Anti-IL23 and anti-TNFα antibodies: compositions and veterinary uses

Abstract

Various embodiments are provided relating to caninized, feline, or equine antibodies that bind canine, feline, and / or equine IL23 and / or TNFα, including bispecific antibodies that bind both IL23 and TNFα. Such antibodies may be used alone or in combination in methods for treating canine, feline, and / or equine subjects with inflammatory conditions, such as inflammatory bowel disease (IBD), osteoarthritis, and inflammatory conditions such as gastroenteritis in dogs and cats.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 387,777, filed December 16, 2022, which is incorporated herein by reference in its entirety.

[0002] Field The present disclosure relates to caninized, felineized, and equineized anti-IL23 and anti-TNFα antibodies and their uses, for example, to treat IL23-mediated and / or TNFα-mediated disorders in dogs, cats, or horses, such as chronic bowel diseases - inflammatory bowel disease (IBD), psoriasis, rheumatoid arthritis, osteoarthritis, sepsis, and multiple sclerosis (MS).

[0003] Sequence Listing Reference A copy of the official sequence listing has been submitted contemporaneously herewith via the USPTO Patent Center as an XML file in WIPO standard ST.26 format with the filename "20136-001WO1.xml," created on December 13, 2023, and 53,419 bytes in size. This sequence listing submitted via the USPTO Patent Center is a part of the present specification and is incorporated herein by reference in its entirety. [Background technology]

[0004] background 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 a proinflammatory cytokine that is a member of the IL-12 cytokine family. It has been shown to be a key cytokine in the maintenance and proliferation of type 17 helper T cells (Th17 cells). IL23 is associated with many chronic inflammatory disorders in humans, including diseases such as psoriasis, rheumatoid arthritis, psoriatic arthritis, osteoarthritis, multiple sclerosis, and inflammatory bowel disease (IBD).

[0005] Like humans, companion animals, such as felines, canines, and equines, suffer from chronic inflammatory disorders. However, to date, the use of anti-IL23 antibodies to treat such diseases in companion animals has not been demonstrated. Furthermore, proteins containing significant amounts of human-derived amino acid sequences may be immunogenic in non-human animals and may not bind to companion animal IL23 in a manner that provides an equally beneficial therapeutic effect in companion animals. See Mauldin et al., Aug. 2010, 21(4):373-382.

[0006] Inflammatory bowel disease (IBD) is a general term used to describe disorders associated with chronic inflammation of the gastrointestinal tract. The exact etiology of companion animal IBD remains unknown, but changes in the immune system's tolerance to dietary antigens, the gut microbiome, and genetic susceptibility may all play a role. IBD in dogs, cats, and horses may share a similar etiology with disorders occurring in humans (also known as Crohn's disease and ulcerative colitis), although the clinical syndrome and histological changes may differ slightly. Canine IBD is characterized by persistent or recurrent symptoms such as vomiting, diarrhea, abdominal pain, weight loss, and / or changes in appetite, as well as inflammation of the gastrointestinal tract. (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." J Anim Sci. 2010; 89(5): 1520-1530. Feline IBD is characterized by inflammation of the gastrointestinal tract (most commonly the small intestine), as well as vomiting, diarrhea, and weight loss. (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).Equine IBD 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).

[0007] Accurate diagnosis of companion animal IBD can be difficult, and the term "idiopathic" is often used when the exact causative agent cannot be identified. A medical history and physical examination, followed by laboratory tests, imaging, and intestinal biopsies (to demonstrate the presence of inflammation) are typically recommended. The prognosis and response to conventional therapy for IBD in dogs, cats, and horses varies and can range from moderate to poor. However, studies evaluating cytokines in companion animals with IBD have shown similar upregulated cytokines to those seen in humans, such as IL-23 and TNFα (Jergens, AE, & Simpson, KW (2012). Inflammatory bowel disease in veterinary medicine. Frontiers in bioscience (Elite edition), 4(4), 1404-1419. https: / / doi.org / 10.2741 / e470, 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, Cerquetella, M., Spaterna, A., Laus, F., Tesei, B., Rossi, G., Antonelli, E., Villanacci, V., & Bassotti, G. (2010). Inflammatory bowel disease in the dog: differences and similarities with humans. World journal of gastroenterology, 16(9), 1050-1056. https: / / doi.org / 10.3748 / wjg.v16.i9.1050, Olofsson, KM, Hjertner, B., Fossum, C., Press, CM, & 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 biologics frequently prescribed and successfully used to treat IBD in humans are unavailable for companion animals, and treatment options remain limited.

[0008] Gastroenteritis in humans and companion animals is associated with inflammation of the gastrointestinal tract. Canine and feline gastroenteritis often results in diarrhea and vomiting. Similar to IBD, studies evaluating gastroenteritis have found the involvement of cytokines involved in inflammation, such as IL-23 and TNFα (Godinez, I., Keestra, AM, Spees, A., & Baumler, AJ (2011). The IL-23 axis in Salmonella gastroenteritis. Cellular microbiology, 13(11), 1639-1647; Munoz-Cruz, S., Gomez-Garcia, A., Millan-Ibarra, J., Giono-Cerezo, S., & Yepez-Mulia, L. (2010). Giardia lamblia: interleukin 6 and tumor necrosis factor-alpha release from mast cells induced through an Ig-independent pathway. Experimental parasitology, 126(3), 298-303. https: / / doi.org / 10.1016 / j.exppara.2010.06.013), suggesting that therapeutic agents targeting IL-23 and TNFα cytokines may aid in the treatment of gastroenteritis (Zhou, P., Li, E., SHEA-DONOHUE, T., & Singer, SM (2007). Tumor necrosis factor α contributes to protection against Giardia lamblia infection in mice. Parasite immunology, 29(7), 367-374). Current therapeutic agents for gastroenteritis in companion animals are limited to electrolytes and antibiotics, and additional compounds that address inflammation of the gastrointestinal tract may be beneficial for treatment.

[0009] In neonatal equine foals, sepsis is a systemic inflammatory response syndrome (SIRS) resulting from a failure of passive transmission, and foals may present 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). Survival rates for neonatal sepsis are poor, with a survival rate of 45-60% (Taylor S. (2015). A review of equine sepsis. Equine veterinary education, 27(2), 99-109. https: / / doi.org / 10.1111 / eve.12290). Inflammatory cytokines such as TNFα and other cytokines downstream of IL-23 have been found to be present at elevated levels in septic foals and are directly associated with the development of multiple organ dysfunction syndrome (Taylor S. (2015). A review of equine sepsis. Equine veterinary education, 27(2), 99-109. https: / / doi.org / 10.1111 / eve.12290). To date, there are no compounds for treating sepsis, and compounds targeting IL-23 and TNFα cytokines may be beneficial in treating sepsis.

[0010] It contains a large amount of TNFα In the case of IL-23 snowflakes (Smo len , JS , Agarwal , SK , Ilivanova , E. , Xu , XL , Miao , Y. , Zhuang , Y. ,& Baker,D.(2017).A randomized 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, MS, Talamonti, M., Novelli, L., Teoli, M., Galluzzo, M., Triggianese, P., & Perricone, R. (2015). Avila Machado, MA, Maciel, AA, de Lemos, LLP, Costa, JO, Kakehasi, AM, Andrade, EIG,...& de Assis Acurcio,F.(2013).Adalimumab in the treatment of rheumatoid arthritis:a systematic review and meta-analysis of randomized clinical trials.Revista Brasileira de Reumatologia (English Edition), 53(5), 419-430., Schiff, MH, Burmester, GR, Kent, JD, Pangan, AL, Kupper, H., Fitzpatrick, SB, & Donovan, C. (2006). Safety analyzes of adalimumab (HUMIRA) in global clinical trials and US postmarketing surveillance of patients with rheumatoid arthritis. Annals of the rheumatic diseases, 65(7), 889-894. https: / / doi.org / 10.1136 / ard.2005.043166). However, recent studies of osteoarthritis in humans have shown that both TNFα and IL-23 are present in the synovial fluid of patients with osteoarthritis (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, Homayounfar, R., Shahi, A., Afsarian, MH, Paknahad, A., Kennedy, D., & Ataollahi, MR (2016). Increased Serum Levels of IL-17A and IL-23 Are Associated with Decreased Vitamin D3 and Increased Pain in Osteoarthritis. PloS one,11(11),e0164757.https: / / doi.org / 10.1371 / journal.pone.0164757, Scanzello, C. R., Umoh, E., Pessler, F., Diaz-Torne, C., Miles, T., Dicarlo, E., & Crow, M. K. (2009).Local cytokine profiles in knee osteoarthritis: Elevated synovial fluid interleukin-15 differentiates early from end-stage disease. Osteoarthritis and cartilage, 17(8), 1040-1048. Because osteoarthritis is characterized by joint inflammation, evidence of these pro-inflammatory cytokines in synovial fluid is not surprising. Like humans, companion animals often develop osteoarthritis as they age. As in humans, osteoarthritis results in pain, joint swelling, laxity, stiffness, or refusal to move, but only a few symptoms (Brown, DC (2017). What can we learn from osteoarthritis pain in companion animals. Clin. Exp. Rheumatol, 35(Suppl 107), 53-58). Medications for treating companion animals with osteoarthritis include anti-inflammatory drugs; however, these can put a strain on the kidneys of older animals. Compounds that target IL-23 and TNFα cytokines may be safer and more effective treatments for osteoarthritis in companion animals. Therefore, there remains a need for methods and compounds that can be used to bind companion animal IL23 in companion animals to treat IL23-associated conditions in companion animals. Ideally, such compounds would specifically bind to companion animal IL23, have a plasma half-life long enough to make therapy feasible, but not be highly immunogenic in companion animals. [Prior art documents] [Non-patent literature]

[0011] [Non-Patent Document 1] Mauldin et al.,Aug.2010,21(4):373-382 [Non-patent document 2] 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 [Non-patent document 3] Suchodolski JS. “Companion-animals symposium: Microbes and gastrointestinal health of dogs and cats.” J Anim Sci.2010;89(5):1520-1530 [Non-patent document 4] 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 [Non-patent document 5] 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 [Non-patent document 6] Jergens,AE,&Simpson,KW(2012).Frontiers in Bioscience(Elite Edition),4(4),1404-1419.https: / / doi.org / 10.2741 / e470

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[0012] The present disclosure relates generally to caninized, felineized, and equineized antibodies that specifically bind IL23 and / or TNFα, and the use of these antibodies in compositions and methods for treating inflammatory conditions, such as inflammatory bowel disease (IBD), osteoarthritis, and gastroenteritis, in dogs, cats, and horses. This Summary is intended to introduce the subject matter of the present disclosure, but does not exhaustively describe each and every embodiment, combination, or variation contemplated and described within the scope of the present disclosure. Further embodiments are contemplated and described by the disclosure in the detailed description, drawings, and claims.

[0013] 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 a bispecific antibody capable of binding to both canine IL23 and canine TNFα (or both feline IL23 and feline TNFα), as well as methods of using such bispecific antibodies in treatment methods in dogs or cats (e.g., treating IBD, sepsis, and gastroenteritis). The present disclosure provides methods and compositions for combination treatment with anti-TNFα antibodies for IBD in companion animals, and thus relates to the fields of biology, molecular biology, and veterinary medicine.

