα light chain (LC) comprising a VL domain having an HVR-L1 region comprising the amino acid sequence of RASQGIRNYLA (SEQ ID NO: 27), an HVR-L2 region comprising the amino acid sequence of AASTLQ (SEQ ID NO: 28), and an HVR-L3 region comprising the amino acid sequence of QRYNRAPYT (SEQ ID NO: 29), and

[0178] (ii) an anti-TNFα heavy chain (HC) comprising a VH domain having an HVR-H1 region comprising the amino acid sequence of FTFDDYAMH (SEQ ID NO: 31), an HVR-H2 region comprising the amino acid sequence of AITWNSGHIDYADSVEGR (SEQ ID NO: 32), and an HVR-H3 region comprising the amino acid sequence of AKVSYLSTASSLDY (SEQ ID NO: 33), and

[0179] (iii) an anti-IL23 scFv antibody fused to the HC, wherein the anti-IL23 scFv antibody comprises a VL domain and a VH domain, the VL domain having an HVR-L1 region comprising the amino acid sequence of RASQGISSWLA (SEQ ID NO: 4), an HVR-L2 region comprising the amino acid sequence of YAASSLQS (SEQ ID NO: 5), and an HVR-L3 region comprising the amino acid sequence of QQYNIYPYT (SEQ ID NO: 6); and the VH domain having an HVR-H1 region comprising the amino acid sequence of KGSGYSFTTYWLG (SEQ ID NO: 8), an HVR-H2 region comprising the amino acid sequence of IMSPVDSDIR (SEQ ID NO: 9), and an HVR-H3 region comprising the amino acid sequence of ARRRPGQGYFDF (SEQ ID NO: 10).

[0180] In some embodiments, the antibody comprises a label or is conjugated to a second portion. The terms "label" and "detectable label" mean a portion attached to an antibody or its analyte so that the reaction (e.g., binding) between members of a specific binding pair can be detected. The labeled member of a specific binding pair is referred to as "detectably labeled." Therefore, the term "labeled binding protein" refers to a protein that is incorporated with a label that is provided for identifying the binding protein. In some embodiments, the label is a detectable marker that can produce a signal that can be detected by visual or instrumental means, for example, a polypeptide that incorporates a radiolabeled amino acid or is attached to a biotin portion that can be detected by labeled avidin (e.g., streptavidin containing a fluorescent marker or enzymatic activity that can be detected by optical or colorimetric methods). Examples of polypeptide labels include, but are not limited to, the following: a radioisotope or radionuclide (e.g., 3 H. 14 C. 35 S. 90 Y. 99 Tc, 111 In, 125 I. 131 I. 177 Lu, 166 Ho or 153Sm); chromophores, fluorescent labels (e.g., FITC, rhodamine, lanthanide phosphors), enzyme labels (e.g., horseradish peroxidase, luciferase, alkaline phosphatase); chemiluminescent markers; biotin groups; predetermined polypeptide epitopes recognized by secondary reporter genes (e.g., leucine zipper pair sequences, secondary antibody binding sites, metal binding domains, epitope tags); and magnetic agents such as gadolinium chelates. Representative examples of labels commonly used in immunoassays include light-generating moieties (e.g., acridine compounds) and fluorescence-generating moieties (e.g., fluorescein). In this regard, a moiety itself may not be detectably labeled, but may become detectable after reacting with another moiety.

[0181] "Amino acid sequence" means the sequence of amino acid residues in a peptide or protein. The terms "polypeptide" and "protein" are used interchangeably to refer to polymers of amino acid residues and are not limited to a minimum length. Such polymers of amino acid residues may contain natural or non-natural amino acid residues and include, but are not limited to, peptides, oligopeptides, dimers, trimers, and multimers of amino acid residues. The definition encompasses both full-length proteins and fragments thereof. The term also includes post-expression modifications of the polypeptide, for example, glycosylation, sialylation, acetylation, phosphorylation, etc. In addition, for the purposes of this disclosure, "polypeptide" refers to a protein comprising modifications to the native sequence, such as deletions, additions, and substitutions (generally conservative in nature), as long as the protein maintains the desired activity. These modifications may be intentional, such as by site-directed mutagenesis, or may be accidental, such as by mutations in the host producing the protein or errors caused by PCR amplification.

[0182] As used herein, the term "epitope" refers to the site at which an antigen binding molecule (e.g., an antibody, an antibody fragment, or a scaffold protein containing an antibody binding region) binds to a target molecule (e.g., an antigen, such as a protein, nucleic acid, carbohydrate, or lipid). An epitope typically includes a chemically active surface grouping of a molecule (such as an amino acid, a polypeptide, or a sugar side chain) and has specific three-dimensional structural characteristics and specific charge characteristics. An epitope can be formed by both adjacent or juxtaposed non-continuous residues (e.g., amino acids, nucleotides, sugars, lipid moieties) of a target molecule. An epitope formed by adjacent residues (e.g., amino acids, nucleotides, sugars, lipid moieties) is typically retained when exposed to a denaturing solvent, while an epitope formed by tertiary folding is typically lost when treated with a denaturing solvent. An epitope may include, but is not limited to, at least 3, at least 5, or 8-10 residues (e.g., amino acids or nucleotides). In some instances, the length of an epitope is less than 20 residues (e.g., amino acids or nucleotides), less than 15 residues, or less than 12 residues. If two antibodies exhibit competitive binding to an antigen, they may bind to the same epitope within the antigen. In some embodiments, epi-position can be identified by a certain minimum distance with the CDR residues on the antigen binding molecules. In some embodiments, epi-position can be identified by the above-mentioned distance, and is further limited to those residues that participate in the key (for example, hydrogen bond) between the antibody residue and the antigen residue. Epi-position can also be identified by various scans, and for example alanine or arginine scan can represent one or more residues that the antigen binding molecules can interact with. Unless clearly stated, one group of residues does not exclude other residues from becoming a part for a specific antibody epi-position as epi-position. On the contrary, the existence of this set shows the minimum series (or species set) of epi-position. Therefore, in some embodiments, a group of residues identified as epi-position specifies the minimum epi-position related to the antigen, rather than the exclusive list of the residue of the epi-position on the antigen.

[0183] In at least one embodiment, an anti-IL23 antibody of the present disclosure comprises a light chain variable domain (VL) comprising HVR-L1 comprising the amino acid sequence of SEQ ID NO: 4, HVR-L2 comprising the amino acid sequence of SEQ ID NO: 5, and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 6, and a heavy chain variable domain (VH) comprising HVR-H1 comprising the amino acid sequence of SEQ ID NO: 8, HVR-H2 comprising the amino acid sequence of SEQ ID NO: 9, and HVR-H3 comprising the amino acid sequence of SEQ ID NO: 10.

[0184] In at least one embodiment, the anti-IL23 antibody of the present disclosure comprises (i) a light chain variable domain (VL) comprising the amino acid sequence of SEQ ID NO: 3, 11, 13 or 15, or a variant thereof, wherein 1, 2, 3, 4, 5 or 6 amino acids of the light chain variable domain (VL) are substituted with different amino acids; (ii) a heavy chain variable domain (VH) comprising the amino acid sequence of SEQ ID NO: 7, 12, 14 or 16, or a variant thereof, wherein 1, 2, 3, 4, 5 or 6 amino acids of the heavy chain variable domain (VH) are substituted with different amino acids; or (iii) a light chain variable domain (VL) as in (i) and a heavy chain variable domain (VH) as in (ii).

[0185] In at least one embodiment, an anti-IL23 antibody of the present disclosure comprises (i) a light chain variable domain (VL) comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 3, 11, 13, or 15, (ii) a heavy chain variable domain (VH) comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 7, 12, 14, or 16, or (iii) a light chain variable domain (VL) as in (i) and a heavy chain variable domain (VH) as in (ii).

[0186] In at least one embodiment, an anti-TNFα antibody of the present disclosure comprises a light chain variable domain (VL) comprising HVR-L1 comprising the amino acid sequence of SEQ ID NO: 27, HVR-L2 comprising the amino acid sequence of SEQ ID NO: 28, and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 29, and a heavy chain variable domain (VH) comprising HVR-H1 comprising the amino acid sequence of SEQ ID NO: 31, HVR-H2 comprising the amino acid sequence of SEQ ID NO: 32, and HVR-H3 comprising the amino acid sequence of SEQ ID NO: 33.

[0187] In at least one embodiment, an anti-TNFα antibody of the present disclosure comprises (i) a light chain variable domain (VL) comprising the amino acid sequence of SEQ ID NO: 26, 34 or 36, or a variant thereof, wherein 1, 2, 3, 4, 5 or 6 amino acids of the light chain variable domain (VL) are substituted with different amino acids; (ii) a heavy chain variable domain (VH) comprising the amino acid sequence of SEQ ID NO: 30, 35 or 37, or a variant thereof, wherein 1, 2, 3, 4, 5 or 6 amino acids of the heavy chain variable domain (VH) are substituted with different amino acids; or (iii) a light chain variable domain (VL) as in (i) and a heavy chain variable domain (VH) as in (ii).

[0188] In at least one embodiment, the anti-TNFα antibody comprises (i) a light chain variable domain (VL) comprising an amino acid sequence that is at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 26, 34, or 36; (ii) a heavy chain variable domain (VH) comprising an amino acid sequence that is at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 30, 35, or 37; or (iii) a heavy chain variable domain (VH) as in (i) and a heavy chain variable domain (VH) as in (ii).

[0189] As used herein, the term "constant region" or "constant domain" refers to a region comprising at least three constant domains.

[0190] The terms "heavy chain constant region" or "constant heavy chain" are used interchangeably to refer to a region comprising at least three heavy chain constant domains, CH1, CH2, and CH3. Non-limiting exemplary heavy chain constant regions include γ, δ, α, ε, and μ. Each heavy chain constant region corresponds to an antibody isotype. For example, an antibody comprising a γ constant region is an IgG antibody, an antibody comprising a δ constant region is an IgD antibody, an antibody comprising an α constant region is an IgA antibody, an antibody comprising a μ constant region is an IgM antibody, and an antibody comprising an ε constant region is an IgE antibody. Certain isotypes can be further subdivided into subclasses. For example, IgG antibodies include, but are not limited to, IgG1 (comprising a γ1 constant region), IgG2 (comprising a γ2 constant region), IgG3 (comprising a γ3 constant region), and IgG4 (comprising a γ4 constant region) antibodies; IgA antibodies include, but are not limited to, IgA1 (comprising an α1 constant region) and IgA2 (comprising an α2 constant region) antibodies; and IgM antibodies include, but are not limited to, IgM1 and IgM2.

[0191] The terms "light chain constant region" or "constant light chain" are used interchangeably to refer to the region comprising the light chain constant domain, CL. Non-limiting exemplary light chain constant regions include λ and κ. Unless otherwise indicated, deletions and alterations in the domain that do not function are encompassed within the term "constant region." Dogs, cats, and horses have antibodies classified as IgG, IgA, IgD, IgE, and IgM. Canine IgG antibody classes are IgG-A, IgG-B, IgG-C, and IgG-D.

