Multispecific antibodies specific for TNFα and IL-17A, antibodies targeting IL-17A and methods of using the same

Abstract

The present invention relates to an isolated multispecific antibody comprising a first domain that specifically binds to TNFα and a second domain that specifically binds to IL-17A and optionally a third domain that specifically binds to human serum albumin. The present invention also relates to an isolated antibody that specifically binds to human IL-17A. In addition, the present invention also relates to a method of using the antibody, a pharmaceutical composition and a method of using the same, and a kit comprising the antibody. The present invention also relates to a nucleic acid comprising a nucleotide sequence encoding the antibody, a vector comprising the nucleic acid, a host cell comprising the nucleic acid or the vector, and a method for producing the antibody.

Description

Technical Field

[0001] The present invention relates to an isolated multispecific antibody comprising a first domain that specifically binds to TNFα, a second domain that specifically binds to IL-17A, and optionally a third domain that specifically binds to human serum albumin. The present invention also relates to methods of using the antibody, pharmaceutical compositions and methods of using the same, and kits comprising the antibody. The present invention also relates to nucleic acids comprising a nucleotide sequence encoding the antibody, vectors comprising the nucleic acids, host cells comprising the nucleic acids or the vectors, and methods of producing the antibodies.

[0002] The present invention also relates to isolated antibodies that specifically bind to human IL-17A, multispecific molecules comprising the isolated antibodies of the present invention, pharmaceutical compositions, and methods of using the same. The present invention also relates to kits comprising the antibodies, nucleic acids comprising nucleotide sequences encoding the antibodies, vectors comprising the nucleic acids, host cells comprising the nucleic acids or vectors, and methods of producing the antibodies. Background Art

[0003] TNFα is a homotrimeric proinflammatory cytokine released by and interacting with cells of the immune system. TNFα exists as a soluble protein and a precursor, termed transmembrane TNFα, which is expressed as a type II polypeptide on the cell surface. Transmembrane TNFα is processed between residues Ala76 and Val77 by metalloproteases such as TNFα-converting enzyme (TACE), releasing the soluble form of TNFα consisting of 157 amino acid residues. Soluble TNFα is a homotrimer of a 17-kDa cleaved monomer. Transmembrane TNFα also exists as a homotrimer of a 26-kDa uncleaved monomer. Both membrane and soluble TNFα are biologically active. TNFα binds to two receptors, TNF receptor 1 (TNFR1) and TNFR2, with transmembrane TNFα exerting its effects primarily through TNFR2. TNFR1 is ubiquitously expressed on various cells, and its engagement triggers a proinflammatory response. TNFR2 expression is almost exclusively restricted to immune cells, where its binding promotes cell survival and proliferation (Bazzoni F, Beutler B, N Engl J Med (1996) 334(26): 1717–25; Locksley RM et al., Cell (2001) 104(4): 487–501; Cabal-Hierro L, Lazo PS, Cell Signal (2012) 24(6): 1297–305; Brenner D et al., Nat Rev Immunol (2015) 15(6): 362–74).

[0004] TNFα has been shown to be upregulated in many human diseases, such as inflammatory and autoimmune diseases, including chronic diseases such as rheumatoid arthritis, Crohn's disease, ulcerative colitis, and multiple sclerosis. Antibodies against TNFα have been proposed for the prevention and treatment of endotoxic shock (Beutler et al., Science, 234, 470-474, 1985). Bodmer et al. (Critical Care Medicine, 21, S441-S446, 1993) and Wherry et al. (Critical Care Medicine, 21, S436-S440, 1993) discussed the therapeutic potential of anti-TNFα antibodies in the treatment of septic shock. Kirschenbaum et al. (Critical Care Medicine, 26, 1625-1626, 1998) also discussed the use of anti-TNFα antibodies in the treatment of septic shock. Anti-TNFα monoclonal antibodies can be used to effectively treat collagen-induced arthritis (Williams et al. (Proc. Natl. Acad. Sci. USA 89, 9784-9788, 1992)). Feldman et al. (Transplantation Proceedings, 30, 4126-4127, 1998), Adorini et al. (Trends in Immunology Today, 18, 209-211, 1997), and Feldman et al. (Advances in Immunology, 64, 283-350, 1997) discuss the use of anti-TNFα antibodies in the treatment of rheumatoid arthritis and Crohn's disease. TNFα antibodies previously used in such treatments are generally chimeric antibodies, such as those described in U.S. Pat. No. 5,919,452.

[0005] Monoclonal antibodies against TNFα have been described in the prior art. Meager et al. (Hybridoma, 6, 305-311, 1987) described mouse monoclonal antibodies against recombinant TNFα. Fendly et al. (Hybridoma, 6, 359-370, 1987) described the use of mouse monoclonal antibodies against recombinant TNFα in defining neutralizing epitopes on TNFα. In addition, in international patent application WO 92 / 11383, TNFα-specific recombinant antibodies, including CDR-grafted antibodies, are disclosed. Rankin et al. (British J. Rheumatology, 34, 334-342, 1995) described the use of such CDR-grafted antibodies in the treatment of rheumatoid arthritis. U.S. Patent No. 5,919,452 discloses anti-TNFα chimeric antibodies and their use in the treatment of pathologies associated with the presence of TNFα. Other anti-TNFα antibodies are disclosed by Stephens et al. (Immunology, 85, 668-674, 1995), GB-A-2 246570, GB-A-2 297 145, US 8,673,310, US 2014 / 0193400, EP 2 390 267 Bl, US 8,293,235, US 8,697,074, WO 2009 / 155723 A2 and WO 2006 / 131013 A2.

[0006] Currently approved anti-TNFα biotherapeutics include: (i) infliximab, a chimeric IgG anti-human monoclonal antibody ( Wiekowski M et al.: “Infliximab (Remicade)”, Handbook of Therapeutic Antibodies, WILEY-VCH; Weinheim, 2007-01-01, p.885-904); (ii) Etanercept, a TNFR2 dimer fusion protein with IgG1 Fc (iii) Adalimumab, a fully human monoclonal antibody (mAb) ( Kupper H et al.: “Adalimumab (Humira)”, Handbook of Therapeutic Antibodies, WILEY-VCH; Weinheim, 2007-01-01, p.697-732); (iv) certolizumab, a PEGylated Fab fragment ( Melmed GY et al.: “Certolizumab pegol”, Nature Reviews. Drug Discovery, Nature Publishing Group, GB, Vol. 7, No. 8, 2008-08-01, p. 641-642); (v) golimumab, a human IgG1K monoclonal antibody ( Mazumdar S et al.: "Golimumab", mAbs, Landes Bioscience, US, Vol. 1, No. 5, 2009-09-01, p. 422-431).

[0007] However, anti-TNFα therapy has been shown to have certain limitations. Not all patients achieve an adequate clinical response to anti-TNFα or maintain a long-term clinical response, leading to the need to switch to new therapies to control their disease. For example, in rheumatoid arthritis (RA) patients treated with anti-TNFα, approximately 40% never respond, and only 20% experience a significant reduction in disease activity. Therefore, there remains a considerable unmet clinical need for more effective inhibition of the progression of diseases such as inflammatory and autoimmune disorders.

[0008] The activation of complementary biological pathways in patients after anti-TNFα treatment may be one of the reasons why many patients do not respond or only partially respond to anti-TNFα treatment. Recently, a large amount of evidence has emerged that IL-17A plays a role in the pathogenesis of inflammatory and autoimmune diseases, such as RA.

[0009] The interleukin-17 (IL-17) family in humans and mice consists of six cytokines: IL-17A, IL-17B, IL-17C, IL-17D, IL-17E (also known as IL-25), and IL-17F, which play a role in acute and chronic inflammatory responses. The interleukin-17 receptor (IL-17R) family includes five members: IL-17RA, IL-17RB, IL-17RC, IL-17RD, and IL-17RE.

[0010] Interleukin-17A (IL-17A or IL17A, synonymous with IL-17 or cytotoxic T lymphocyte-associated antigen-8 (CTLA-8)) is a homodimeric proinflammatory cytokine. IL-17A is produced by a subset of memory CD4+ T cells (termed Th17), CD8+ T cells (Tc17), invariant NKT cells, γδ T cells, non-T, non-B lymphocytes (termed type 3 innate lymphocytes), and neutrophils. IL-17A and IL-17F form a unique subset within the IL-17 family. They have the greatest sequence homology and identity within the IL-17 family, while other members of the IL-17 family have significantly lower sequence identity with IL-17A (Starnes, T. et al., J Immunol. 167(8):4137-40 (2001); Aggarwal, S. and Gurney, AL, J. Leukoc Biol, 71(1):1-8 (2002)). Both IL-17A and IL-17F signal through a heterodimeric receptor complex composed of IL-17RA and IL-17RC (Toy D. et al., J Immunol. 2006; Wright JF. et al., J Immunol. 2008; 181(4):2799–2805). IL-17A and IL-17F can form IL-17A / A or IL-17F / F disulfide-linked homodimers and IL-17A / F disulfide-linked heterodimers (Wright JF. et al., J Immunol. 2008; 181(4):2799–2805; Liang SC. et al., J Immunol. 2007; 179(11):7791–7799). IL-17A and IL-17F induce the expression of proinflammatory cytokines and antimicrobial peptides.

[0011] Human IL-17A (CTLA-8, Swiss Prot Q 16552, also known as IL-17 or IL17; SEQ ID NO: 33) is associated with various inflammatory diseases, such as autoimmune diseases, metabolic disorders and cancer (Ouyang W. et al., Immunity. 2008; 28(4): 454–467; Milner JD., Curr Opin Immunol. 2011; 23(6): 784–788; Kuchroo VK. et al., Nat Med. 2012; 18(1): 42–47; Ahmed M. and Gaffen SL., Cytokine Growth Factor Rev. 2010; 21(6): 449–453; Trinchieri G., Annu Rev Immunol. 2012; 30: 677–706; Gallimore AM, Godkin A., N Engl J Med. 2013; 368(3): 282–284; Ye P. et al., J Exp Med. 2001; 194(4): 519–527; Chung DR. et al., J Immunol. 2003; 170(4): 1958–1963; Huang W. et al., J Infect Dis. 2004; 190(3): 624–631; Ishigame H. et al., Immunity. 2009; 30(1): 108–119; see for review Gu C. et al., Cytokine. 2013 Nov 64(2). IL-17A plays a role in inducing other inflammatory cytokines and chemokines to recruit neutrophils, acute phase proteins, antimicrobial peptides, mucins, matrix metalloproteinases, and adhesion molecules. IL-17A also cooperates with other cytokines including TNFα and IL-1β to further induce the expression of chemokines (Chabaud M. et al., J. Immunol. 161(1):409-14 (1998)).

[0012] Pathological production of IL-17A leads to excessive inflammation and tissue damage (see for review Gu et al., Cytokine. 2013 November; 64(2)). High IL-17A levels are found in patients with multiple sclerosis (MS), psoriasis, asthma, Crohn's disease, and rheumatoid arthritis. Treatment of animals with IL-17A neutralizing antibodies can reduce the incidence and severity of autoimmune encephalomyelitis (Komiyama, Y. et al., J. Immunol. 177 (2006) 566-573). In addition, IL-17A neutralizing antibodies can reduce the severity and incidence of rheumatoid arthritis models in mice, and high levels of IL-17A can be detected in the synovial fluid of inflamed joints of rheumatoid arthritis patients (Ziolkowska, M. et al., J. Immunol. 164 (2000) 2832-2838; Kotake, S. et al., J. Clin. Invest. 103 (1999) 1345-1352; Hellings P. W. et al., Am. J. Resp. Cell Mol. Biol. 28 (2003) 42-50).

[0013] A large amount of experimental evidence from humans and animal models supports the development of IL-17A-targeted therapies. Several anti-IL-17 antibodies have been developed, including AIN457 (secukinumab; see U.S. Patent No. 7,807,155 and WO2006 / 013107), LY2439821 (ixekizumab; see U.S. Patent Nos. 7,838,638 and 8,110,191 and WO2007 / 070750), SCH900117 (Merck), and RG4943 (Roche). WO 2006 / 013107, WO 2006 / 054059, WO2007 / 070750, WO 2007 / 149032, WO 2008 / 001063, WO 2008 / 021156, WO 2010 / 034443, WO2010 / 102251, WO 2012 / 018767、WO 2014 / 161570、WO 2014 / 001368、WO 2014 / 122613、WO2015 / 070697、WO 2015 / 137843、WO 2016 / 048188、WO 2016 / 113557、WO 2016 / 138842 and WO2017 / 068472 disclose examples of anti-IL-17A antibodies.

[0014] Several clinical trials have been conducted or are ongoing for various molecules that block IL-17A signaling. Biologic agents that target IL-17A or its receptor and their efficacy are being evaluated in inflammatory or autoimmune conditions such as rheumatoid arthritis, ankylosing spondyloarthritis, Crohn's disease, psoriasis, multiple sclerosis, and ozone-induced neutrophilia.

[0015] For example, secukinumab is a fully human IgG1κ anti-IL-17A monoclonal antibody (U.S. Patent No. 7,807,155 and WO 2006 / 013107) that is now approved for the treatment of psoriasis, psoriatic arthritis, and ankylosing spondylitis (see for review Wang et al., Eur J Rheumatol 2017(4)272-7). In Phase III studies (FUTURE I and FUTURE II) evaluating the long-term efficacy and safety of secukinumab in subjects with psoriatic arthritis, secukinumab was significantly more effective than placebo in improving the symptoms and signs of psoriatic arthritis (Mease PJ. et al. N Engl J Med 2015; 373: 1329-39; McInnes IB. et al. The Lancet; 386: 1137-46).

[0016] Ixekizumab is a humanized anti-IL-17A monoclonal antibody (U.S. Patent Nos. 7,838,638 and 8,110,191 and WO 2007 / 070750) that was studied in a 24-week Phase III trial (SPIRIT-P1) in patients with active psoriatic arthritis who had not received prior biologic therapy (Mease PJ. et al., Ann Rheum Dis. 2017 Jan;76(1):79-87). The study showed that ixekizumab treatment resulted in improvements in disease activity and physical function, as well as inhibition of the progression of structural damage, in patients with active psoriatic arthritis who had not received prior biologic therapy. Ixekizumab was also effective in treating patients with moderate to severe plaque psoriasis (Griffiths CEM et al., The Lancet;386:541-51).

[0017] Brodalumab is a fully human IL-17 receptor (IL-17RA) monoclonal antibody (see U.S. Patent No. 7,767,206) that has been shown to be effective in treating psoriasis (Papp KA. et al. N Engl J Med 2012;366:1181-9). In a placebo-controlled phase II study, it also showed significant and sustained responses in patients with psoriatic arthritis (Mease PJ. et al., N Engl J Med 2014;370:2295-306). However, treatment with brodalumab has also been associated with strong adverse events such as upper respiratory tract infections, fatigue, diarrhea, and reported suicidal thoughts and behaviors (see for review Wang et al., Eur J Rheumatol 2017(4)272-7).

[0018] Therefore, IL-17A is a promising target for the treatment of inflammatory and autoimmune diseases. Although some anti-IL-17A antibodies have been discovered so far, there is still a need to develop improved therapeutic antibodies that can effectively reduce or eliminate IL-17A activity in inflammatory responses and autoimmune diseases, while having better safety and being suitable for development. In addition to having beneficial affinity, efficacy and immunogenicity, therapeutic antibodies should also have improved biophysical properties, leading to better developability, high-yield manufacturability and protein stability. A therapeutic approach that simultaneously blocks the biological pathways of TNFα and IL-17A has the potential to significantly improve response rates and address unmet needs in the treatment of diseases mediated by TNFα and IL-17A. So far, several biotherapeutic agents that specifically bind to IL-17 and TNFα have been proposed. WO 2010 / 102251 (Abbvie Inc.) discloses a bispecific tetravalent antibody that binds to TNFα and IL-17. WO 2013 / 063110 (Abbvie Inc.) discloses a multivalent DVD-Ig binding protein capable of binding to TNF and IL-17. WO 2014 / 044758 (Covagen ACJ) discloses a fusion construct capable of inhibiting glycosylated IL-17A and binding to TNFα. In addition, WO 2014 / 137961 (Eli Lilly and Company) and WO 2017 / 132457 (Janssen Biotech) disclose bispecific antibodies against TNF and IL-17A. WO 2017 / 102830 (UCB Biopharma) discloses a multispecific antibody capable of inhibiting TNFα, IL-17A, and IL-17F, particularly comprising a binding domain specific for human TNFα and a binding domain specific for human IL-17A and human IL-17F. Xu et al., Oncotarget, 8 (2017) 81860-81872 produced an IgG-like bispecific antibody (bsAb), in which the two arms of the IgG-like molecule are directed against TNFα and IL-17A, respectively. Interestingly, WO 2015 / 014979 (Roche, see also Fischer et al., Arthritis & Rheumatology 67 (2015) 51–62) discloses a bispecific tetravalent IL-17AxTNF antibody construct that is either bivalent ("2+2" construct) or monovalent ("2+2" construct) for each of the two antigens. According to WO 2015 / 014979, bivalent constructs are superior to monovalent alternatives.