[0014] Various embodiments provided by the present disclosure include, but are not limited to: 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 RASQGISSWLA (SEQ ID NO: 4); the HVR-L2 region comprises the amino acid sequence YAASSLQS (SEQ ID NO: 5) and the HVR-L3 region comprises the amino acid sequence QQYNIYPYT (SEQ ID NO: 6); and / or (b) the HVR-H1 region comprises the amino acid sequence KGSGYSFTTYWLG (SEQ ID NO: 8), the HVR-H2 region comprises the amino acid sequence IMSPVDSDIR (SEQ ID NO: 9), and the HVR-H3 region comprises the amino acid sequence ARRRPGQGYFDF (SEQ ID NO: 10). Embodiment 2. The antibody of embodiment 1, wherein the antibody is caninized, felineized, or equineized. Embodiment 3. The antibody of any one of embodiments 1-2, wherein the antibody comprises a light chain variable domain (VL) amino acid sequence having at least 90% identity to a sequence selected from SEQ ID NOs: 3, 11, 13, and 15, and / or a heavy chain variable domain (VH) amino acid sequence having at least 90% identity to a sequence selected from SEQ ID NOs: 7, 12, 14, and 16, and optionally (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. Embodiment 4. The antibody of any one of embodiments 1-3, wherein the antibody comprises a light chain variable domain (VL) comprising an amino acid sequence selected from SEQ ID NOs: 3, 11, 13, and 15, and / or a heavy chain variable domain (VH) comprising an amino acid sequence selected from SEQ ID NOs: 7, 12, 14, and 16, optionally wherein (i) the antibody comprises a light chain variable domain (VL) comprising an amino acid sequence selected from SEQ ID NO: 3 and a heavy chain variable domain (VH) comprising an amino acid sequence selected from SEQ ID NO: 7, (ii) the antibody comprises a light chain variable domain (VL) comprising an amino acid sequence selected from SEQ ID NO: 11 and a heavy chain variable domain (VH) 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) comprising an amino acid sequence selected from SEQ ID NO: 15 and a heavy chain variable domain (VH) comprising an amino acid sequence selected from SEQ ID NO: 16. Embodiment 5. The antibody of any one of embodiments 1-4, wherein the antibody comprises a light chain (LC) amino acid sequence having at least 90% identity to an amino acid sequence selected from SEQ ID NOs: 17, 19, 22, and 24, and / or a heavy chain (HC) amino acid sequence having at least 90% identity to an amino acid sequence selected from SEQ ID NOs: 18, 20, 21, 23, and 25, and optionally 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. Embodiment 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 RASQGIRNYLA (SEQ ID NO: 27). the HVR-L2 region comprises the amino acid sequence AASTLQ (SEQ ID NO: 28) and the HVR-L3 region comprises the amino acid sequence QRYNRAPYT (SEQ ID NO: 29); and / or (a) the HVR-H1 region comprises the amino acid sequence FTFDDYAMH (SEQ ID NO: 31), the HVR-H2 region comprises the amino acid sequence AITWNSGHIDYADSVEGR (SEQ ID NO: 32), and the HVR-H3 region comprises the amino acid sequence AKVSYLSTASSLDY (SEQ ID NO: 33). Embodiment 7. The antibody of embodiment 6, wherein the antibody is caninized, felineized, or equineized. Embodiment 8. The antibody of any one of embodiments 6-7, wherein the antibody comprises a light chain variable domain (VL) amino acid sequence having at least 90% identity to a sequence selected from SEQ ID NOs: 26, 34, and 36, and / or a heavy chain variable domain (VH) amino acid sequence having at least 90% identity to a sequence selected from SEQ ID NOs: 30, 35, and 37, and optionally (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. Embodiment 9. The antibody comprises a light chain variable domain (VL) comprising an amino acid sequence selected from SEQ ID NOs: 26, 34, and 36, and / or a heavy chain variable domain (VH) comprising an amino acid sequence selected from SEQ ID NOs: 30, 35, and 37, and optionally the antibody comprises (i) a light chain variable domain (VL) comprising an amino acid sequence selected from SEQ ID NO: 26 and a heavy chain variable domain (VH) comprising an amino acid sequence selected from SEQ ID NO: 30, (ii) a light chain variable domain (VL) comprising an amino acid sequence selected from SEQ ID NO: 34 and a heavy chain variable domain (VH) comprising an amino acid sequence selected from SEQ ID NO: 35, (iii) a light chain variable domain (VL) comprising an amino acid sequence selected from SEQ ID NO: 36. 9. The antibody of any one of embodiments 6-8, comprising: (i) a light chain variable domain (VL) comprising an amino acid sequence selected from SEQ ID NO: 26 and a heavy chain variable domain (VH) comprising an amino acid sequence selected from SEQ ID NO: 37; (ii) a light chain variable domain (VL) comprising an amino acid sequence selected from SEQ ID NO: 36 and a heavy chain variable domain (VH) comprising an amino acid sequence selected from SEQ ID NO: 37; (iii) a light chain variable domain (VL) comprising an amino acid sequence selected from SEQ ID NO: 36 and a heavy chain variable domain (VH) comprising an amino acid sequence selected from SEQ ID NO: 30; or (iv) a light chain variable domain (VL) comprising an amino acid sequence selected from SEQ ID NO: 26 and a heavy chain variable domain (VH) comprising an amino acid sequence selected from SEQ ID NO: 37. Embodiment 10. The antibody comprises a light chain (LC) amino acid sequence having at least 90% identity to a sequence selected from SEQ ID NOs: 38, 40, and 41, and / or a heavy chain (HC) amino acid sequence having at least 90% identity to a sequence selected from SEQ ID NOs: 39, 42, 43, and 44, and optionally 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 antibody of any one of embodiments 6-9, comprising: (i) the LC amino acid sequence of SEQ ID NO: 41 and the HC amino acid sequence of SEQ ID NO: 42; (ii) the LC amino acid sequence of SEQ ID NO: 40 and the HC amino acid sequence of SEQ ID NO: 43; (iii) the LC amino acid sequence of SEQ ID NO: 41 and the HC amino acid sequence of SEQ ID NO: 43; (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. Embodiment 11. The antibody of any one of embodiments 6-10, wherein the antibody is an scFv antibody, and optionally 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. Embodiment 12. The antibody of any one of embodiments 1-11, wherein the antibody is an antibody fragment selected from Fv, scFv, Fab, Fab', F(ab')2, and Fab'-SH. Embodiment 13. The antibody of any one of embodiments 1-12, wherein the antibody comprises a canine heavy chain constant region selected from an IgG-A, IgG-B, IgG-C, and IgG-D constant region. Embodiment 14. The antibody of any one of embodiments 1-13, 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. Embodiment 15. The antibody of any one of embodiments 1-14, wherein the antibody comprises a heavy chain constant region having a "Y" mutation at position 252 (EU numbering). Embodiment 16. A bispecific antibody that binds to canine IL23 and canine TNFα, said antibody comprising: a light chain (LC) comprising a VL domain having an HVR-L1 region comprising the amino acid sequence RASQGISSWLA (SEQ ID NO: 4), an HVR-L2 region comprising the amino acid sequence YAASSLQS (SEQ ID NO: 5), and an HVR-L3 region comprising the amino acid sequence QQYNIYPYT (SEQ ID NO: 6); and a heavy chain (HC) comprising a VH domain having an HVR-H1 region comprising the amino acid sequence KGSGYSFTTYWLG (SEQ ID NO: 8), an HVR-H2 region comprising the amino acid sequence IMSPVDSDIR (SEQ ID NO: 9), and an HVR-H3 region comprising the amino acid sequence ARRRPGQGYFDF (SEQ ID NO: 10). and an scFv antibody fused to the HC, the scFv antibody comprising a VL domain having an HVR-L1 region comprising the amino acid sequence RASQGIRNYLA (SEQ ID NO: 27), an HVR-L2 region comprising the amino acid sequence AASTLQ (SEQ ID NO: 28), and an HVR-L3 region comprising the amino acid sequence QRYNRAPYT (SEQ ID NO: 29), and a VH domain having an HVR-H1 region comprising the amino acid sequence FTFDDYAMH (SEQ ID NO: 31), an HVR-H2 region comprising the amino acid sequence AITWNSGHIDYADSVEGR (SEQ ID NO: 32), and an HVR-H3 region comprising the amino acid sequence AKVSYLSTASSLDY (SEQ ID NO: 33). Embodiment 17. The bispecific antibody of embodiment 16, wherein the antibody comprises a light chain (LC) comprising a light chain variable domain (VL) amino acid sequence having at least 90% identity to a sequence selected from SEQ ID NOs: 3, 11, 13, and 15, and a heavy chain (HC) comprising a heavy chain variable domain (VH) amino acid sequence having at least 90% identity to a sequence selected from SEQ ID NOs: 7, 12, 14, and 16, wherein the HC is fused to an scFv antibody comprising a light chain variable domain (VL) amino acid sequence having at least 90% identity to a sequence selected from SEQ ID NOs: 26, 34, and 36, and a heavy chain variable domain (VH) amino acid sequence having at least 90% identity to a sequence selected from SEQ ID NOs: 30, 35, and 37. Embodiment 18. The bispecific antibody of any one of embodiments 16-17, 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, and optionally 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. Embodiment 19. A bispecific antibody that binds to canine IL23 and canine TNFα, said antibody comprising a light chain (LC) comprising a VL domain having an HVR-L1 region comprising the amino acid sequence RASQGIRNYLA (SEQ ID NO: 27), an HVR-L2 region comprising the amino acid sequence AASTLQ (SEQ ID NO: 28), and an HVR-L3 region comprising the amino acid sequence QRYNRAPYT (SEQ ID NO: 29), a VH domain having an HVR-H1 region comprising the amino acid sequence FTFDDYAMH (SEQ ID NO: 31), an HVR-H2 region comprising the amino acid sequence AITWNSGHIDYADSVEGR (SEQ ID NO: 32), and an HVR-H3 region comprising the amino acid sequence AKVSYLSTASSLDY (SEQ ID NO: 33). and an scFv antibody fused to the HC, the scFv antibody comprising a VL domain having an HVR-L1 region comprising the amino acid sequence RASQGISSWLA (SEQ ID NO: 4), an HVR-L2 region comprising the amino acid sequence YAASSLQS (SEQ ID NO: 5), and an HVR-L3 region comprising the amino acid sequence QQYNIYPYT (SEQ ID NO: 6), and a VH domain having an HVR-H1 region comprising the amino acid sequence KGSGYSFTTYWLG (SEQ ID NO: 8), an HVR-H2 region comprising the amino acid sequence IMSPVDSDIR (SEQ ID NO: 9), and an HVR-H3 region comprising the amino acid sequence ARRRPGQGYFDF (SEQ ID NO: 10). 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 having at least 90% identity to a sequence selected from SEQ ID NOs: 26, 34, and 36, and a heavy chain variable domain (VH) amino acid sequence having at least 90% identity to a sequence selected from SEQ ID NOs: 30, 35, and 37, wherein the LC is fused to an scFv antibody comprising a light chain variable domain (VL) amino acid sequence having at least 90% identity to a sequence selected from SEQ ID NOs: 3, 11, 13, and 15, and a heavy chain variable domain (VH) amino acid sequence having at least 90% identity to a sequence selected from SEQ ID NOs: 7, 12, 14, and 16. Embodiment 21. A bispecific antibody that binds to canine, feline, and / or equine IL23 and canine, feline, and / or equine TNFα, wherein said antibody: (i) an anti-IL23 light chain (LC) comprising a VL domain having an HVR-L1 region comprising the amino acid sequence RASQGISSWLA (SEQ ID NO: 4), an HVR-L2 region comprising the amino acid sequence YAASSLQS (SEQ ID NO: 5), and an HVR-L3 region comprising the amino acid sequence 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 KGSGYSFTTYWLG (SEQ ID NO: 8), an HVR-H2 region comprising the amino acid sequence IMSPVDSDIR (SEQ ID NO: 9), and an HVR-H3 region comprising the amino acid sequence 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 RASQGIRNYLA (SEQ ID NO: 27), an HVR-L2 region comprising the amino acid sequence AASTLQ (SEQ ID NO: 28), and an HVR-L3 region comprising the amino acid sequence 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 FTFDDYAMH (SEQ ID NO: 31), an HVR-H2 region comprising the amino acid sequence AITWNSGHIDYADSVEGR (SEQ ID NO: 32), and an HVR-H3 region comprising the amino acid sequence AKVSYLSTASSLDY (SEQ ID NO: 33); 2. The bispecific antibody comprising: Embodiment 22. (i) the anti-IL23 light chain (LC) comprises a light chain variable domain (VL) amino acid sequence having at least 90% identity to a sequence selected from 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 having at least 90% identity to a sequence selected from 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 having at least 90% identity to a sequence selected from 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 having at least 90% identity to a sequence selected from SEQ ID NOs: 30, 35, and 37; 22. The bispecific antibody of embodiment 21. Embodiment 23. (i) the anti-IL23 light chain (LC) comprises an amino acid sequence having at least 90% identity to a sequence selected from 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 having at least 90% identity to a sequence selected from 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 to SEQ ID NOs: 42, 43, and 44; 23. The bispecific antibody of any one of embodiments 21-22. Embodiment 24. An isolated nucleic acid(s) encoding the antibody of any one of embodiments 1 to 23. Embodiment 25. A host cell comprising the nucleic acid(s) of embodiment 24. Embodiment 26. A method for producing an antibody, comprising culturing the host cell of embodiment 25 and isolating the antibody. Embodiment 27. A pharmaceutical composition comprising the antibody of any one of embodiments 1 to 23 and a pharmaceutically acceptable carrier. Embodiment 28. A method of treating a dog, cat, or horse having a condition associated with IL23, 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. Embodiment 29. A method of maintaining remission of an IL23-associated condition 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. Embodiment 30. The method of any one of embodiments 28-29, wherein the IL23-associated condition is an inflammatory disease. Embodiment 31. The method of any one of embodiments 28-30, wherein the IL23-associated condition is an inflammatory disease of the gastrointestinal system. Embodiment 32. The method of any one of embodiments 28-31, wherein the IL23-associated condition is inflammatory bowel disease. Embodiment 33. The method of any one of embodiments 28-32, 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. Embodiment 34. A method of treating a dog, cat, or horse having a condition associated with IL23 and TNFα, comprising administering to the dog, cat, or horse a therapeutically effective amount of an IL23 antibody of embodiments 1-5 or 12-23, and an anti-TNFα antibody of any one of embodiments 6-23, or the pharmaceutical composition of embodiment 24. 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, and the IL23- or IL23 / TNFα-associated condition is an inflammatory disease of the gastrointestinal system. Embodiment 36. The method of any one of embodiments 34-35, wherein the IL23 or IL23 / TNFα associated condition is ankylosing spondylitis, asthma, cancer, Crohn's disease, idiopathic arthritis, psoriasis, plaque psoriasis, psoriatic arthritis, rheumatoid arthritis, osteoarthritis, or ulcerative colitis. Embodiment 37. The method of any one of embodiments 28-36, wherein the antibody or pharmaceutical composition is administered parenterally. Embodiment 38. The method of any one of embodiments 28-37, wherein the antibody or pharmaceutical composition is administered by intramuscular, intraperitoneal, intracerebrospinal, subcutaneous, intra-arterial, intrasynovial, intrathecal, or inhalation route. Embodiment 39. The method of any one of embodiments 28-38, wherein the method further comprises administration of 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 alpha4-integrin antibody, a beta9-integrin, an IL12 antibody, an IL1β antibody, or an anti-BlyS antibody. Embodiment 40. The method of any one of embodiments 28-39, wherein the antibody is administered in an amount ranging from 0.01 mg / kg body weight to 100 mg / kg body weight per dose. Embodiment 41. A method of reducing canine, feline, or equine IL23 and / or TNFα signaling function in a cell, comprising reducing binding of IL23 and / or TNFα to the signaling function by the cell by exposing the cell to the antibody of any one of Embodiments 1 to 23 under conditions permissive for binding of the antibody to IL23 and / or TNFα. Embodiment 42. The method of embodiment 41, wherein the cells are exposed to the antibody or pharmaceutical composition ex vivo. Embodiment 43. The method of embodiment 41, wherein the cell is exposed to the antibody or pharmaceutical composition in vivo. Embodiment 44. The method of any one of embodiments 41-43, wherein the cells are canine, feline, or equine cells. Embodiment 45. A method for detecting IL23 and / or TNFα in a sample from a companion animal species, 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. Embodiment 46. The method of embodiment 45, wherein the sample is a biological sample obtained from a dog, cat, or horse. DETAILED DESCRIPTION OF THE INVENTION

[0015] The present disclosure provides a detailed description, 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, and uses of these antibodies, including methods for treating various diseases and disorders mediated by or associated with the binding activity of IL23 and / or TNFα. Methods for designing, producing, or purifying bispecific antibodies to canine IL23 and canine TNFα are also provided. Methods for detecting IL23 and / or TNFα in samples derived from companion animal species are provided.

[0016] As used herein and in the appended claims, the singular forms "a" and "an" include plural referents unless the context clearly dictates otherwise. Thus, for example, a reference to "a protein" includes a plurality of proteins, and a reference to "a compound" refers to a plurality of compounds. It is further noted that the claims may be drafted to exclude any element. Accordingly, this statement is intended to serve as a precedent standard for the use of exclusive terminology such as "solely," "only," or the use of "negative" limitations in connection with the recitation of claim elements. The use of "comprise," "comprises," "comprising," "include," "includes," and "including" are interchangeable and not intended to be limiting. Where the description of various embodiments uses the term "comprising," those skilled in the art will further understand that in some specific instances, an embodiment may alternatively be described using the language "consisting essentially of" or "consisting of."

[0017] Where a range of values ​​is provided, unless the context clearly dictates otherwise, it is understood that each intervening integer and tenth of each intervening integer between the upper and lower limits of that range, as well as any other stated or intervening value in that stated range, is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included within the smaller ranges and are also encompassed herein, subject to any specific excluded limits in the stated range. When a stated range includes one or both of those upper and lower limits, ranges excluding (i) either one or (ii) both of those included upper and lower limits are also encompassed within the invention. For example, "1 to 50" includes "2 to 25," "5 to 20," "25 to 50," "1 to 10," etc.

[0018] Generally, the nomenclature used herein, and the techniques and procedures described herein, include those well understood and commonly used by those of skill in the art, such as, for example, the general techniques and methodology described in Green and Sambrook, Molecular Cloning: A Laboratory Manual (Fourth Edition), Vols. 1-3, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, 2012 (hereinafter "Sambrook"), and Current Protocols in Molecular Biology, edited by FMA Ausubel, first published in book form by Greene Publishing Associates, Inc. and John Wiley & Sons, Inc. in 1987, supplemented periodically until 2011, and now available in journal form online as Current Protocols in Molecular Biology, Vols. 00-130, (1987-2020), published by Wiley & Sons, Inc. in the Wiley Online Library (hereinafter "Ausubel").

[0019] All publications, patents, patent applications, and other documents referenced in this disclosure are incorporated herein by reference in their entirety for all purposes to the same extent as if each individual publication, patent, patent application, or other document was individually indicated to be incorporated herein by reference for all purposes.

[0020] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. It is to be understood that the terms used herein are for the purpose of describing particular embodiments and are not intended to be limiting. For purposes of interpreting this disclosure, the following terminology explanations apply, and where appropriate, terms used in the singular also include the plural and vice versa.

[0021] As used herein, "IL23" or "IL-23" refers to the cytokine protein interleukin 23 (or IL-23) and, unless otherwise indicated, encompasses IL23 protein sourced from any vertebrate, including mammals such as primates (e.g., humans and cynomolgus monkeys) and rodents (e.g., mice and rats), as well as companion animals (e.g., canines, felines, and horses). The term also includes naturally occurring IL23 variants (e.g., splice variants or allelic variants). The amino acid sequences of recombinant forms of exemplary canine IL23 and feline IL23 proteins are set forth in Table 1 below and in the accompanying sequence listing.