[0192] In at least one embodiment, the present disclosure provides a chimeric anti-IL23 antibody comprising: (a) (i) a light chain amino acid sequence of SEQ ID NO: 17; (ii) a heavy chain amino acid sequence of SEQ ID NO: 18; or (iii) a light chain amino acid sequence as in (i) and a heavy chain sequence as in (ii).

[0193] In some embodiments, the anti-IL23 and anti-TNFα antibodies of the present disclosure may comprise a canine heavy chain constant region selected from IgG-A, IgG-B, IgG-C, and IgG-D constant regions.

[0194] In some embodiments, at least one amino acid residue in a portion of a mouse heavy chain variable domain (VH) or a mouse light chain variable domain (VL) has been replaced by the corresponding amino acid from a canine variable region. In some embodiments, the modified chain is fused to a canine constant heavy chain or a canine constant light chain.

[0195] In some embodiments, the caninized TNFα antibody comprises: (i) a light chain VL domain sequence of SEQ ID NO: 11 or 13, or a variant thereof that retains the same HVR sequence and has at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 11 or 13, (ii) a heavy chain VH domain sequence of SEQ ID NO: 12 or 14, or a variant thereof that retains the same HVR sequence and has at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 12 or 14, or (iii) a light chain VL domain sequence as in (i) and a heavy chain VL domain sequence as in (ii).

[0196] In at least one embodiment, the present disclosure provides a caninized anti-IL23 antibody comprising: (a) (i) a light chain amino acid sequence of SEQ ID NO: 19 or 22; (ii) a heavy chain amino acid sequence of SEQ ID NO: 20, 21 or 23; or (iii) a light chain amino acid sequence as in (i) and a heavy chain sequence as in (ii).

[0197] In at least one embodiment, the present disclosure provides a bispecific antibody that binds to canine IL23 and canine TNFα and comprises (i) a light chain variable domain (VL) comprising the amino acids of SEQ ID NO: 19; and (ii) a heavy chain variable domain (VH) comprising the amino acid sequence of SEQ ID NO: 47 or 48.

[0198] In some embodiments, the biological activity of an Fc polypeptide is the ability to bind to FcRn. In some embodiments, the biological activity of an Fc polypeptide is the ability to bind to C1q. In some embodiments, the biological activity of an Fc polypeptide is the ability to bind to CD16. In some embodiments, the biological activity of an Fc polypeptide is the ability to bind to Protein A.

[0199] In some embodiments, the variant IgG Fc polypeptide comprises a variant IgG Fc polypeptide of a companion animal species. In some embodiments, the variant IgG Fc polypeptide comprises a variant canine IgG Fc polypeptide. In some embodiments, the variant IgG Fc polypeptide (e.g., a variant canine IgG-AFc polypeptide, a variant canine IgG-C Fc polypeptide, or a variant canine IgG-D Fc polypeptide, a variant feline IgG1a Fc polypeptide) has an activity that is substantially lacking in a reference (e.g., wild-type) polypeptide.

[0200] Antibodies can be modified to extend or shorten their half-life. In some embodiments involving higher doses of antibodies, a shorter half-life may be desirable for acute treatment. In some embodiments involving lower doses of antibodies, a longer half-life may be desirable for extended treatment. For example, as discussed below, IgG Fc mutations that affect FcRn interactions can be introduced.

[0201] In some embodiments, the IL23 and / or TNFα antibody comprises a wild-type or variant IgG Fc having complement fixation activity (or complement dependent cytotoxicity (CDC)). In some embodiments, the IL23 and / or TNFα antibody comprises a wild-type or variant IgG Fc having antibody dependent cellular cytotoxicity (ADCC) activity. In some embodiments, the IL23 and / or TNFα antibody comprises a wild-type or variant IgG Fc having antibody dependent cellular phagocytosis (ADCP) activity. In some embodiments, the IL23 and / or TNFα antibody comprises a wild-type or variant IgG Fc having complement fixation activity and / or ADCC activity and / or ADCP activity. The IgG Fc polypeptide may be modified to have effector function or to have enhanced effector function.

[0202] In some embodiments, the IL23 and / or TNFα antibody comprises a wild-type or variant IgG Fc that binds to canine FcRn at low pH.

[0203] In some embodiments, the variant IgG Fc (e.g., a variant canine IgG Fc polypeptide) has a modified FcRn binding affinity compared to a reference polypeptide. In some embodiments, the variant IgG Fc has increased FcRn binding affinity at acidic pH (e.g., at a pH in the range of about 5.0 to about 6.5, such as at a pH of about 5.0, at a pH of about 5.5, at a pH of about 6.0, or at a pH of about 6.5) compared to a reference polypeptide.

[0204] In some embodiments, the anti-IL23 antibody binds to canine IL23, feline IL23, or equine IL23 with a dissociation constant (Kd) of less than 5 x 10 -6 M, less than 1x 10 -6 M, less than 5x 10 -7 M, less than 1x 10 -7 M, less than 5x 10 -8 M, less than 1x 10 -8 M, less than 5x 10 -9 M, less than 1x 10 -9 M, less than 5x 10 -10 M, less than 1x 10 -10 M, less than 5x 10 -11 M, less than 1x 10 -11 M, less than 5x 10 -12 M or less than 1x 10 -12 M, as measured by biosensor.

[0205] In some embodiments, the anti-IL23 antibody binds to canine IL23, human IL23, feline IL23, or equine IL23 with a Kd between 5 x 10 -6 M and 1x 10 -6 Between M, 5x 10 -6 M and 5x 10 -7 Between M, 5x 10 -6 M and 1x 10 -7 Between M, 5x 10 -6 M and 5x10 -8 Between M, 5x 10 -6 M and 1x 10 -8 Between M, 5x 10 -6 M and 5x 10 -9 Between M, 5x 10 -6 M and 1x 10 -9 Between M, 5x10 -6 M and 5x 10 -10 Between M, 5x 10 -6 M and 1x 10 -10 Between M, 5x 10 -6 M and 5x 10 -11 Between M, 5x 10 -6 M and 1x10 -11 Between M, 5x 10 -6 M and 5x 10 - 12M, 5x 10 -6 M and 1x 10 -12 Between M, 1x 10 -6 M and 5x 10 -7 Between M, 1x 10 -6 M and 1x 10 -7 Between M, 1x 10 -6 M and 5x 10 -8 Between M, 1x 10 -6 M and 1x 10 -8 Between M, 1x10 -6 M and 5x10 -9 Between M, 1x 10 -6 M and 1x 10 -9 Between M, 1x 10 -6 M and 5x 10 -10 Between M, 1x 10 -6 M and 1x 10 -10 Between M, 1x 10 -6 M and 5x 10 -11 Between M, 1x 10 -6 M and 1x 10-11 Between M, 1x 10 -6 M and 5x 10 -12 Between M, 1x 10 -6 M and 1x 10 -12 Between M, 5x 10 -7 M and 1x 10 -7 Between M, 5x 10 -7 M and 5x 10 -8 Between M, 5x 10 -7 M and 1x 10 -8 Between M, 5x 10 -7 M and 5x 10 -9 Between M, 5x 10 -7 M and 1x 10 -9 Between M, 5x 10 -7 M and 5x 10 -10 Between M, 5x 10 -7 M and 1x 10 -10 Between M, 5x 10 -7 M and 5x 10 -11 Between M, 5x 10 -7 M and 1x 10 -11 Between M, 5x 10 -7 M and 5x 10 -12 Between M, 5x 10 -7 M and 1x 10 -12 Between M, 1x 10 -7 M and 5x 10 -8 Between M, 1x 10 -7 M and 1x 10 -8 Between M, 1x 10 - 7 M and 5x 10 -9 Between M, 1x 10 -7 M and 1x 10 -9 Between M, 1x 10 -7 M and 5x 10 -10 Between M, 1x 10 -7 M and 1x 10 -10 Between M, 1x 10 -7 M and 5x 10 -11 Between M, 1x 10 -7 M and 1x 10 -11 Between M, 1x 10 -7 M and 5x 10 -12 Between M, 1x10 -7 M and 1x 10 -12 Between M, 5x 10-8 M and 1x 10 -8 Between M, 5x 10 -8 M and 5x 10 -9 Between M, 5x 10 -8 M and 1x10 - 9 Between M, 5x 10 -8 M and 5x 10 -10 Between M, 5x 10 -8 M and 1x 10 -10 Between M, 5x 10 -8 M and 5x 10 -11 Between M, 5x10 -8 M and 1x 10 -11 Between M, 5x 10 -8 M and 5x 10 -12 Between M, 5x 10 -8 M and 1x 10 -12 Between M, 1x 10 -8 M and 5x10 -9 Between M, 1x 10 -8 M and 1x 10 -9 Between M, 1x 10 -8 M and 5x 10 -10 Between M, 1x 10 -8 M and 1x 10 -10 Between M, 1x 10 -8 M and 5x 10 -11 Between M, 1x 10 -8 M and 1x 10 -11 Between M, 1x 10 -8 M and 5x 10 -12 Between M, 1x 10 -8 M and 1x 10 -12 Between M, 5x 10 -9 M and 1x 10 -9 Between M, 5x10 -9 M and 5x 10 -10 Between M, 5x 10 -9 M and 1x 10 -10 Between M, 5x 10 -9 M and 5x 10 -11 Between M, 5x 10 -9 M and 1x 10 -11 Between M, 5x 10 -9 M and 5x10 -12 Between M, 5x 10 -9 M and 1x 10-12 Between M, 1x 10 -9 M and 5x 10 -10 Between M, 1x 10 -9 M and 1x 10 -10 Between M, 1x 10 -9 M and 5x 10 -11 Between M, 1x 10 -9 M and 1x 10 -11 Between M, 1x 10 -9 M and 5x 10 -12 Between M, 1x 10 -9 M and 1x 10 -12 Between M, 5x10 -10 M and 1x 10 -10 Between M, 5x 10 -10 M and 5x 10 -11 Between M, 1x 10 -10 M and 5x 10 -11 Between M, 1x 10 -10 M and 1x 10 -11 Between M, 1x 10 -10 M and 5x 10 -12 Between M, 1x 10 -10 M and 1x 10 -12 Between M, 5x 10 -11 M and 1x 10 -12 Between M, 5x 10 -11 M and 5x 10 -12 Between M, 5x 10 -11 M and 1x 10 -12 Between M, 1x 10 -11 M and 5x 10 -12 Between M or 1x10 -11 M and 1x 10 -12 Between M, as measured by biolayer interferometry.

[0206] In some embodiments, the anti-IL23 antibody binds to canine IL23, human IL23, feline IL23, or equine IL23 as determined by immunoblot analysis.