[0019] However, there remains a need for improved anti-inflammatory drugs that can effectively neutralize the activity of IL-17A and TNFα for the treatment of conditions where a significant portion of patients remain inadequately responsive to treatment, such as inflammatory and autoimmune diseases, such as rheumatoid arthritis. There remains a need to develop improved therapeutic antibodies that effectively neutralize the activity of IL-17A and TNFα, have favorable affinity and efficacy, and have an improved safety profile, such as reduced immunogenicity. Furthermore, therapeutic antibodies should have improved biophysical properties, resulting in better developability, high-yield manufacturability, and superior antibody stability. Summary of the Invention

[0020] It is an object of the present invention to provide a medicament for improving the treatment of inflammatory and autoimmune diseases.

[0021] The antibodies of the present invention provide a new treatment option for patients with unmet medical needs. The present invention provides a new multispecific antibody that can simultaneously inhibit IL-17A and TNFa and has further improved properties that are beneficial for treatment, such as higher affinity, improved efficacy, selectivity, safety (such as low immunogenicity) and improved biophysical properties (such as developability and stability).

[0022] In one aspect, the present disclosure relates to an isolated multispecific antibody comprising a first domain that specifically binds IL-17A, a second domain that specifically binds TNFα, and optionally a third domain that specifically binds human serum albumin.

[0023] In one aspect, the present disclosure relates to a pharmaceutical composition comprising a multispecific antibody of the present invention and a pharmaceutically acceptable carrier.

[0024] In another aspect, the present disclosure provides a multispecific antibody of the invention or a pharmaceutical composition of the invention for use as a medicament.

[0025] In another aspect, the present disclosure provides a multispecific antibody of the invention or a pharmaceutical composition of the invention for use in treating a disorder mediated by IL-17A and / or TNFα or a disorder treatable by inhibiting Gro-α secretion, in particular for treating an inflammatory condition or an autoimmune disease.

[0026] In one aspect, the present disclosure provides the use of the multispecific antibodies of the invention or the pharmaceutical compositions of the invention for treating disorders mediated by IL-17A and / or TNFα or disorders treatable by inhibiting Gro-α secretion, in particular for treating inflammatory conditions or autoimmune diseases.

[0027] In one aspect, the present disclosure provides use of a multispecific antibody of the present invention or a pharmaceutical composition of the present invention in the preparation of a medicament for treating a disorder mediated by IL-17A and / or TNFα or a disorder that can be treated by inhibiting Gro-α secretion, in particular for treating inflammatory conditions or autoimmune diseases.

[0028] In yet another aspect, the present disclosure provides a method for treating a disorder mediated by IL-17A and / or TNFα, comprising administering an effective amount of the multispecific antibody of the present invention or the pharmaceutical composition of the present invention to a subject in need thereof.

[0029] In one aspect, the present disclosure provides a kit comprising a multispecific antibody of the present invention, or a pharmaceutical composition of the present invention.

[0030] In another aspect, the present disclosure provides a nucleic acid comprising a nucleotide sequence encoding a multispecific antibody of the present invention, a vector comprising the nucleic acid, or a host cell comprising the nucleic acid or the vector.

[0031] In another aspect, the present disclosure provides a method for producing the multispecific antibody or its binding domain or fragment thereof of the present invention, the method comprising the following steps: culturing a host cell containing a nucleic acid or vector encoding the multispecific antibody or its binding domain or fragment thereof of the present invention.

[0032] The aspects, advantageous features and preferred embodiments of the present disclosure outlined in the following, individually or in combination, further contribute to solving the objects of the present invention:

[0033] 1. An isolated multispecific antibody comprising a first domain that specifically binds to IL-17A and a second domain that specifically binds to TNFα.

[0034] 2. The multispecific antibody according to claim 1, wherein the antibody comprises only one domain that specifically binds to IL-17A and / or only one domain that specifically binds to TNFα.

[0035] 3. The multispecific antibody according to item 1 or 2, wherein the antibody is capable of neutralizing the biological activities of human TNFα and human IL-17A.

[0036] 4. The multispecific antibody according to any of the preceding items, wherein the antibody selectively binds human IL-17A relative to human IL-17B, IL-17C, IL-17D, IL-17E and IL-17F as measured by ELISA.

[0037] 5. The multispecific antibody according to any one of the preceding items, further comprising a third domain specific for an antigen other than IL-17A or TNFα.

[0038] 6. The multispecific antibody according to item 5, wherein the antibody comprises a third domain that specifically binds to human serum albumin (HSA), preferably, the antibody comprises only one domain that specifically binds to human serum albumin.

[0039] 7. The multispecific antibody according to any of the preceding items, wherein the domains are capable of binding simultaneously to their respective antigens or receptors.

[0040] 8. The multispecific antibody according to any of the preceding items, wherein the first domain and the second domain, and, optionally, the third domain, are independently selected from the group consisting of: Fab, Fv, scFv, dsFv, scAb, STAB, single domain antibodies (sdAb or dAb), single domain heavy chain antibodies, and single domain light chain antibodies, VHH, VNAR, single domain antibodies based on shark VNAR structures, and binding domains based on alternative scaffolds including but not limited to ankyrin-based domains, fynomers, avimers, anticalins, fibronectin, and binding sites built into antibody constant regions (e.g., Modular Antibody Technology of F-star). TM ), preferably, selected from the group consisting of: Fab, Fv and scFv, more preferably, wherein the first domain and / or the second domain and / or the third domain is Fv or scFv.

[0041] 9. The multispecific antibody according to any of the preceding items, wherein the format of the multispecific antibody is selected from the group consisting of: single-chain diabody (scDb), tandem scDb (Tandab), linear dimer scDb (LD-scDb), cyclic dimer scDb (CD-scDb), bispecific T cell engager (BiTE; tandem bi-scFv), tandem tri-scFv, tribody (Fab-(scFv)2) or bibody (Fab-(scFv)1), Fab, Fab-Fv2, Morrison (IgGCH3-scFv fusion (Morrison L) or IgG CL-scFv fusion (Morrison H)), triabodies, scDb-scFv, bispecific Fab2, diabodies, tetrabodies, scFv-Fc-scFv fusions, scFv-HSA-scFv fusions, diabodies, DVD-Ig, COVD, IgG-scFab, scFab-dsscFv, Fv2-Fc, IgG-scFv fusions such as bsAb (scFv linked to the C-terminus of the light chain), Bs1Ab (scFv linked to the N-terminus of the light chain), Bs2Ab (scFv linked to the N-terminus of the heavy chain), Bs3Ab (scFv linked to the C-terminus of the heavy chain), Ts1Ab (scFv linked to the C-terminus of the heavy chain), The invention also provides the present invention relates to bispecific antibodies based on heterodimeric Fc domains, such as Knob-into-Hole antibodies (KiH); Fv, scFv, scDb, tandem bi-scFv, tandem tri-scFv, Fab-(scFv)2, Fab-(scFv)1, Fab, Fab-Fv2, COVD, MATCH and DuoBody fused to the N- and / or C-terminus of any chain of a heterodimeric Fc domain or any other heterodimeric domain, preferably a triabody or scDb-scFv.

[0042] 10. The multispecific antibody according to any of the preceding items, wherein the antibody does not comprise an immunoglobulin Fc region polypeptide, and optionally, does not comprise a CH1 and / or CL region.

[0043] 11. The multispecific antibody of any of the preceding items, wherein the antibody is a triabody.

[0044] 12. The multispecific antibody of claim 11 , wherein the first domain, the second domain and the third domain are independently selected from the group consisting of: Fab and scFv, preferably, wherein the second domain is Fab and the first domain and the third domain are scFv.

[0045] 13. The multispecific antibody of claim 9, wherein the antibody is scDb-scFv, preferably wherein the scFv portion is fused to scDb at the C-terminus, more preferably wherein the first domain and the second domain form scDb and the third domain is scFv.

[0046] 14. The multispecific antibody according to item 13, wherein the antibody is represented by the following formula:

[0047] VLA-L1-VHC-L2-VLC-L3-VHA-L4-VLB-L5–VHB or

[0048] VLB-L1-VHA-L2-VLA-L3-VHB-L4-VLC-L5–VHC or

[0049] VLC-L1-VHB-L2-VLB-L3-VHC-L4-VLA-L5–VHA or

[0050] VLA-L1-VHB-L2-VLB-L3-VHA-L4-VLC-L5–VHC,

[0051] Preferably VLB-L1-VHA-L2-VLA-L3-VHB-L4-VLC-L5–VHC or VLA-L1-VHB-L2-VLB-L3-VHA-L4-VLC-L5–VHC,

[0052] More preferably VLB-L1-VHA-L2-VLA-L3-VHB-L4-VLC-L5–VHC,

[0053] wherein VLA and VHA are the light chain variable region and the heavy chain variable region of the first domain, respectively; and VLB and VHB are the light chain variable region and the heavy chain variable region of the second domain, respectively; and VLC and VHC are the light chain variable region and the heavy chain variable region of the third domain, respectively, and wherein L1, L2, L3, L4 and L5 are polypeptide linkers.

[0054] 15. The multispecific antibody according to item 14, wherein L1 and L3 are as shown in SEQ ID NO: 132.

[0055] 16. The multispecific antibody according to any one of items 14 to 15, wherein L2, L4 and L5 are as shown in SEQ ID NO: 23.

[0056] 17. The multispecific antibody according to any one of the preceding items, wherein the antibody has the following characteristics

[0057] (a) having an ability to neutralize IL-17A with a potency relative to secukinumab (relative potency) of greater than 2, e.g., greater than 5, greater than 10, greater than 15, greater than 20, greater than 25, greater than 30, greater than 35, greater than 40, greater than 45, preferably greater than 50, as determined by measuring Gro-α secretion in the HT-29 assay, and wherein the relative potency is the IC of secukinumab in ng / mL as measured in the HT-29 assay 50 The values ​​are comparable to the IC values ​​in ng / mL of the multispecific antibodies measured in the HT-29 assay. 50 The ratio of the values; and

[0058] (b) has the ability to neutralize TNFα with a potency (relative potency) of at least 1, e.g., greater than 1, greater than 1.5, greater than 2, greater than 2.5, greater than 3, greater than 3.5, preferably greater than 4, more preferably greater than 4.5 relative to the scDb according to SEQ ID NO: 149 (A13) as determined by measuring Gro-α secretion in the HT-29 assay, and wherein the relative potency is the IC in nM of the scDb according to SEQ ID NO: 149 measured in the HT-29 assay 50 The values ​​are comparable to the IC values ​​in nM of the multispecific antibodies measured in the HT-29 assay. 50 The ratio of the values; and

[0059] (c) optionally, having the ability to block the interaction between IL-17A and IL-17RA, with a potency relative to secukinumab (relative potency) as determined in an ELISA assay of greater than 2, e.g., greater than 3, greater than 4, greater than 5, greater than 6, greater than 7, greater than 8, greater than 9, preferably greater than 10, and wherein the relative potency is the IC of secukinumab in ng / mL as measured by ELISA 50 The values ​​were compared with the IC values ​​of the multispecific antibodies in ng / mL measured by ELISA. 50 The ratio of the values; and

[0060] (d) optionally, has the ability to neutralize TNFα with a potency (relative potency) of at least 0.4, e.g. at least 0.5, preferably at least 1 relative to the scDb according to SEQ ID NO: 149 (A13) as determined in the L929 assay, and wherein said relative potency is the IC in nM of said scDb according to SEQ ID NO: 149 measured in the L929 assay 50 The values ​​are comparable to the IC values ​​in nM for the multispecific antibodies measured in the L929 assay. 50 and / or

[0061] (e) Binding to human IL-17A, dissociation constant (KD ) is less than 5 nM, e.g., less than 4 nM, less than 3 nM, less than 2 nM, less than 1 nM, preferably less than 0.5 nM, as measured by surface plasmon resonance; and optionally, binds to cynomolgus monkey IL-17A, K D Less than 5 nM, e.g., less than 4 nM, less than 3 nM, less than 2 nM, less than 1 nM, preferably less than 0.5 nM, as measured by surface plasmon resonance;

[0062] (f) Binds to human TNFα, dissociation constant (K D ) is less than 5 nM, e.g., less than 4 nM, less than 3 nM, less than 2 nM, less than 1 nM, preferably less than 0.5 nM, more preferably less than 0.25 nM, as measured by surface plasmon resonance; and

[0063] (g) optionally binds to human serum albumin with a dissociation constant (K D ) is less than 5 nM, e.g., less than 4 nM, less than 3 nM, preferably less than 2 nM, as measured by surface plasmon resonance; and optionally binds to cynomolgus monkey serum albumin with a dissociation constant (K D ) is less than 5 nM, e.g., less than 4 nM, less than 3 nM, preferably less than 2 nM, as measured by surface plasmon resonance.

[0064] 18. The multispecific antibody according to any one of the preceding items, wherein the antibody has the following characteristics

[0065] (a) a melting temperature (Tm) of at least 55°C, preferably at least 58°C, more preferably at least 60°C, as measured by differential scanning fluorimetry in phosphate-citrate buffer, pH 6.4, 150 mM NaCl;

[0066] (b) when the multispecific antibody is at a starting concentration of 10 mg / ml in phosphate buffered saline (PBS) at pH 7.4, after five consecutive freeze-thaw cycles, the monomer content loss is less than 5%, e.g., less than 4%, less than 3%, less than 2%, preferably 1% or less;

[0067] (c) when the multispecific antibody is stored at an initial concentration of 10 mg / ml in phosphate buffered saline (PBS) at pH 7.4 at 4°C for at least two weeks, in particular at least four weeks, the monomer content loss is less than 10%, preferably less than 5%; and / or

[0068] (d) when the multispecific antibody is stored at an initial concentration of 10 mg / ml in phosphate buffered saline (PBS) at pH 7.4 at 37°C for at least two weeks, particularly at least four weeks, the monomer content loss is less than 20%, preferably less than 15%.

[0069] 19. The multispecific antibody of any one of the preceding items, wherein each domain comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein:

[0070] (a) the VH comprises three complementary determining regions HCDR1, HCDR2 and HCDR3 in sequence, and

[0071] (b) The VL comprises three complementary determining regions LCDR1, LCDR2 and LCDR3 in sequence.

[0072] 20. The multispecific antibody of any of the preceding items, wherein the first domain that specifically binds to IL-17A comprises a set of CDRs: HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3, wherein the set of CDRs has 10 or fewer amino acid substitutions relative to the set of CDRs: (i) HCDR1′ as set forth in SEQ ID NO: 1; HCDR2′ as set forth in SEQ ID NO: 2; HCDR3′ as set forth in SEQ ID NO: 3; LCDR1′ as set forth in SEQ ID NO: 12; LCDR2′ as set forth in SEQ ID NO: 13; LCDR3′ as set forth in SEQ ID NO: 14, or (ii) HCDR1′ as set forth in SEQ ID NO: 39; HCDR2′ as set forth in SEQ ID NO: 40; HCDR3′ as set forth in SEQ ID NO: 41; LCDR1′ as set forth in SEQ ID NO: 50; LCDR2′ as set forth in SEQ ID NO: 51; LCDR3′ as set forth in SEQ ID NO: 52.