[0022] As used herein, unless otherwise indicated, "TNF" or "TNFa" or "TNFα" refers to the cytokine protein, tumor necrosis factor, TNFα protein, sourced from any vertebrate, including mammals such as primates (e.g., humans and cynomolgus monkeys) and rodents (e.g., mice and rats), and companion animals (e.g., canines, felines, and horses). The term also includes naturally occurring TNFα variants (e.g., splice variants or allelic variants).

[0023] As used herein, an "IL23-mediated condition" or "IL23-mediated disease" encompasses any disease or disorder associated with the specific binding of IL23 to the IL23 receptor or other specific IL23 binding target, and may include diseases associated with, caused by, or characterized by elevated levels or gradients of IL23 concentration. For example, specific binding of IL23 stimulates the production of Th17 cells, which are involved in the immune response. Thus, an IL23-mediated disease can include, but is not limited to, any disease or condition mediated by and / or responsive to an antagonist or inhibitor of IL23 binding to the IL23 receptor or other IL23 target. Specific 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.

[0024] As used herein, a "TNFα-mediated condition" or "TNFα-mediated disease" encompasses any disease or disorder associated with the specific binding of TNFα to a TNFα receptor or other specific TNFα binding target, and can include diseases associated with, caused by, or characterized by elevated levels or gradients of TNFα concentration. Thus, a TNFα-mediated disease can include, but is not limited to, any disease or condition mediated by and / or responsive to antagonists or inhibitors of TNFα binding to a TNFα receptor or other specific TNFα binding target. Specific exemplary diseases are provided elsewhere herein.

[0025] As used herein, "antibody" refers to a molecule comprising one or more polypeptide chains that specifically binds to or is immunologically reactive with a particular antigen. Exemplary antibodies of the present disclosure include monoclonal antibodies, polyclonal antibodies, chimeric antibodies, caninized antibodies, felineized antibodies, multispecific (or heteroconjugate) antibodies (e.g., bispecific antibodies), monovalent antibodies (e.g., single-arm 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).

[0026] An "anti-IL23 antibody" or "antibody that binds IL23" refers to an antibody that binds to IL23 with sufficient affinity so that the antibody is useful as a diagnostic and / or therapeutic agent in targeting IL23. An anti-IL23 antibody may further be specified to refer to an antibody that binds to a specific form of IL23, such as canine IL23 (e.g., "anti-canine IL23" or "anti-IL23 that binds canine IL23"). In some embodiments, the extent to which an anti-IL23-specific antibody binds 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 antibody's binding to IL23, as measured, for example, by radioimmunoassay (RIA) or surface plasmon resonance (SPR). In some embodiments, an antibody that binds IL23 has an affinity of <1 μM, <100 nM, <10 nM, <1 nM, <0.1 nM, <0.01 nM, or <1 pM (e.g., 10 -8 M or less, e.g., 10 -8 M~10 -13 M, e.g., 10 -9 M~10 -13 Dissociation constant (K D )

[0027] An "anti-TNFα antibody" or "antibody that binds TNFα" refers to an antibody that binds to TNFα with sufficient affinity so that the antibody is useful as a diagnostic and / or therapeutic agent in targeting TNFα. An anti-TNFα antibody may further be specified to refer to an antibody that binds to a specific form of TNFα, such as canine TNFα (e.g., "anti-canine TNFα" or "anti-TNFα that binds canine TNFα"). In some embodiments, the extent to which an anti-TNFα specific antibody binds 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 antibody's binding to TNFα, as measured, for example, by radioimmunoassay (RIA) or surface plasmon resonance (SPR). In some embodiments, an antibody that binds to TNFα has an affinity of <1 μM, <100 nM, <10 nM, <1 nM, <0.1 nM, <0.01 nM, or <1 pM (e.g., 10 -8 M or less, e.g., 10 -8 M~10 -13 M, e.g., 10 -9 M~10 -13 Dissociation constant (K D )

[0028] "Full-length antibody," "intact antibody," or "whole antibody" are used interchangeably herein to refer to an antibody having a structure substantially similar to a native antibody structure, or an antibody having a heavy chain containing an Fc region as defined herein.

[0029] The "class" of an antibody refers to the type of constant domain or constant region present in its heavy chain. There are five major classes of human antibodies: IgA, IgD, IgE, IgG, and IgM, some of which are further divided into subclasses (isotypes) (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2). The heavy chain constant domains corresponding to the various immunoglobulin classes are called α, δ, ε, γ, and μ, respectively. Canine, feline, and equine species have various classes of antibodies shared by many other mammalian species. For example, canine species have the antibody classes IgGA, IgGB, IgGC, and IgGD, while feline species have the antibody classes IgGA1, IgGA2, and IgGB.

[0030] "Variable region" or "variable domain" refers to the domain of an antibody heavy or light chain that is involved in binding the antibody to an antigen. The heavy and light chain variable domains (V, V ... H and V L ) generally have a similar structure, with each domain containing four conserved framework regions (FR) and three hypervariable regions (HVR). (See, e.g., Kindt et al., Kuby Immunology, 6 th (See, e.g., W.H. Freeman and Co., p. 91). To confer antigen-binding specificity, a single V H or V L Furthermore, an antibody that binds to a particular antigen may have a V domain derived from the antibody that binds that antigen. H or V L Each complementary V domain is used L or V H They can be isolated by screening libraries of domains (see, for example, Portolano et al., J. Immunol. 150:880-887 (1993); Clarkson et al., Nature 352:624-628 (1991)).

[0031] As used herein, "hypervariable region" or "HVR" refers to each region of an antibody variable domain that is hypervariable in sequence and / or forms a structurally defined loop ("hypervariable loop"). Generally, naturally occurring antibodies contain six HVRs: heavy chain variable domain (V H ) and three (HVR-H1, HVR-H2, HVR-H3) in the light chain variable domain (V L ), three of which (HVR-L1, HVR-L2, HVR-L3). HVRs generally contain amino acid residues from hypervariable loops and / or "complementarity-determining regions" (CDRs). Several descriptions of hypervariable regions are in use and are encompassed herein. 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 alternatives refer to the location of structural loops (Chothia and Lesk J. Mol. Biol. 196:901-917 (1987)). AbM hypervariable regions represent a compromise between Kabat CDRs and Chothia structural loops and are used in Oxford Molecular's AbM antibody modeling software. The "contact" hypervariable regions are based on an analysis of the available complex crystal structures. Residues from each of these hypervariable regions are shown in the table below. [Table 14]

[0032] Unless otherwise indicated, HVR residues and other residues in the variable domain (e.g., FR residues) are numbered herein according to Kabat et al., supra.

[0033] As used herein, hypervariable regions can include extended or alternative hypervariable regions such as: L Domains 24-36 or 24-34 (L1), 46-56 or 50-56 (L2) and 89-97 or 89-96 (L3), and V H In domains 26-35 or 30-35 (H1), 50-61, 50-65 or 49-65 (H2) and 93-102, 94-102, or 95-102 (H3), the variable domain residues are numbered according to Kabat et al., supra, for each of these definitions.

[0034] As used herein, "complementarity determining region," or "CDR," refers to the region within the HVR of a variable domain that is most highly sequence variable and / or involved in antigen recognition. Generally, a naturally occurring antibody contains six CDRs: a heavy chain variable domain (V H ) and the light chain variable domain (V L ), three (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 (L1), 50-56 (L2), 89-97 (L3), 31-35 (H1), 50-61 (H2), and 95-102 (H3) (numbering according to Kabat et al., supra).

[0035] "Framework" or "FR" refers to variable domain residues other than hypervariable region (HVR) residues. The FR of a variable domain generally consists of four FR domains: FR1, FR2, FR3, and FR4. Thus, the HVR and FR sequences generally correspond to the V H (or V L ) in the following sequence: FR1-H1(L1)-FR2-H2(L2)-FR3-H3(L3)-FR4.

[0036] "Native antibody" refers to an immunoglobulin molecule of natural origin. For example, a native IgG antibody is a heterotetrameric glycoprotein of approximately 150,000 daltons, composed of two identical light chains and two identical heavy chains linked by disulfides. Each heavy chain contains, from the N-terminus to the C-terminus, a variable region (V), also called a variable heavy domain or heavy chain variable domain. H ), followed by three constant domains (CH1, CH2, and CH3). Similarly, each light chain contains, from the N-terminus to the C-terminus, a variable region (V), also called a variable light domain or a light chain variable domain. L ), followed by a constant light domain (CL). The light chain of an antibody can be assigned to one of two types, called kappa (κ) and lambda (λ), based on the amino acid sequence of its constant domain.

[0037] As used herein, a "monoclonal antibody" refers to an antibody obtained from a substantially homogeneous antibody population. That is, the individual antibodies comprising the population are identical and / or bind to the same epitope, except for possible antibody variants (e.g., antibody variants contain naturally occurring mutations or mutations that arise during the production of the monoclonal antibody and are generally present in minor amounts). In contrast to polyclonal antibody preparations, which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody of a monoclonal antibody preparation is directed against a single determinant on an antigen. Thus, the term "monoclonal" indicates the character of the antibody as being obtained from a substantially homogeneous antibody population and should not be construed as requiring production of the antibody by any particular method. For example, monoclonal antibodies can be produced by a variety of techniques, including, but not limited to, hybridoma methods, recombinant DNA methods, phage display methods, and methods utilizing transgenic animals containing all or part of the human immunoglobulin loci; such methods, as well as other exemplary methods for producing monoclonal antibodies, are described herein.

[0038] A "chimeric antibody" refers to an antibody in which a portion of the heavy and / or light chain is derived from a particular source or species, with the remainder of the heavy and / or light chain 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 canine. In some embodiments, a canine chimeric antibody may contain 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 mouse heavy chain variable domain (VH) framework region or a mouse light chain variable domain (VL) framework region.

[0039] A "caninized antibody" refers to a chimeric antibody comprising amino acid sequences derived from non-canine HVRs and amino acid sequences derived from canine FRs. In certain embodiments, a caninized antibody will comprise substantially all of at least one, and typically two, variable domains, with all or substantially all of its HVRs corresponding to HVRs of a non-canine antibody and all or substantially all of its FRs corresponding to FRs of a canine antibody. A caninized antibody may optionally 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.

[0040] A "felineized antibody" refers to a chimeric antibody comprising amino acid sequences derived from non-feline HVRs and amino acid sequences derived from feline FRs. In certain embodiments, a felineized antibody will comprise substantially all of at least one, and typically two, variable domains, with all or substantially all of its HVRs corresponding to HVRs of a non-feline antibody and all or substantially all of its FRs corresponding to FRs of a feline antibody. A felineized antibody may optionally comprise at least a portion of an antibody constant region derived from a feline antibody. A "felineized form" of an antibody (e.g., a non-feline antibody) refers to an antibody that has undergone felineization.

[0041] A "canine antibody" refers to an antibody possessing an amino acid sequence that corresponds to the amino acid sequence of an antibody produced by a canine or canine cell, or an antibody from a non-canine source that utilizes the canine antibody repertoire, or that corresponds to the sequence encoding another canine antibody. This definition of canine antibody specifically excludes caninized antibodies that contain non-canine antigen-binding residues.

[0042] The "consensus framework" is based on immunoglobulin V L or V H In selecting framework sequences, the framework represents the most commonly occurring amino acid residues. L or V H The sequences are selected from a subgroup of variable domain sequences. Generally, the subgroup of sequences is a subgroup as found in Kabat et al., Sequences of Proteins of Immunological Interest, Fifth Edition, NIH Publication 91-3242, Bethesda, MD (1991), vols. 1-3. In some embodiments, the V L With respect to V, the subgroup is subgroup kappa I as in Kabat et al., supra. In some embodiments, V H With respect to the subgroup, the subgroup is subgroup III as in Kabat et al., supra.

[0043] As used herein, an "acceptor framework" refers to a light chain variable domain (V) derived from an immunoglobulin framework or a consensus framework. L ) framework or heavy chain variable domain (V H) framework. An acceptor framework "derived from" an immunoglobulin framework or consensus framework may contain the same amino acid sequence or may contain altered amino acid sequences. 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, a V L The acceptor framework is V L It is identical to an immunoglobulin framework sequence or a consensus framework sequence.

[0044] "Fc region" refers to a dimeric complex comprising the C-terminal polypeptide sequence of an immunoglobulin heavy chain, the C-terminal polypeptide sequence being obtained by papain digestion of an intact antibody. The Fc region may comprise a native Fc sequence or an Fc sequence variant. The Fc sequence of an immunoglobulin generally comprises two constant domains (a CH2 domain and a CH3 domain), and optionally a CH4 domain. The boundaries of an immunoglobulin heavy chain Fc sequence may vary depending on the immunoglobulin class and species.

[0045] The term "IgX Fc" denotes that the Fc region is derived from a particular antibody isotype (e.g., IgG, IgA, IgD, IgE, IgM, etc.), where "X" represents the antibody isotype. Thus, for example, "IgG Fc" denotes the Fc region of the gamma chain, "IgA Fc" denotes the Fc region of the alpha chain, "IgD Fc" denotes the Fc region of the delta chain, "IgE Fc" denotes the Fc region of the epsilon chain, and "IgM Fc" denotes the Fc region of the mu chain. In some embodiments, an IgG Fc region comprises a CH1, hinge, CH2, CH3, and CL1. "IgX-N-Fc" denotes that the Fc region is derived from a particular subclass of antibody isotype (e.g., 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 canine). 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 the Fc region of IgG-A, IgG-B, IgG-C, or IgG-D may be expressed at higher levels in recombinant production systems. "IgX Fc" and "IgX Fc polypeptide" are intended to include wild-type IgX Fc polypeptide and IgX Fc polypeptide variants.

[0046] "Effector function" refers to a biological activity attributable to the Fc region of an antibody, which varies depending on the antibody isotype. Examples of antibody effector functions include C1q binding and complement-dependent cytotoxicity (CDC), Fc receptor binding, antibody-dependent cellular cytotoxicity (ADCC), phagocytosis, down-regulation of cell surface receptors (e.g., B cell receptors), and B cell activation.

[0047] An "immunoconjugate" refers to an antibody conjugated to one or more heterologous molecule(s), including but not limited to, a cytotoxic agent.

[0048] A "multispecific antibody" is an antibody that has at least two different binding sites, each site having a different binding specificity. Multispecific antibodies can be full-length antibodies or antibody fragments, and the various binding sites can each bind to a different antigen, or the various binding sites can bind to two different epitopes of the same antigen.

[0049] "Fv fragment" refers to an antibody fragment containing a complete antigen recognition and binding site. This region naturally consists of a dimer of one heavy- and one light-chain variable domain in tight association, which may be covalent (e.g., in scFv). This configuration allows the three HVRs of each variable domain to interact to form a V H -V L An antigen-binding site is defined 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 containing only three antigen-specific HVRs) has the ability to recognize and bind antigen, although usually with lower affinity than the entire binding site.

[0050] "Fab fragment" refers to an antibody fragment containing the variable and constant domains of the light chain and the variable and first constant domain (CH1) of the heavy chain. "F(ab')2 fragment" comprises a pair of Fab fragments, which are generally covalently linked near their carboxy termini by hinge cysteines between them. Other chemical couplings of antibody fragments are known in the art.

[0051] As used herein, "antigen-binding arm" refers to the 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 (e.g., immunoglobulin light and heavy chain HVR and / or variable domain sequences).

[0052] "Single-chain Fv" or "scFv" refers to the V of an antibody. H and V L scFv antibodies generally refer to antibody fragments containing V domains, where the V domains are present in a single polypeptide chain. H Polypeptides containing domain sequences and V L The V domains are fused via a polypeptide linker. H and V L The domains enable the hypervariable regions of the scFv to form a desired antigen-binding structure.

[0053] As used herein, "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 serving to conjugate or fuse various polypeptides.