[0207] In some embodiments, the anti-TNFα antibody binds to canine TNFα, human TNFα, feline TNFα, or equine TNFα with a dissociation constant (Kd) of less than 5 x 10 -6 M, less than 1x 10 -6 M, less than 5x 10 -7 M, less than 1x 10 -7 M, less than 5x 10-8 M, less than 1x 10 -8 M, less than 5x 10 -9 M, less than 1x 10 -9 M, less than 5x 10 -10 M, less than 1x 10 -10 M, less than 5x 10 -11 M, less than 1x 10 -11 M, less than 5x 10 -12 M or less than 1x 10 -12 M, as measured by biosensor.

[0208] In some embodiments, the anti-TNFα antibody binds to canine TNFα, human TNFα, feline TNFα, or equine TNFα with a Kd between 5 x 10 -6 M and 1x 10 -6 Between M, 5x 10 -6 M and 5x 10 -7 Between M, 5x 10 -6 M and 1x 10 -7 Between M, 5x 10 -6 M and 5x10 -8 Between M, 5x 10 -6 M and 1x 10 -8 Between M, 5x 10 -6 M and 5x 10 -9 Between M, 5x 10 -6 M and 1x 10 -9 Between M, 5x 10 -6 M and 5x 10 -10 Between M, 5x10 -6 M and 1x 10 -10 Between M, 5x 10 -6 M and 5x 10 -11 Between M, 5x 10 -6 M and 1x10 -11 Between M, 5x 10 -6 M and 5x 10 -12 Between M, 5x 10 -6 M and 1x10 -12 Between M, 1x 10 -6 M and 5x 10 -7 Between M, 1x10 -6 M and 1x 10 -7 Between M, 1x 10 -6 M and 5x 10 -8 Between M, 1x 10 -6 M and 1x 10 -8Between M, 1x 10 -6 M and 5x 10 -9 Between M, 1x 10 -6 M and 1x 10 -9 Between M, 1x 10 -6 M and 5x 10 -10 Between M, 1x 10 -6 M and 1x 10 -10 Between M, 1x10 -6 M and 5x 10 -11 Between M, 1x 10 -6 M and 1x 10 -11 Between M, 1x 10 -6 M and 5x 10 -12 Between M, 1x 10 -6 M and 1x10 -12 Between M, 5x 10 -7 M and 1x 10 -7 Between M, 5x 10 -7 M and 5x 10 -8 Between M, 5x 10 -7 M and 1x 10 -8 Between M, 5x10 -7 M and 5x 10 -9 Between M, 5x 10 -7 M and 1x 10 -9 Between M, 5x 10 -7 M and 5x 10 -10 Between M, 5x 10 -7 M and 1x10 -10 Between M, 5x 10 -7 M and 5x 10 -11 Between M, 5x 10 -7 M and 1x 10 -11 Between M, 5x 10 -7 M and 5x 10 -12 Between M, 5x 10 -7 M and 1x 10 -12 Between M, 1x 10 -7 M and 5x 10 -8 Between M, 1x10 -7 M and 1x 10 -8 Between M, 1x 10 -7 M and 5x10 -9 Between M, 1x 10 -7 M and 1x 10 -9 Between M, 1x 10 -7 M and 5x 10 -10Between M, 1x 10 -7 M and 1x 10 -10 Between M, 1x 10 -7 M and 5x 10 -11 Between M, 1x 10 -7 M and 1x 10 -11 Between M, 1x 10 -7 M and 5x 10 -12 Between M, 1x 10 -7 M and 1x 10 -12 Between M, 5x 10 -8 M and 1x 10 -8 Between M, 5x 10 -8 M and 5x 10 -9 Between M, 5x 10 -8 M and 1x 10 -9 Between M, 5x 10 -8 M and 5x 10 -10 Between M, 5x 10 -8 M and 1x10 -10 Between M, 5x 10 -8 M and 5x 10 -11 Between M, 5x 10 -8 M and 1x 10 -11 Between M, 5x 10 -8 M and 5x 10 -12 Between M, 5x 10 -8 M and 1x 10 -12 Between M, 1x 10 -8 M and 5x 10 -9 Between M, 1x 10 -8 M and 1x 10 -9 Between M, 1x 10 -8 M and 5x 10 -10 Between M, 1x 10 -8 M and 1x 10 -10 Between M, 1x 10 -8 M and 5x 10 -11 Between M, 1x 10 -8 M and 1x 10 -11 Between M, 1x 10 -8 M and 5x 10 -12 Between M, 1x 10 -8 M and 1x 10 -12 Between M, 5x 10 -9 M and 1x 10 -9 Between M, 5x 10 -9 M and 5x 10-10 Between M, 5x 10 -9 M and 1x 10 -10 Between M, 5x10 -9 M and 5x 10 -11 Between M, 5x 10 -9 M and 1x 10 -11 Between M, 5x 10 -9 M and 5x 10 -12 Between M, 5x 10 -9 M and 1x10 -12 Between M, 1x 10 -9 M and 5x 10 -10 Between M, 1x 10 -9 M and 1x 10 -10 Between M, 1x 10 -9 M and 5x 10 -11 Between M, 1x 10 -9 M and 1x 10 -11 Between M, 1x 10 -9 M and 5x 10 -12 Between M, 1x 10 -9 M and 1x 10 -12 Between M, 5x 10 -10 M and 1x 10 -10 Between M, 5x 10 -10 M and 5x 10 -11 Between M, 1x 10 -10 M and 5x 10 -11 Between M, 1x 10 -10 M and 1x 10 -11 Between M, 1x 10 -10 M and 5x 10 -12 Between M, 1x 10 -10 M and 1x 10 -12 Between M, 5x 10 -11 M and 1x 10 -12 Between M, 5x10 -11 M and 5x10 -12 Between M, 5x 10 -11 M and 1x 10 -12 Between M, 1x 10 -11 M and 5x 10 -12 Between M or 1x 10 -11 M and 1x 10 -12 M, as measured by a biosensor.

[0209] In some embodiments, the anti-TNFα antibody binds to canine TNFα, human TNFα, feline TNFα, or equine TNFα as determined by immunoblot analysis.

[0210] In some embodiments, the variant has at least 1, 2, 3, 4, 5, or 6 amino acids substituted with different amino acids.

[0211] In some embodiments, after aligning the sequences and introducing spaces (if necessary) to achieve maximum percentage sequence identity, and without considering any conservative substitutions as part of sequence identity, the variant has at least about 50% sequence identity with the reference nucleic acid molecule or polypeptide. Such variants include, for example, polypeptides in which one or more amino acid residues are added or deleted at the N-terminus or C-terminus of the polypeptide. In some embodiments, the variant has at least about 50% sequence identity, at least about 60% sequence identity, at least about 65% sequence identity, at least about 70% sequence identity, at least about 75% sequence identity, at least about 80% sequence identity, at least about 85% sequence identity, at least about 90% sequence identity, at least about 95% sequence identity, at least about 97% sequence identity, at least about 98% sequence identity, or at least about 99% sequence identity with the sequence of the reference nucleic acid or polypeptide.

[0212] In some embodiments, the anti-IL23 antibody can reduce IL23 signaling function in a companion animal species by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or 100% compared to IL23 signaling function in the absence of the antibody.In some embodiments, the reduction in IL23 signaling function is between 10% and 15%, between 10% and 20%, between 10% and 25%, between 10% and 30%, between 10% and 35%, between 10% and 40%, between 10% and 45%, between 10% and 50%, between 10% and 60%, between 10% and 70%, between 10% and 80%, between 10% and 90%, between 10% and 100%, between 15% and 20%, between 15% and 25%, between 15% and 30%, between 15% and 35%, between 15% and 40%, between 15% and 45%, between 15% and 50%, between 15% and 60%, between 10% and 70%, between 10% and 80%, between 10% and 90%, between 10% and 100%, between 15% and 20%, between 15% and 25%, between 15% and 30%, between 15% and 35%, between 15% and 40%, between 15% and 45%, between 15% and 50%, between 15% and 60%, between 15 ...5% and 80%, between 15% and 90%, between 15% and 100%, between Between 5% and 70%, between 15% and 80%, between 15% and 90%, between 15% and 100%, between 20% and 25%, between 20% and 30%, between 20% and 35%, between 20% and 40%, between 20% and 45%, between 20% and 50%, between 20% and 60%, between 20% and 70%, between 20% and 80%, between 20% and 90%, between 20% and 100%, between 25% and 30%, between 25% and 35%, between 25% and 40%, between 25% and 45%, between 25% and 50%, between 25% and 60%, between 25% and 70%, between 25% and 80%, between 25% and 90 %, between 25% and 100%, between 30% and 35%, between 30% and 40%, between 30% and 45%, between 30% and 50%, between 30% and 60%, between 30% and 70%, between 30% and 80%, between 30% and 90%, between 30% and 100%, between 35% and 40%, between 35% and 45%, between 35% and 50%, between 35% and 60%, between 35% and 70%, between 35% and 80%, between 35% and 90%, between 35% and 100%, between 40% and 45%, between 40% and 50%, between 40% and 60%, between 40% and 70%, between 40% and 80%, Between 40% and 90%, between 40% and 100%, between 45% and 50%, between 45% and 60%, between 45% and 70%, between 45% and 80%, between 45% and 90%, between 45% and 100%, between 50% and 60%, between 50% and 70%, between 50% and 80%, between 50% and 90%, between 50% and 100%, between 60% and 70%, between 60% and 80%, between 60% and 90%, between 60% and 100%, between 70% and 80%, between 70% and 90%, between 70% and 100%, between 80% and 90%, between 80% and 100%, or between 90% and 100%.

[0213] In some embodiments, the anti-TNFα antibody can reduce TNFα signaling function in a companion animal species by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90% or 100% compared to NGF signaling function in the absence of the antibody.In some embodiments, the reduction in NGF signaling function is between 10% and 15%, between 10% and 20%, between 10% and 25%, between 10% and 30%, between 10% and 35%, between 10% and 40%, between 10% and 45%, between 10% and 50%, between 10% and 60%, between 10% and 70%, between 10% and 80%, between 10% and 90%, between 10% and 100%, between 15% and 20%, between 15% and 25%, between 15% and 30%, between 15% and 35%, between 15% and 40%, between 15% and 45%, between 15% and 50%, between 15% and 60%, between 15% and 15 ... Between % and 70%, Between 15% and 80%, Between 15% and 90%, Between 15% and 100%, Between 20% and 25%, Between 20% and 30%, Between 20% and 35%, Between 20% and 40%, Between 20% and 45%, Between 20% and 50%, Between 20% and 60%, Between 20% and 70%, Between 20% and 80%, Between 20% and 90%, Between 20% and 100%, Between 25% and 30%, Between 25% and 35%, Between 25% and 40%, Between 25% and 45%, Between 25% and 50%, Between 25% and 60%, Between 25% and 70%, Between 25% and 80%, Between 25% and 90% between 25% and 100%, between 30% and 35%, between 30% and 40%, between 30% and 45%, between 30% and 50%, between 30% and 60%, between 30% and 70%, between 30% and 80%, between 30% and 90%, between 30% and 100%, between 35% and 40%, between 35% and 45%, between 35% and 50%, between 35% and 60%, between 35% and 70%, between 35% and 80%, between 35% and 90%, between 35% and 100%, between 40% and 45%, between 40% and 50%, between 40% and 60%, between 40% and 70%, between 40% and 80%, Between 40% and 90%, between 40% and 100%, between 45% and 50%, between 45% and 60%, between 45% and 70%, between 45% and 80%, between 45% and 90%, between 45% and 100%, between 50% and 60%, between 50% and 70%, between 50% and 80%, between 50% and 90%, between 50% and 100%, between 60% and 70%, between 60% and 80%, between 60% and 90%, between 60% and 100%, between 70% and 80%, between 70% and 90%, between 70% and 100%, between 80% and 90%, between 80% and 100%, or between 90% and 100%.