[0073] 21. The multispecific antibody according to item 20, wherein

[0074] (i)

[0075] (a) the HCDR1 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 1, 4, and 7;

[0076] (b) the HCDR2 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 2, 5 and 8;

[0077] (c) the HCDR3 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 3, 6, and 9;

[0078] (d) the LCDR1 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 12, 15, and 18;

[0079] (e) the LCDR2 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 13, 16, and 19; and

[0080] (f) the LCDR3 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 14, 17, and 20; or

[0081] (ii)

[0082] (a) the HCDR1 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 39, 42, and 45;

[0083] (b) the HCDR2 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 40, 43, and 46;

[0084] (c) the HCDR3 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 41, 44, and 47;

[0085] (d) the LCDR1 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 50, 53, and 56;

[0086] (e) the LCDR2 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 51, 54, and 57; and

[0087] (f) The LCDR3 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 52, 55 and 58.

[0088] 22. The multispecific antibody of claim 21 , comprising (i) HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs: 1, 2, and 3, respectively, and LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs: 12, 13, and 14, respectively, or (ii) (i) HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs: 39, 40, and 41, respectively, and LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs: 50, 51, and 52, respectively.

[0089] 23. The multispecific antibody according to any of the preceding items, wherein the first domain that specifically binds to IL-17A comprises a heavy chain variable region VHA and wherein the VHA is VH3 or VH4, preferably VH3.

[0090] 24. The multispecific antibody according to any one of the preceding items, wherein the first domain that specifically binds to IL-17A comprises a light chain variable region VLA and wherein the VLA comprises Vκ framework FR1, FR2 and FR3, in particular Vκ1 or Vκ3FR1 to FR3, preferably Vκ1 FR1 to FR3, and a framework FR4, wherein the framework FR4 is selected from VκFR4, in particular Vκ1 FR4, Vκ3FR4, and VλFR4, in particular VλFR4 comprising an amino acid sequence that is at least 60, 70, 80, 90% identical to an amino acid sequence selected from any one of SEQ ID NO: 26 to SEQ ID NO: 32, preferably VλFR4 as shown in any one of SEQ ID NO: 26 to SEQ ID NO: 32, preferably VλFR4 as shown in SEQ ID NO: 26 or 27, more preferably VλFR4 as shown in SEQ ID NO: 27.

[0091] 25. The multispecific antibody of any one of items 23 to 24, wherein the VHA comprises (i) an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 10; and / or the VLA comprises an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 21, or the VHA comprises (ii) an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 48; and / or the VLA comprises an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 59.

[0092] 26. The multispecific antibody of claim 25, wherein the VHA comprises (i) an amino acid sequence selected from the group consisting of SEQ ID NOs: 10 and 11; and / or the VLA comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 21 and 22, or the VHA comprises (ii) an amino acid sequence selected from the group consisting of SEQ ID NOs: 48 and 49; and / or the VLA comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 59 and 60.

[0093] 27. The multispecific antibody according to item 26, comprising (i) the VHA sequence of SEQ ID NO: 10 and / or the VLA sequence of SEQ ID NO: 21, or (ii) the VHA sequence of SEQ ID NO: 48 and / or the VLA sequence of SEQ ID NO: 59.

[0094] 28. A multispecific antibody according to any of the preceding items, wherein the second domain that specifically binds to TNFα comprises a set of CDRs: HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, wherein the set of CDRs has 10 or fewer amino acid substitutions relative to a set of CDRs: HCDR1' as shown in SEQ ID NO: 63; HCDR2' as shown in SEQ ID NO: 64; HCDR3' as shown in SEQ ID NO: 65; LCDR1' as shown in SEQ ID NO: 76; LCDR2' as shown in SEQ ID NO: 77; LCDR3' as shown in SEQ ID NO: 78.

[0095] 29. The multispecific antibody according to item 28, wherein

[0096] (a) the HCDR1 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 63, 66 and 69;

[0097] (b) the HCDR2 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 64, 67, and 70;

[0098] (c) the HCDR3 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 65, 68 and 71;

[0099] (d) the LCDR1 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 76, 79, and 82;

[0100] (e) the LCDR2 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 77, 80, and 83; and

[0101] (f) The LCDR3 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 78, 81 and 84.

[0102] 30. The multispecific antibody of claim 29, comprising the HCDR1, HCDR2 and HCDR3 sequences of SEQ ID NOs: 63, 64, and 65, respectively, and the LCDR1, LCDR2 and LCDR3 sequences of SEQ ID NOs: 76, 77, and 78, respectively.

[0103] 31. The multispecific antibody according to any one of the preceding items, wherein the second domain that specifically binds to TNFα comprises a heavy chain variable region VHB and wherein the VHB is VH3 or VH4, preferably VH3.

[0104] 32. A multispecific antibody according to any of the preceding items, wherein the second domain that specifically binds to TNFα comprises a light chain variable region VLB and wherein the VLB comprises a Vκ framework FR1, FR2 and FR3, in particular Vκ1 or Vκ3 FR1 to FR3, preferably Vκ1 FR1 to FR3, and a framework FR4, wherein the framework FR4 is selected from VκFR4, in particular Vκ1 FR4, Vκ3FR4, and VλFR4, in particular a VλFR4 comprising an amino acid sequence that is at least 60, 70, 80, 90% identical to an amino acid sequence selected from any one of SEQ ID NO: 26 to SEQ ID NO: 32, preferably a VλFR4 as shown in any one of SEQ ID NO: 26 to SEQ ID NO: 32, preferably a VλFR4 as shown in SEQ ID NO: 26 or 27, more preferably a VλFR4 as shown in SEQ ID NO: 27.

[0105] 33. A multispecific antibody according to any one of items 31 to 32, wherein the VHB comprises an amino acid sequence that is at least 90% identical to an amino acid sequence selected from the group consisting of SEQ ID NO: 72, 73, 74 and 75, preferably an amino acid sequence that is at least 90% identical to SEQ ID NO: 72; and / or the VLB comprises an amino acid sequence that is at least 90% identical to an amino acid sequence selected from the group consisting of SEQ ID NO: 85, 86, 87, 88 and 89, preferably an amino acid sequence that is at least 90% identical to SEQ ID NO: 85.

[0106] 34. The multispecific antibody of claim 33, wherein the VHB comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 72, 73, 74, and 75; and / or the VLB comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 85, 86, 87, 88, and 89.

[0107] 35. The multispecific antibody according to claim 34, comprising (i) the VHB sequence of SEQ ID NO: 72 and / or the VLB sequence of SEQ ID NO: 85; or (ii) the VHB sequence of SEQ ID NO: 75 and / or the VLB sequence of SEQ ID NO: 88; or (iii) the VHB sequence of SEQ ID NO: 75 and / or the VLB sequence of SEQ ID NO: 89.

[0108] 36. The multispecific antibody according to any one of items 6 to 35, wherein the third domain that specifically binds to HSA comprises a set of CDRs: HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, wherein the set of CDRs has 10 or fewer amino acid substitutions relative to such a set of CDRs:

[0109] (a) HCDR1′ is set forth in SEQ ID NO:90; HCDR2′ is set forth in SEQ ID NO:91; HCDR3′ is set forth in SEQ ID NO:92; LCDR1′ is set forth in SEQ ID NO:100; LCDR2′ is set forth in SEQ ID NO:101; and LCDR3′ is set forth in SEQ ID NO:102; or

[0110] (b) HCDR1' is shown in SEQ ID NO: 111; HCDR2' is shown in SEQ ID NO: 112; HCDR3' is shown in SEQ ID NO: 113; LCDR1' is shown in SEQ ID NO: 120; LCDR2' is shown in SEQ ID NO: 121; and LCDR3' is shown in SEQ ID NO: 122.

[0111] 37. The multispecific antibody according to item 36, wherein

[0112] (a) the HCDR1 is an amino acid sequence selected from any one of SEQ ID NOs: 90, 93 and 96; the HCDR2 is an amino acid sequence selected from any one of SEQ ID NOs: 91, 94 and 97; the HCDR3 is an amino acid sequence selected from any one of SEQ ID NOs: 92, 95 and 98; the LCDR1 is an amino acid sequence selected from any one of SEQ ID NOs: 100, 103 and 106; the LCDR2 is an amino acid sequence selected from any one of SEQ ID NOs: 101, 104 and 107; and the LCDR3 is an amino acid sequence selected from any one of SEQ ID NOs: 102, 105 and 108; or

[0113] (b) the HCDR1 is set forth as an amino acid sequence selected from any one of SEQ ID NOs: 111, 114 and 117; the HCDR2 is set forth as an amino acid sequence selected from any one of SEQ ID NOs: 112, 115 and 118; the HCDR3 is set forth as an amino acid sequence selected from any one of SEQ ID NOs: 113, 116 and 119; the LCDR1 is set forth as an amino acid sequence selected from any one of SEQ ID NOs: 121, 124 and 127; the LCDR2 is set forth as an amino acid sequence selected from any one of SEQ ID NOs: 122, 125 and 128; and the LCDR3 is set forth as an amino acid sequence selected from any one of SEQ ID NOs: 123, 126 and 129.

[0114] 38. The multispecific antibody according to item 37, comprising

[0115] (a) the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs: 90, 91, and 92, respectively, and the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs: 100, 101, and 102, respectively; or

[0116] (b) HCDR1, HCDR2 and HCDR3 sequences of SEQ ID NOs: 111, 112, and 113, respectively, and LCDR1, LCDR2 and LCDR3 sequences of SEQ ID NOs: 121, 122, and 123, respectively.

[0117] 39. The multispecific antibody according to any one of items 6 to 38, wherein the third domain that specifically binds to HSA comprises a heavy chain variable region VHC and wherein the VHC is VH3 or VH4, preferably VH3.

[0118] 40. The multispecific antibody of any one of items 6 to 39, wherein the third domain that specifically binds to HSA comprises a light chain variable region VLC and wherein the VLC comprises Vκ framework FR1, FR2 and FR3, in particular Vκ1 or Vκ3 FR1 to FR3, preferably Vκ1 FR1 to FR3, and a framework FR4, wherein the framework FR4 is selected from VκFR4, in particular Vκ1 FR4, Vκ3 FR4, and VλFR4, in particular VλFR4 comprising an amino acid sequence that is at least 60, 70, 80, 90% identical to an amino acid sequence selected from any one of SEQ ID NO: 26 to SEQ ID NO: 32, preferably VλFR4 as shown in any one of SEQ ID NO: 26 to SEQ ID NO: 32, preferably VλFR4 as shown in SEQ ID NO: 26 or 27, more preferably VλFR4 as shown in SEQ ID NO: 27.

[0119] 41. A multispecific antibody according to any one of items 39 to 40, wherein the VHC comprises an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 99; and / or the VLC comprises an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 109.

[0120] 42. The multispecific antibody according to item 41, comprising the VHC sequence of SEQ ID NO: 99 and / or the VLC sequence of SEQ ID NO: 109.

[0121] 43. A multispecific antibody according to any one of items 39 to 40, wherein the VHC comprises an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 110; and / or the VLC comprises an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 120.

[0122] 44. The multispecific antibody according to claim 43, comprising the VHC sequence of SEQ ID NO: 110 and / or the VLC sequence of SEQ ID NO: 120.

[0123] 45. The multispecific antibody according to any of the preceding items, wherein the antibody is humanized.

[0124] 46. ​​A multispecific antibody according to any of the preceding items, wherein the antibody comprises an amino acid sequence that is at least 80% identical, preferably at least 90% identical, to a sequence selected from SEQ ID NO: 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, and 148, preferably 143, wherein the CDRs have sequences according to items 22, 30 and 38(a).

[0125] 47. The multispecific antibody according to any of the preceding items, wherein the antibody comprises an amino acid sequence selected from any one of SEQ ID NOs: 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, and 148, preferably 143.

[0126] 48. A pharmaceutical composition comprising the multispecific antibody according to any one of the preceding items and a pharmaceutically acceptable carrier.

[0127] 49. The multispecific antibody of any one of items 1 to 47, or the pharmaceutical composition of item 48, for use as a medicament.

[0128] 50. The multispecific antibody of any one of items 1 to 47, or the pharmaceutical composition of item 48, for use in treating a disorder mediated by IL-17A and / or TNFα or a disorder treatable by inhibiting Gro-α secretion.

[0129] 51. The multispecific antibody of any one of items 1 to 47, or the pharmaceutical composition of item 48, for use in treating an inflammatory condition or an autoimmune disease.

[0130] 52. The multispecific antibody of any one of items 1 to 47, or the pharmaceutical composition of item 48, for use in treating cancer, arthritis, rheumatoid arthritis, osteoarthritis, reactive arthritis, psoriasis, chronic obstructive pulmonary disease, systemic lupus erythematosus (SLE), lupus nephritis, autoimmune inflammatory bowel disease, asthma, multiple sclerosis, or cystic fibrosis, bone loss, airway hypersensitivity, demyelinating disorders, skin hypersensitivity, acute transplant rejection, allograft rejection, graft-versus-host disease, systemic sclerosis, urological inflammatory disorders, cardiovascular disease, vasculitis, recurrent fevers, glucose metabolism disorders, lung disease, peridontitis, stromal keratitis of the liver, allergies, inflammatory pain, spondyloarthropathies, sepsis, septic or endotoxic shock, meningitis, surgical trauma, autoimmune blood disorders, Alzheimer's disease, sarcoidosis, cirrhosis, hepatitis, glomerulonephritis, or dislipidemia.

[0131] 53. Use of the multispecific antibody of any one of items 1 to 47, or the pharmaceutical composition of item 48, in the preparation of a medicament for treating a disorder mediated by IL-17A and / or TNFα or a disorder treatable by inhibiting Gro-α secretion.

[0132] 54. Use of the multispecific antibody of any one of items 1 to 47, or the pharmaceutical composition of item 48, in the preparation of a medicament for treating an inflammatory condition or an autoimmune disease.

[0133] 55. Use of the multispecific antibody of any one of items 1 to 47, or the pharmaceutical composition of item 48, in the preparation of a medicament for treating cancer, arthritis, rheumatoid arthritis, osteoarthritis, reactive arthritis, psoriasis, chronic obstructive pulmonary disease, systemic lupus erythematosus (SLE), lupus nephritis, autoimmune inflammatory bowel disease, asthma, multiple sclerosis, or cystic fibrosis, bone loss, airway hypersensitivity, demyelinating disorders, skin hypersensitivity, acute transplant rejection, allogeneic transplant rejection, graft-versus-host disease, systemic sclerosis, urological inflammatory disorders, cardiovascular disease, vasculitis, periodic fevers, glucose metabolism disorders, lung disease, peridontitis, stromal keratitis of the liver, allergy, inflammatory pain, spondyloarthropathy, sepsis, septic or endotoxic shock, meningitis, surgical trauma, autoimmune blood disorders, Alzheimer's disease, sarcoidosis, cirrhosis, hepatitis, glomerulonephritis, or dislipidemia.

[0134] 56. A method for treating a disorder mediated by IL-17A and / or TNFα, the method comprising administering an effective amount of the multispecific antibody of any one of items 1 to 47, or the pharmaceutical composition of item 48, such that the disorder is alleviated.

[0135] 57. The method of item 56, wherein the disorder mediated by IL-17A and / or TNFα is an inflammatory condition or an autoimmune disease.

[0136] 58. The method of claim 56, wherein the disorder mediated by IL-17A and / or TNFα is cancer, arthritis, rheumatoid arthritis, osteoarthritis, reactive arthritis, psoriasis, chronic obstructive pulmonary disease, systemic lupus erythematosus (SLE), lupus nephritis, autoimmune inflammatory bowel disease, asthma, multiple sclerosis cystic fibrosis, bone loss, airway hypersensitivity, demyelinating disorders, skin hypersensitivity, acute transplant rejection, allogeneic transplant rejection, graft-versus-host disease, systemic sclerosis, urological inflammatory disorders, cardiovascular disease, vasculitis, recurrent fevers, glucose metabolism disorders, lung disease, peridontitis, stromal keratitis of the liver, allergy, inflammatory pain, spondyloarthropathies, sepsis, septic or endotoxic shock, meningitis, surgical trauma, autoimmune blood disorders, Alzheimer's disease, sarcoidosis, cirrhosis, hepatitis, glomerulonephritis, or dislipidemia.