[0054] "Diabody" refers to a small antibody fragment with two antigen-binding sites, which are bound to the same polypeptide chain (V H and V L ) in the light chain variable domain (V L ) connected 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.

[0055] "Linear antibody" refers 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.

[0056] A "naked antibody" refers to an antibody that is not conjugated to a heterologous moiety (e.g., a cytotoxic moiety) or radiolabel.

[0057] "Affinity" refers to the sum total of the strength of 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, which 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 is generally determined by 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.

[0058] "Specifically binds" or "specific binding" means a binding activity of approximately 1 x 10 -7 "Secondary affinity" refers to antibody binding to an antigen with an affinity value of about 10 nM or less. In some embodiments, an antibody may have a secondary affinity for an antigen other than the one to which it specifically binds, and "secondary affinity" generally refers to antibody binding to a secondary antigen with an affinity value of greater than about 10 nM, as described elsewhere herein. Even though an antibody may have a secondary affinity for a secondary antigen, such an antibody will still specifically bind to the primary antigen.

[0059] An "isolated antibody" refers to an antibody that has been separated from components of its natural environment. In some embodiments, the antibody is purified to greater than 95% or greater than 99% purity, for example, as measured by electrophoretic methods (e.g., SDS-PAGE, isoelectric focusing (IEF), capillary electrophoresis) or chromatographic methods (e.g., ion exchange or reverse-phase HPLC). For a review of methods for assessing antibody purity, see, for example, Flatman et al., J. Chromatogr. B 848:79-87.

[0060] With respect to polypeptide or antibody sequences, "percent (%) amino acid sequence identity" and "homology," as used herein, are defined as the percentage of amino acid residues in a candidate sequence that are identical with those in a specific peptide or polypeptide sequence, after aligning the sequences and introducing gaps as necessary to maximize the percent sequence identity, not counting any conservative substitutions as part of the sequence identity. Alignment for purposes of determining percent amino acid sequence identity can be accomplished in a variety of ways within the skill of the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN, or MEGALINE™ (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.

[0061] An "amino acid substitution" refers to the replacement of one amino acid in a polypeptide with 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 products can be screened for a desired activity (e.g., retained / improved antigen binding, reduced immunogenicity, or improved ADCC or CDC, or enhanced pharmacokinetics).

[0062] The term "vector" is used to describe a polynucleotide that can be manipulated to contain cloned polynucleotide(s) that can be propagated in a host cell. A vector can contain one or more of the following elements: an origin of replication, one or more regulatory sequences (e.g., promoters or enhancers) that control the expression of a polypeptide of interest, or one or more selectable marker genes (e.g., antibiotic resistance genes and genes that can be used in colorimetric assays, e.g., β-galactosidase). The term "expression vector" refers to a vector used to express a polypeptide of interest in a host cell.

[0063] A "host cell" refers to a cell that can be or has been the recipient of a vector or isolated polynucleotide. Host cells can be prokaryotic or eukaryotic. Exemplary eukaryotic cells include mammalian cells, such as primate or non-primate animal cells, fungal cells, such as yeast, plant cells, and insect cells. Non-limiting exemplary mammalian cells include, but are not limited to, NS0 cells, PER.C6® cells (Crucell), 293 cells, and CHO cells, as well as their derivatives (such as 293-6E, DG44, CHO-S, and CHO-K cells). A host cell includes the progeny of a single host cell, which may not necessarily be completely identical (in morphology or in overall composition of genomic DNA) to the original parent cell due to natural, accidental, or deliberate mutation. A host cell includes cells transfected in vivo with polynucleotide(s) encoding the amino acid sequence(s) provided herein.

[0064] The term "companion animal species" refers to animals suitable for human companionship. In some embodiments, companion animal species are small mammals, such as dogs, cats, canines, felines, horses, rabbits, ferrets, guinea pigs, rodents, etc. In some embodiments, companion animal species are larger animals, such as camels or livestock (e.g., equines, cows, pigs, etc.).

[0065] "Reduce" or "inhibit" means to decrease, reduce, or suppress an activity, function, or amount compared to a reference. In some embodiments, "reduce" or "inhibit" refers to the ability to cause an overall decrease of more than 20%. In some embodiments, "reduce" or "inhibit" refers to the ability to cause an overall decrease of more than 50%. In some embodiments, "reduce" or "inhibit" refers to the ability to cause an overall decrease of more than 75%, 85%, 90%, or 95%. In some embodiments, the amount is suppressed or decreased over a period of time compared to a control dose (such as a placebo) over the same period of time. 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 examples, a reference is obtained from a non-diseased or untreated sample from a companion animal. In some examples, a reference is obtained from one or more healthy animals of a particular species that are not the animals being tested or treated.

[0066] As used herein, "substantially similar" or "substantially the same" refers to the difference between two values ​​(e.g., one associated with a test antibody and the other associated with a reference antibody) that one of skill in the art would recognize as the difference between the values ​​(e.g., K D The degree of similarity between two values ​​is sufficiently high that the two values ​​are deemed to have little or no biological and / or statistical significance within the context of the biological feature measured by the two values.

[0067] As used herein, "substantially different" means that one of skill in the art would recognize the difference between two values ​​(typically, one associated with a molecule and the other associated with a reference molecule) as being within the range of the values ​​(e.g., K D The difference between two values ​​is sufficiently high that it is considered statistically significant within the context of the biological feature measured by the two values.

[0068] "Treatment," "treat," or "treating" refers to an intervention in an attempt to alter the natural history of a disorder in the individual being treated and may be performed for prophylaxis or during the course of clinical pathology. Desired results of treatment may include, but are not limited to, prevention of the onset or recurrence of the disorder, alleviation of symptoms, reduction of any direct or indirect pathological consequences of the disorder, prevention of metastasis, slowing the rate of progression, amelioration or palliation of the disease state, and a tendency toward remission or improved prognosis. For example, treatment may include administering to a subject a therapeutically effective amount of a pharmaceutical formulation comprising an anti-IL23 antibody to delay the onset or slow the progression of an IL23-mediated disease or condition, or a disease or condition in which IL23 may play a role in the pathogenesis and / or progression. Treatment does not require 100 percent elimination of all aspects of the disorder.

[0069] A "pharmaceutical formulation" refers to a preparation of an active ingredient(s) in a form that allows the biological activity of the active ingredient(s) 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 contain one or more active agents. For example, a pharmaceutical formulation may contain an anti-IL23 antibody as the only active agent in the formulation, or may contain an anti-IL23 antibody and one or more additional active agents.

[0070] A "pharmaceutically acceptable carrier" refers to an ingredient in a pharmaceutical formulation, other than an active ingredient, that is non-toxic to a subject to which it is administered. Pharmaceutically acceptable carriers include, but are not limited to, buffers, excipients, stabilizers, or preservatives.

[0071] A "therapeutically effective amount" refers to the amount of an active ingredient or agent (e.g., a pharmaceutical formulation) to achieve a desired therapeutic or prophylactic result (e.g., to treat or prevent a disease, disorder, or condition in a subject). In the case of an IL23-mediated disease or condition, a therapeutically effective amount of a therapeutic agent is an amount that will reduce, prevent, inhibit, and / or alleviate to some extent one or more of the symptoms associated with the disease, disorder, or condition.

[0072] "Individual" or "subject" refers to mammals, including, but not limited to, domesticated or companion animals (e.g., cattle, sheep, felines, canines, and equines), primates (e.g., humans and non-human primates such as monkeys), rabbits, and rodents (e.g., mice and rats).

[0073] Anti-IL23 and anti-TNFα antibodies

[0074] Table 1 below outlines the sequences referred to in this disclosure, including the canine IL23 protein, the feline IL23 protein, and the various anti-IL23 and anti-TNFα antibodies of the disclosure, and their sequence identifiers. The sequences are also included in the attached sequence listing.

[0075] [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6] [Table 1-7]

[0076] Provided herein are novel antibodies directed against canine, feline, or equine IL23 and / or canine, feline, or equine TNFα. The anti-IL23 and / or anti-TNFα antibodies provided herein include, but are not limited to, monoclonal antibodies, chimeric antibodies, caninized, feline, or equine antibodies, scFv antibodies, and bispecific antibodies that bind to both IL23 and TNFα. Also provided herein are the amino acid sequences of the monoclonal antibodies. For example, the 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 for the monoclonal antibodies described herein are provided. In addition, the amino acid sequences of the HVRs, framework sequences, light chain variable domain (VL) sequences, and heavy chain variable domain (VH) sequences of various caninized and felineized light and heavy chains are provided.

[0077] The present disclosure provides a novel anti-IL23 antibody of the present disclosure, which is a monoclonal antibody 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 of the C340 VL domain, HVR-L1, HVR-L2, and HVR-L3, are provided as SEQ ID NOs: 4, 5, and 6, respectively. The corresponding hypervariable regions of the C340 VH domain, HVR-H1, HVR-H2, and HVR-H3, are provided as SEQ ID NOs: 8, 9, and 10, respectively.

[0078] In at least one embodiment, the present disclosure provides chimeric anti-IL23 antibodies derived from C340. In one embodiment of the chimeric anti-IL23 antibodies, the antibodies comprise 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. As described in the Examples, the tight binding affinity of the anti-IL23 chimeric antibodies to canine IL23 was measured.

[0079] In at least one embodiment, the disclosure provides caninized versions of the C340 anti-IL23 antibody. The VL domains of two different caninized versions are provided as SEQ ID NOs: 11 and 13. The VH domains of two different caninized versions are provided as SEQ ID NOs: 12 and 14. The VL domain of the caninized version comprises the C340 VL domain, HVR-L1, HVR-L2, HVR-L3 sequences of SEQ ID NOs: 4, 5, and 6, respectively. The VH domain of the caninized version comprises the C340 VH domain, HVR-H1, HVR-H2, HVR-H3 sequences of SEQ ID NOs: 8, 9, and 10, respectively.

[0080] The light chain (LC) and heavy chain (HC) sequences of exemplary caninized anti-IL23 antibodies, 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 a canine kappa light chain constant region are SEQ ID NOs: 19 and 22. Exemplary caninized anti-IL23 heavy chains comprising a canine Fc IgG-B constant region are provided as SEQ ID NOs: 20, 21, and 23. It is contemplated that various exemplary caninized anti-IL23 antibodies may 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.

[0081] In at least one embodiment, the present disclosure also provides a felinized version of the C340 anti-IL23 antibody. An exemplary VL domain of the felinized version is provided as SEQ ID NO: 15, and the VH domain of the felinized version is provided as SEQ ID NO: 16. The felinized version VL domain of SEQ ID NO: 15 comprises the C340 HVR-L1, HVR-L2, HVR-L3 sequences of SEQ ID NOs: 4, 5, and 6, respectively. Similarly, the felinized version VH domain of SEQ ID NO: 16 comprises the C340 HVR-H1, HVR-H2, HVR-H3 sequences of SEQ ID NOs: 8, 9, and 10, respectively.

[0082] 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 C340 heavy chain variable domain (VH) is provided as SEQ ID NO: 30. The corresponding hypervariable regions of the D2E7 VL domain, HVR-L1, HVR-L2, and HVR-L3, are provided as SEQ ID NOs: 27, 28, and 29, respectively. The corresponding hypervariable regions of the D2E7 VH domain, HVR-H1, HVR-H2, and HVR-H3, are provided as SEQ ID NOs: 31, 32, and 33, respectively.

[0083] In at least one embodiment, the present disclosure provides a chimeric anti-TNFα antibody derived from an 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, respectively. The light chain (LC) and heavy chain (HC) amino acid sequences of the exemplary chimeric anti-TNFα D2E7 antibody are presented in Table 1 as SEQ ID NOs: 38 and 39, respectively.

[0084] In at least one embodiment, the present disclosure provides caninized versions of the anti-TNFα D2E7 antibody VL and VH domains as SEQ ID NOs: 34 and 35, respectively. The light chain (LC) and heavy chain (HC) sequences of an exemplary caninized anti-TNFα antibody 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 various exemplary caninized anti-TNFα antibodies may be prepared using any combination of the LC sequences of SEQ ID NOs: 40 and 41 and the HC sequences of SEQ ID NOs: 42, 43, and 44.

[0085] In at least one embodiment, the present disclosure also provides a felinized version of the D2E7 anti-TNFα antibody. An exemplary VL domain of the felinized version is provided as SEQ ID NO: 36, and an exemplary VH domain of the felinized version is provided as SEQ ID NO: 37. The felinized version 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 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.

[0086] In at least one embodiment, the 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 VL domain of the scFv antibody comprises the amino acid sequence of SEQ ID NO: 26, 34, or 40, and the VL domain of the scFv antibody comprises the amino acid sequence of SEQ ID NO: 30, 35, or 41. In at least one embodiment, the polypeptide linker fusing the VL and VH domains of the scFv antibody comprises the amino acid sequence GGGGGSGGGGSGGGGGS (SEQ ID NO: 49). In at least one embodiment, an 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.

[0087] Although the polypeptide of SEQ ID NO:49 is exemplified as the scFv antibody in Table 1, one of skill in the art will appreciate that a wide 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. Generally, polypeptides comprising a polypeptide chain of 5 to 30 amino acids can be used to fuse the polypeptide components of the scFv antibody and bispecific antibody structures of the present disclosure.

[0088]

[0010] 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 the six HVR sequences of the C340 anti-IL23 antibody and the 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" bispecific antibody structure of anti-IL23 LC / HC + anti-TNFα LC / HC that binds to canine, feline, and / or equine IL23 and canine, feline, and / or equine TNFα, wherein the antibody (i) an anti-IL23 light chain (LC) comprising a VL domain having an HVR-L1 region comprising the amino acid sequence RASQGISSWLA (SEQ ID NO: 4), an HVR-L2 region comprising the amino acid sequence YAASSLQS (SEQ ID NO: 5), and an HVR-L3 region comprising the amino acid sequence 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 KGSGYSFTTYWLG (SEQ ID NO: 8), an HVR-H2 region comprising the amino acid sequence IMSPVDSDIR (SEQ ID NO: 9), and an HVR-H3 region comprising the amino acid sequence 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 RASQGIRNYLA (SEQ ID NO: 27), an HVR-L2 region comprising the amino acid sequence AASTLQ (SEQ ID NO: 28), and an HVR-L3 region comprising the amino acid sequence 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 FTFDDYAMH (SEQ ID NO: 31), an HVR-H2 region comprising the amino acid sequence AITWNSGHIDYADSVEGR (SEQ ID NO: 32), and an HVR-H3 region comprising the amino acid sequence AKVSYLSTASSLDY (SEQ ID NO: 33); Includes.

[0089] In some embodiments, the bispecific antibody binds to canine IL23 and canine TNFα and comprises: (i) an anti-IL23 light chain (LC) comprising a light chain variable domain (VL) amino acid sequence having at least 90% identity to a sequence selected from SEQ ID NOs: 3, 11, 13, and 15; (ii) an anti-IL23 heavy chain (HC) comprising a heavy chain variable domain (VH) amino acid sequence having at least 90% identity to a sequence selected from SEQ ID NOs: 7, 12, 14, and 16; (iii) an anti-TNFα light chain (LC) comprising a light chain variable domain (VL) amino acid sequence having at least 90% identity to a sequence selected from SEQ ID NOs: 26, 34, and 36; and (iv) an anti-TNFα heavy chain (HC) comprising a heavy chain variable domain (VH) amino acid sequence having at least 90% identity to a sequence selected from SEQ ID NOs: 30, 35, and 37.