[0214] Pharmaceutical composition

[0215] The terms "pharmaceutical formulation" and "pharmaceutical composition" refer to a preparation that is in a form that permits the biological activity of the active ingredient to be effective, and that contains no additional components that are unacceptably toxic to a subject to which the formulation would be administered.

[0216] The pharmaceutically acceptable carrier is nontoxic to the recipient at the dosages and concentrations employed and is compatible with the other ingredients of the formulation. The pharmaceutically acceptable carrier is suitable for the formulation employed. Examples of pharmaceutically acceptable carriers include alumina; aluminum stearate; lecithin; serum proteins such as human serum albumin, canine or other animal albumin; buffers such as phosphate, citrate, tromethamine, or HEPES buffer; glycine; sorbic acid; potassium sorbate; partial glyceride mixtures of saturated vegetable fatty acids; water; salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silicon dioxide, or magnesium trisilicate; polyvinyl pyrrolidone, cellulose-based substances; polyethylene glycol; sucrose; mannitol; or amino acids, including but not limited to arginine.

[0217] The pharmaceutical composition can be stored in a lyophilized form. Therefore, in some embodiments, the preparation process includes a lyophilization step. Then, before administration to a dog, the lyophilized composition can be reconstituted, typically as an aqueous composition suitable for parenteral administration. In other embodiments, particularly when the antibody is highly stable to heat and oxidative denaturation, the pharmaceutical composition can be stored as a liquid (i.e., as an aqueous composition), which can be administered to a dog directly or in an appropriate dilution. The lyophilized composition can be reconstituted with sterile water for injection (WFI). Antibacterial agents, such as benzyl alcohol, may be included. Therefore, the present invention provides pharmaceutical compositions in solid or liquid form.

[0218] When administered, the pH of the pharmaceutical composition may be in the range of about pH 5 to about pH 8. If the compositions of the present invention are used for therapeutic purposes, they are sterile. Sterility can be achieved by any of several means known in the art, including filtration through a sterile filtration membrane (e.g., a 0.2 micron membrane). Sterility can be maintained with or without the use of an antimicrobial agent.

[0219] Exemplary Uses of Antibodies and Pharmaceutical Compositions

[0220] The antibodies of the present invention or pharmaceutical compositions comprising the antibodies can be used to treat IL23-mediated conditions, disorders or diseases in a subject, wherein the subject can be a companion animal, including but not limited to a dog or a cat.

[0221] In some embodiments, anti-IL23 antibodies or pharmaceutical compositions comprising the same, with or without anti-TNFα antibodies, can be used according to the methods herein to treat IL23-related conditions. In some embodiments, anti-IL23 and / or anti-TNFα antibodies or pharmaceutical compositions are administered to companion animals (such as dogs or cats) to treat IL23- or TNFα-related conditions. In some embodiments, anti-IL23 and / or anti-TNFα antibodies or pharmaceutical compositions are administered to companion animals (such as dogs or cats) to maintain remission of IL23- or TNFα-related conditions.

[0222] A therapeutically effective amount is an amount in which any toxic or deleterious effects of a substance / molecule, agonist, or antagonist are offset by the therapeutically beneficial effects. Thus, a therapeutically effective amount can be delivered in one or more administrations. A therapeutically effective amount is an amount that is effective to achieve the desired therapeutic or preventive result at a dosage and over a desired time period.

[0223] In some embodiments, the anti-IL23 and / or anti-TNFα antibody or a pharmaceutical composition comprising an IL23 and / or TNFα antibody is administered parenterally by subcutaneous administration, intravenous infusion, or intramuscular injection. In some embodiments, the anti-IL23 and / or anti-TNFα antibody or a pharmaceutical composition comprising an anti-IL23 and / or anti-TNFα antibody is administered as a bolus or by continuous infusion over a period of time. In some embodiments, the anti-IL23 and / or anti-TNFα antibody or a pharmaceutical composition comprising an anti-IL23 and / or anti-TNFα antibody is administered intramuscularly, intraperitoneally, intracerebrospinal fluid, subcutaneously, intraarterially, intrasynovially, intrathecally, or by inhalation.

[0224] As described herein, anti-IL23 antibodies can be administered alone, in combination with anti-TNFα antibodies, or in combination with the TNFα binding region of a bispecific antibody in an amount ranging from 0.01 mg / kg to 100 mg / kg body weight per dose. In some embodiments, IL23 can be administered alone or in combination with a TNFα antibody in an amount ranging from 0.5 mg / kg to 50 mg / kg body weight per dose. In some embodiments, IL23 and / or TNFα antibodies can be administered in an amount ranging from 0.1 mg / kg to 10 mg / kg body weight per dose. In some embodiments, IL23 and / or TNFα antibodies can be administered in an amount ranging from 0.1 mg / kg to 100 mg / kg body weight per dose. In some embodiments, IL23 and / or TNFα antibodies can be administered in an amount ranging from 1 mg / kg to 10 mg / kg body weight per dose. In some embodiments, the IL23 and / or TNFα antibody can be administered in an amount ranging from 0.5 mg / kg to 100 mg / kg body weight, 1 mg / kg to 100 mg / kg body weight, 5 mg / kg to 100 mg / kg body weight, 10 mg / kg to 100 mg / kg body weight, 20 mg / kg to 100 mg / kg body weight, 50 mg / kg to 100 mg / kg body weight, 1 mg / kg to 10 mg / kg body weight, 5 mg / kg to 10 mg / kg body weight, 0.5 mg / kg to 10 mg / kg body weight, 0.01 mg / kg to 0.5 mg / kg body weight, 0.01 mg / kg to 0.1 mg / kg body weight, or 5 mg / kg to 50 mg / kg body weight. In some embodiments, the IL23 and / or TNFα antibody can be administered in an amount of 0.5 mg / kg body weight. In some embodiments, IL23 and / or TNFα antibodies may be administered in an amount of 2 mg / kg body weight.

[0225] IL23 can be administered to a companion animal alone or in combination with a TNFα antibody or a pharmaceutical composition comprising an IL23 and / or TNFα antibody, at one time or over a series of treatments. For example, IL23 and / or TNFα antibody or a pharmaceutical composition comprising an IL23 and / or TNFα antibody can be administered at least once, more than once, at least twice, at least three times, at least four times, or at least five times.

[0226] In some embodiments, the dose is administered once a week for at least two or three consecutive weeks, and in some embodiments, this treatment cycle is repeated two or more times, optionally interspersed with one or more weeks without treatment. In other embodiments, the therapeutically effective dose is administered once a day for two to five consecutive days, and in some embodiments, this treatment cycle is repeated two or more times, optionally interspersed with one or more days or one or more weeks without treatment.

[0227] Administration that can be "combined" with one or more other therapeutic agents includes simultaneous (parallel) administration and continuous administration or sequential administration in any order. The term "simultaneously" as used herein refers to the administration of two or more therapeutic agents, wherein at least part of the administration overlaps in time, or wherein the administration of one therapeutic agent is in a short time period relative to the administration of the other therapeutic agents. For example, two or more therapeutic agents are administered at a time interval of no more than about a specified number of minutes. The term "sequentially" as used herein refers to the administration of two or more therapeutic agents, wherein one or more agents are continued to be administered after stopping the administration of one or more other agents, or wherein one or more agents are started before the administration of one or more other agents. For example, the administration of two or more therapeutic agents is an administration with a time interval of more than about a specific number of minutes. As used herein, "combination" refers to the administration of another treatment modality in addition to a treatment modality. Therefore, "combination" refers to the administration of another treatment modality before, during, or after the administration of a treatment modality to an animal.

[0228] In some embodiments, the method comprises administering in combination with an IL23 and / or TNFα antibody, or a pharmaceutical composition comprising an IL23 and / or TNFα antibody, an IL17 antibody, an IL-5 antibody, an IL-10 antibody, an IL-31 antibody, an IL4 antibody, an IL13 antibody, an IgE antibody, a CD11α antibody, an IL6 antibody, an IL6R antibody, an α4-integrin antibody, a β7-integrin antibody, an IL12 antibody, an IL1β antibody, or an anti-BlyS antibody.

[0229] Provided herein are methods of exposing an IL23 and / or TNFα antibody or a pharmaceutical composition comprising an IL23 and / or TNFα antibody to a cell under conditions that allow the antibody to bind to IL23 and / or TNFα. In some embodiments, the cell is exposed to the antibody or pharmaceutical composition ex vivo. In some embodiments, the cell is exposed to the antibody or pharmaceutical composition in vivo. In some embodiments, the cell is exposed to an IL23 and / or TNFα antibody or pharmaceutical composition under conditions that allow the antibody to bind to IL23 and / or TNFα.

[0230] In some embodiments, cells can be exposed to IL23 and / or TNFα antibodies or pharmaceutical compositions in vivo by any one or more of the administration methods described herein, including but not limited to intraperitoneal injection, intramuscular injection, and intravenous injection into a subject. In some embodiments, cells can be exposed to IL23 and / or TNFα antibodies or pharmaceutical compositions in vitro by exposing them to a culture medium comprising the antibody or pharmaceutical composition. In some embodiments, before exposing the cells to a culture medium comprising the antibody or pharmaceutical composition, the permeability of the cell membrane can be affected by using any number of methods understood by those skilled in the art, such as electroporating the cells or exposing the cells to a solution containing calcium chloride.