[0137] 59. A nucleic acid encoding the multispecific antibody or fragment thereof according to any one of items 1 to 47.

[0138] 60. A vector comprising the nucleic acid of item 59.

[0139] 61. A host cell comprising the nucleic acid of item 59 or the vector of item 60.

[0140] 62. A method for producing the multispecific antibody of any one of items 1 to 47, the method comprising the following steps: culturing a host cell comprising a nucleic acid or vector encoding the multispecific antibody or fragment thereof of any one of items 1 to 47.

[0141] 63. A kit comprising the multispecific antibody of any one of items 1 to 47, or the pharmaceutical composition of item 48.

[0142] Another object of the present invention is to provide anti-IL-17A antibodies with improved affinity, efficacy, and improved biophysical properties, such as improved solubility, developability, and stability.

[0143] The anti-IL-17A antibodies of the present invention have improved properties that are advantageous for use in therapy, such as higher affinity, improved efficacy, selectivity, improved biophysical properties such as solubility, developability, and stability.

[0144] Thus, in one aspect, the present disclosure provides an isolated antibody having binding specificity for human IL-17A, in particular, comprising a set of CDRs: HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3, wherein the set of CDRs has 10 or fewer, preferably 0, amino acid substitutions relative to such a set of CDRs: HCDR1′ is an amino acid sequence selected from any one of SEQ ID NOs: 1, 4, and 7; HCDR2′ is an amino acid sequence selected from any one of SEQ ID NOs: 2, 5, and 8; HCDR3′ is an amino acid sequence selected from any one of SEQ ID NOs: 3, 6, and 9; LCDR1′ is an amino acid sequence selected from any one of SEQ ID NOs: 12, 15, and 18; LCDR2′ is an amino acid sequence selected from any one of SEQ ID NOs: 13, 16, and 19; and LCDR3′ has an amino acid sequence selected from any one of SEQ ID NOs: 14, 17, and 20. In one aspect, the present disclosure relates to a multispecific molecule comprising the isolated antibody of the present disclosure.

[0145] In another aspect, the present disclosure provides an isolated antibody having binding specificity to human IL-17A, particularly comprising a set of CDRs: HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3, wherein the set of CDRs has 10 or fewer, preferably 0, amino acid substitutions relative to such a set of CDRs: HCDR1′ is an amino acid sequence selected from any one of SEQ ID NOs: 39, 42, and 45; HCDR2′ is an amino acid sequence selected from any one of SEQ ID NOs: 40, 43, and 46; HCDR3′ is an amino acid sequence selected from any one of SEQ ID NOs: 41, 44, and 47; LCDR1′ is an amino acid sequence selected from any one of SEQ ID NOs: 50, 53, and 56; LCDR2′ is an amino acid sequence selected from any one of SEQ ID NOs: 51, 54, and 57; and LCDR3′ has an amino acid sequence selected from any one of SEQ ID NOs: 52, 55, and 58. In one aspect, the disclosure relates to a multispecific molecule comprising an isolated antibody of the disclosure.

[0146] In one aspect, the present disclosure relates to a pharmaceutical composition comprising an isolated antibody of the present disclosure, or a multispecific molecule comprising an isolated antibody of the present disclosure, and a pharmaceutically acceptable carrier.

[0147] In another aspect, the present disclosure relates to an antibody of the present disclosure or a multispecific molecule comprising the isolated antibody, or a pharmaceutical composition of the present disclosure, for use as a medicament.

[0148] In one aspect, the present disclosure relates to antibodies, or multispecific molecules comprising the isolated antibodies, or pharmaceutical compositions of the present disclosure for treating a disorder mediated by IL-17A or a disorder treatable by inhibiting GRO-α secretion.

[0149] In one aspect, the present disclosure relates to the use of an antibody of the present disclosure, or a multispecific molecule comprising the isolated antibody of the present disclosure, or a pharmaceutical composition of the present disclosure, in the preparation of a medicament for treating a disorder mediated by IL-17A or a disorder treatable by inhibiting GRO-α secretion.

[0150] In another aspect, the present disclosure relates to a method for treating a condition mediated by IL-17A, comprising administering to a subject in need thereof an effective amount of an antibody, multispecific molecule, or pharmaceutical composition of the present disclosure. In yet another aspect, the present disclosure relates to nucleic acids encoding the antibodies of the present disclosure. In another aspect, the present disclosure relates to vectors comprising the nucleic acids. In another aspect, the present disclosure relates to host cells comprising the nucleic acids or vectors.

[0151] In another aspect, the present disclosure relates to a method for producing an antibody of the present disclosure, comprising the steps of culturing a host cell comprising a nucleic acid or a vector encoding the antibody of the present disclosure.

[0152] The aspects, advantageous features and preferred embodiments of the present disclosure outlined in the following, individually or in combination, further contribute to solving the objects of the present invention:

[0153] 1. An isolated antibody having binding specificity for human IL-17A, comprising a set of CDRs: HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3, wherein the set of CDRs has 10 or fewer amino acid substitutions relative to such a set of CDRs:

[0154] (i) HCDR1′ is an amino acid sequence selected from any one of SEQ ID NOs: 1, 4, and 7;

[0155] HCDR2′ is an amino acid sequence selected from any one of SEQ ID NOs: 2, 5, and 8;

[0156] HCDR3' is an amino acid sequence selected from any one of SEQ ID NOs: 3, 6, and 9;

[0157] LCDR1′ is an amino acid sequence selected from any one of SEQ ID NOs: 12, 15, and 18;

[0158] LCDR2′ is an amino acid sequence selected from any one of SEQ ID NOs: 13, 16, and 19;

[0159] LCDR3' has an amino acid selected from any one of SEQ ID NOs: 14, 17, and 20; or

[0160] (ii) HCDR1′ is an amino acid sequence selected from any one of SEQ ID NOs: 39, 42, and 45;

[0161] HCDR2′ is an amino acid sequence selected from any one of SEQ ID NOs: 40, 43, and 46;

[0162] HCDR3′ is an amino acid sequence selected from any one of SEQ ID NOs: 41, 44, and 47;

[0163] LCDR1′ is an amino acid sequence selected from any one of SEQ ID NOs: 50, 53, and 56;

[0164] LCDR2′ is an amino acid sequence selected from any one of SEQ ID NOs: 51, 54, and 57;

[0165] LCDR3′ has an amino acid sequence selected from any one of SEQ ID NOs: 52, 55, and 58.

[0166] 2. The antibody of claim 1, comprising a set of CDRs: HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3, wherein the set of CDRs has 10 or fewer amino acid substitutions relative to such a set of CDRs:

[0167] (i) HCDR1′ is represented by SEQ ID NO: 1;

[0168] HCDR2' is shown in SEQ ID NO: 1;

[0169] HCDR3' is shown in SEQ ID NO: 3;

[0170] LCDR1' is shown in SEQ ID NO: 12;

[0171] LCDR2' is shown in SEQ ID NO: 13;

[0172] LCDR3' is shown in SEQ ID NO: 14; or

[0173] (ii) HCDR1′ is represented by SEQ ID NO: 39;

[0174] HCDR2' is shown in SEQ ID NO:40;

[0175] HCDR3' is shown in SEQ ID NO:41;

[0176] LCDR1' is shown in SEQ ID NO:50;

[0177] LCDR2' is shown in SEQ ID NO:51;

[0178] LCDR3' is shown in SEQ ID NO:52.

[0179] 3. The antibody according to item 1 or 2, comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein:

[0180] (c) the VH comprises three complementary determining regions HCDR1, HCDR2 and HCDR3 in sequence, and

[0181] (d) The VL comprises three complementary determining regions LCDR1, LCDR2 and LCDR3 in sequence.

[0182] 4. The antibody according to item 3, wherein

[0183] (i)

[0184] (a) the HCDR1 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 1, 4, and 7;

[0185] (b) the HCDR2 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 2, 5 and 8;

[0186] (c) the HCDR3 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 3, 6, and 9;

[0187] (d) the LCDR1 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 12, 15, and 18;

[0188] (e) the LCDR2 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 13, 16, and 19; and

[0189] (f) the LCDR3 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 14, 17, and 20; or

[0190] (ii)

[0191] (a) the HCDR1 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 39, 42, and 45;

[0192] (b) the HCDR2 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 40, 43, and 46;

[0193] (c) the HCDR3 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 41, 44, and 47;

[0194] (d) the LCDR1 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 50, 53, and 56;

[0195] (e) the LCDR2 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 51, 54, and 57; and

[0196] (f) The LCDR3 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 52, 55 and 58.

[0197] 5. The antibody of claim 4, wherein the antibody comprises (i) HCDR1, HCDR2 and HCDR3 sequences of SEQ ID NOs: 1, 2 and 3, respectively, and LCDR1, LCDR2 and LCDR3 sequences of SEQ ID NOs: 12, 13 and 14, respectively; or (ii) HCDR1, HCDR2 and HCDR3 sequences of SEQ ID NOs: 39, 40 and 41, respectively, and LCDR1, LCDR2 and LCDR3 sequences of SEQ ID NOs: 50, 51 and 52, respectively.

[0198] 6. The antibody of any one of items 3 to 5, wherein the VH is VH3 or VH4, preferably VH3.

[0199] 7. The antibody according to any one of items 3 to 6, wherein the VL comprises a Vκ framework FR1, FR2 and FR3, in particular Vκ1 or Vκ3 FR1 to FR3, preferably Vκ1 FR1 to FR3, and a framework FR4, wherein the framework FR4 is selected from VκFR4, in particular Vκ1 FR4, Vκ3 FR4, and VλFR4, in particular a VλFR4 comprising an amino acid sequence that is at least 60, 70, 80, 90% identical to an amino acid sequence selected from any one of SEQ ID NO: 26 to SEQ ID NO: 32, preferably a VλFR4 as shown in any one of SEQ ID NO: 26 to SEQ ID NO: 32, preferably a VλFR4 as shown in SEQ ID NO: 26 or 27, more preferably a VλFR4 as shown in SEQ ID NO: 27.

[0200] 8. An antibody according to any one of items 3 to 7, wherein (i) the VH comprises an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 10; and / or the VL comprises an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 21; or (i) the VH comprises an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 48; and / or the VL comprises an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 59.

[0201] 9. The antibody of claim 8, wherein (i) the VH comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 10 and 11; and / or the VL comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 21 and 22; or (ii) the VH comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 48 and 49; and / or the VL comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 59 and 60.

[0202] 10. The antibody according to item 9, comprising (i) the VH sequence of SEQ ID NO: 10 and / or the VL sequence of SEQ ID NO: 21; or (ii) the VH sequence of SEQ ID NO: 48 and / or the VL sequence of SEQ ID NO: 59.

[0203] 11. The antibody according to item 9, comprising (i) the VH sequence of SEQ ID NO: 11 and / or the VL sequence of SEQ ID NO: 49; or (ii) the VH sequence of SEQ ID NO: 11 and / or the VL sequence of SEQ ID NO: 60.

[0204] 12. The antibody of any preceding item, wherein the antibody has binding specificity for cynomolgus monkey IL-17A.

[0205] 13. The antibody according to any of the preceding items, wherein the antibody selectively binds human IL-17A relative to human IL-17B, IL-17C, IL-17D, IL-17E and IL-17F as measured by ELISA.

[0206] 14. The antibody according to any one of the preceding items, which binds to IL-17A

[0207] (a) inhibiting or blocking the binding between IL-17A and its receptor (IL-17RA), and

[0208] (b) Reduce or neutralize IL-17A activity.

[0209] 15. The antibody of claim 14, wherein the antibody is capable of inhibiting GRO-α secretion when assessed in vitro in the HT-29 assay.

[0210] 16. The antibody according to any of the preceding items, wherein the antibody:

[0211] (h) having the ability to block the interaction between IL-17A and IL-17RA, with a potency relative to secukinumab (relative potency) of greater than 5, preferably greater than 10, more preferably greater than 15, even more preferably greater than 20 as determined in an ELISA assay, wherein the relative potency is the IC value of secukinumab in ng / mL as measured by ELISA. 50 The values ​​are similar to the IC values ​​in ng / mL of the scFv form of the present invention measured by ELISA. 50 and / or

[0212] (i) having the ability to neutralize IL-17A with a potency relative to secukinumab (relative potency) of greater than 50, preferably greater than 100, more preferably greater than 150 as determined by measuring GRO-α secretion in the HT-29 assay, and wherein the relative potency is the IC of secukinumab in ng / mL as measured in the HT-29 assay 50 The values ​​are comparable to the IC values ​​in ng / mL of the scFv forms of the antibodies of the invention measured in the HT-29 assay. 50 and / or

[0213] (j) capable of inhibiting the activity of 1 ng of human IL-17A by 50% at a concentration of 1 ng / ml or less, preferably 0.5 ng / ml or less, more preferably 0.2 ng / ml or less, as determined by measuring GRO-α secretion induced by human IL-17A in the presence of 50 pg / ml TNFα in an HT-29 assay.

[0214] 17. The antibody according to any of the preceding items, wherein the antibody:

[0215] (a) Binding to human IL-17A, dissociation constant (K D ) is less than 5 nM, in particular less than 1 nM, less than 0.5 nM, less than 0.2 nM, more particularly less than 100 pM, more particularly less than 50 pM, as measured by surface plasmon resonance, in particular by surface plasmon resonance in a direct setting; and

[0216] (b) optionally, in combination with cynomolgus monkey IL-17A, K DLess than 10 nM, in particular less than 7 nM, less than 5 nM, less than 2 nM, less than 1 nM, more in particular less than 0.5 nM as measured by surface plasmon resonance, in particular as measured by surface plasmon resonance in a trapping setup.

[0217] 18. The antibody according to item 17, wherein the antibody binds to human IL-17A with a dissociation constant (K D ) is less than 0.5 nM, less than 0.2 nM, less than 100 pM, in particular less than 50 pM, as measured by surface plasmon resonance, in particular by surface plasmon resonance in a direct setup.

[0218] 19. The antibody according to any of the preceding items, wherein the antibody:

[0219] (e) when in scFv format, a melting temperature (Tm) of at least 60°C, particularly at least 62°C, at least 65°C, more particularly at least 70°C, as determined by differential scanning fluorimetry, particularly wherein the antibody is in phosphate-citrate buffer, pH 6.4, 150 mM NaCl;

[0220] (f) when in scFv format, the antibody of the invention loses less than 5%, particularly less than 3%, more particularly less than 1% of monomer content after 5 consecutive freeze-thaw cycles when the starting concentration is 10 mg / ml, particularly wherein the antibody is in phosphate buffered saline (PBS) at pH 7.4;

[0221] (g) when in scFv format, the antibody of the invention has a monomer content loss of 5% or less, particularly less than 4%, less than 3%, less than 2%, more particularly less than 1%, after storage at 4°C for at least two weeks, particularly at least 4 weeks, when the initial concentration is 10 mg / ml, particularly wherein the antibody is in phosphate buffered saline (PBS) at pH 7.4; and / or

[0222] (h) When the initial concentration of the antibody of the present invention is 10 mg / ml, the monomer content loss is less than 5% after storage at 37°C for at least two weeks, especially at least four weeks.

[0223] 20. The antibody of any of the preceding items, wherein the antibody is selected from the group consisting of: a monoclonal antibody, a chimeric antibody, a Fab, a Fv, a scFv, a dsFv, a scAb, a STAB, a single domain antibody (sdAb or dAb), a single domain heavy chain antibody, and a single domain light chain antibody, a VHH, a VNAR, a single domain antibody based on the VNAR structure of a shark, and binding domains based on alternative scaffolds including but not limited to ankyrin-based domains, fynomers, avimers, anticalins, fibronectin, and binding sites built into antibody constant regions (e.g., Modular Antibody Technology of F-star). TM ), preferably scFv.