[0090] In some embodiments, the present disclosure provides a bispecific antibody that binds to canine IL23 and canine TNFα, comprising: (i) an anti-IL23 light chain (LC) comprising an amino acid sequence having at least 90% identity to a sequence selected from SEQ ID NOs: 19 and 22; (ii) an anti-IL23 heavy chain (HC) comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NOs: 20, 21, and 23; (iii) an anti-TNFα light chain (LC) comprising an amino acid sequence having at least 90% identity to a sequence selected from SEQ ID NOs: 40 and 41; and (iv) an anti-TNFα heavy chain (HC) comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NOs: 42, 43, and 44.

[0091] In addition to anti-IL23 LC / HC + anti-TNFα LC / HC bispecific antibody structures, the present disclosure provides bispecific antibodies 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: (i) an anti-IL23 light chain (LC) comprising a VL domain having an HVR-L1 region comprising the amino acid sequence RASQGISSWLA (SEQ ID NO: 4), an HVR-L2 region comprising the amino acid sequence YAASSLQS (SEQ ID NO: 5), and an HVR-L3 region comprising the amino acid sequence 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 KGSGYSFTTYWLG (SEQ ID NO: 8), an HVR-H2 region comprising the amino acid sequence IMSPVDSDIR (SEQ ID NO: 9), and an HVR-H3 region comprising the amino acid sequence ARRRPGQGYFDF (SEQ ID NO: 10); (iii) an anti-TNFα scFv antibody fused to the HC, comprising a VL domain having an HVR-L1 region comprising the amino acid sequence RASQGIRNYLA (SEQ ID NO: 27), an HVR-L2 region comprising the amino acid sequence AASTLQ (SEQ ID NO: 28), and an HVR-L3 region comprising the amino acid sequence QRYNRAPYT (SEQ ID NO: 29), and a VH domain having an HVR-H1 region comprising the amino acid sequence FTFDDYAMH (SEQ ID NO: 31), an HVR-H2 region comprising the amino acid sequence AITWNSGHIDYADSVEGR (SEQ ID NO: 32), and an HVR-H3 region comprising the amino acid sequence AKVSYLSTASSLDY (SEQ ID NO: 33); Includes.

[0092] 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, the antibody comprising: (i) an anti-TNFα light chain (LC) comprising a VL domain having an HVR-L1 region comprising the amino acid sequence RASQGIRNYLA (SEQ ID NO: 27), an HVR-L2 region comprising the amino acid sequence AASTLQ (SEQ ID NO: 28), and an HVR-L3 region comprising the amino acid sequence QRYNRAPYT (SEQ ID NO: 29); (ii) an anti-TNFα heavy chain (HC) comprising a VH domain having an HVR-H1 region comprising the amino acid sequence FTFDDYAMH (SEQ ID NO: 31), an HVR-H2 region comprising the amino acid sequence AITWNSGHIDYADSVEGR (SEQ ID NO: 32), and an HVR-H3 region comprising the amino acid sequence AKVSYLSTASSLDY (SEQ ID NO: 33); and (iii) an anti-IL23 scFv antibody fused to the HC, comprising a VL domain having an HVR-L1 region comprising the amino acid sequence RASQGISSWLA (SEQ ID NO: 4), an HVR-L2 region comprising the amino acid sequence YAASSLQS (SEQ ID NO: 5), and an HVR-L3 region comprising the amino acid sequence QQYNIYPYT (SEQ ID NO: 6), and a VH domain having an HVR-H1 region comprising the amino acid sequence KGSGYSFTTYWLG (SEQ ID NO: 8), an HVR-H2 region comprising the amino acid sequence IMSPVDSDIR (SEQ ID NO: 9), and an HVR-H3 region comprising the amino acid sequence ARRRPGQGYFDF (SEQ ID NO: 10); Includes.

[0093] In some embodiments, the antibody comprises a label or is conjugated to a second moiety. The terms "label" and "detectable label" refer to a moiety attached to an antibody or its analyte that renders the reaction (e.g., binding) between members of a specific binding pair detectable. A labeled member of a specific binding pair is referred to as "detectably labeled." Thus, the term "labeled binding protein" refers to a protein that incorporates a label that renders the binding protein identifiable. In some embodiments, the label is a detectable marker that can produce a signal detectable by visual or instrumental means, such as the incorporation of a radiolabeled amino acid or the attachment of a biotinyl moiety to a polypeptide that can be detected by labeled avidin (e.g., streptavidin containing a fluorescent marker or enzymatic activity that can be detected by optical or colorimetric methods). Examples of labels for polypeptides include, but are not limited to, radioisotopes or radionuclides (e.g., 3 H, 14 C. 35 S, 90 Y, 99 Tc, 111 In, 125 I, 131 I, 177 Lu, 166 Ho, or 153Sm), chromogens, fluorescent labels (e.g., FITC, rhodamine, lanthanide phosphors), enzyme labels (e.g., horseradish peroxidase, luciferase, alkaline phosphatase), chemiluminescent markers, biotinyl groups, predetermined polypeptide epitopes recognized by secondary reporters (e.g., leucine zipper pair sequences, binding sites for secondary antibodies, metal binding domains, epitope tags), and magnetic agents such as gadolinium chelates. Representative examples of labels commonly utilized in immunoassays include light-emitting moieties (e.g., acridinium compounds) and fluorescence-emitting moieties (e.g., fluorescein). In this regard, the moiety itself need not be detectably labeled but may become detectable upon reaction with yet another moiety.

[0094] An "amino acid sequence" refers to 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, with no minimum length restriction. Such polymers of amino acid residues may contain natural or unnatural amino acid residues, including, but not limited to, peptides, oligopeptides, dimers, trimers, and multimers of amino acid residues. Both full-length proteins and fragments thereof are encompassed by the definition. The term also includes post-expression modifications of the polypeptide (e.g., glycosylation, sialylation, acetylation, phosphorylation, and the like). Furthermore, for purposes of this disclosure, "polypeptide" refers to a protein containing modifications such as deletions, additions, and substitutions (generally conservative in nature) to the native sequence, so long as the desired activity is maintained. These modifications may be deliberate, such as through site-directed mutagenesis, or may be accidental, such as through mutations of hosts producing the protein or errors due to PCR amplification.

[0095] As used herein, the term "epitope" refers to a site on a target molecule (e.g., an antigen, such as a protein, nucleic acid, carbohydrate, or lipid) to which an antigen-binding molecule (e.g., an antibody, antibody fragment, or scaffold protein containing an antibody-binding region) binds. Epitopes comprise chemically active surface groups of molecules, such as amino acids, polypeptides, or sugar side chains, and often have specific three-dimensional structural characteristics as well as specific charge characteristics. Epitopes can be formed from both contiguous residues of a target molecule or juxtaposed noncontiguous residues (e.g., amino acids, nucleotides, sugars, or lipid moieties). Epitopes formed from contiguous residues (e.g., amino acids, nucleotides, sugars, or lipid moieties) are typically retained upon exposure to denaturing solvents, whereas epitopes formed by tertiary folding are typically lost upon treatment with denaturing solvents. Epitopes can include, but are not limited to, at least three, at least five, or 8-10 residues (e.g., amino acids or nucleotides). In some examples, 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 that antigen. In some embodiments, an epitope can be identified by a certain minimum distance to CDR residues on an antigen-binding molecule. In some embodiments, an epitope can be identified by the above distance and further limited to residues involved in binding (e.g., hydrogen bonding) between antibody and antigen residues. Epitopes can also be identified by various scanning methods; for example, alanine or arginine scanning can indicate one or more residues with which an antigen-binding molecule can interact. Unless explicitly stated, the identification of a set of residues as an epitope does not exclude other residues from being part of the epitope for a particular antibody. Rather, the existence of such a set represents a minimal system (or species) of epitopes. Thus, in some embodiments, the set of residues identified as an epitope is not an exclusive list of residues for the epitope on the antigen, but rather represents the minimal epitope of relevance to the antigen.

[0096] 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.

[0097] In at least one embodiment, an anti-IL23 antibody of the 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) such as (i) and a heavy chain variable domain (VH) such as (ii).

[0098] In at least one embodiment, an anti-IL23 antibody of the 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) such as (i) and a heavy chain variable domain (VH) such as (ii).

[0099] 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.

[0100] In at least one embodiment, the anti-TNFα antibody of the 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, in which 1, 2, 3, 4, 5, or 6 amino acids of the light chain variable domain (VL) are replaced by 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, in which 1, 2, 3, 4, 5, or 6 amino acids of the heavy chain variable domain (VH) are replaced by different amino acids; or (iii) a light chain variable domain (VL) such as (i) and a heavy chain variable domain (VH) such as (ii).

[0101] In at least one embodiment, the anti-TNFα antibody 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 the amino acid sequence of SEQ ID NO: 26, 34, or 36; (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 the amino acid sequence of SEQ ID NO: 30, 35, or 37; or (iii) a heavy chain variable domain (VH) such as (i) and a heavy chain variable domain (VH) such as (ii).

[0102] As used herein, the term "constant region" or "constant domain" refers to a region comprising at least three constant domains.

[0103] 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 gamma, delta, alpha, epsilon, and mu. Each heavy chain constant region corresponds to an antibody's isotype. For example, an antibody comprising a gamma constant region is an IgG antibody, a delta constant region is an IgD antibody, an alpha constant region is an IgA antibody, a mu constant region is an IgM antibody, and an epsilon constant region is an IgE antibody. A particular isotype can be further subdivided into subclasses. For example, IgG antibodies include, but are not limited to, IgG1 (containing a γ1 constant region), IgG2 (containing a γ2 constant region), IgG3 (containing a γ3 constant region), and IgG4 (containing a γ4 constant region) antibodies; IgA antibodies include, but are not limited to, IgA1 (containing an α1 constant region) and IgA2 (containing an α2 constant region) antibodies; and IgM antibodies include, but are not limited to, IgM1 and IgM2.

[0104] 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. Deletions and modifications within the domain that do not alter function are encompassed within the term "constant region" unless otherwise specified. Dogs, cats, and horses have antibody classes such as IgG, IgA, IgD, IgE, and IgM. Within the canine IgG antibody class are IgG-A, IgG-B, IgG-C, and IgG-D.

[0105] In at least one embodiment, the disclosure provides (a) a chimeric anti-IL23 antibody comprising: (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 such as (i) and a heavy chain sequence such as (ii).

[0106] In some embodiments, the anti-IL23 and anti-TNFα antibodies of the disclosure may comprise a canine heavy chain constant region selected from an IgG-A, IgG-B, IgG-C, and IgG-D constant region.

[0107] In some embodiments, at least one amino acid residue in a portion of the mouse heavy chain variable domain (VH) or the mouse light chain variable domain (VL) is replaced with the corresponding amino acid from a canine variable region, hi some embodiments, the modified chain is fused to a canine constant heavy chain or a canine constant light chain.

[0108] In some embodiments, the caninized TNFα antibody comprises (i) the light chain VL domain sequence of SEQ ID NO: 11 or 13, or a variant thereof which retains the same HVR sequences 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) the heavy chain VH domain sequence of SEQ ID NO: 12 or 14, or a variant thereof which retains the same HVR sequences 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).

[0109] In at least one embodiment, the disclosure provides (a) a caninized anti-IL23 antibody comprising (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 such as (i) and a heavy chain sequence such as (ii).

[0110] In at least one embodiment, the present disclosure provides a bispecific antibody that binds to canine IL23 and canine TNFα, and that comprises (i) a light chain variable domain (VL) comprising the amino acid sequence 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.

[0111] 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.

[0112] In some embodiments, the IgG Fc polypeptide variant includes a companion animal species IgG Fc polypeptide variant. In some embodiments, the IgG Fc polypeptide variant includes a canine IgG Fc polypeptide variant. In some embodiments, the IgG Fc polypeptide variant (e.g., a canine IgG-A Fc polypeptide variant, a canine IgG-C Fc polypeptide variant, or a canine IgG-D Fc polypeptide variant, a feline IgG1a Fc polypeptide variant) has a capability that the reference (e.g., wild-type) polypeptide substantially lacks.

[0113] The antibody can be modified to extend or shorten its half-life. In some embodiments requiring a higher dose of antibody, a shorter half-life may be desirable for acute treatment. In some embodiments requiring a lower dose of antibody, a longer half-life may be desirable for prolonged treatment. For example, as discussed below, IgG Fc mutations that affect FcRn interaction can be introduced.

[0114] In some embodiments, the IL23 and / or TNFα antibody comprises a wild-type IgG Fc or an IgG Fc variant that has complement fixation activity (or complement dependent cytotoxicity (CDC)). In some embodiments, the IL23 and / or TNFα antibody comprises a wild-type IgG Fc or an IgG Fc variant that has antibody-dependent cellular cytotoxicity (ADCC) activity. In some embodiments, the IL23 and / or TNFα antibody comprises a wild-type IgG Fc or an IgG Fc variant that has antibody-dependent cellular phagocytosis (ADCP) activity. In some embodiments, the IL23 and / or TNFα antibody comprises a wild-type IgG Fc or an IgG Fc variant that has 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.

[0115] In some embodiments, the IL23 and / or TNFα antibody comprises a wild-type IgG Fc or an IgG Fc variant that binds to canine FcRn at low pH.

[0116] In some embodiments, the IgG Fc variants (e.g., canine IgG Fc polypeptide variants) have modified FcRn binding affinity relative to a reference polypeptide, hi some embodiments, the IgG Fc variants have 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 about pH 5.0, about pH 5.5, about pH 6.0, or about pH 6.5) relative to the reference polypeptide.

[0117] In some embodiments, the anti-IL23 antibody is at least 5x10 as measured by a biosensor. -6 Under M, 1x10 -6 Under M, 5x10 -7 Under M, 1x10 -7 Under M, 5x10 -8 Under M, 1x10 -8 Under M, 5x10 -9 Under M, 1x10 -9 Under M, 5x10 -10 Under M, 1x10 -10 Under M, 5x10 -11 Under M, 1x10 -11 Under M, 5x10 -12 Less than M or 1x10 -12 It binds to canine IL23, feline IL23, or equine IL23 with a dissociation constant (Kd) less than M.