[0231] In some embodiments, binding results in a decrease in IL23 and / or TNFα or IL12 signaling function of the cell. In some embodiments, the IL23 and / or TNFα antibody can reduce IL23 and / or TNFα or IL12 signaling function in the cell by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or 100% compared to IL23 and / or TN or IL12 signaling function in the absence of the antibody.In some embodiments, the reduction in IL23 and / or TNFα or IL12 signaling function is between 10% and 15%, between 10% and 20%, between 10% and 25%, between 10% and 30%, between 10% and 35%, between 10% and 40%, between 10% and 45%, between 10% and 50%, between 10% and 60%, between 10% and 70%, between 10% and 80%, between 10% and 90%, between 10% and 100%, between 15% and 20%, between 15% and 25%, between 15% and 30%, between 15% and 35%, between 15% and 40%, between 15% and 45%, between 15% and 50%, between 15% and 1 ... Between % and 60%, between 15% and 70%, between 15% and 80%, between 15% and 90%, between 15% and 100%, between 20% and 25%, between 20% and 30%, between 20% and 35%, between 20% and 40%, between 20% and 45%, between 20% and 50%, between 20% and 60%, between 20% and 70%, between 20% and 80%, between 20% and 90%, between 20% and 100%, between 25% and 30%, between 25% and 35%, between 25% and 40%, between 25% and 45%, between 25% and 50%, between 25% and 60%, between 25% and 70%, between 25% and 80%, Between 25% and 90%, between 25% and 100%, between 30% and 35%, between 30% and 40%, between 30% and 45%, between 30% and 50%, between 30% and 60%, between 30% and 70%, between 30% and 80%, between 30% and 90%, between 30% and 100%, between 35% and 40%, between 35% and 45%, between 35% and 50%, between 35% and 60%, between 35% and 70%, between 35% and 80%, between 35% and 90%, between 35% and 100%, between 40% and 45%, between 40% and 50%, between 40% and 60%, between 40% and 70%, between 40% and 80% or between 90% and 100%, between 45% and 50%, between 45% and 60%, between 45% and 70%, between 45% and 80%, between 45% and 90%, between 45% and 100%, between 50% and 60%, between 50% and 70%, between 50% and 80%, between 50% and 90%, between 50% and 100%, between 60% and 70%, between 60% and 80%, between 60% and 90%, between 60% and 100%, between 70% and 80%, between 70% and 90%, between 70% and 100%, between 80% and 90%, between 80% and 100%, or between 90% and 100%.

[0232] Provided herein are methods for detecting, diagnosing and monitoring diseases associated with IL23 and / or TNFα using IL23 and / or TNFα antibodies, polypeptides and polynucleotides. Provided herein are methods for determining whether a companion animal will respond to IL23 and / or TNFα antibody therapy. In some embodiments, the method includes detecting whether an animal has cells expressing IL23 and / or TNFα using IL23 and / or TNFα antibodies. In some embodiments, the detection method includes contacting the sample with an antibody, polypeptide or polynucleotide, and determining whether the binding level is different from the binding level of a reference sample or a comparison sample (such as a control). In some embodiments, the method can be used to determine whether the antibody or polypeptide described herein is suitable for treatment of a subject animal.

[0233] In some embodiments, the sample is a biological sample. The term "biological sample" means a certain amount of material from an organism or a former organism. In some embodiments, the biological sample is a cell or cell / tissue lysate. In some embodiments, biological samples include but are not limited to blood (e.g., whole blood), plasma, serum, urine, synovial fluid, and epithelial cells.

[0234] In some embodiments, cells or cell / tissue lysates are contacted with an IL23 and / or TNFα antibody and binding between the antibody and the cell is determined. When the test cell shows binding activity compared to a reference cell of the same tissue type, it may indicate that the subject will benefit from treatment with the IL23 and / or TNFα antibody. In some embodiments, the test cell is from a companion animal's tissue.

[0235] Various methods known in the art for detecting specific antibody-antigen binding can be used. Exemplary immunoassays that can be performed include fluorescence polarization immunoassay (FPIA), fluorescence immunoassay (FIA), enzyme immunoassay (EIA), turbidity inhibition immunoassay (NIA), enzyme-linked immunosorbent assay (ELISA), and radioimmunoassay (RIA). Indicator moieties or labeling groups can be attached to the subject antibody and selected to meet the needs of the various uses of the method, which is generally determined by the availability of assay equipment and compatible immunoassay procedures. Suitable labels include, but are not limited to, radionuclides (e.g., 125 I. 131 I. 35 S. 3 H or 32 P), an enzyme (e.g., alkaline phosphatase, horseradish peroxidase, luciferase, or β-galactosidase), a fluorescent moiety or protein (e.g., fluorescein, rhodamine, phycoerythrin, GFP, or BFP), or a luminescent moiety (e.g., Qdot supplied by QuantumDot Corporation, Palo Alto, Calif. TMNanoparticles). General techniques for performing the various immunoassays described above are known to those of ordinary skill in the art.

[0236] For diagnostic purposes, polypeptides comprising antibodies can be labeled with a detectable portion, including but not limited to radioisotopes, fluorescent labels, and various enzyme-substrate labels known in the art. Methods of conjugating labels to antibodies are known in the art. In some embodiments, IL23 and / or TNFα antibodies do not require labeling, and a second labeled antibody that binds to the first IL23 and / or TNFα antibody can be used to detect its presence. In some embodiments, IL23 and / or TNFα antibodies can be used in any known assay method, such as competitive binding assays, direct and indirect sandwich assays, and immunoprecipitation assays. Zola, Monoclonal Antibodies: A Manual of Techniques, pp. 147-158 (CRC Press, Inc. 1987). IL23 and / or TNFα antibodies and polypeptides can also be used in in vivo diagnostic assays, such as in vivo imaging. In general, antibodies or polypeptides are labeled with radionuclides (such as 111 In, 99 Tc, 14 C. 131 I. 125 I. 3 H or any other radionuclide label, including those outlined herein) labels, so that cells or tissues of interest can be located using immunoscintigraphy. Antibodies can also be used as staining reagents in pathology using techniques well known in the art.

[0237] In some embodiments, the first antibody is used for diagnosis and the second antibody is used for treatment. In some embodiments, the first antibody and the second antibody are different. In some embodiments, the first antibody and the second antibody can be combined with the antigen simultaneously by combining with an independent epitope.

[0238] Example

[0239] The various features and embodiments of the present disclosure are illustrated in the following representative examples, which are intended to be illustrative and not limiting. Those skilled in the art will readily appreciate that the specific examples are merely illustrative of the present invention, as more fully described in the claims that follow. Each embodiment and feature described in this application should be understood to be interchangeable and combinable with each embodiment contained therein.

[0240] Example 1: Recombinant expression of canine IL23 or p40

[0241] The sequence of the heterodimeric canine IL23 polypeptide designed for recombinant expression is shown in SEQ ID NO: 1. This canine IL23 polypeptide (SEQ ID NO: 1) contains a linker that stabilizes the two subunits of the complex, a polyHis tag for purification, and a tag for in vitro site-specific biotinylation. This single-chain canine IL23 polypeptide was expressed as a monomeric protein in CHO cells. The expressed protein was purified using a Ni-NTA column.

[0242] The sequence of the heterodimeric feline IL23 polypeptide for recombinant expression was also designed as SEQ ID NO: 2. This feline IL23 polypeptide (SEQ ID NO: 2) contains a linker between the two subunits that stabilizes the complex. Canine p40 polypeptide was also expressed in mammalian CHO cells.

[0243] Example 2: Anti-IL23 Antibody Gene Synthesis, CHO Cell Expression and Purification

[0244] DNA sequences encoding the anti-IL23 (clone C340) VL domain of SEQ ID NO: 3 and the VH domain of SEQ ID NO: 7 were fused to DNA sequences encoding canine constant CLκ and IgG B regions. These anti-IL23 (clone C340) genes were chemically synthesized and inserted into expression vectors suitable for transfection into CHO host cells. These expression vectors were transfected into CHO cells. Chimeric anti-IL23 antibodies were expressed in CHO cells, and the expressed antibodies were purified by one or more of various steps, including protein A column chromatography.

[0245] result: The chimeric antibody was well expressed and appeared as a monodisperse peak on gel filtration chromatography.

[0246] Example 3: Demonstration of canine IL23 binding activity

[0247] Most monoclonal anti-IL23 antibodies are unable to bind to non-primate homologous targets. Surprisingly, as shown in the results of the study in this example, the chimeric anti-IL23 antibodies described herein (e.g., the anti-IL23 antibodies of Table 1 above) were found to have high binding affinity for both canine IL23 and canine p40. This study was conducted to measure the binding affinity of anti-IL23 antibodies to IL23 and p40 antigens using Biacore 8K.

[0248] Material

[0249] Table 2: Samples used

[0250]

[0251] Table 3: Instruments and reagents used

[0252]

[0253] method

[0254] 1. Antigen-antibody affinity measurement

[0255] Preparation of running buffer: Dilute one volume of 10× buffer with 9 volumes of degassed, filtered MilliQ water.

[0256] Preparation of regeneration buffer (10 mM glycine): Weigh a certain amount of glycine and dissolve it in MilliQ water. Adjust the pH to 1.5-1.7 to obtain regeneration buffer.

[0257] The assay was performed at 25°C and the running buffer was HBS-EP+.

[0258] The antibody was injected onto the S-series sensor chip Protein A as the capture agent.

[0259] Antigen was diluted to various concentrations and injected onto the surface of flow cells 1 and 2 as the association phase, followed by injection of running buffer as the dissociation phase.

[0260] The binding model is shown below. The run configurations used are listed in Table 4 below.

[0261] Table 4: Run Configuration

[0262]

[0263]

[0264] result

[0265] All data were processed using Biacore 8K Evaluation Software version 3.0. Flow cell 1 and buffer blank injections in each cycle were used as a double reference for response unit subtraction. Biacore binding affinity values ​​are provided in Table 5 below.

[0266] Table 5: Affinity measurements of antibodies and antigens

[0267]

[0268] Example 4: Caninization of anti-IL23 C340 antibody

[0269] This example illustrates caninization studies of an anti-IL23 C340 antibody having a VL domain of SEQ ID NO: 3 and a VH domain of SEQ ID NO: 7. Preliminary analysis of the anti-IL23 C340 VL and VH amino acid sequences identified candidate amino acid substitutions at various positions that could be used to caninize the sequences. Table 6 below provides a list of these candidate amino acid substitutions.

[0270] Table 6: Candidate VL (SEQ ID NO: 3) and VH (SEQ ID NO: 7) amino acid substitutions

[0271]

[0272]

[0273]

[0274]

[0275] Based on the candidate amino acid substitutions, genes encoding various caninized variants of VL (SEQ ID NO: 3) and VH (SEQ ID NO: 7) were expressed using human IgG1 and κ frameworks. The amino acid sequences of the caninized variants of VL and VH have been aligned and are listed in Tables 7 and 8 below (the "-" sign indicates the position of the amino acid change).

[0276] Table 7: Alignment of caninized VL domains based on SEQ ID NO: 3

[0277]

[0278] Table 8: Alignment of caninized VH domains based on SEQ ID NO: 7

[0279]

[0280]

[0281] The genes encoding each of the caninized VL and VH variants listed above in Tables 7 and 8 were expressed in a mammalian cell system. Various combinations of anti-IL23 VL and VH polypeptides were tested for expression and canine IL23 binding.