[0224] 21. The antibody according to claim 20, wherein the scFv has an amino acid sequence selected from (i) the group consisting of SEQ ID NO: 24 and SEQ ID NO: 25, preferably, wherein the scFv has the amino acid sequence of SEQ ID NO: 24; or the scFv has an amino acid sequence selected from (ii) the group consisting of SEQ ID NO: 61 and SEQ ID NO: 62, preferably, wherein the scFv has the amino acid sequence of SEQ ID NO: 61.

[0225] 22. The isolated antibody according to item 20, wherein the antibody is an IgG selected from the group consisting of IgG1, IgG2, IgG3 and IgG4, preferably, wherein the antibody is IgG1 or IgG4.

[0226] 23. The isolated antibody of any preceding item, wherein the antibody is humanized.

[0227] 24. The antibody of any one of items 1 to 23, which is a multispecific molecule.

[0228] 25. The antibody of claim 24, wherein the antibody format is selected from the group consisting of: single-chain diabody (scDb), tandem scDb (Tandab), linear dimer scDb (LD-scDb), cyclic dimer scDb (CD-scDb), bispecific T cell engager (BiTE; tandem bi-scFv), tandem tri-scFv, triabody (Fab-(scFv)2) or diabody (Fab-(scFv)1), Fab, Fab-Fv2, Morrison (IgG CH3-scFv fusion (Morrison L) or IgG CL-scFv fusion (Morrison H)), triabodies, scDb-scFv, bispecific Fab2, diabodies, tetrabodies, scFv-Fc-scFv fusion, scFv-HSA-scFv fusion, diabodies, DVD-Ig, COVD, IgG-scFab, scFab-dsscFv, Fv2-Fc, IgG-scFv fusions such as bsAb (scFv linked to the C-terminus of the light chain), Bs1Ab (scFv linked to the N-terminus of the light chain), Bs2Ab (scFv linked to the N-terminus of the heavy chain), Bs3Ab (scFv linked to the C-terminus of the heavy chain), Ts1Ab (scFv linked to the N-terminus of the heavy and light chains), Ts2Ab (dsscFv linked to the C-terminus of the heavy chain), heterodimeric Fc domain-based bispecific antibodies such as Knob-into-Hole antibodies (KiH); Fv, scFv, scDb, tandem bi-scFv, tandem tri-scFv, Fab-(scFv)2, Fab-(scFv)1, Fab, Fab-Fv2, COVD, MATCH and DuoBody fused to the N- and / or C-terminus of any one chain of the heterodimeric Fc domain or any other heterodimeric domain.

[0229] 26. A pharmaceutical composition comprising the antibody according to any one of items 1 to 25 and a pharmaceutically acceptable carrier.

[0230] 27. The antibody according to any one of items 1 to 25, or the pharmaceutical composition according to item 26, for use as a medicament.

[0231] 28. The antibody of any one of items 1 to 25, or the pharmaceutical composition of item 26, for use in treating a disorder mediated by IL-17A or a disorder treatable by inhibiting GRO-α secretion.

[0232] 29. The antibody of any one of items 1 to 25, or the pharmaceutical composition of item 26, for use in treating an inflammatory condition or an autoimmune disease.

[0233] 30. The antibody of any one of items 1 to 25, or the pharmaceutical composition of item 26, for use in treating cancer, arthritis, rheumatoid arthritis, osteoarthritis, reactive arthritis, psoriasis, chronic obstructive pulmonary disease, systemic lupus erythematosus (SLE), lupus nephritis, autoimmune inflammatory bowel disease, asthma, multiple sclerosis, or cystic fibrosis, bone loss, airway hypersensitivity, demyelinating disorders, skin hypersensitivity, acute transplant rejection, allograft rejection, graft-versus-host disease, systemic sclerosis, urological inflammatory disorders, cardiovascular disease, vasculitis, recurrent fevers, glucose metabolism disorders, lung disease, peridontitis, hepatic stromal keratitis, allergy, inflammatory pain, spondyloarthropathies, sepsis, septic or endotoxic shock, meningitis, surgical trauma, autoimmune blood disorders, Alzheimer's disease, sarcoidosis, cirrhosis, hepatitis, glomerulonephritis, or dislipidemia.

[0234] 31. Use of the antibody of any one of items 1 to 25, or the pharmaceutical composition of item 26, in the preparation of a medicament for treating a disorder mediated by IL-17A or a disorder treatable by inhibiting GRO-α secretion.

[0235] 32. Use of the antibody of any one of items 1 to 25, or the pharmaceutical composition of item 26, in the preparation of a medicament for treating an inflammatory condition or an autoimmune disease.

[0236] 33. Use of the antibody of any one of items 1 to 25, or the pharmaceutical composition of item 26, in the preparation of a medicament for treating cancer, arthritis, rheumatoid arthritis, osteoarthritis, reactive arthritis, psoriasis, chronic obstructive pulmonary disease, systemic lupus erythematosus (SLE), lupus nephritis, autoimmune inflammatory bowel disease, asthma, multiple sclerosis, or cystic fibrosis, bone loss, airway hypersensitivity, demyelinating disorders, skin hypersensitivity, acute transplant rejection, allograft rejection, graft-versus-host disease, systemic sclerosis, urological inflammatory disorders, cardiovascular disease, vasculitis, recurrent fevers, glucose metabolism disorders, lung disease, peridontitis, hepatic stromal keratitis, allergy, inflammatory pain, spondyloarthropathies, sepsis, septic or endotoxic shock, meningitis, surgical trauma, autoimmune blood disorders, Alzheimer's disease, sarcoidosis, cirrhosis, hepatitis, glomerulonephritis, or dislipidemia.

[0237] 34. A method for treating a disorder mediated by IL-17A, the method comprising administering an effective amount of the antibody of any one of items 1 to 25, or the pharmaceutical composition of item 26, such that the disorder is alleviated.

[0238] 35. The method of item 34, wherein the disorder mediated by IL-17A is an inflammatory condition or an autoimmune disease.

[0239] 36. The method of claim 34, wherein the disease mediated by IL-17A is cancer, arthritis, rheumatoid arthritis, osteoarthritis, reactive arthritis, psoriasis, chronic obstructive pulmonary disease, systemic lupus erythematosus (SLE), lupus nephritis, autoimmune inflammatory bowel disease, asthma, multiple sclerosis cystic fibrosis, bone loss, airway hypersensitivity, demyelinating disorders, skin hypersensitivity, acute transplant rejection, allograft rejection, graft-versus-host disease, systemic sclerosis, urological inflammatory disorders, cardiovascular disease, vasculitis, recurrent fevers, glucose metabolism disorders, lung disease, peridontitis, hepatic stromal keratitis, allergy, inflammatory pain, spondyloarthropathies, sepsis, septic or endotoxic shock, meningitis, surgical trauma, autoimmune blood disorders, Alzheimer's disease, sarcoidosis, cirrhosis, hepatitis, glomerulonephritis, or dislipidemia.

[0240] 37. A nucleic acid encoding the antibody according to items 1-25.

[0241] 38. A vector comprising the nucleic acid of item 37.

[0242] 39. A host cell comprising the nucleic acid of item 37 or the vector of item 38.

[0243] 40. A method for producing the antibody of any one of items 1-25, the method comprising the following steps: culturing a host cell, the host cell comprising a nucleic acid or a vector encoding the antibody of items 1-25.

[0244] 41. A kit comprising the antibody of any one of items 1 to 25, or the pharmaceutical composition of item 26.

[0245] The present disclosure contemplates all combinations of any one or more of the aspects and / or embodiments described above, as well as combinations with any one or more of the embodiments presented in the detailed description and examples.

[0246] Other features, objects, and advantages of the compositions and methods herein will be apparent from the description and drawings, and from the claims. BRIEF DESCRIPTION OF THE DRAWINGS

[0247] Figure 1The results show that labeled IL-17A is suitable for the sorting process. The biological activity of labeled IL-17A in the HT-29 assay. Three-fold serial dilutions of labeled and unlabeled IL-17A were tested in parallel for their potential to induce GRO-α secretion in HT-29 cells. The EC values ​​of labeled IL-17A (IL-17A-RPE) and unlabeled IL-17A (IL-17A) were 0. 50 The values ​​were 82.8ng / ml and 55ng / ml respectively.

[0248] Figure 2 Shown are the potencies of 27-07-G02 rabbit IgG (A) and 27-31-C04 IgG (B) in neutralizing IL-17A in the HT-29 assay.

[0249] Figure 3 Shown are the potencies of 27-07-G02 rabbit IgG (A) and 27-31-C04 IgG (B) to inhibit the interaction between IL-17A and IL-17RA.

[0250] Figure 4 The potency of humanized scFvs A1 and A2 (A) and PRO571 and PRO592 (B) in neutralizing IL-17A in the HT-29 assay is shown.

[0251] Figure 5 The potency of anti-IL-17A scFvs A1 and A2 (A) and PRO571 and PRO592 (B) to inhibit the interaction between human IL-17A and IL-17RA (ELISA) is shown.

[0252] Figure 6 The target specificity of scFv A1 (A) and scFv PRO571 and PRO592 (B) is shown. The potential of IL-17B to IL-17F to inhibit the interaction of biotinylated IL-17A with the scFv was analyzed by competition ELISA. The dose-dependent effects of IL-17A and IL-17B to IL-17F are shown.

[0253] Figure 7 DSF-measured thermal unfolding curves are shown for scFvs A1 and A2 (A) and scFvs PRO571 and PRO592 (B). The resulting Tm values ​​were determined by fitting the data to the Boltzmann equation to obtain the midpoint of the transition.

[0254] Figure 8Shown are storage stability studies of scFv A1 (A) and scFv PRO571 and PRO592 (B) at concentrations greater than 10 mg / mL at three temperatures (37°C, 4°C, and -80°C) for 4 weeks. The change in monomer content over time at different storage temperatures is shown on the left; the change in protein concentration over time at different storage temperatures (4°C (left) and 37°C (right)) is shown on the right. Monomer content was determined by integration of SE-HPLC peak areas and by UV 280 Measure to calculate protein concentration.

[0255] Figure 9 Shown are the monitoring of monomer content of scFv A1 (A) and scFv PRO571 and PRO592 (B) during five repeated freeze and thaw cycles.

[0256] Figure 10 Figure 2 shows a trispecific format. A, Three domain arrangements of the Fab-(scFv)2 molecule were designed. ScFv fusions at the CL and CH1 positions of the trispecific format were considered equivalent, resulting in three variants of this format. B, Three variants of the scDb-scFv domain arrangement were designed. The structures are shown at the bottom with specificity. The Gly-Ser linker connecting the variable domains and the disulfide bonds between the domains are indicated in gray brackets.

[0257] Figure 11 The general process of lead manufacturing is shown. A, Overlay of SE-HPLC traces of the final A3-A5 (left panel) and A6-A8 (right panel) samples. Peaks with retention times of 7-8 minutes correspond to the apparent molecular weight of the monomers of the respective molecules; peaks with retention times >10 minutes are artifacts related to buffer and salts. B, SDS-PAGE analysis of A3-A8 under non-reducing (left) and reducing (right) buffer conditions. A molecular weight reference was loaded in the middle lane. Under non-reducing conditions, the bands for A3-A5 and A6-A8 correspond to the expected molecular weights of ∼100 kDa and ∼75 kDa, respectively. As expected, for the heterodimeric Fab-(scFv)2 antibody format (A3-A5), the band shifted to ∼50 kDa under reducing conditions, while for A6-A8, a band with a molecular weight of ∼75 kDa was also observed under reducing conditions.

[0258] Figure 12 The efficacy of neutralizing TNFα in the L929 assay is shown. The absorbance measured using the Cell Counting Kit-8 is shown as a function of the concentration (nM) of the trispecific molecules A3-A8. A13 (parent bispecific, HSA binder and TNFα blocker) was used as a reference.

[0259] Figure 13Comparison of the potency to neutralize human and cynomolgus monkey TNF-α is shown. Absorbance measured using a Cell Counting Kit-8 in the presence of human or cynomolgus monkey TNF-α is expressed as a function of A5 and A7 concentrations.

[0260] Figure 14 Simultaneous blockade of TNF-α and IL-17A in the presence of HSA in the HT-29 assay is shown. The simultaneous in vitro blocking effect of six trispecific molecules, A3-A8, on TNFα and IL-17A was analyzed using an HT-29 cell-based assay in the presence of 1 mg / ml HSA. Secukinumab (IL-17A blocker) and A13 (parent bispecific, HSA binder and TNFα blocker) were used as references. The obtained GRO-α secretion data are presented as a function of molecular concentration in nM (A, C, and E) and ng / ml (B, D, and F). "Without TNF" represents GRO-α secretion with the addition of IL-17A alone, corresponding to the maximal effect of TNFα blockade. "Without IL17a" represents GRO-α secretion after the addition of TNFα alone, corresponding to the maximal effect of IL-17A blockade. "No IL17a, no TNFa" represents background GRO-α secretion in the absence of added IL-17A and TNFα, corresponding to the maximal effect of simultaneous blockade of TNFα and IL-17A.

[0261] Figure 15 Neutralization of IL-17A binding to IL-17RA in a competitive ELISA is shown. Absorbance measured in a competitive ELISA evaluating IL-17A binding to IL-17RA is expressed as a function of increasing concentrations of six trispecific molecules (A3-A8). Secukinumab (IL-17A blocker) was used as a reference.

[0262] Figure 16 Simultaneous binding to human TNFα, human IL-17A, and HSA by SPR analysis is shown. Six possible injection sequences of different analytes (human TNFα, human IL-17A, and HSA) were performed on the MASS-1 SPR device, and the resulting sensorgrams are shown. The trispecific molecule was immobilized on the sensor chip (A3 in channel 1B, Ch1B, A4 in channel 2B, Ch2B, A5 in channel 3B, Ch3B, A6 in channel 4B, Ch4B, A7 in channel 5B, Ch5B, A8 in channel 6B, Ch6B) and the antigens were injected sequentially.

[0263] Figure 17 Shown is a storage stability study conducted at 10 mg / mL protein concentration for 4 weeks at 37°C, 4°C, and -80°C. The % monomer content and % monomer loss over time were recorded at d0, d2, d7, d14, d21, and d28.

[0264] Figure 18 Shown are overlays of SE-HPLC traces of d0 (black, shaded) and d28 (grey) stability samples of A5 (left), A7 (center), and A8 (right).

[0265] Figure 19 Shown are the pharmacokinetic profiles and ADA analysis of A7 following intravenous (n=3) and subcutaneous (n=3) injection in cynomolgus monkeys.

[0266] Figure 20 Schematic representation of the structures of Morrison L constructs A14 and A15 is shown.

[0267] Figure 21 Shown are the average sizes of soluble complexes of scDb-scFvA7 determined by dynamic light scattering.

[0268] Figure 22 Shown are the average sizes of soluble complexes of A14 determined by dynamic light scattering.

[0269] Figure 23 Shown are the average sizes of soluble complexes of A15 determined by dynamic light scattering.

[0270] Figure 24 The protein concentration recovery of the soluble complex of scDb-scFv A7 is shown.

[0271] Figure 25 The protein concentration recovery of the soluble complex of A14 is shown.

[0272] Figure 26 The protein concentration recovery of the soluble complex of A15 is shown. DETAILED DESCRIPTION

[0273] The present disclosure is based on the discovery of multispecific antibody molecules that specifically bind to human IL-17A and TNFα and have improved affinity, efficacy and selectivity. In addition, the multispecific antibodies of the present disclosure have better safety because the inventors have demonstrated that the antibodies do not form immune complexes with TNFα and therefore have potential low immunogenicity. Due to the bivalent binding of other multispecific antibodies (such as Covagen, Abbvie, etc.), there is a high probability of forming immune complexes, which may lead to immunogenicity or other adverse reactions. In contrast, the monovalent bispecific and trispecific constructs of the present disclosure have a reduced probability of forming such complexes, thereby resulting in a lower probability of drug-resistant antibodies and immune-related adverse reactions. In addition, the multispecific antibodies of the present disclosure have better biophysical properties, such as developability and high-yield manufacturability, relatively few impurities, and excellent stability.