[0118] In some embodiments, the anti-IL23 antibody has a titer of 5x10 as measured by biolayer interferometry. -6 M~1x10 -6 Medium, 5x10 -6 M~5x10 -7 Medium, 5x10 -6 M~1x10 -7 Medium, 5x10 -6 M~5x10 -8 Medium, 5x10 -6 M~1x10 -8M、5x10 -6 M~5x10 -9 M、5x10 -6 M~1x10 -9 M、5x10 -6 M~5x10 -10 M、5x10 -6 M~1x10 -10 M、5x10 -6 M~5x10 -11 M、5x10 -6 M~1x10 -11 M、5x10 -6 M~5x10 - 12M、5x10 -6 M~1x10 -12 M、1x10 -6 M~5x10 -7 M、1x10 -6 M~1x10 -7 M、1x10 -6 M~5x10 -8 M、1x10 -6 M~1x10 -8 M、1x10 -6 M~5x10 -9 M、1x10 -6 M~1x10 -9 M、1x10 -6 M~5x10 -10 M、1x10 -6 M~1x10 -10 M、1x10 -6 M~5x10 -11 M、1x10 -6 M~1x10 -11 M、1x10 -6 M~5x10 -12 M、1x10 -6 M~1x10 -12 M、5x10 -7 M~1x10 -7 M、5x10 -7 M~5x10 -8 M、5x10 -7 M~1x10 -8 M、5x10 -7 M~5x10 -9 M、5x10 -7 M~1x10 -9 M、5x10 -7 M~5x10 -10 M、5x10-7 M~1x10 -10 M、5x10 -7 M~5x10 -11 M、5x10 -7 M~1x10 -11 M、5x10 -7 M~5x10 -12 M、5x10 -7 M~1x10 -12 M、1x10 -7 M~5x10 -8 M、1x10 -7 M~1x10 -8 M、1x10 -7 M~5x10 -9 M、1x10 -7 M~1x10 -9 M、1x10 -7 M~5x10 -10 M、1x10 -7 M~1x10 -10 M、1x10 -7 M~5x10 -11 M、1x10 -7 M~1x10 -11 M、1x10 -7 M~5x10 -12 M、1x10 -7 M~1x10 -12 M、5x10 -8 M~1x10 -8 M、5x10 -8 M~5x10 -9 M、5x10 -8 M~1x10 -9 M、5x10 -8 M~5x10 -10 M、5x10 -8 M~1x10 -10 M、5x10 -8 M~5x10 -11 M、5x10 -8 M~1x10 -11 M、5x10 -8 M~5x10 -12 M、5x10 -8 M~1x10 -12 M、1x10 -8 M~5x10 -9 M、1x10 -8 M~1x10 -9 M、1x10 -8M~5x10 -10 M、1x10 -8 M~1x10 -10 M、1x10 -8 M~5x10 -11 M、1x10 -8 M~1x10 -11 M、1x10 -8 M~5x10 -12 M、1x10 -8 M~1x10 -12 M、5x10 -9 M~1x10 -9 M、5x10 -9 M~5x10 -10 M、5x10 -9 M~1x10 -10 M、5x10 -9 M~5x10 -11 M、5x10 -9 M~1x10 -11 M、5x10 -9 M~5x10 -12 M、5x10 -9 M~1x10 -12 M、1x10 -9 M~5x10 -10 M、1x10 -9 M~1x10 -10 M、1x10 -9 M~5x10 -11 M、1x10 -9 M~1x10 -11 M、1x10 -9 M~5x10 -12 M、1x10 -9 M~1x10 -12 M、5x10 -10 M~1x10 -10 M、5x10 -10 M~5x10 -11 M、1x10 -10 M~5x10 -11 M、1x10 -10 M~1x10 -11 M、1x10 -10 M~5x10 -12 M、1x10 -10 M~1x10 -12 M、5x10 -11 M~1x10 -12 M、5x10 -11 M~5x10-12 Medium, 5x10 -11 M~1x10 -12 Medium, 1x10 -11 M~5x10 -12 Medium, or 1x10 -11 M~1x10 -12 It binds to canine IL23, human IL23, feline IL23, or equine IL23 with a Kd of M.

[0119] In some embodiments, the anti-IL23 antibody binds to canine IL23, human IL23, feline IL23, or equine IL23 as determined by immunoblot analysis.

[0120] In some embodiments, the anti-TNFα antibody is at least 5x10 as measured by a biosensor. -6 Under M, 1x10 -6 Under M, 5x10 -7 Under M, 1x10 -7 Under M, 5x10 -8 Under M, 1x10 -8 Under M, 5x10 -9 Under M, 1x10 -9 Under M, 5x10 -10 Under M, 1x10 -10 Under M, 5x10 -11 Under M, 1x10 -11 Under M, 5x10 -12 Less than M or 1x10 -12 It binds to canine TNFα, human TNFα, feline TNFα, or equine TNFα with a dissociation constant (Kd) of less than M.

[0121] In some embodiments, the anti-TNFα antibody is at least 5x10 as measured by a biosensor. -6 M~1x10 -6 Medium, 5x10 -6 M~5x10 -7 Medium, 5x10 -6 M~1x10 -7 Medium, 5x10 -6 M~5x10 -8 Medium, 5x10 -6 M~1x10 -8 Medium, 5x10 -6 M~5x10-9 M、5x10 -6 M~1x10 -9 M、5x10 -6 M~5x10 -10 M、5x10 -6 M~1x10 -10 M、5x10 -6 M~5x10 -11 M、5x10 -6 M~1x10 -11 M、5x10 -6 M~5x10 - 12M、5x10 -6 M~1x10 -12 M、1x10 -6 M~5x10 -7 M、1x10 -6 M~1x10 -7 M、1x10 -6 M~5x10 -8 M、1x10 -6 M~1x10 -8 M、1x10 -6 M~5x10 -9 M、1x10 -6 M~1x10 -9 M、1x10 -6 M~5x10 -10 M、1x10 -6 M~1x10 -10 M、1x10 -6 M~5x10 -11 M、1x10 -6 M~1x10 -11 M、1x10 -6 M~5x10 -12 M、1x10 -6 M~1x10 -12 M、5x10 -7 M~1x10 -7 M、5x10 -7 M~5x10 -8 M、5x10 -7 M~1x10 -8 M、5x10 -7 M~5x10 -9 M、5x10 -7 M~1x10 -9 M、5x10 -7 M~5x10 -10 M、5x10 -7 M~1x10 -10M、5x10 -7 M~5x10 -11 M、5x10 -7 M~1x10 -11 M、5x10 -7 M~5x10 -12 M、5x10 -7 M~1x10 -12 M、1x10 -7 M~5x10 -8 M、1x10 -7 M~1x10 -8 M、1x10 -7 M~5x10 -9 M、1x10 -7 M~1x10 -9 M、1x10 -7 M~5x10 -10 M、1x10 -7 M~1x10 -10 M、1x10 -7 M~5x10 -11 M、1x10 -7 M~1x10 -11 M、1x10 -7 M~5x10 -12 M、1x10 -7 M~1x10 -12 M、5x10 -8 M~1x10 -8 M、5x10 -8 M~5x10 -9 M、5x10 -8 M~1x10 -9 M、5x10 -8 M~5x10 -10 M、5x10 -8 M~1x10 -10 M、5x10 -8 M~5x10 -11 M、5x10 -8 M~1x10 -11 M、5x10 -8 M~5x10 -12 M、5x10 -8 M~1x10 -12 M、1x10 -8 M~5x10 -9 M、1x10 -8 M~1x10 -9 M、1x10 -8 M~5x10 -10 M、1x10-8 M~1x10 -10 M、1x10 -8 M~5x10 -11 M、1x10 -8 M~1x10 -11 M、1x10 -8 M~5x10 -12 M、1x10 -8 M~1x10 -12 M、5x10 -9 M~1x10 -9 M、5x10 -9 M~5x10 -10 M、5x10 -9 M~1x10 -10 M、5x10 -9 M~5x10 -11 M、5x10 -9 M~1x10 -11 M、5x10 -9 M~5x10 -12 M、5x10 -9 M~1x10 -12 M、1x10 -9 M~5x10 -10 M、1x10 -9 M~1x10 -10 M、1x10 -9 M~5x10 -11 M、1x10 -9 M~1x10 -11 M、1x10 -9 M~5x10 -12 M、1x10 -9 M~1x10 -12 M、5x10 -10 M~1x10 -10 M、5x10 -10 M~5x10 -11 M、1x10 -10 M~5x10 -11 M、1x10 -10 M~1x10 -11 M、1x10 -10 M~5x10 -12 M、1x10 -10 M~1x10 -12 M、5x10 -11 M~1x10 -12 M、5x10 -11 M~5x10 -12 M、5x10 -11M~1x10 -12 Medium, 1x10 -11 M~5x10 -12 Medium, or 1x10 -11 M~1x10 -12 It binds to canine TNFα, human TNFα, feline TNFα, or equine TNFα with a Kd of M.

[0122] In some embodiments, the anti-TNFα antibody binds to canine TNFα, human TNFα, feline TNFα, or equine TNFα as determined by immunoblot analysis.

[0123] In some embodiments, the variant has at least 1, 2, 3, 4, 5, or 6 amino acids substituted with different amino acids.

[0124] In some embodiments, a variant has at least about 50% sequence identity with a reference nucleic acid molecule or polypeptide, after aligning the sequences and introducing gaps as necessary to maximize the percent sequence identity, without considering any conservative substitutions as part of the sequence identity. 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, a 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.

[0125] In some embodiments, the anti-IL23 antibody may 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%, 10% and 20%, 10% and 25%, 10% and 30%, 10% and 35%, 10% and 40%, 10% and 45%, 10% and 50%, 10% and 60%, 10% and 70%, 10% and 80%, 10% and 90%, 10% and 100%, 15% and 20%, 15% and 25%, 15% and 30%, 15% and 35%, 15% and 40%, 15% and 45%, 15 ... 5%~60%, 15%~70%, 15%~80%, 15%~90%, 15%~100%, 20%~25%, 20%~30%, 20%~35%, 20%~40%, 20%~45%, 20%~50%, 20%~60%, 20%~70%, 20%~80%, 20%~90%, 20%~100%, 25%~30%, 25%~35%, 25%~40%, 25%~45%, 25%~50%, 25%~60%, 25%~70%, 25%~80%, 25 %~90%, 25%~100%, 30%~35%, 30%~40%, 30%~45%, 30%~50%, 30%~60%, 30%~70%, 30%~80%, 30%~90%, 30%~100%, 35%~40%, 35%~45%, 35%~50%, 35%~60%, 35%~70%, 35%~80%, 35%~90%, 35%~100%, 40%~45%, 40%~50%, 40%~60%, 40%~70%, 40%~80%, 40 %~90%, 40%~100%, 45%~50%, 45%~60%, 45%~70%, 45%~80%, 45%~90%, 45%~100%, 50%~60%, 50%~70%, 50%~80%, 50%~90%, 50%~100%, 60%~70%, 60%~80%, 60%~90%, 60%~100%, 70%~80%, 70%~90%, 70%~100%, 80%~90%, 80%~100%, or 90%~100%.

[0126] In some embodiments, the anti-TNFα antibody may 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 decrease in NGF signaling function is between 10% and 15%, 10% and 20%, 10% and 25%, 10% and 30%, 10% and 35%, 10% and 40%, 10% and 45%, 10% and 50%, 10% and 60%, 10% and 70%, 10% and 80%, 10% and 90%, 10% and 100%, 15% and 20%, 15% and 25%, 15% and 30%, 15% and 35%, 15% and 40%, 15% and 45%, 15% and 50%, 15 ... %~60%, 15%~70%, 15%~80%, 15%~90%, 15%~100%, 20%~25%, 20%~30%, 20%~35%, 20%~40%, 20%~45%, 20%~50%, 20%~60%, 20%~70%, 20%~80%, 20%~90%, 20%~100%, 25%~30%, 25%~35%, 25%~40%, 25%~45%, 25%~50%, 25%~60%, 25%~70%, 25%~80%, 25% ~90%, 25%~100%, 30%~35%, 30%~40%, 30%~45%, 30%~50%, 30%~60%, 30%~70%, 30%~80%, 30%~90%, 30%~100%, 35%~40%, 35%~45%, 35%~50%, 35%~60%, 35%~70%, 35%~80%, 35%~90%, 35%~100%, 40%~45%, 40%~50%, 40%~60%, 40%~70%, 40%~80%, 40 %~90%, 40%~100%, 45%~50%, 45%~60%, 45%~70%, 45%~80%, 45%~90%, 45%~100%, 50%~60%, 50%~70%, 50%~80%, 50%~90%, 50%~100%, 60%~70%, 60%~80%, 60%~90%, 60%~100%, 70%~80%, 70%~90%, 70%~100%, 80%~90%, 80%~100%, or 90%~100%.

[0127] Pharmaceutical Composition

[0128] The terms "pharmaceutical formulation" and "pharmaceutical composition" refer to a preparation that is in a form that allows the biological activity of the active ingredient(s) to be effective and that does not contain additional components that are unacceptably toxic to the subject to which the formulation is administered.

[0129] Pharmaceutically acceptable carriers are non-toxic to recipients at the dosages and concentrations used and are compatible with the other ingredients of the formulation. Pharmaceutically acceptable carriers are appropriate for the formulation used. 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 silica, or magnesium trisilicate), polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sucrose, mannitol, or amino acids (including but not limited to arginine).

[0130] The pharmaceutical composition may be stored in lyophilized form. Thus, in some embodiments, the preparation process includes a lyophilization step. The lyophilized composition may then be reconstituted, typically as an aqueous composition suitable for parenteral administration, before being administered to a canine. In other embodiments, particularly when the antibody is highly stable against thermal and oxidative denaturation, the pharmaceutical composition may be stored as a liquid (i.e., as an aqueous composition), which may be administered directly or together with a suitable diluent to a canine. The lyophilized composition may be reconstituted with sterile water for injection (WFI). A bacteriostatic agent, such as benzyl alcohol, may also be included. Thus, the present invention provides pharmaceutical compositions in solid or liquid form.

[0131] The pH of the pharmaceutical composition may range from about pH 5 to about pH 8 at the time of administration. The compositions of the present invention are sterile when used for therapeutic purposes. 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 may be maintained with or without antibacterial agents.

[0132] Exemplary Uses of Antibodies and Pharmaceutical Compositions

[0133] The antibodies of the invention or pharmaceutical compositions comprising the antibodies of the invention may be useful for treating an IL23-mediated condition, disorder, or disease in a subject, which may be a companion animal, including but not limited to, a dog or cat.

[0134] In some embodiments, anti-IL23 antibodies, with or without anti-TNFα antibodies, or pharmaceutical compositions comprising them, may be utilized in accordance with the methods herein to treat conditions associated with IL23. 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 conditions associated with IL23 or TNFα. 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 a trend toward remission of conditions associated with IL23 and / or TNFα.

[0135] A therapeutically effective amount can be an amount in which the therapeutically beneficial effects outweigh any toxic or detrimental effects of a substance / molecule, agonist, or antagonist. A therapeutically effective amount can be delivered in one or more administrations. A "therapeutically effective amount" refers to an amount that is effective, at dosages and for periods of time necessary, to achieve the desired therapeutic or prophylactic result.

[0136] In some embodiments, pharmaceutical compositions comprising anti-IL23 and / or anti-TNFα antibodies or IL23 and / or TNFα antibodies are administered parenterally by subcutaneous administration, intravenous infusion, or intramuscular injection. In some embodiments, pharmaceutical compositions comprising anti-IL23 and / or anti-TNFα antibodies or anti-IL23 and / or anti-TNFα antibodies are administered as a bolus injection or by continuous infusion over a period of time. In some embodiments, pharmaceutical compositions comprising anti-IL23 and / or anti-TNFα antibodies or anti-IL23 and / or anti-TNFα antibodies are administered intramuscularly, intraperitoneally, intracerebrospinal, subcutaneously, intra-arterially, intraspinal fluid, intrathecally, or by inhalation routes.

[0137] The anti-IL23 antibody alone, the combination of an anti-IL23 antibody and an anti-TNFα antibody, or the combination of an anti-IL23 antibody and a TNFα binding region in a bispecific antibody described herein may be administered in an amount ranging from 0.01 mg / kg to 100 mg / kg of body weight per dose. In some embodiments, the IL23 antibody alone or the combination of an IL23 antibody and a TNFα antibody may be administered in an amount ranging from 0.5 mg / kg to 50 mg / kg of body weight per dose. In some embodiments, the IL23 and / or TNFα antibody may be administered in an amount ranging from 0.1 mg / kg to 10 mg / kg of body weight per dose. In some embodiments, the IL23 and / or TNFα antibody may be administered in an amount ranging from 0.1 mg / kg to 100 mg / kg of body weight per dose. In some embodiments, the IL23 and / or TNFα antibody may be administered in an amount ranging from 1 mg / kg to 10 mg / kg of body weight per dose. In some embodiments, the IL23 and / or TNFα antibody may 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 may be administered in an amount of 0.5 mg / kg body weight. In some embodiments, the IL23 and / or TNFα antibody may be administered in an amount of 2 mg / kg body weight.

[0138] The IL23 antibody alone or the combination of the IL23 antibody and the TNFα antibody, or the pharmaceutical composition comprising the IL23 and / or TNFα antibody, can be administered to the companion animal at one time or over a series of treatments. For example, the IL23 and / or TNFα antibody or the pharmaceutical composition comprising the IL23 and / or TNFα antibody can be administered at least once, more than once, at least two times, at least three times, at least four times, or at least five times.