[0282] As shown in the results listed in Table 9 (below), "wild-type" anti-IL23 C340 VL and VH domain polypeptides expressed well. Surprisingly, some versions of the caninized VH domain polypeptide (e.g., variant V1) were found to pair well with "wild-type" C340 VL (SEQ ID NO: 3). Similarly, caninized VL (e.g., variant V2) paired well with wild-type C340 VH (SEQ ID NO: 7). However, caninized VH domain variant V1 failed to pair with caninized VL domain variant V2. Further results from these studies are provided in Table 9.

[0283] Table 9

[0284]

[0285]

[0286] As an exemplary caninized anti-IL23, a variant comprising VL domain V2.1 (SEQ ID NO: 11) and VH domain V1.1 (SEQ ID NO: 12) expressed well and was observed to maintain full canine IL23 binding activity using Biacore binding affinity assay.

[0287] A mutation was also introduced into the Fc region at amino acid position 252 (EU numbering), located at the AB turn. This mutation, described in U.S. Patent No. 7,658,921B2, changes the amino acid at position 252 to a Y (L252Y) and results in an increased in vivo half-life of IgG antibodies. This half-life-extending Fc region mutation was introduced as L252Y into a caninized anti-IL23 heavy chain IgG-B, generating the "long-acting" caninized anti-IL23 of SEQ ID NO: 20.

[0288] Example 5: Felinization of anti-IL23 (C340) antibody

[0289] This example illustrates felinization studies of anti-IL23 (C340) having the VL domain of SEQ ID NO: 3 and the VH domain of SEQ ID NO: 7.

[0290] Felinization of the VL and VH domains was performed using the same general method as for caninization described in Example 4. Preliminary analysis of the anti-IL23 C340 VL and VH amino acid sequences was performed to identify candidate amino acid substitutions at various positions that could be used to felinize the sequences. Genes encoding felinized variants of the VL (SEQ ID NO: 3) and VH (SEQ ID NO: 7) using a feline IgG framework were expressed in a mammalian cell system. The feline IL23 binding activity of the expressed felinized variants was measured for identification. Various combinations of anti-IL23 VL and VH polypeptides were tested for expression and canine IL23 binding.

[0291] Felinized anti-IL23 antibodies can also be expressed using a half-life-extended Fc by replacing the amino acid position 252 in the feline Fc region with a "Y" by EU numbering. This half-life-extending Fc region mutation is introduced as L252Y into the caninized anti-IL23 heavy chain IgG-B, resulting in a "long-acting" felinized anti-IL23.

[0292] Example 6: Caninization of anti-TNFαD2E7 antibody

[0293] This example illustrates caninization studies of anti-TNFα D2E7 having a VL domain of SEQ ID NO: 26 and a VH domain of SEQ ID NO: 30.

[0294] The resulting caninized anti-TNFα D2E7 VL domain sequence of SEQ ID NO: 34 and VH domain sequence of SEQ ID NO: 35 were used to generate a caninized anti-TNFα D2E7 antibody. A caninized anti-TNFα D2E7 antibody was generated comprising a caninized VL domain of SEQ ID NO: 34 linked to a canine kappa constant region to provide a light chain sequence of SEQ ID NO: 40, and a caninized VH domain of SEQ ID NO: 35 linked to a canine IgG-B constant region to provide a heavy chain sequence of SEQ ID NO: 41. This caninized anti-TNFα was expressed using mammalian Expi293F cells.

[0295] This example demonstrates that the caninized anti-TNFα antibodies described herein have high binding affinity to canine TNFα (Sino Biologicals).This study was performed to measure the binding affinity of the antibodies to the antigen using Biacore 8K.

[0296] Material

[0297] Table 10: Samples used

[0298]

[0299] Table 11: Instruments and reagents used

[0300]

[0301]

[0302] method

[0303] 1. Antigen-antibody affinity measurement

[0304] Preparation of running buffer: Dilute one volume of 10× buffer with 9 volumes of degassed, filtered MilliQ water.

[0305] Preparation of regeneration buffer (10 mM glycine): Weigh a certain amount of glycine and dissolve it in MilliQ water. Adjust the pH to 1.5-1.7 to obtain regeneration buffer.

[0306] The assay was performed at 25°C and the running buffer was HBS-EP+.

[0307] The antibody was injected onto the S-series sensor chip Protein A as the capture agent.

[0308] Antigen was diluted to various concentrations and injected onto the surface of flow cells 1 and 2 as the association phase, followed by injection of running buffer as the dissociation phase.

[0309] The binding model is shown below. The run configurations used are listed in Table 12 below.

[0310] Table 12: Run Configuration

[0311]

[0312] result

[0313] All data were processed using Biacore 8K Evaluation Software version 3.0. Flow cell 1 and buffer blank injections in each cycle were used as a double reference for response unit subtraction. Biacore binding affinity values ​​are provided in Table 13 below.

[0314] Table 13: Affinity measurements of antibodies and antigens

[0315]

[0316] The caninized version of the anti-TNFα D2E7 antibody comprises a light chain (LC) of SEQ ID NO: 40 having a caninized variable domain (VL) linked to a canine kappa constant region, and a heavy chain of SEQ ID NO: 43 having a caninized variable domain (VH) linked to a canine IgG-B constant region, further comprising an Fc region with a "long-acting" 252Y variant in the Fc region by EU numbering. A caninized anti-TNFα with long-acting Fc variants in the IgG-A, IgG-C, and IgG-D constant regions was also designed.

[0317] A feline version of the anti-TNFα D2E7 antibody was also designed, comprising the feline light chain variable domain (VL) of D2E7 (SEQ ID NO: 36) feline κ and the feline heavy chain variable domain (VH) of D2E7 (SEQ ID NO: 37), and feline IgGa or IgGb, wherein the Fc of canine IgG can be modified by replacing the "Y" in the Fc at amino acid position 252 according to EU numbering.

[0318] Example 7: Expression and purification of bispecific anti-canine IL23 and anti-canine TNFα molecules in CHO cells

[0319] Caninized versions of the anti-TNFα VH and VL domains provided in Table 1 can be used to form scFv antibodies. The sequence structures of two exemplary scFv anti-TNFα antibodies are illustrated in Table 1 by the amino acid sequences of SEQ ID NO: 45 or 46. The scFv antibody sequence of SEQ ID NO: 46 also includes a double Cys variant that allows disulfide bond formation.

[0320] Using the anti-TNFα scFv molecules and the caninized anti-IL23 antibodies described in Table 1, a first exemplary bispecific fusion molecule was designed that specifically binds to both canine IL23 and canine TNFα, having the following fusion structure: (1) a caninized anti-IL23 light chain of SEQ ID NO: 19 comprising a caninized variable domain (VL) linked to a canine kappa light chain constant region; (2) a caninized anti-IL23 heavy chain of SEQ ID NO: 20 comprising a caninized variable domain (VH) linked to a canine IgG-B constant region with a long-acting 252Y Fc region; and (3) a scFv caninized anti-TNFα antibody of SEQ ID NO: 45. The complete bispecific anti-IL23 / anti-TNFα antibody fusion structure is provided in Table 1 as the amino acid sequences of SEQ ID NOs: 47 and 48. The bispecific anti-IL23 / anti-TNFα antibody of SEQ ID NO: 48 includes the scFv anti-TNFα antibody of SEQ ID NO: 46, which has two cysteines to allow intramolecular SS bond formation. The resulting complete bispecific antibody fusion structure with the scFv of SEQ ID NO: 46 is provided as the amino acid sequence of SEQ ID NO: 48. The bispecific antibody molecules of SEQ ID NOs: 47 and 48 were expressed from mammalian cells and purified by single-step protein A column chromatography.

[0321] A second exemplary bispecific anti-IL23 / anti-TNFα antibody was designed that specifically binds to both canine IL23 and canine TNFα, having the following four-chain structure formed by combining: (1) an anti-IL23 antibody structure having a light chain of SEQ ID NO: 19 containing a caninized VH domain and a heavy chain of SEQ ID NO: 21 containing a caninized VH, which contains a "knob" in its heavy chain; and (2) an anti-TNFα antibody structure formed having a light chain of SEQ ID NO: 41 containing a caninized VH domain and a heavy chain of SEQ ID NO: 44 containing a caninized VH, which contains a "hole" in its heavy chain. In addition, as shown in the sequences in Table 1, the anti-TNFα LC of SEQ ID NO: 41 and the anti-TNFα HC of SEQ ID NO: 44 were engineered as follows: (1) in the anti-TNFα light chain of SEQ ID NO: 41, one cysteine ​​was removed from the CL region and one cysteine ​​was added to the VL domain; and (2) in the anti-TNFα heavy chain of SEQ ID NO: 44, one cysteine ​​was removed from the CH1 region and one cysteine ​​was added to the VH domain. After co-transfection into CHO cells, the plasmids encoding the four chains of the bispecific anti-IL23 / anti-TNFα antibody expressed very well and monodisperse. Mass spectrometry analysis of the obtained deglycosylated and DTT-reduced proteins indicated the expected 1:1:1:1 ratio.

[0322] Example 8: Study on the Treatment of Canine IBD with Anti-IL23 Antibodies

[0323] Various doses ranging from 0.01 mg / kg to 100 mg / kg can be used to evaluate the efficacy and safety of anti-IL23 in managing inflammatory bowel disease (IBD) in dogs.

[0324] All dogs received doses of caninized IL23 antibody or caninized long-acting antibody, with the first dose administered on day 0. Subsequent doses were administered weekly, biweekly, monthly, or every 2 months, every 3 months, or up to every 6 months.

[0325] Histopathology of endoscopic gastrointestinal biopsies can be used to determine the effectiveness of canine IL23 antibodies in the management of IBD.

[0326] Measurable IBD biomarkers include, but are not limited to, IL-1β, IL6, IL8, IL9, IFN-γ, TNFα, CCL2, IL22, CRP, LL37, TFF3, and OSM.

[0327] The Canine Inflammatory Bowel Disease Activity Index (CIBDAI) score can be used to screen therapeutic candidates.

[0328] Control conditions may include diet.

[0329] Example 9: Study of Canine IBD Treated with Combination Therapy Using Anti-IL23 and Anti-TNFα Antibodies

[0330] Various doses of IL23 mAb (ranging from 0.01 mg / kg to 100 mg / kg) in any combination with TNFα mAb (ranging from 0.01 mg / kg to 100 mg / kg) can be used to evaluate the efficacy and safety of IL23 mAb in combination with TNFα mAb for the management of inflammatory bowel disease (IBD) in dogs.

[0331] All dogs received doses of a combination of caninized IL23 antibody or caninized IL23 antibody long-acting antibody and caninized TNFα antibody or caninized IL23 antibody long-acting antibody, with the first dose administered on day 0. Subsequent doses were administered weekly, biweekly, monthly, or every 2 months, every 3 months, for up to six months or longer.

[0332] Alternatively, various doses ranging from 0.01 mg / kg to 100 mg / kg can be used to evaluate the efficacy and safety of the IL23 / TNFα bispecific antibody for treating or managing inflammatory bowel disease (IBD) in dogs.