[0274] The present disclosure also provides antibodies that specifically bind to human IL-17A protein, as well as pharmaceutical compositions, methods of producing, and methods of using such antibodies and pharmaceutical compositions.

[0275] Unless defined otherwise, 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.

[0276] Unless otherwise indicated, the terms "including" and "comprising" are used herein in an open and non-limiting sense. With respect to these latter embodiments, the term "comprising" therefore encompasses the narrower term "consisting of."

[0277] Unless otherwise indicated herein or clearly contradicted by context, in the context of describing the present invention (particularly in the context of the following claims), the terms "a," "an," "the," and similar references should be construed to encompass both the singular and the plural. For example, the term "a cell" includes a plurality of cells, including mixtures thereof. When compounds, salts, etc. are used in the plural, this is also considered to refer to a single compound, salt, etc.

[0278] The multispecific antibodies of the present disclosure

[0279] In one aspect, the present disclosure provides an isolated multispecific antibody comprising a first domain that specifically binds IL-17A and a second domain that specifically binds TNFα.

[0280] The term "TNFα" or "TNF-α" or "tumor necrosis factor" refers specifically to human TNFα. TNFα exists as a soluble protein and as a precursor, which is called transmembrane TNFα, expressed as a type II polypeptide on the cell surface. Transmembrane TNFα is processed between residues Ala76 and Val77 by metalloproteinases such as TNFα converting enzyme (TACE), thereby releasing a soluble form of TNFα of 157 amino acid residues. Soluble TNFα is a homotrimer of a 17-kDa cleaved monomer. Transmembrane TNFα also exists as a homotrimer of a 26-kD uncleaved monomer. As used herein, the term "TNFα" includes both soluble and transmembrane forms. The term "TNFa" refers specifically to human transmembrane TNFα having UniProt ID No. P01375, reproduced herein as SEQ ID NO: 134. The term "TNFa" specifically refers to the soluble transmembrane TNFa having UniProt ID No. P01375, reproduced herein as SEQ ID NO: 135.

[0281] Suitably, the antibodies of the present disclosure are isolated antibodies. As used herein, the term "isolated antibody" refers to an antibody that is substantially free of other antibodies with different antigenic specificities (e.g., an isolated antibody that specifically binds only to IL-17A and TNFα is substantially free of antibodies that specifically bind to antigens other than IL-17A and TNFα). However, isolated antibodies that specifically bind to IL-17A and TNFα may have cross-reactivity with other antigens (e.g., IL-17A and TNFα molecules from other species). In addition, the isolated antibody may be substantially free of other cellular material and / or chemicals.

[0282] Suitably, the antibodies of the present disclosure are monoclonal antibodies. As used herein, the term "monoclonal antibody" or "monoclonal antibody composition" refers to antibodies that have substantially identical amino acid sequences or are derived from the same genetic source. A monoclonal antibody composition exhibits binding specificity and affinity for a particular epitope, or exhibits binding specificity and affinity for multiple specific epitopes.

[0283] Antibodies of the present disclosure include, but are not limited to, chimeric and humanized.

[0284] The term "chimeric antibody" is an antibody molecule in which (a) the constant region or a portion thereof is changed, replaced or exchanged so that the antigen binding site (variable region) is linked to a constant region of a different or changed class, effector function and / or type, or is linked to a completely different molecule that gives the chimeric antibody new properties, such as enzymes, toxins, hormones, growth factors, drugs, etc.; or (b) the variable region or a portion thereof is changed, replaced or exchanged for a variable region with a different or changed antigen specificity. For example, a mouse antibody can be modified by replacing its constant region with that of a human immunoglobulin. Due to the replacement of the human constant region, the chimeric antibody can retain its specificity for recognizing the antigen while having reduced human antigenicity compared to the original mouse antibody.

[0285] As used herein, a "humanized" antibody is an antibody that retains the reactivity of a non-human antibody while having low immunogenicity to humans. For example, this can be achieved by retaining the non-human CDRs and replacing the rest of the antibody with human counterparts (i.e., the framework portions of the constant and variable regions). Additional framework region modifications can be made within human framework sequences and CDR sequences derived from the germline of another mammalian species. The humanized antibodies of the present disclosure may comprise amino acid residues that are not encoded by human sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutations in vivo, or conservative substitutions to promote stability or preparation). See, for example, Morrison et al., Proc. Natl. Acad. Sci. USA, 81:6851-6855, 1984; Morrison and Oi, Adv. Immunol., 44:65-92, 1988; Verhoeyen et al., Science, 239:1534-1536, 1988; Padlan, Molec. Immun., 28:489-498, 1991; and Padlan, Molec. Immun., 31:169-217, 1994. Other examples of human engineering technology include, but are not limited to, the Xoma technology disclosed in U.S. Patent No. 5,766,886.

[0286] As used herein, the term "recombinant humanized antibody" includes all humanized antibodies of the present disclosure that are prepared, expressed, created, or isolated by recombinant means, such as antibodies isolated from animals (such as rabbits); antibodies expressed by transfection into host cells using recombinant expression vectors; antibodies isolated from a recombinant combinatorial human antibody library; or antibodies prepared, expressed, created, or isolated by any other means involving splicing human immunoglobulin gene sequences with other DNA sequences. Such recombinant human antibodies have variable and constant regions (if present) derived from human germline immunoglobulin sequences. However, such antibodies can be subjected to in vitro mutagenesis (or, when transgenic animals for human Ig sequences are used, to in vivo somatic mutagenesis), and therefore, the amino acid sequences of the VH (antibody heavy chain variable region) and VL (antibody light chain variable region) of the recombinant antibodies, although derived from and related to human germline VH and VL sequences, may not naturally occur within the human antibody germline.

[0287] Suitably, the antibodies or binding domains thereof of the present disclosure are humanized. Suitably, the antibodies or binding domains thereof of the present disclosure are humanized and comprise CDRs derived from rabbit.

[0288] As used herein, the term "multispecific antibody" refers to an antibody that binds to two or more different epitopes on at least two or more different targets (e.g., IL-17A and TNFα), or an antibody that binds to two or more different epitopes on the same target. The "multispecific antibody" of the present disclosure has two or more binding domains, such as two or three binding domains. The term "multispecific antibody" includes bispecific, trispecific, tetraspecific, pentaspecific and hexaspecific. As used herein, the term "bispecific antibody" refers to an antibody that binds to, for example, two different targets (e.g., IL-17A and TNFα) or two different epitopes on the same target. As used herein, the term "trispecific antibody" refers to an antibody that binds to, for example, three different targets (e.g., IL-17A, TNFα and HSA) or three different epitopes on the same target.

[0289] The term "multivalent antibody" refers to a single binding molecule with more than one valency, where "valency" is described as the number of antigen binding moieties that bind to an epitope on the same target molecule. "Valent" refers to the presence of a specific number of binding domains that are specific for one antigen in one molecule. Thus, the terms "monovalent", "bivalent", "tetravalent" and "hexavalent" refer to the presence of one, two, four and six binding domains that are specific for one antigen in one molecule, respectively. As used herein, the term "monovalent antibody" refers to an antibody with a single antigen binding portion that binds to a single epitope on a target molecule, such as IL-17A or TNFα. As used herein, the term "bivalent antibody" refers to an antibody with two antigen binding moieties, each of which binds to the same epitope.

[0290] The multispecific antibodies disclosed herein can be monovalent or multivalent for binding to IL-17A, such as bivalent, trivalent, or tetravalent, preferably monovalent.

[0291] The multispecific antibodies disclosed herein can be monovalent or multivalent for binding to TNFα, for example, bivalent, trivalent or tetravalent, preferably monovalent. Since TNFα forms a trimer, it may be trivalent and can form a three-dimensional immune complex in which the antibody has several domains that specifically bind to TNFα, for example, a bivalent, trivalent or multivalent antibody for binding to TNFα. To illustrate, at different antigen / antibody ratios, size studies of the immune complexes formed between TNF and infliximab (a chimeric TNFα IgG antibody) and etanercept (a TNFR2 dimer fusion protein with IgG1 Fc) showed that each antibody produced an immune complex with a unique size characteristic (Kim MS et al., J Mol Biol. 2007; 374: 1374–1388). Therefore, potential high immunogenicity is one of the concerns of therapeutic antibodies targeting TNFα. Therefore, in a preferred embodiment, the multispecific antibodies disclosed herein are monovalent for binding to TNFα.

[0292] Suitably, the multispecific antibodies of the present disclosure comprise only one domain that specifically binds to IL-17A and / or only one domain that specifically binds to TNFα. In a preferred embodiment, the multispecific antibodies of the present disclosure comprise only one domain that specifically binds to TNFα. Suitably, the multispecific antibodies of the present disclosure comprise only one domain that specifically binds to IL-17A and only one domain that specifically binds to TNFα. In a specific embodiment, the multispecific antibodies of the present disclosure consist of: a first domain that specifically binds to IL-17A, a second domain that specifically binds to TNFα, and optionally, a polypeptide linker between the two domains. In a specific embodiment, the optional polypeptide linker is present and consists of a polypeptide having 4 to 25 amino acid residues.

[0293] Suitably, the multispecific antibodies disclosed herein are advantageously capable of neutralizing the biological activity of human TNFα and human IL-17A. It will be understood that the term "neutralization," as used herein, refers to a reduction in biological signaling activity, which may be partial or complete. Suitable methods for determining neutralization are known in the art, and certain such assays are provided in the Examples herein.

[0294] In one embodiment, the antibodies or first domains thereof of the present disclosure selectively bind to human IL-17A relative to human IL-17B, IL-17C, IL-17D, IL-17E, and IL-17F, particularly as measured by ELISA. As used herein, the term "selectively binds" refers to an antibody, composition, formulation, etc. that does not significantly bind to IL-17B / C / D / E / F but does bind to IL-17A. Selective binding is characterized by high affinity (or low K D) and low to moderate IC 50 , as distinguished from nonspecific binding, which typically has low affinity (or high K D ) and moderate to high IC 50 Typically, when the antibody binds to the K D Less than 10 -7 Suitably, the antibodies or first domains thereof of the present disclosure bind to human IL-17A with a higher affinity or K than they bind to human IL-17B, IL-17C, IL-17D, IL-17E and IL-17F. D Suitably, the IC values ​​of the antibodies or first domains thereof for IL-17B, IL-17C, IL-17D, IL-17E and IL-17F are lower than those for IL-17B, IL-17C, IL-17D, IL-17E and IL-17F. 50 The IC values ​​of IL-17A were compared 50 At least 100 times higher, e.g., at least 200 times higher, at least 300 times higher, at least 400 times higher, as measured by ELISA. In one embodiment, the antibody of the present disclosure or its first domain binds to human IL-17A but not to human IL-17B, IL-17C, IL-17D, IL-17E and IL-17F, in particular as measured by SPR and / or ELISA.

[0295] In another embodiment, the multispecific antibodies of the present disclosure further comprise a third domain having specificity for an antigen different from IL-17A and TNFα. Suitably, the multispecific antibodies of the present disclosure are trispecific antibodies. As used herein, "trispecific antibody" refers to an antibody molecule having three antigen-binding domains, for example, one binding domain that binds to human TNFα, another binding domain that binds to human IL-17A, and another binding domain that binds to an antigen that can extend the half-life of the antibody molecule, such as human serum albumin.

[0296] In particular, the multispecific antibodies of the present disclosure further comprise a third domain that specifically binds to human serum albumin (HSA).

[0297] The present inventors unexpectedly discovered that adding a third domain that specifically binds to human serum albumin to the multispecific antibody of the present disclosure comprising a first domain that specifically binds to IL-17A and a second domain that specifically binds to TNFα has the following beneficial effects:

[0298] (i) the serum half-life of a multispecific antibody of the present disclosure comprising at least one domain that specifically binds to human serum albumin is increased; and

[0299] (ii) The addition of a human serum albumin binding domain to the multispecific antibodies of the present disclosure is compatible with the functions of other binding domains, such as the neutralizing activity of the Il-17A and TNFα binding domains.

[0300] The term "HSA" specifically refers to human serum albumin with UniProt ID number P02768. Human serum albumin (HSA) is an abundant protein of 66.4 kDa in human serum (accounting for 50% of total protein) composed of 585 amino acids (Sugio, Protein Eng, Vol. 12, 1999, 439-446). The multifunctional HSA protein is associated with its structure, which allows it to bind and transport many metabolites, such as fatty acids, metal ions, bilirubin, and certain drugs (Fanali, Molecular Aspects of Medicine, Vol. 33, 2012, 209-290). The concentration of HSA in serum is approximately 3.5-5 g / dL. Albumin-binding antibodies can be used, for example, to extend the in vivo serum half-life of drugs or proteins conjugated thereto.

[0301] Suitably, in one embodiment, the multispecific antibodies of the present disclosure comprise a first domain that specifically binds to IL-17A, a second domain that specifically binds to TNFα, and a third domain that specifically binds to human serum albumin. The multispecific antibodies of the present disclosure may be monovalent or multivalent, for example, bivalent, trivalent, or tetravalent, with monovalent being preferred, for binding to human serum albumin. Suitably, the multispecific antibodies of the present disclosure comprise only one domain that specifically binds to IL-17A, only one domain that specifically binds to TNFα, and only one domain that specifically binds to human serum albumin. In a specific embodiment, the multispecific antibodies of the present disclosure consist of: a first domain that specifically binds to IL-17A, a second domain that specifically binds to TNFα, a third domain that specifically binds to human serum albumin, and optionally, a polypeptide linker between the two domains. In a specific embodiment, the optional polypeptide linker is present and consists of a polypeptide having 4 to 25 amino acid residues.

[0302] Advantageously, the multispecific antibodies of the present disclosure comprise a first domain that specifically binds to IL-17A and a second domain that specifically binds to TNFα, and optionally, a third domain that specifically binds to human serum albumin, wherein the domains are capable of simultaneously binding to their respective antigens or receptors.

[0303] The domains of the multispecific antibodies of the present disclosure, e.g., the first domain, the second domain, the third domain are independently selected from the group consisting of: Fab, Fv, scFv, dsFv, scAb, STAB, single domain antibodies (sdAb or dAb), single domain heavy chain antibodies, and single domain light chain antibodies, VHH, VNAR, single domain antibodies based on shark VNAR structures, and binding domains based on alternative scaffolds (including but not limited to ankyrin-based domains, fynomers, avimers, anticalins, fibronectin, and binding sites constructed in antibody constant regions (e.g., Modular Antibody Technology of F-star) TM )), preferably, is selected from the group consisting of: Fab, Fv and scFv, more preferably, wherein the first domain and / or the second domain and / or the third domain is Fv or scFv.

[0304] The multispecific antibodies of the present disclosure may be in any suitable format. In one embodiment, the format of the multispecific antibodies of the present disclosure is selected from the group consisting of single-chain diabodies (scDb), tandem scDb (Tandab), linear dimer scDb (LD-scDb), cyclic dimer scDb (CD-scDb), bispecific T cell engagers (BiTE; tandem bi-scFv), tandem tri-scFv, triabody (Fab-(scFv)2) or diabody (Fab-(scFv)1), Fab, Fab-Fv2, Morrison (IgG CH3-scFv fusion (Morrison L) or IgG CL-scFv fusion (Morrison H)), triabodies, scDb-scFv, bispecific Fab2, diabodies, tetrabodies, scFv-Fc-scFv fusions, scFv-HSA-scFv fusions, diabodies, DVD-Ig, COVD, IgG-scFab, scFab-dsscFv, Fv2-Fc, IgG-scFv fusions such as bsAb (scFv linked to the C-terminus of the light chain), Bs1Ab (scFv linked to the N-terminus of the light chain), Bs2Ab (scFv linked to the N-terminus of the heavy chain), Bs3Ab (scFv linked to the C-terminus of the heavy chain), Ts1Ab (scFv linked to the C-terminus of the heavy chain), The invention also provides the present invention relates to bispecific antibodies based on heterodimeric Fc domains, such as Knob-into-Hole antibodies (KiH); Fv, scFv, scDb, tandem bi-scFv, tandem tri-scFv, Fab-(scFv)2, Fab-(scFv)1, Fab, Fab-Fv2, COVD, MATCH and DuoBody fused to the N- and / or C-terminus of any chain of a heterodimeric Fc domain or any other heterodimeric domain, preferably a triabody or scDb-scFv.