[0139] In some embodiments, the dose is administered once weekly for at least two or three consecutive weeks, and in some embodiments, this treatment cycle is repeated two or more times, optionally with one or more weeks off treatment between the cycles. In other embodiments, the therapeutically effective dose is administered once daily for two to five consecutive days, and in some embodiments, this treatment cycle is repeated two or more times, optionally with one or more days or one or more weeks off treatment between the cycles.

[0140] Administration can be "in conjunction with" one or more additional therapeutic agents, including simultaneous (concurrent) administration and sequential or consecutive administration in any order. The term "concurrent" is used herein to refer to the administration of two or more therapeutic agents, where at least a portion of the administration overlaps in time, or the administration period of one therapeutic agent is short compared to the administration period of another therapeutic agent. For example, two or more therapeutic agents are administered within a specified approximate number of minutes of each other. The term "sequentially" is used herein to refer to the administration of two or more therapeutic agents, where the administration of one or more agent(s) follows the discontinuation of the administration of one or more other agent(s), or where the administration of one or more agent(s) is initiated before the administration of one or more other agent(s). For example, the administration of two or more therapeutic agents is administered more than a specified approximate number of minutes apart. As used herein, "in conjunction with" refers to the administration of one treatment modality in addition to another treatment modality. Thus, "in conjunction with" refers to the administration of one treatment modality to an animal before, during, or after the administration of the other treatment modality.

[0141] In some embodiments, the methods comprise administering an IL23 and / or TNFα antibody or a pharmaceutical composition comprising an IL23 and / or TNFα antibody in combination with 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 beta7-integrin antibody, an IL12 antibody, an IL1β antibody, or an anti-BlyS antibody.

[0142] Provided herein are methods of exposing cells to an IL23 and / or TNFα antibody or a pharmaceutical composition comprising an IL23 and / or TNFα antibody under conditions that allow for binding of the antibody to IL23 and / or TNFα. In some embodiments, the cells are exposed to the antibody or pharmaceutical composition ex vivo. In some embodiments, the cells are exposed to the antibody or pharmaceutical composition in vivo. In some embodiments, the cells are exposed to the IL23 and / or TNFα antibody or pharmaceutical composition under conditions that allow for binding of the antibody to extracellular IL23 and / or TNFα.

[0143] In some embodiments, cells may be exposed to an IL23 and / or TNFα antibody or pharmaceutical composition in vivo by any one or more of the administration methods described herein, including, but not limited to, intraperitoneal, intramuscular, or intravenous injection into a subject. In some embodiments, cells may be exposed to an IL23 and / or TNFα antibody or pharmaceutical composition ex vivo by exposing the cells to a culture medium containing the antibody or pharmaceutical composition. In some embodiments, prior to exposing the cells to a culture medium containing the antibody or pharmaceutical composition, the permeability of the cell membrane may be affected using any number of methods understood by those of skill in the art, such as electroporating the cells or exposing the cells to a solution containing calcium chloride.

[0144] In some embodiments, binding reduces IL23 and / or TNFα or IL12 signaling function by the cell. In some embodiments, the IL23 and / or TNFα antibody may 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 TNFα or IL12 signaling function in the cell 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%, 10% and 20%, 10% and 25%, 10% and 30%, 10% and 35%, 10% and 40%, 10% and 45%, 10% and 50%, 10% and 60%, 10% and 70%, 10% and 80%, 10% and 90%, 10% and 100%, 15% and 20%, 15% and 25%, 15% and 30%, 15% and 35%, 15% and 40%, 15 ... 5%, 15%~50%, 15%~60%, 15%~70%, 15%~80%, 15%~90%, 15%~100%, 20%~25%, 20%~30%, 20%~35%, 20%~40%, 20%~45%, 20%~50%, 20%~60%, 20%~70%, 20%~80%, 20%~90%, 20%~100%, 25%~30%, 25%~35%, 25%~40%, 25%~45%, 25%~50%, 25%~60%, 25%~70%, 25 %~80%, 25%~90%, 25%~100%, 30%~35%, 30%~40%, 30%~45%, 30%~50%, 30%~60%, 30%~70%, 30%~80%, 30%~90%, 30%~100%, 35%~40%, 35%~45%, 35%~50%, 35%~60%, 35%~70%, 35%~80%, 35%~90%, 35%~100%, 40%~45%, 40%~50%, 40%~60%, 40%~70%, 40%~80 %, 40%-90%, 40%-100%, 45%-50%, 45%-60%, 45%-70%, 45%-80%, 45%-90%, 45%-100%, 50%-60%, 50%-70%, 50%-80%, 50%-90%, 50%-100%, 60%-70%, 60%-80%, 60%-90%, 60%-100%, 70%-80%, 70%-90%, 70%-100%, 80%-90%, 80%-100%, or 90%-100%.

[0145] Provided herein are methods using IL23 and / or TNFα antibodies, polypeptides, and polynucleotides for the detection, diagnosis, and monitoring of pathologies associated with IL23 and / or TNFα. Provided herein are methods for determining whether a companion animal will respond to IL23 and / or TNFα antibody therapy. In some embodiments, the methods involve using an IL23 and / or TNFα antibody to detect whether the animal has cells that express IL23 and / or TNFα. In some embodiments, the detection method involves contacting a sample with an antibody, polypeptide, or polynucleotide and determining whether the level of binding differs from that of a reference or comparison sample (e.g., a control). In some embodiments, the methods may be useful for determining whether an antibody or polypeptide described herein is an appropriate therapeutic agent for a subject animal.

[0146] In some embodiments, the sample is a biological sample. The term "biological sample" refers to a quantity of material derived from an organism or originally a living 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.

[0147] In some embodiments, cells or cell / tissue lysates are contacted with an IL23 and / or TNFα antibody and binding between the antibody and the cells is determined. If the test cells exhibit binding activity compared to reference cells of the same tissue type, this may indicate that the subject would benefit from treatment with an IL23 and / or TNFα antibody. In some embodiments, the test cells are derived from tissues of companion animals.

[0148] 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), immunonephrotic inhibition assay (NIA), enzyme-linked immunosorbent assay (ELISA), and radioimmunoassay (RIA). Indicator moieties, or label groups, can be attached to the test antibody and are selected to meet the needs of the various uses of the method, which are often required to have assay equipment and compatible immunoassay procedures available. Suitable labels include, but are not limited to, radionuclides (e.g., 125 I, 131 I, 35 S, 3 H, or 32 P), enzymes (e.g., alkaline phosphatase, horseradish peroxidase, luciferase, or p-galactosidase), fluorescent moieties or proteins (e.g., fluorescein, rhodamine, phycoerythrin, GFP, or BFP), or luminescent moieties (e.g., Qdot™ nanoparticles supplied by Quantum Dot Corporation, Palo Alto, Calif.). General techniques to be used in performing the various immunoassays described above are known to those of skill in the art.

[0149] For diagnostic purposes, polypeptides, including antibodies, can be labeled with a detectable moiety, including, but not limited to, radioisotopes, fluorescent labels, and various enzyme-substrate labels known in the art. Methods for conjugating labels to antibodies are known in the art. In some embodiments, IL23 and / or TNFα antibodies need not be labeled, and their presence can be detected using a second, labeled antibody that binds to the first IL23 and / or TNFα antibody. 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. Generally, antibodies or polypeptides are conjugated to radionuclides ( 111 In, 99 Tc, 14 C. 131 I, 125 I, 3 H, or any other radionuclide label, including those outlined herein. Antibodies may also be used as staining reagents in pathology, using techniques well known in the art.

[0150] In some embodiments, the first antibody is used for diagnostic purposes and the second antibody is used as a therapeutic agent. In some embodiments, the first and second antibodies are different. In some embodiments, the first and second antibodies can both bind to the antigen simultaneously by binding to distinct epitopes. [Example]

[0151] Various features and embodiments of the present disclosure are illustrated in the following representative examples, which are intended to be illustrative and not limiting. As one skilled in the art will readily appreciate, the specific examples are merely illustrative of the invention, which is more fully described in the claims that follow. It should be understood that all embodiments and features described in this application are interchangeable and combinable with all embodiments contained herein.

[0152] Example 1: Recombinant expression of canine IL23 or p40

[0153] The heterodimeric canine IL23 polypeptide sequence for recombinant expression was designed as SEQ ID NO: 1. This canine IL23 polypeptide (SEQ ID NO: 1) contains a linker between the two subunits to stabilize the complex, a poly-His tag for purification, and a tag for in vitro site-specific biotinylation. This single-chain canine IL23 polypeptide was expressed in CHO cells as a monomeric protein. The expressed protein was purified using a Ni-NTA column.

[0154] The heterodimeric feline IL23 polypeptide sequence 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 to stabilize the complex. The canine p40 polypeptide was also expressed in mammalian CHO cells.

[0155] Example 2: Anti-IL23 antibody gene synthesis, expression from CHO cells, and purification

[0156] The DNA sequence encoding the anti-IL23 (clone C340) VL domain of SEQ ID NO: 3 and the VH domain of SEQ ID NO: 7 was fused to a DNA sequence encoding the canine constant CL kappa 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 vector(s) were transfected into CHO cells. The chimeric anti-IL23 antibodies were expressed in CHO cells, and the expressed antibodies were purified by one or more of a variety of steps, including protein A column chromatography.

[0157] Results: The chimeric antibody was well expressed and presented as a monodisperse peak in gel filtration chromatography.

[0158] Example 3: Demonstration of canine IL23 binding activity

[0159] Most monoclonal anti-IL23 antibodies fail to bind to their non-primate cognate targets. Surprisingly, as shown by the results of the study in this example, the chimeric anti-IL23 antibodies described herein (e.g., the anti-IL23 antibodies in Table 1 above) were found to have high binding affinity for both canine IL23 and canine p40. This study was performed using Biacore 8K to measure the binding affinity of the anti-IL23 antibodies for the IL23 and p40 antigens.

[0160] material

[0161] [Table 2]

[0162] [Table 3]

[0163] method

[0164] 1. Measurement of antigen affinity for antibody

[0165] Preparation of running buffer: Dilute 1 volume of 10x buffer with 9 volumes of degassed, filtered MilliQ water.

[0166] Preparation of renaturation buffer (10 mM glycine): Weigh out a certain amount of glycine and dissolve it in MilliQ water. Adjust the pH to 1.5-1.7 to obtain the renaturation buffer.

[0167] The assay was carried out at 25°C and the running buffer was HBS-EP+.

[0168] The antibodies were injected as capture on a Series S Sensor Chip Protein A.

[0169] The antigen was diluted to several concentrations and injected over the surfaces of flow cells 1 and 2 as the association phase, followed by injection of running buffer as the dissociation phase.

[0170] The coupling model is shown below: The execution configurations used are listed in Table 4 below.

[0171] [Table 4]

[0172] result

[0173] All data were processed using Biacore 8K Evaluation software version 3.0. At each cycle, flow cell 1 and a buffer blank injection were used as double references for response unit subtraction. Biacore binding affinity values ​​are presented in Table 5 below.

[0174] [Table 5]

[0175] Example 4: Caninization of anti-IL23 C340 antibody

[0176] 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. Initial 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. A list of these candidate amino acid substitutions is provided below in Table 6.

[0177] [Table 6-1] [Table 6-2] [Table 6-3]

[0178] Based on the amino acid substitution candidates, genes encoding several caninized VL (SEQ ID NO: 3) and VH (SEQ ID NO: 7) variants were expressed using human IgG1 and kappa frameworks. The amino acid sequences of the caninized VL and VH variants are aligned and listed in Tables 7 and 8 below (the "-" symbol indicates the position where the amino acid was changed).

[0179] [Table 7]

[0180] [Table 8]

[0181] Genes encoding each of the caninized VL and VH variants listed above in Tables 7 and 8 were expressed in mammalian cell lines. Various combinations of anti-IL23 VL and VH polypeptides were tested for expression and canine IL23 binding.

[0182] As shown by the results listed in Table 9 (below), the "wild-type" anti-IL23 C340 VL and VH domain polypeptides were well expressed. Surprisingly, some versions of the caninized VH domain polypeptide (e.g., variant V1) were found to pair well with the "wild-type" C340 VL (SEQ ID NO: 3). Similarly, the caninized VL (e.g., variant V2) paired well with the 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 of these studies are presented in Table 9.

[0183] [Table 9]

[0184] As an exemplary caninized anti-IL23, a variant comprising the VL domain V2.1 (SEQ ID NO: 11) and the VH domain V1.1 (SEQ ID NO: 12) was found to be well expressed and retain full canine IL23 binding activity using Biacore binding affinity assays.

[0185] A mutation was also introduced at amino acid position 252 (EU numbering), located in the AB turn of the Fc region. This mutation changes the amino acid at position 252 to Y (L252Y), resulting in an extended in vivo half-life of the IgG antibody. This is described in U.S. Patent No. 7,658,921 B2. This half-life-extending Fc region mutation, L252Y, was introduced into caninized anti-IL23 heavy chain IgG-B to generate the "long-acting" caninized anti-IL23 of SEQ ID NO: 20.

[0186] Example 5: Fetinylation of anti-IL23 (C340) antibody

[0187] This example illustrates felinization studies of anti-IL23(C340) having a VL domain of SEQ ID NO:3 and a VH domain of SEQ ID NO:7.

[0188] The VL and VH domains are felineized using the same general methodology as described for caninization in Example 4. An initial analysis of the anti-IL23 C340 VL and VH amino acid sequences was performed to identify potential amino acid substitutions at various positions that could be used to felineize the sequences. Genes encoding felineized VL (SEQ ID NO: 3) and VH (SEQ ID NO: 7) variants using a feline IgG framework are expressed in a mammalian cell line. The feline IL23-binding activity of the expressed felineized variants is measured and identified. Various combinations of anti-IL23 VL and VH polypeptides were tested for expression and canine IL23 binding.

[0189] Alternatively, a felinized anti-IL23 antibody can be expressed using a feline Fc with a half-life extended by substituting "Y" for amino acid 252 (EU numbering) in the feline Fc. This half-life extending Fc region mutation, L252Y, is introduced into a felinized anti-IL23 heavy chain IgG-B, generating a "long-acting" felinized anti-IL23.

[0190] Example 6: Caninization of anti-TNFα D2E7 antibody

[0191] This example illustrates caninization studies of anti-TNFα D2E7, which has a VL domain of SEQ ID NO:26 and a VH domain of SEQ ID NO:30.

[0192] The resulting caninized anti-TNFα D2E7 VL domain sequence (SEQ ID NO: 34) and VH domain sequence (SEQ ID NO: 35) were used to generate a caninized anti-TNFα D2E7 antibody. A caninized anti-TNFα D2E7 antibody was generated, comprising the caninized VL domain of SEQ ID NO: 34 linked to a canine kappa constant region to yield the light chain sequence of SEQ ID NO: 40, and the caninized VH domain of SEQ ID NO: 35 linked to a canine IgG-B constant region to yield the heavy chain sequence of SEQ ID NO: 41. This caninized anti-TNFα antibody was expressed using mammalian Expi293F cells.

[0193] This example demonstrates that the caninized anti-TNFα antibodies described herein have high binding affinity to canine TNFα (Sino Biologicals). This study was performed using Biacore 8K to measure the binding affinity of the antibody to the antigen.

[0194] material

[0195] [Table 10]

[0196] [Table 11]

[0197] method

[0198] 1. Measurement of antigen affinity for antibody

[0199] Preparation of running buffer: Dilute 1 volume of 10x buffer with 9 volumes of degassed, filtered MilliQ water.

[0200] Preparation of renaturation buffer (10 mM glycine): Weigh out a certain amount of glycine and dissolve it in MilliQ water. Adjust the pH to 1.5-1.7 to obtain the renaturation buffer.