[0333] Histopathology of endoscopic gastrointestinal biopsies can be used to determine the effectiveness of canine IL23 antibodies in the management of IBD.

[0334] Measurable IBD biomarkers include, but are not limited to, IL-1β, IL6, IL8, IL9, IFN-γ, TNFα, CCL2, IL22, CRP, LL37, TFF3, and OSM.

[0335] The Canine Inflammatory Bowel Disease Activity Index (CIBDAI) score can be used to screen therapeutic candidates.

[0336] Control conditions may include diet.

Claims

1. An anti-IL23 antibody that binds to canine, feline and / or equine IL23, comprising (i) a first light chain hypervariable region (HVR-L1), a second light chain hypervariable region (HVR-L2), and a third light chain hypervariable region (HVR-L3), and / or (ii) a first heavy chain hypervariable region (HVR-H1), a second heavy chain hypervariable region (HVR-H2), and a third heavy chain hypervariable region (HVR-H3), wherein: (a) the HVR-L1 region comprises the amino acid sequence of RASQGISSWLA (SEQ ID NO:4), the HVR-L2 region comprises the amino acid sequence of YAASSLQS (SEQ ID NO:5), and the HVR-L3 region comprises the amino acid sequence of QQYNIYPYT (SEQ ID NO:6); and / or (b) the HVR-H1 region comprises the amino acid sequence of KGSGYSFTTYWLG (SEQ ID NO: 8), the HVR-H2 region comprises the amino acid sequence of IMSPVDSDIR (SEQ ID NO: 9), and the HVR-H3 region comprises the amino acid sequence of ARRRPGQGYFDF (SEQ ID NO: 10).

2. The antibody of claim 1, wherein the antibody is caninized, felineized, or equinized.

3. The antibody of claim 1, wherein the antibody comprises a light chain variable domain (VL) amino acid sequence that is at least 90% identical to a sequence selected from SEQ ID NOs: 3, 11, 13, and 15; and / or a heavy chain variable domain (VH) amino acid sequence that is at least 90% identical to a sequence selected from SEQ ID NOs: 7, 12, 14, and 16; optionally, wherein: (i) the light chain variable domain (VL) comprises a variant of SEQ ID NO: 3, 11, 13 and 15, wherein 1 to 6 amino acids of the light chain variable domain (VL) are substituted by different amino acids; and / or (ii) the heavy chain variable domain (VH) comprises a variant of SEQ ID NO: 7, 12, 14 and 16, wherein 1 to 6 amino acids of the heavy chain variable domain (VH) are substituted by different amino acids.

4. The antibody of claim 1, wherein the antibody comprises a light chain variable domain (VL) and / or a heavy chain variable domain (VH), wherein the light chain variable domain comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 3, 11, 13 and 15, and the heavy chain variable domain comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 7, 12, 14 and 16; optionally, wherein: (i) the antibody comprises a light chain variable domain (VL) and a heavy chain variable domain (VH), the light chain variable domain comprising an amino acid sequence selected from SEQ ID NO: 3, and the heavy chain variable domain comprising an amino acid sequence selected from SEQ ID NO: 7; (ii) the antibody comprises a light chain variable domain (VL) and a heavy chain variable domain (VH), the light chain variable domain comprising an amino acid sequence selected from SEQ ID NO: 11, and the heavy chain variable domain comprising an amino acid sequence selected from SEQ ID NO: 12; (iii) the antibody comprises a light chain variable domain (VL) comprising an amino acid sequence selected from SEQ ID NO: 13 and a heavy chain variable domain (VH) comprising an amino acid sequence selected from SEQ ID NO: 14; or (iv) the antibody comprises a light chain variable domain (VL) and a heavy chain variable domain (VH), the light chain variable domain comprising an amino acid sequence selected from SEQ ID NO: 15, and the heavy chain variable domain comprising an amino acid sequence selected from SEQ ID NO:

16.

5. The antibody of any one of claims 1 to 4, wherein the antibody comprises: a light chain (LC) amino acid sequence that is at least 90% identical to an amino acid sequence selected from SEQ ID NOs: 17, 19, 22, and 24, and / or a heavy chain (HC) amino acid sequence that is at least 90% identical to SEQ ID NOs: 18, 20, 21, 23, and 25; optionally, wherein the antibody comprises: (i) the LC amino acid sequence of SEQ ID NO: 17 and the HC amino acid sequence of SEQ ID NO: 18; (ii) the LC amino acid sequence of SEQ ID NO: 19 and the HC amino acid sequence of SEQ ID NO: 20; (iii) the LC amino acid sequence of SEQ ID NO: 19 and the HC amino acid sequence of SEQ ID NO: 21; (iv) the LC amino acid sequence of SEQ ID NO: 22 and the HC amino acid sequence of SEQ ID NO: 23; or (v) The LC amino acid sequence of SEQ ID NO: 24 and the HC amino acid sequence of SEQ ID NO:

25.

6. An anti-TNFα antibody that binds to canine, feline and / or equine TNFα, comprising (i) a first light chain hypervariable region (HVR-L1), a second light chain hypervariable region (HVR-L2), and a third light chain hypervariable region (HVR-L3), and / or (ii) a first heavy chain hypervariable region (HVR-H1), a second heavy chain hypervariable region (HVR-H2), and a third heavy chain hypervariable region (HVR-H3), wherein: (a) the HVR-L1 region comprises the amino acid sequence of RASQGIRNYLA (SEQ ID NO: 27), the HVR-L2 region comprises the amino acid sequence of AASTLQ (SEQ ID NO: 28), and the HVR-L3 comprises the amino acid sequence of QRYNRAPYT (SEQ ID NO: 29); and / or (a) The HVR-H1 region comprises the amino acid sequence of FTFDDYAMH (SEQ ID NO: 31), the HVR-H2 region comprises the amino acid sequence of AITWNSGHIDYADSVEGR (SEQ ID NO: 32), and the HVR-H3 region comprises the amino acid sequence of AKVSYLSTASSLDY (SEQ ID NO: 33).

7. The antibody of claim 6, wherein the antibody is caninized, felineized, or equinized.

8. The antibody of claim 6, wherein the antibody comprises a light chain variable domain (VL) amino acid sequence that is at least 90% identical to a sequence selected from SEQ ID NOs: 26, 34, and 36; and / or a heavy chain variable domain (VH) amino acid sequence that is at least 90% identical to a sequence selected from SEQ ID NOs: 30, 35, and 37; optionally, wherein: (i) the light chain variable domain (VL) comprises a variant of SEQ ID NO: 26, 34 and 36, wherein 1 to 6 amino acids of the light chain variable domain (VL) are substituted by different amino acids; and / or (ii) the heavy chain variable domain (VH) comprises a variant of SEQ ID NO: 30, 35 and 37, wherein 1 to 6 amino acids of the heavy chain variable domain (VH) are substituted by different amino acids.

9. The antibody of claim 6, wherein the antibody comprises a light chain variable domain (VL) and / or a heavy chain variable domain (VH), the light chain variable domain comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 26, 34, and 36, and the heavy chain variable domain comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 30, 35, and 37; optionally, wherein the antibody comprises: (i) a light chain variable domain (VL) comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 26 and a heavy chain variable domain (VH) comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 30; (ii) a light chain variable domain (VL) comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 34 and a heavy chain variable domain (VH) comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 35; (iii) a light chain variable domain (VL) comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 36 and a heavy chain variable domain (VH) comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 37; (iv) a light chain variable domain (VL) comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 36 and a heavy chain variable domain (VH) comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 37; (v) a light chain variable domain (VL) comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 36 and a heavy chain variable domain (VH) comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 30; or (vi) a light chain variable domain (VL) comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 26 and a heavy chain variable domain (VH) comprising an amino acid sequence selected from the group consisting of SEQ ID NO:

37.

10. The antibody of any one of claims 6 to 9, wherein the antibody comprises: a light chain (LC) amino acid sequence that is at least 90% identical to a sequence selected from SEQ ID NOs: 38, 40, and 41 and / or a heavy chain (HC) amino acid sequence that is at least 90% identical to a sequence selected from SEQ ID NOs: 39, 42, 43, and 44; optionally, wherein the antibody comprises: (i) the LC amino acid sequence of SEQ ID NO: 38 and the HC amino acid sequence of SEQ ID NO: 39; (ii) the LC amino acid sequence of SEQ ID NO:40 and the HC amino acid sequence of SEQ ID NO:42; (iii) the LC amino acid sequence of SEQ ID NO:41 and the HC amino acid sequence of SEQ ID NO:42; (iv) the LC amino acid sequence of SEQ ID NO:40 and the HC amino acid sequence of SEQ ID NO:43; (v) the LC amino acid sequence of SEQ ID NO:41 and the HC amino acid sequence of SEQ ID NO:43; (vi) the LC amino acid sequence of SEQ ID NO:40 and the HC amino acid sequence of SEQ ID NO:44; or (vi) the LC amino acid sequence of SEQ ID NO:41 and the HC amino acid sequence of SEQ ID NO:

44.

11. The antibody of any one of claims 6 to 9, wherein the antibody is a scFv antibody; optionally, wherein the scFv antibody comprises an amino acid sequence having at least 90% sequence identity to a sequence selected from SEQ ID NOs: 45 and 46.

12. The antibody of any one of claims 1 to 11, wherein the antibody is an antibody fragment selected from the group consisting of Fv, scFv, Fab, Fab', F(ab')2 and Fab'-SH.

13. The antibody of any one of claims 1 to 11, wherein the antibody comprises a canine heavy chain constant region selected from the group consisting of IgG-A, IgG-B, IgG-C, and IgG-D constant regions.

14. The antibody of any one of claims 1 to 13, wherein the antibody comprises: (i) a canine light chain constant region and / or a canine heavy chain constant region; (ii) a feline light chain constant region and / or a feline heavy chain constant region; or (iii) equine light chain constant region and / or equine heavy chain constant region.

15. The antibody of any one of claims 1 to 14, wherein the antibody comprises a heavy chain constant region having a "Y" mutation at position 252 according to EU numbering.