[0305] The term "diabody" refers to an antibody fragment with two antigen-binding sites, which comprises a VH (VH-VL) connected to a VL in the same polypeptide chain. By using a linker that is too short to allow pairing between the two domains on the same chain, these domains are forced to pair with the complementary domains of another chain to produce two antigen-binding sites. Diabodies can be bivalent or bispecific. Diabodies are more fully described in, for example, EP404097, WO 93 / 01161, Hudson et al., Nat. Med. 9: 129-134 (2003) and Holliger et al., Proc. Natl. Acad. Sci. USA 90: 6444-6448 (1993). Tribodies and tetrabodies are also described in Hudson et al., Nat. Med. 9: 129-134 (2003). The bispecific scDb, in particular the bispecific monomeric scDb, in particular comprises two variable heavy chain domains (VH) or fragments thereof and two variable light chain domains (VL) or fragments thereof, which are connected by linkers L1, L2 and L3 in the following order: VHA-L1-VLB-L2-VHB-L3-VLA, VHA-L1-VHB-L2-VLB-L3-VLA, VLA-L1-VLB-L2-VHB-L3-VHA, VLA-L1-V HB-L2-VLB-L3-VHA, VHB-L1-VLA-L2-VHA-L3-VLB, VHB-L1-VHA-L2-VLA-L3-VLB, VLB-L1-VLA-L2-VHA-L3-VHB, or VLB-L1-VHA-L2-VLA-L3-VHB, wherein the VLA and VHA domains together form an antigen-binding site for a first antigen, and VLB and VHB together form an antigen-binding site for a second antigen. Linker L1 is particularly a peptide of 2-10 amino acids, more particularly 3-7 amino acids, and most particularly 5 amino acids, and linker L3 is particularly a peptide of 1-10 amino acids, more particularly 2-7 amino acids, and most particularly 5 amino acids. The intermediate linker L2 is particularly a peptide of 10-40 amino acids, more particularly 15-30 amino acids, and most particularly 20-25 amino acids.

[0306] In one embodiment, the multispecific antibodies of the present disclosure comprise an immunoglobulin Fc region polypeptide. The term "Fc region" herein is used to define the C-terminal region of an immunoglobulin heavy chain, including native sequence Fc regions and variant Fc regions. Suitable native sequence Fc regions include human IgG1, IgG2 (IgG2A, IgG2B), IgG3, and IgG4. "Fc receptor" or "FcR" describes a receptor that binds to the Fc region of an antibody. Preferred FcRs are native sequence human FcRs. In addition, preferred FcRs are FcRs (gamma receptors) that bind to IgG antibodies, and include receptors of the FcγRI, FcγRII, and FcγRIII subclasses, including allelic variants and splice forms of these receptors. FcγRII receptors include FcγRIIA ("activating receptor") and FcγRIIB ("inhibitory receptor"), which have similar amino acid sequences, differing primarily in their cytoplasmic domains. The activating receptor FcγRIIA contains an immunoreceptor tyrosine-based activation motif (ITAM) in its cytoplasmic domain. The inhibitory receptor FcγRIIB contains an immunoreceptor tyrosine-based inhibition motif (ITIM) in its cytoplasmic domain (see M. Daeron, Annu. Rev. Immunol. 5:203-234 (1997)). For an overview of FcRs, see Ravetch and Kinet, Annu. Rev. Immunol. 9:457-92 (1991); Capet et al., Immunomethods 4:25-34 (1994); and de Haas et al., J. Lab. Clin. Med. 126:330-41 (1995). The term "FcR" as used herein includes other FcRs, including those to be identified in the future. The term "Fc receptor" or "FcR" also includes the neonatal receptor FcRn, which is responsible for the transfer of maternal IgG to the fetus. Guyer et al., J. Immunol. 117: 587 (1976) and Kim et al., J. Immunol. 24: 249 (1994). Methods for measuring binding to FcRn are known (see, for example, Ghetie and Ward, Immunol. Today 18: (12): 592-8 (1997); Ghetie et al., Nature Biotechnology 15 (7): 637-40 (1997); Hinton et al., J. Biol. Chem. TJI (8): 6213-6 (2004); WO 2004 / 92219 (Hinton et al.)).In vivo binding to FcRn and the serum half-life of human FcRn high-affinity binding polypeptides can be determined, for example, in transgenic mice expressing human FcRn or transfected human cell lines, or in primates administered polypeptides having variant Fc regions. WO 2004 / 42072 (Presta) describes antibody variants that have improved or reduced FcR binding. See also, for example, Shields et al., J. Biol. Chem. 9(2):6591-6604 (2001).

[0307] In order to increase the amount of specificity / functionality at the same or lower molecular weight, it is advantageous to use antibodies comprising antibody fragments, such as Fv, Fab, Fab' and F(ab')2 fragments and other antibody fragments. These smaller molecules retain the antigen binding activity of intact antibodies and also exhibit improved tissue penetration and pharmacokinetic properties compared to intact immunoglobulin molecules. Although such fragments appear to show many advantages over intact immunoglobulins, they are also plagued by high serum clearance rates because they lack the Fc domain, which has a long half-life in vivo (Medasan et al., 1997, J. Immunol. 158: 2211-2217). Molecules with lower molecular weights can more effectively penetrate target tissues (e.g., solid cancers) and therefore have the potential to improve efficacy at the same or lower doses. Suitably, the antibodies of the present disclosure do not comprise an immunoglobulin Fc region polypeptide and, optionally, do not include a CH1 and CL region, particularly when the multispecific antibody comprises a third domain that specifically binds to human serum albumin.

[0308] Suitably, the antibodies of the present disclosure may be in trimeric form (Fab-(scFv)2). Suitably, the first domain and / or the second domain and / or the third domain are Fab or scFv domains. In particular, the multispecific antibodies of the present disclosure have one Fab domain and two scFv domains, in particular wherein the scFv domain is fused to the carboxyl terminus of each chain of the Fab domain. The inventors tested the optimal relative positions of the various binding domains in the trispecific format in terms of pharmacodynamics and biophysical properties and surprisingly found that, in a trimeric form, the multispecific antibodies of the present disclosure have advantageous properties when the second domain that specifically binds to TNFα is a Fab domain, and the first and third domains that specifically bind to IL-17A and HSA, respectively, are scFv domains fused to the Fab domains.

[0309] Preferably, the antibodies of the present disclosure are in the form of scDb-scFv. The term "scDb-scFv" refers to an antibody format in which a single-chain Fv (scFv) fragment is fused to a single-chain diabody (scDb) via a flexible Gly-Ser linker. Suitably, the first domain and / or the second domain and / or the third domain are Fv or scFv domains. In particular, the multispecific antibodies of the present disclosure, when in the form of scDb-scFv, have one scFv domain fused at the C-terminus to an scDb consisting of two other domains. The multispecific antibodies of the present disclosure, when in the form of scDb-Fv, can be represented by the following formula:

[0310] VLA-L1-VHC-L2-VLC-L3-VHA-L4-VLB-L5–VHB or

[0311] VLB-L1-VHA-L2-VLA-L3-VHB-L4-VLC-L5–VHC or

[0312] VLC-L1-VHB-L2-VLB-L3-VHC-L4-VLA-L5–VHA or

[0313] VLA-L1-VHB-L2-VLB-L3-VHA-L4-VLC-L5–VHC,

[0314] Preferred VLB-L1-VHA-L2-VLA-L3-VHB-L4-VLC-L5–VHC or VLA-

[0315] L1-VHB-L2-VLB-L3-VHA-L4-VLC-L5–VHC,

[0316] More preferably VLB-L1-VHA-L2-VLA-L3-VHB-L4-VLC-L5–VHC,

[0317] Among them, VLA and VHA are the light chain variable region and heavy chain variable region of the first domain (specifically binding to IL-17A), respectively; VLB and VHB are the light chain variable region and heavy chain variable region of the second domain (specifically binding to TNFα), respectively; VLC and VHC are the light chain variable region and heavy chain variable region of the third domain (specifically binding to HSA), respectively, among which L1, L2, L3, L4 and L5 are polypeptide linkers.

[0318] In the context of the present disclosure, the term " polypeptide linker " refers to a joint consisting of an amino acid residue chain connected by a peptide bond connecting two domains, and each domain is connected to one end of a joint. The length of a polypeptide linker should be enough to connect two molecules so that they have correct conformation each other, thereby maintaining required activity. In a specific embodiment, a polypeptide linker has a continuous chain of 2 to 30 amino acid residues (such as 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 amino acid residues). In addition, the amino acid residue selected to be included in the polypeptide linker should show a characteristic that will not significantly interfere with polypeptide activity. Therefore, the linker peptide should not show an inconsistency with polypeptide activity as a whole, or interfere with internal folding, or form a key with the amino acid residues in one or more monomers or will seriously hinder other interacting charges of the combination of monomeric domains. In a specific embodiment, a polypeptide linker is an unstructured polypeptide. Useful linkers include glycine-serine or GS linkers. As will be appreciated by those skilled in the art, "Gly-Ser" or "GS" linkers refer to polymers of glycine and serine in series (including, for example, (Gly-Ser)n, (GSGGS)n, (GGGGS)n, and (GGGS)n, wherein n is an integer of at least one), glycine-alanine polymers, alanine-serine polymers, and other flexible linkers, such as tethers for shaker potassium channels, and a variety of other flexible peptide linkers. Glycine-serine polymers are preferred because these two amino acids are relatively unstructured and can therefore be used as neutral tethers between components. Secondly, serine is hydrophilic and can therefore dissolve substances that may be globular glycine chains. Thirdly, similar chains have been shown to effectively connect the subunits of recombinant proteins (such as single-chain antibodies).

[0319] Suitably, said L1 and L3 of the present disclosure are as shown in SEQ ID NO: 132. Suitably, said L2, L4 and L5 of the present disclosure are as shown in SEQ ID NO: 23.

[0320] The present inventors tested the optimal relative positions of the various binding domains in a trispecific format in terms of pharmacodynamics and biophysical properties and surprisingly found that, in the scDb-scFv format, when the first domain and the second domain that specifically bind to IL-17A and TNFα, respectively, form an scDb, and the third domain that specifically binds to HSA is an scFv, the multispecific antibody of the present disclosure has favorable properties.

[0321] The multispecific antibodies of the present disclosure have the following advantageous properties:

[0322] (a) having an ability to neutralize IL-17A with a potency relative to secukinumab (relative potency) of greater than 2, e.g., greater than 5, greater than 10, greater than 15, greater than 20, greater than 25, greater than 30, greater than 35, greater than 40, greater than 45, preferably greater than 50, as determined by measuring Gro-α secretion in the HT-29 assay, and wherein the relative potency is the IC of secukinumab in ng / mL as measured in the HT-29 assay 50 The values ​​are comparable to the IC values ​​in ng / mL of the multispecific antibodies measured in the HT-29 assay. 50 The ratio of the values; and

[0323] (b) has the ability to neutralize TNFα with a potency (relative potency) of at least 1, e.g., greater than 1, greater than 1.5, greater than 2, greater than 2.5, greater than 3, greater than 3.5, preferably greater than 4, more preferably greater than 4.5 relative to the scDb according to SEQ ID NO: 149 (A13) as determined by measuring Gro-α secretion in the HT-29 assay, and wherein the relative potency is the IC in nM of the scDb according to SEQ ID NO: 149 measured in the HT-29 assay 50 The values ​​are comparable to the IC values ​​in nM of the multispecific antibodies measured in the HT-29 assay. 50 The ratio of the values; and

[0324] (c) optionally, having the ability to block the interaction between IL-17A and IL-17RA, with a potency relative to secukinumab (relative potency) as determined in an ELISA assay of greater than 2, e.g., greater than 3, greater than 4, greater than 5, greater than 6, greater than 7, greater than 8, greater than 9, preferably greater than 10, and wherein the relative potency is the IC of secukinumab in ng / mL as measured by ELISA 50 The values ​​were compared with the IC values ​​of the multispecific antibodies in ng / mL measured by ELISA. 50 The ratio of the values; and

[0325] (d) optionally, has the ability to neutralize TNFα with a potency (relative potency) of at least 0.4, e.g. at least 0.5, preferably at least 1 relative to the scDb according to SEQ ID NO: 149 (A13) as determined in the L929 assay, and wherein said relative potency is the IC in nM of said scDb according to SEQ ID NO: 149 measured in the L929 assay 50 The values ​​are comparable to the IC values ​​in nM for the multispecific antibodies measured in the L929 assay. 50 and / or

[0326] (e) Dissociation constant (K) for binding to human IL-17AD ) is less than 5 nM, e.g., less than 4 nM, less than 3 nM, less than 2 nM, less than 1 nM, preferably less than 0.5 nM, as measured by surface plasmon resonance; and optionally, a K for binding to cynomolgus monkey IL-17A D Less than 5 nM, e.g., less than 4 nM, less than 3 nM, less than 2 nM, less than 1 nM, preferably less than 0.5 nM, as measured by surface plasmon resonance;

[0327] (f) Dissociation constant (K) for binding to human TNFα D ) is less than 5 nM, e.g., less than 4 nM, less than 3 nM, less than 2 nM, less than 1 nM, preferably less than 0.5 nM, more preferably less than 0.25 nM, as measured by surface plasmon resonance; and

[0328] (g) Optionally, the dissociation constant (K) for binding to human serum albumin D ) is less than 5 nM, e.g., less than 4 nM, less than 3 nM, preferably less than 2 nM, as measured by surface plasmon resonance; and optionally, a dissociation constant (K) for binding to cynomolgus monkey serum albumin D ) is less than 5 nM, e.g., less than 4 nM, less than 3 nM, preferably less than 2 nM, as measured by surface plasmon resonance.

[0329] As used herein, the term "affinity" refers to the strength of the interaction between an antibody and an antigen at a single antigenic site. Within each antigenic site, the variable regions of the antibody "arms" interact with the antigen at many sites through weak non-covalent forces; the greater the interaction, the stronger the affinity.

[0330] "Binding affinity" generally refers to the strength of the sum of non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless otherwise indicated, as used herein, "binding affinity" or "binding to" refers to intrinsic binding affinity that reflects a 1:1 interaction between members of a binding pair (e.g., an antibody and an antigen). The affinity of a molecule X for its partner Y can generally be expressed in terms of the dissociation constant (K D ) represents. Affinity can be measured by conventional methods known in the art, including the methods described herein. Low-affinity antibodies generally bind antigen slowly and tend to dissociate easily, while high-affinity antibodies generally bind antigen faster and tend to remain bound longer. Various methods for measuring binding affinity are known in the art, any of which can be used for the purposes of the present disclosure. Specific illustrative and exemplary embodiments for measuring binding affinity, i.e., binding strength, are described below.

[0331] The term "K assoc ”, “Ka” or “Kon ” refers to the on-rate of a particular antibody-antigen interaction, and the term “K dis ”, “Kd” or “K off " refers to the off-rate of a particular antibody-antigen interaction. In one embodiment, the term "K" as used herein D " refers to the dissociation constant, which is obtained from the ratio of Kd to Ka (i.e., Kd / Ka) and is expressed as a molar concentration (M). In one embodiment, the "Kd" according to the present disclosure is D ” or “K D The binding affinity of an antibody can be determined, for example, by the dissociation constant (KD) or KD value, by using a surface plasmon resonance assay using a MASS-1 SPR instrument (Sierra Sensors), as described in the Examples. D ) OK. Lower K D indicates stronger affinity, while higher K D Indicates weaker affinity.