[0201] The assay was carried out at 25°C and the running buffer was HBS-EP+.

[0202] The antibodies were injected as capture on a Series S Sensor Chip Protein A.

[0203] The antigen was diluted to several concentrations and injected over the surfaces of flow cells 1 and 2 as the association phase, followed by injection of running buffer as the dissociation phase.

[0204] The combined model is shown below: The execution configurations used are listed in Table 12 below.

[0205] [Table 12]

[0206] result

[0207] All data were processed using Biacore 8K Evaluation software version 3.0. At each cycle, flow cell 1 and a buffer blank injection were used as double references for response unit subtraction. Biacore binding affinity values ​​are presented in Table 13 below.

[0208] [Table 13]

[0209] The caninized version of the anti-TNFα D2E7 antibody comprises a light chain (LC) of SEQ ID NO: 40, which has a caninized variable domain (VL) linked to a canine kappa constant region, and a heavy chain of SEQ ID NO: 43, which has a caninized variable domain (VH) linked to a canine IgG-B constant region, and further comprises an Fc region with the EU numbering 252Y "long-acting" variant in the Fc. Caninized anti-TNFα have also been engineered with long-acting Fc variants of the IgG-A, IgG-C, and IgG-D constant regions.

[0210] A felineized version of the anti-TNFα D2E7 antibody has also been designed, comprising the felineized light chain variable domain (VL) of D2E7 feline kappa (SEQ ID NO: 36) and the felineized heavy chain variable domain (VH) of D2E7 (SEQ ID NO: 37), and feline IgGa, or IgGb (the Fc of canine IgG can be modified by replacing amino acid 252 (EU numbering) in the Fc with "Y").

[0211] Example 7: Expression and purification of bispecific anti-canine IL23 and anti-canine TNFα molecules from CHO cells

[0212] 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 for disulfide bond formation.

[0213] Using the anti-TNFα scFv molecules and caninized anti-IL23 antibodies described in Table 1, a first exemplary bispecific fusion molecule that specifically binds both canine IL23 and canine TNFα was designed with 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) an scFv caninized anti-TNFα antibody of SEQ ID NO: 45. The fusion structure of the complete bispecific anti-IL23 / anti-TNFα antibody is presented 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 comprises the scFv anti-TNFα antibody of SEQ ID NO: 46, which has two cysteines that allow for the formation of intramolecular SEQ ID NOs. The resulting complete bispecific antibody fusion structure with the scFv of SEQ ID NO: 46 is presented as the amino acid sequence of SEQ ID NO: 48. The bispecific antibody molecules of SEQ ID NOs: 47 and 48 are expressed from mammalian cells and purified by single-step Protein A column chromatography.

[0214] A second exemplary bispecific anti-IL23 / anti-TNFα antibody that specifically binds both canine IL23 and canine TNFα was designed with a four-chain structure formed by combining the following structures: (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 (the heavy chain includes a "knob"), and (2) an anti-TNFα antibody structure formed with 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 (the heavy chain includes a "hole"). Additionally, as shown by 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, a cysteine ​​was removed from the CL region and added to the VL domain, and (2) in the anti-TNFα heavy chain of SEQ ID NO: 44, a cysteine ​​was removed from the CH1 region and 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 were monodisperse. Mass spectrometry analysis of the proteins obtained after deglycosylation and reduction with DTT indicated a 1:1:1:1 ratio, as expected.

[0215] Example 8: Study on the treatment of canine IBD with anti-IL23 antibodies

[0216] The efficacy and safety of anti-IL23 for managing inflammatory bowel disease (IBD) in canines can be evaluated using various doses ranging from 0.01 mg / kg to 100 mg / kg.

[0217] All canines received either caninized IL23 antibody or caninized long-acting antibody. The first dose was administered on day 0. Subsequent doses were administered weekly, biweekly, monthly, or every two, three, or up to six months.

[0218] Histopathological examination of endoscopic gastrointestinal biopsies can be used to determine the efficacy of caninized IL23 antibodies in managing IBD.

[0219] IBD biomarkers that can be measured include, but are not limited to, IL-1 beta, IL6, IL8, IL9, IFN-gamma, TNF alpha, CCL2, IL22, CRP, LL37, TFF3, OSM.

[0220] The Canine Inflammatory Bowel Disease Activity Index (CIBDAI) score can be used to screen candidates for treatment.

[0221] Controlled conditions may include diet.

[0222] Example 9: Study of the treatment of canine IBD with combination therapy using anti-IL23 and anti-TNFα antibodies

[0223] The efficacy and safety of IL23 monoclonal antibody in combination with TNFα monoclonal antibody for managing inflammatory bowel disease (IBD) in canines can be evaluated using any combination of IL23 monoclonal antibody at various doses ranging from 0.01 mg / kg to 100 mg / kg and TNFα monoclonal antibody ranging from 0.01 mg / kg to 100 mg / kg.

[0224] All canines received a combination of either a caninized IL23 antibody or a caninized long-acting IL23 antibody with either a caninized TNFα antibody or a caninized long-acting IL23 antibody. The first dose was administered on day 0. Subsequent doses were administered weekly, biweekly, monthly, or every two, three, or up to six months or more.

[0225] Alternatively, the efficacy and safety of IL23 / TNFα bispecific antibodies that can be used to treat or manage inflammatory bowel disease (IBD) in canines can be evaluated using various doses ranging from 0.01 mg / kg to 100 mg / kg.

[0226] Histopathological examination of endoscopic gastrointestinal biopsies can be used to determine the efficacy of caninized IL23 antibodies in managing IBD.

[0227] IBD biomarkers that can be measured include, but are not limited to, IL-1 beta, IL6, IL8, IL9, IFN-gamma, TNF alpha, CCL2, IL22, CRP, LL37, TFF3, OSM.

[0228] The Canine Inflammatory Bowel Disease Activity Index (CIBDAI) score can be used to screen candidates for treatment.

[0229] Controlled 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 RASQGISSWLA (SEQ ID NO: 4), the HVR-L2 region comprises the amino acid sequence YAASSLQS (SEQ ID NO: 5), and the HVR-L3 region comprises the amino acid sequence QQYNIYPYT (SEQ ID NO: 6); and / or (b) the HVR-H1 region comprises the amino acid sequence KGSGYSFTTYWLG (SEQ ID NO: 8), the HVR-H2 region comprises the amino acid sequence IMSPVDSDIR (SEQ ID NO: 9), and the HVR-H3 region comprises the amino acid sequence ARRRPGQGYFDF (SEQ ID NO: 10); The anti-IL23 antibody.

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

3. The antibody comprises a light chain variable domain (VL) amino acid sequence having at least 90% identity to a sequence selected from SEQ ID NOs: 3, 11, 13, and 15, and / or a heavy chain variable domain (VH) amino acid sequence having at least 90% identity to a sequence selected from SEQ ID NOs: 7, 12, 14, and 16, and optionally (i) the light chain variable domain (VL) comprises a variant of SEQ ID NOs: 3, 11, 13, and 15, in which 1 to 6 amino acids of the light chain variable domain (VL) are replaced 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 replaced by different amino acids; The antibody described in claim 1.

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

5. The antibody comprises a light chain (LC) amino acid sequence having at least 90% identity to an amino acid sequence selected from SEQ ID NOs: 17, 19, 22, and 24, and / or a heavy chain (HC) amino acid sequence having at least 90% identity to an amino acid sequence selected from SEQ ID NOs: 18, 20, 21, 23, and 25, and optionally 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; The antibody of any one of claims 1 to 4, comprising:

6. 1. 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 RASQGIRNYLA (SEQ ID NO:27), the HVR-L2 region comprises the amino acid sequence AASTLQ (SEQ ID NO:28), and the HVR-L3 region comprises the amino acid sequence QRYNRAPYT (SEQ ID NO:29); and / or (a) The anti-TNFα antibody, wherein the HVR-H1 region comprises the amino acid sequence FTFDDYAMH (SEQ ID NO: 31), the HVR-H2 region comprises the amino acid sequence AITWNSGHIDYADSVEGR (SEQ ID NO: 32), and the HVR-H3 region comprises the amino acid sequence AKVSYLSTASSLDY (SEQ ID NO: 33).

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

8. The antibody comprises a light chain variable domain (VL) amino acid sequence having at least 90% identity to a sequence selected from SEQ ID NOs: 26, 34, and 36, and / or a heavy chain variable domain (VH) amino acid sequence having at least 90% identity to a sequence selected from SEQ ID NOs: 30, 35, and 37, and optionally (i) the light chain variable domain (VL) comprises a variant of SEQ ID NOs: 26, 34, and 36, in which 1 to 6 amino acids of the light chain variable domain (VL) are replaced 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 replaced by different amino acids; The antibody described in claim 6.

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

10. The antibody comprises a light chain (LC) amino acid sequence having at least 90% identity to a sequence selected from SEQ ID NOs: 38, 40, and 41, and / or a heavy chain (HC) amino acid sequence having at least 90% identity to a sequence selected from SEQ ID NOs: 39, 42, 43, and 44, and optionally 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; The antibody of any one of claims 6 to 9, comprising:

11. The antibody of any one of claims 6-9, wherein the antibody is an scFv antibody, and optionally 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 according to any one of claims 1 to 11, wherein the antibody is an antibody fragment selected from Fv, scFv, Fab, Fab', F(ab')2, and Fab'-SH.

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

14. The antibody (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 The antibody of any one of claims 1 to 13, comprising:

15. The antibody according to 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. 1. A bispecific antibody that binds to canine, feline, and / or equine IL23 and canine, feline, and / or equine TNFα, said antibody comprising: (a) an anti-IL23 light chain (LC) comprising a VL domain having an HVR-L1 region comprising the amino acid sequence RASQGISSWLA (SEQ ID NO: 4), an HVR-L2 region comprising the amino acid sequence YAASSLQS (SEQ ID NO: 5), and an HVR-L3 region comprising the amino acid sequence QQYNIYPYT (SEQ ID NO: 6); an anti-IL23 heavy chain (HC) comprising a VH domain having an HVR-H1 region comprising the amino acid sequence KGSGYSFTTYWLG (SEQ ID NO: 8), an HVR-H2 region comprising the amino acid sequence IMSPVDSDIR (SEQ ID NO: 9), and an HVR-H3 region comprising the amino acid sequence ARRRPGQGYFDF (SEQ ID NO: 10); and an anti-TNFα heavy chain (HC) fused to the HC. an anti-TNFα scFv antibody, comprising a VL domain having an HVR-L1 region comprising the amino acid sequence RASQGIRNYLA (SEQ ID NO: 27), an HVR-L2 region comprising the amino acid sequence AASTLQ (SEQ ID NO: 28), and an HVR-L3 region comprising the amino acid sequence QRYNRAPYT (SEQ ID NO: 29), and a VH domain having an HVR-H1 region comprising the amino acid sequence FTFDDYAMH (SEQ ID NO: 31), an HVR-H2 region comprising the amino acid sequence AITWNSGHIDYADSVEGR (SEQ ID NO: 32), and an HVR-H3 region comprising the amino acid sequence 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 RASQGIRNYLA (SEQ ID NO: 27), an HVR-L2 region comprising the amino acid sequence AASTLQ (SEQ ID NO: 28), and an HVR-L3 region comprising the amino acid sequence 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 FTFDDYAMH (SEQ ID NO: 31), an HVR-H2 region comprising the amino acid sequence AITWNSGHIDYADSVEGR (SEQ ID NO: 32), and an HVR-H3 region comprising the amino acid sequence AKVSYLSTASSLDY (SEQ ID NO: 33), and an anti-IL23 antibody fused to the HC. an anti-IL23 scFv antibody comprising a VL domain having an HVR-L1 region comprising the amino acid sequence RASQGISSWLA (SEQ ID NO: 4), an HVR-L2 region comprising the amino acid sequence YAASSLQS (SEQ ID NO: 5), and an HVR-L3 region comprising the amino acid sequence QQYNIYPYT (SEQ ID NO: 6), and a VH domain having an HVR-H1 region comprising the amino acid sequence KGSGYSFTTYWLG (SEQ ID NO: 8), an HVR-H2 region comprising the amino acid sequence IMSPVDSDIR (SEQ ID NO: 9), and an HVR-H3 region comprising the amino acid sequence ARRRPGQGYFDF (SEQ ID NO: 10); The bispecific antibody comprising:

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

18. 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; and optionally 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; 18. The bispecific antibody of claim 16, comprising:

19. 1. A bispecific antibody that binds to canine, feline, and / or equine IL23 and canine, feline, and / or equine TNFα, said antibody comprising: (i) an anti-IL23 light chain (LC) comprising a VL domain having an HVR-L1 region comprising the amino acid sequence RASQGISSWLA (SEQ ID NO: 4), an HVR-L2 region comprising the amino acid sequence YAASSLQS (SEQ ID NO: 5), and an HVR-L3 region comprising the amino acid sequence 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 KGSGYSFTTYWLG (SEQ ID NO: 8), an HVR-H2 region comprising the amino acid sequence IMSPVDSDIR (SEQ ID NO: 9), and an HVR-H3 region comprising the amino acid sequence 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 RASQGIRNYLA (SEQ ID NO: 27), an HVR-L2 region comprising the amino acid sequence AASTLQ (SEQ ID NO: 28), and an HVR-L3 region comprising the amino acid sequence 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 FTFDDYAMH (SEQ ID NO: 31), an HVR-H2 region comprising the amino acid sequence AITWNSGHIDYADSVEGR (SEQ ID NO: 32), and an HVR-H3 region comprising the amino acid sequence AKVSYLSTASSLDY (SEQ ID NO: 33); 2. The bispecific antibody comprising:

20. (i) the anti-IL23 light chain (LC) comprises a light chain variable domain (VL) amino acid sequence having at least 90% identity to a sequence selected from 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 having at least 90% identity to a sequence selected from 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 having at least 90% identity to a sequence selected from 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 having at least 90% identity to a sequence selected from SEQ ID NOs: 30, 35, and 37; 20. The bispecific antibody of claim 19.

21. (i) the anti-IL23 light chain (LC) comprises an amino acid sequence having at least 90% identity to a sequence selected from 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 having at least 90% identity to a sequence selected from 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 to SEQ ID NOs: 42, 43, and 44; A bispecific antibody according to any one of claims 19-20.

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

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

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

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

28. The method according to any one of claims 26 to 27, wherein the pathology associated with IL23 is an inflammatory disease.

29. The method according to any one of claims 26 to 28, wherein the pathology associated with IL23 is an inflammatory disease of the gastrointestinal system.

30. The method of any one of claims 26 to 29, wherein the IL23-associated pathology 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 for treating a dog, cat, or horse having a pathology associated with IL23 and TNFα, comprising administering to the dog, cat, or horse a therapeutically effective amount of an IL23 antibody described in any one of claims 1-5 or 12-21, and a TNFα antibody described in any one of claims 6-21, or a pharmaceutical composition described in claim 25.

33. 22. A method of treating a dog, cat, or horse having an IL23 / TNFα associated pathology, comprising administering to said dog, cat, or horse a therapeutically effective amount of a bispecific IL23 / TNFα antibody according to any one of claims 16-21.

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

35. 35. The method of any one of claims 26-34, wherein the antibody or pharmaceutical composition is administered by intramuscular, intraperitoneal, intracerebrospinal, subcutaneous, intra-arterial, intrasynovial, intrathecal, or inhalation route.

36. 36. The method of any one of claims 26-35, wherein the method further comprises administration of 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 beta7-integrin antibody, an IL12 antibody, an IL1β antibody, or an anti-BlyS antibody.

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

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

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

40. 40. The method of any one of claims 38-39, wherein the cell is a canine cell, a feline cell, or an equine cell.

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

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