16. A bispecific antibody that binds to canine, feline and / or equine IL23 and canine, feline and / or equine TNFα, wherein the antibody comprises: (a) an anti-IL23 light chain (LC) comprising a VL domain having an HVR-L1 region comprising the amino acid sequence of RASQGISSWLA (SEQ ID NO: 4), an HVR-L2 region comprising the amino acid sequence of YAASSLQS (SEQ ID NO: 5), and an HVR-L3 region comprising the amino acid sequence of QQYNIYPYT (SEQ ID NO: 6); and an anti-IL23 heavy chain (HC) comprising a VH domain having an HVR-H1 region comprising the amino acid sequence of KGSGYSFTTYWLG (SEQ ID NO: 8), an HVR-H2 region comprising the amino acid sequence of IMSPVDSDIR (SEQ ID NO: 9), and an HVR-H3 region comprising the amino acid sequence of ARRRPGQGYFDF (SEQ ID NO: 10). NO: 10); and an anti-TNFα scFv antibody fused to the HC, wherein the anti-TNFα scFv antibody comprises a VL domain and a VH domain, the VL domain having an HVR-L1 region comprising the amino acid sequence of RASQGIRNYLA (SEQ ID NO: 27), an HVR-L2 region comprising the amino acid sequence of AASTLQ (SEQ ID NO: 28), and an HVR-L3 comprising the amino acid sequence of QRYNRAPYT (SEQ ID NO: 29); and the VH domain having an HVR-H1 region comprising the amino acid sequence of FTFDDYAMH (SEQ ID NO: 31), an HVR-H2 region comprising the amino acid sequence of AITWNSGHIDYADSVEGR (SEQ ID NO: 32), and an HVR-H3 region comprising the amino acid sequence of AKVSYLSTASSLDY (SEQ ID NO: 33); or (b) an anti-TNFα light chain (LC) comprising a VL domain having an HVR-L1 region comprising the amino acid sequence of RASQGIRNYLA (SEQ ID NO: 27), an HVR-L2 region comprising the amino acid sequence of AASTLQ (SEQ ID NO: 28), and an HVR-L3 region comprising the amino acid sequence of QRYNRAPYT (SEQ ID NO: 29), and an anti-TNFα heavy chain (HC) comprising a VH domain having an HVR-H1 region comprising the amino acid sequence of FTFDDYAMH (SEQ ID NO: 31), an HVR-H2 region comprising the amino acid sequence of AITWNSGHIDYADSVEGR (SEQ ID NO: 32), and an HVR-H3 region comprising the amino acid sequence of AKVSYLSTASSLDY (SEQ ID NO: 33), and an anti-IL23 scFv antibody fused to the HC, wherein the anti-IL23 The scFv antibody comprises a VL domain and a VH domain, wherein the VL domain has an HVR-L1 region comprising the amino acid sequence of RASQGISSWLA (SEQ ID NO:4), an HVR-L2 region comprising the amino acid sequence of YAASSLQS (SEQ ID NO:5), and an HVR-L3 region comprising the amino acid sequence of QQYNIYPYT (SEQ ID NO:6); the VH domain has an HVR-H1 region comprising the amino acid sequence of KGSGYSFTTYWLG (SEQ ID NO:8), an HVR-H2 region comprising the amino acid sequence of IMSPVDSDIR (SEQ ID NO:9), and an HVR-H3 region comprising the amino acid sequence of ARRRPGQGYFDF (SEQ ID NO:10).

17. The bispecific antibody according to claim 16, wherein: (a) the anti-IL23 light chain (LC) comprises a light chain variable domain (VL) amino acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 3, 11, 13, and 15, and the anti-IL23 heavy chain (HC) comprises a heavy chain variable domain (VH) amino acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 7, 12, 14, and 16, wherein the HC is fused to an anti-TNFα scFv antibody comprising a light chain variable domain (VL) amino acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 26, 34, and 36, and a heavy chain variable domain (VH) amino acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 30, 35, and 37; or (b) the anti-TNFα light chain (LC) comprises a light chain variable domain (VL) amino acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 26, 34, and 36, and the anti-TNFα heavy chain (HC) comprises a heavy chain variable domain (VH) amino acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 30, 35, and 37, wherein the HC is fused to an anti-IL23 scFv antibody comprising a light chain variable domain (VL) amino acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 3, 11, 13, and 15, and a heavy chain variable domain (VH) amino acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 7, 12, 14, and 16.

18. The bispecific antibody of any one of claims 16 to 17, wherein the light chain (LC) comprises an amino acid sequence that is at least 90% identical to a sequence selected from SEQ ID NOs: 19 and 22; and the heavy chain (HC) comprises an amino acid sequence that is at least 90% identical to a sequence selected from SEQ ID NOs: 47 and 48; optionally, wherein the antibody comprises: (i) the LC amino acid sequence of SEQ ID NO: 19 and the HC amino acid sequence of SEQ ID NO: 47; (ii) the LC amino acid sequence of SEQ ID NO: 19 and the HC amino acid sequence of SEQ ID NO: 48; (iii) the LC amino acid sequence of SEQ ID NO: 22 and the HC amino acid sequence of SEQ ID NO: 47; or (iv) the LC amino acid sequence of SEQ ID NO: 22 and the HC amino acid sequence of SEQ ID NO:

48.

19. A bispecific antibody that binds to canine, feline and / or equine IL23 and canine, feline and / or equine TNFα, wherein the antibody comprises: (i) an anti-IL23 light chain (LC) comprising a VL domain having an HVR-L1 region comprising the amino acid sequence of RASQGISSWLA (SEQ ID NO:4), an HVR-L2 region comprising the amino acid sequence of YAASSLQS (SEQ ID NO:5), and an HVR-L3 region comprising the amino acid sequence of QQYNIYPYT (SEQ ID NO:6); (ii) an anti-IL23 heavy chain (HC) comprising a VH domain having an HVR-H1 region comprising the amino acid sequence of KGSGYSFTTYWLG (SEQ ID NO: 8), an HVR-H2 region comprising the amino acid sequence of IMSPVDSDIR (SEQ ID NO: 9), and an HVR-H3 region comprising the amino acid sequence of ARRRPGQGYFDF (SEQ ID NO: 10); (iii) an anti-TNFα light chain (LC) comprising a VL domain having an HVR-L1 region comprising the amino acid sequence of RASQGIRNYLA (SEQ ID NO:27), an HVR-L2 region comprising the amino acid sequence of AASTLQ (SEQ ID NO:28), and an HVR-L3 comprising the amino acid sequence of QRYNRAPYT (SEQ ID NO:29); and (iv) an anti-TNFα heavy chain (HC) comprising a VH domain having an HVR-H1 region comprising the amino acid sequence of FTFDDYAMH (SEQ ID NO: 31), an HVR-H2 region comprising the amino acid sequence of AITWNSGHIDYADSVEGR (SEQ ID NO: 32), and an HVR-H3 region comprising the amino acid sequence of AKVSYLSTASSLDY (SEQ ID NO: 33).

20. The bispecific antibody of claim 19, wherein: (i) the anti-IL23 light chain (LC) comprises a light chain variable domain (VL) amino acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 3, 11, 13, and 15; (ii) the anti-IL23 heavy chain (HC) comprises a heavy chain variable domain (VH) amino acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 7, 12, 14, and 16; (iii) the anti-TNFα light chain (LC) comprises a light chain variable domain (VL) amino acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 26, 34, and 36; and (iv) the anti-TNFα heavy chain (HC) comprises a heavy chain variable domain (VH) amino acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 30, 35, and 37.

21. The bispecific antibody according to any one of claims 19 to 20, wherein: (i) the anti-IL23 light chain (LC) comprises an amino acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 19 and 22; (ii) the anti-IL23 heavy chain (HC) comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NOs: 20, 21 and 23; (iii) the anti-TNFα light chain (LC) comprises an amino acid sequence that is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 40 and 41; and (iv) the anti-TNFα heavy chain (HC) comprises an amino acid sequence having at least 90% sequence identity with SEQ ID NOs: 42, 43 and 44.

22. An isolated nucleic acid or vector encoding the antibody of any one of claims 1 to 21.

23. An isolated host cell comprising the nucleic acid or vector of claim 22.

24. A method of producing an antibody, comprising culturing the host cell of claim 23 and isolating the antibody.

25. A pharmaceutical composition comprising the antibody of any one of claims 1 to 21 and a pharmaceutically acceptable carrier.

26. A method of treating a dog, cat or horse having a disorder associated with IL23, the method comprising administering to the dog, cat or horse a therapeutically effective amount of the antibody of any one of claims 1 to 21 or the pharmaceutical composition of claim 25.

27. A method of maintaining remission of an IL23-associated disorder in a dog, cat or horse, the method comprising administering to the dog, cat or horse a therapeutically effective amount of the antibody of any one of claims 1 to 21 or the pharmaceutical composition of claim 25.

28. The method of any one of claims 26 to 27, wherein the IL23-related disorder is an inflammatory disease.

29. The method of any one of claims 26 to 28, wherein the IL23-related disorder is a gastrointestinal inflammatory disease.

30. The method of any one of claims 26 to 29, wherein the IL23-related disorder is inflammatory bowel disease.

31. The method of any one of claims 26 to 30, wherein the IL23-associated condition is ankylosing spondylitis, asthma, cancer, Crohn's disease, idiopathic arthritis, psoriasis, plaque psoriasis, psoriatic arthritis, rheumatoid arthritis, osteoarthritis, or ulcerative colitis.

32. A method of treating a dog, cat or horse having a condition associated with IL23 and TNFα, the method comprising administering to the dog, cat or horse a therapeutically effective amount of an IL23 antibody according to any one of claims 1 to 5 or 12 to 21, and a TNFα antibody according to any one of claims 6 to 21, or a pharmaceutical composition according to claim 25.

33. A method of treating a dog, cat or horse having a disorder associated with IL23 / TNFα, said method comprising administering to said dog, cat or horse a therapeutically effective amount of the bispecific IL23 / TNFα antibody of any one of claims 16 to 21.

34. The method of any one of claims 26 to 33, wherein the antibody or the pharmaceutical composition is administered parenterally.

35. The method of any one of claims 26 to 34, wherein the antibody or the pharmaceutical composition is administered by intramuscular, intraperitoneal, cerebrospinal, subcutaneous, intraarterial, intrasynovial, intrathecal, or inhalational routes.

36. The method of any one of claims 26 to 35, wherein the method further comprises administering an IL17 antibody, an IL-5 antibody, an IL-31 antibody, an IL4 antibody, an IL13 antibody, an IL23 antibody, an IgE antibody, a CD11α antibody, an IL6R antibody, an α4-integrin antibody, a β7-integrin antibody, an IL12 antibody, an IL1β antibody, or an anti-BlyS antibody.

37. The method of any one of claims 23 to 33, wherein the antibody is administered in an amount ranging from 0.01 mg / kg to 100 mg / kg body weight per dose.

38. A method for reducing IL23 and / or TNFα signaling function in a cell, the method comprising exposing the antibody according to any one of claims 1 to 21 to the cell under conditions that allow the antibody to bind to IL23 and / or TNFα, thereby reducing the binding of the cell to IL23 and / or TNFα signaling function.

39. The method of claim 38, wherein the cell is exposed to the antibody or the pharmaceutical composition in vivo.

40. The method of any one of claims 38 to 39, wherein the cell is a canine cell, a feline cell, or a horse cell.

41. A method for detecting IL23 and / or TNFα in a sample from a companion animal species, comprising contacting the sample with an antibody as described in any one of claims 1 to 21 under conditions permissive for binding of the antibody to IL23 and / or TNFα, and detecting whether a complex is formed between the antibody and IL23 and / or TNFα in the sample.

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