[0332] Thus, in suitable embodiments, the antibodies of the present disclosure bind to human IL-17A with a dissociation constant (K D ) between 1 pM and 10 nM, between 1 pM and 7 nM, between 1 pM and 5 nM, between 1 pM and 4 nM, between 1 pM and 3 nM, between 1 pM and 2.5 nM, between 1 pM and 2 nM, between 1 pM and 1.5 nM, between 1 pM and 1 nM, preferably between 1 pM and 0.5 nM, as measured by surface plasmon resonance. In suitable embodiments, the antibodies of the present disclosure bind to human IL-17A with a dissociation constant (K D ) is between 1 and 500 pM, as measured by surface plasmon resonance. In suitable embodiments, the antibodies of the present disclosure bind to human IL-17A with a dissociation constant (K D ) is less than 5 nM, less than 4 nM, less than 3 nM, less than 2 nM, less than 1 nM, preferably less than 0.5 nM, as measured by surface plasmon resonance. Suitably, the antibodies of the present disclosure have a dissociation constant (K) of less than 1 nM. D ) binds to human IL-17A. Suitably, the antibodies of the present disclosure bind to human IL-17A with a dissociation constant (K) of less than 0.5 nM. D ) binds to human IL-17A. In another embodiment, the antibodies of the present disclosure bind to cynomolgus monkey IL-17A, K D Less than 10 nM, e.g., less than 7 nM, less than 5 nM, less than 4 nM, less than 3 nM, less than 2 nM, less than 1 nM, preferably less than 0.5 nM, as measured by surface plasmon resonance (SPR).

[0333] Suitably, the antibodies of the present disclosure bind to human TNFα with a dissociation constant (K D ) between 1 pM and 10 nM, between 1 pM and 7 nM, between 1 pM and 5 nM, between 1 pM and 4 nM, between 1 pM and 3 nM, between 1 pM and 2.5 nM, between 1 pM and 2 nM, between 1 pM and 1.5 nM, preferably between 1 pM and 1 nM, preferably between 1 pM and 0.5 nM, more preferably between 1 pM and 0.25 nM, as measured by surface plasmon resonance. In suitable embodiments, the dissociation constant (K) of an antibody of the present disclosure for binding to human TNFα is D ) is between 1 and 500 pM, preferably between 1 and 250 pM, as measured by surface plasmon resonance. In suitable embodiments, the antibodies of the present disclosure bind to human TNFα with a dissociation constant (K D ) is less than 5 nM, less than 4 nM, less than 3 nM, less than 2 nM, less than 1 nM, preferably less than 0.5 nM, more preferably less than 0.25 nM, as measured by surface plasmon resonance. Suitably, the antibodies of the present disclosure have a dissociation constant (K) of less than 0.5 nM. D ) binds to human TNFα. Suitably, the antibodies of the present disclosure bind to human TNFα with a dissociation constant (K) of less than 0.25 nM. D ) binds to human TNFα.

[0334] In suitable embodiments, the dissociation constant (K) of an antibody of the present disclosure for binding to human serum albumin is D ) between 1 pM and 10 nM, between 1 pM and 7 nM, between 1 pM and 5 nM, between 1 pM and 4 nM, between 1 pM and 3 nM, preferably between 1 pM and 2 nM, as measured by surface plasmon resonance. In suitable embodiments, the antibodies of the present disclosure bind to human serum albumin with a dissociation constant (K D ) is between 1 and 2000 pM, as measured by surface plasmon resonance. In suitable embodiments, the antibodies of the present disclosure bind to human serum albumin with a dissociation constant (K D ) is less than 5 nM, less than 4 nM, less than 3 nM, preferably less than 2 nM, as measured by surface plasmon resonance. Suitably, the antibodies of the present disclosure have a dissociation constant (K) of less than 2 nM. D ) binds to human serum albumin. In another embodiment, the antibodies of the present disclosure bind to the K D Less than 10 nM, e.g., less than 7 nM, less than 5 nM, less than 4 nM, less than 3 nM, preferably less than 2 nM, as measured by surface plasmon resonance (SPR).

[0335] Suitably, the antibodies of the present disclosure have advantageous biophysical characteristics.

[0336] (a) a melting temperature (Tm) of at least 55°C, preferably at least 58°C, more preferably at least 60°C, as measured by differential scanning fluorimetry in phosphate-citrate buffer, pH 6.4, 150 mM NaCl;

[0337] (b) when the multispecific antibody is at a starting concentration of 10 mg / ml in phosphate buffered saline (PBS) at pH 7.4, after five consecutive freeze-thaw cycles, the monomer content loss is less than 5%, e.g., less than 4%, less than 3%, less than 2%, preferably 1% or less;

[0338] (c) when the multispecific antibody is stored at an initial concentration of 10 mg / ml in phosphate buffered saline (PBS) at pH 7.4 at 4°C for at least two weeks, in particular at least four weeks, the monomer content loss is less than 10%, preferably less than 5%; and / or

[0339] (d) when the multispecific antibody is stored at an initial concentration of 10 mg / ml in phosphate buffered saline (PBS) at pH 7.4 at 37°C for at least two weeks, particularly at least four weeks, the monomer content loss is less than 20%, preferably less than 15%.

[0340] Melting (Tm) was determined by differential scanning fluorimetry (DSF) as previously described (Egan et al., MAbs, 9(1)(2017), 68-84; Niesen et al., Nature Protocols, 2(9)(2007) 2212-2221). The midpoint of the transition to thermal unfolding was determined by differential scanning fluorimetry (DSF) using a fluorescent dye Orange determined (see Wong & Raleigh, Protein Science 25 (2016) 1834-1840). Samples were prepared in citrate phosphate buffer at pH 6.4 at a final protein concentration of 50 μg / mL and contained 5x The final concentration of Orange was 100 μl in a total volume of 100 μl. 25 microliters of the prepared sample was added in triplicate to a white-walled AB gene PCR plate. The analysis was performed in a qPCR machine used as a thermal cycler, and fluorescence emission was detected using the software's custom dye calibration program. The PCR plate containing the test samples was temperature-changed from 25°C to 96°C in increments of 1°C, with a 30-second pause after each temperature increase. The total assay time was approximately two hours. Tm was calculated by calculating the inflection point of the curve using the mathematical second derivative method using GraphPad Prism software. The reported Tm is the average of three measurements.

[0341] The loss of monomer content was determined by SE-HPLC. SE-HPLC is a separation technique based on a solid stationary phase and a liquid mobile phase, as described in USP Chapter 621. The method utilizes a hydrophobic stationary phase and an aqueous mobile phase to separate molecules based on their size and shape. The separation of molecules occurs between the void volume (V0) and the total permeate volume (VT) of a specific column. SE-HPLC measurements were performed on a Chromaster HPLC system (Hitachi High-Technologies Corporation), which was equipped with automatic sample injection and a UV detector with a detection wavelength set to 280 nm. The equipment was controlled by the software EZChrom Elite (Agilent Technologies, Version 3.3.2SP2), which also supported the analysis of the resulting chromatograms. The protein sample was cleared by centrifugation before injection, and the temperature of the autosampler was maintained at 6°C. For analysis of scFv samples, a Shodex KW403-4F column (Showa Denko Co., Ltd., #F6989202) was used with a standard buffered saline mobile phase (50 mM sodium phosphate pH 6.5, 300 mM sodium chloride) at a recommended flow rate of 0.35 mL / min. The target sample load per injection was 5 μg. Samples were detected by UV detection at a wavelength of 280 nm, and data were recorded using a suitable software suite. The resulting chromatograms were analyzed in the range of V0 to Vt, excluding matrix-related peaks with elution times >10 minutes.

[0342] Exemplary domains that specifically bind to IL-17A

[0343] The multispecific antibodies of the present disclosure comprise a first domain that specifically binds to IL-17A, wherein the domain comprises a heavy chain variable region (VH) and a light chain variable region (VL), and wherein: (a) the VH comprises three complementarity determining regions HCDR1, HCDR2, and HCDR3 in sequence, and (b) the VL comprises three complementarity determining regions LCDR1, LCDR2, and LCDR3 in sequence.

[0344] Suitable domains that specifically bind to IL-17A for use in the multispecific antibodies of the present disclosure include, but are not limited to:

[0345] The humanized monoclonal antibodies or their binding domains described below in the section “Anti-IL-17a Antibodies of the Disclosure,” whose sequences are listed in Table 1;

[0346] AIN457 (also known as secukinumab; disclosed in U.S. Pat. No. 7,807,155 and WO 2006 / 013107, the entire contents of which are incorporated herein by reference) or an antigen-binding fragment thereof;

[0347] LY2439821 (also known as ixekizumab; disclosed in U.S. Pat. Nos. 7,838,638 and 8,110,191 and WO 2007 / 070750, the entire contents of which are incorporated herein by reference) or an antigen-binding fragment thereof;

[0348] SCH900117 or its antigen-binding fragment (Merck);

[0349] RG4943 or its antigen-binding fragment (Roche);

[0350] ·WO 2006 / 013107,WO 2006 / 054059,WO 2007 / 070750,WO 2007 / 149032,WO2008 / 001063,WO 2008 / 021156,WO 2010 / 034443,WO 2010 / 102251,WO 2012 / 018767,WO2014 / 161570,WO 2014 / 001368,WO 2014 / 122613,WO 2015 / 070697,WO 2015 / 137843,WO2016 / 048188,WO 2016 / 113557,WO 2016 / 138842,WO The anti-IL-17A antibodies or antigen-binding fragments thereof disclosed in 2017 / 068472, the entire contents of which are incorporated herein by reference.

[0351] Preferred domains that specifically bind to IL-17A for use in the multispecific antibodies of the present disclosure include, but are not limited to, the humanized monoclonal antibodies or their binding domains presented below, the sequences of which are listed in Table 1.

[0352] Thus, in one embodiment, the present disclosure provides a multispecific antibody comprising a first domain that specifically binds to IL-17A and a second domain that specifically binds to TNFα, wherein the first domain comprises a set of CDRs: HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3, wherein the set of CDRs has 10 or fewer amino acid substitutions, e.g., 9 or fewer amino acid substitutions, 8 or fewer amino acid substitutions, 7 or fewer amino acid substitutions, 6 or fewer amino acid substitutions, 5 or fewer amino acid substitutions, 4 or fewer amino acid substitutions, 3 or fewer amino acid substitutions, 2 or fewer amino acid substitutions, 1 or 0 amino acid substitutions, preferably 0 amino acid substitutions, relative to a set of CDRs: (i) HCDR1′ is an amino acid sequence selected from any one of SEQ ID NOs: 1, 4, and 7, preferably SEQ ID NO: 1; HCDR2′ is an amino acid sequence selected from any one of SEQ ID NOs: 2, 5, and 8, preferably SEQ ID NO: 2; HCDR3′ is an amino acid sequence selected from any one of SEQ ID NOs: 2, 5, and 8, preferably SEQ ID NO: 2; NO: 3, 6 and 9, preferably SEQ ID NO: 3; LCDR1 'is an amino acid sequence selected from any one of SEQ ID NOs: 12, 15 and 18, preferably SEQ ID NO: 12; LCDR2' is an amino acid sequence selected from any one of SEQ ID NOs: 13, 16 and 19, preferably SEQ ID NO: 13; and LCDR3' has an amino acid sequence selected from any one of SEQ ID NOs: 14, 17 and 20, preferably SEQ ID NO: 14; or (ii) wherein HCDR1 'is an amino acid sequence selected from any one of SEQ ID NOs: 39, 42 and 45, preferably SEQ ID NO: 39; HCDR2' is an amino acid sequence selected from any one of SEQ ID NOs: 40, 43 and 46, preferably SEQ ID NO: 40; HCDR3' is an amino acid sequence selected from any one of SEQ ID NOs: 41, 44 and 47, preferably SEQ ID NO: 41; LCDR1 'is an amino acid sequence selected from SEQ ID NOs: 39, 42 and 45, preferably SEQ ID NO: 39; NO:50, 53 and 56, preferably SEQ ID NO:50; LCDR2' is an amino acid sequence selected from any one of SEQ ID NO:51, 54 and 57, preferably SEQ ID NO:51; and LCDR3' has an amino acid sequence selected from any one of SEQ ID NO:52, 55 and 58, preferably SEQ ID NO:52.

[0353] In particular, the present disclosure provides a multispecific antibody comprising a first domain that specifically binds to IL-17A and a second domain that specifically binds to TNFα, wherein the first domain comprises a VH CDR having the amino acid sequence of any one of the VH CDRs listed in Table 1. In particular, the domain comprises (or alternatively consists of) one, two, three or more VH CDRs having the amino acid sequence of any one of the VH CDRs listed in Table 1.

[0354] Suitably, the first domain that specifically binds to IL-17A comprises a heavy chain variable region (VH), wherein the VH comprises, in order: (i) three complementarity determining regions HCDR1, HCDR2 and HCDR3, wherein the HCDR1 has an amino acid sequence selected from any one of SEQ ID NOs: 1, 4 and 7, the HCDR2 has an amino acid sequence selected from any one of SEQ ID NOs: 2, 5 and 8, and the HCDR3 has an amino acid sequence selected from any one of SEQ ID NOs: 3, 6 and 9. In particular, the present disclosure provides antibodies that have binding specificity to human IL-17A and comprise the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs: 1, 2, and 3, respectively; or (ii) three complementarity determining regions HCDR1, HCDR2, and HCDR3, wherein the HCDR1 has an amino acid sequence selected from any one of SEQ ID NOs: 39, 42, and 45, the HCDR2 has an amino acid sequence selected from any one of SEQ ID NOs: 40, 43, and 46, and the HCDR3 has an amino acid sequence selected from any one of SEQ ID NOs: 41, 44, and 47. In particular, the present disclosure provides antibodies that have binding specificity to human IL-17A and comprise the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs: 39, 40, and 41, respectively.

[0355] The present disclosure also provides a multispecific antibody comprising a first domain that specifically binds to IL-17A and a second domain that specifically binds to TNFα, wherein the first domain comprises a VL CDR having the amino acid sequence of any one of the VL CDRs listed in Table 1. In particular, the first domain that specifically binds to IL-17A comprises (or alternatively consists of) one, two, three or more VL CDRs having the amino acid sequence of any of the VL CDRs listed in Table 1.

[0356] Suitably, the first domain that specifically binds to IL-17A comprises a light chain variable region (VL), wherein the VL comprises, in sequence: (i) three complementarity determining regions LCDR1, LCDR2 and LCDR3, wherein the LCDR1 has an amino acid sequence selected from any one of SEQ ID NOs: 12, 15 and 18, the LCDR2 has an amino acid sequence selected from any one of SEQ ID NOs: 13, 16 and 19, and the LCDR3 has an amino acid sequence selected from any one of SEQ ID NOs: 14, 17 and 20. In particular, the present disclosure provides antibodies that have binding specificity to human IL-17A and comprise the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs: 12, 13, and 14, respectively; or (ii) three complementarity determining regions, LCDR1, LCDR2, and LCDR3, wherein LCDR1 has an amino acid sequence selected from any one of SEQ ID NOs: 50, 53, and 56, LCDR2 has an amino acid sequence selected from any one of SEQ ID NOs: 51, 54, and 57, and LCDR3 has an amino acid sequence selected from any one of SEQ ID NOs: 52, 55, and 58. In particular, the present disclosure provides antibodies that have binding specificity to human IL-17A and comprise the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs: 50, 51, and 52, respectively.

[0357] Suitably, the present disclosure provides a multispecific antibody comprising a first domain that specifically binds to IL-17A and a second domain that specifically binds to TNFα, wherein the first domain comprises a heavy chain variable region (VH) and a light chain variable region (VL),

[0358] (i)

[0359] (a) the VH comprises three complementary determining regions HCDR1, HCDR2, and HCDR3 in sequence, wherein the HCDR1 has an amino acid sequence selected from any one of SEQ ID NOs: 1, 4, and 7, the HCDR2 has an amino acid sequence selected from any one of SEQ ID NOs: 2, 5, and 8, and the HCDR3 has an amino acid sequence selected from any one of SEQ ID NOs: 3, 6, and 9; and

[0360] (b) the VL comprises three complementarity determining regions, LCDR1, LCDR2 and LCDR3, in sequence, wherein the LCDR1 has an amino acid sequence selected from any one of SEQ ID NOs: 12, 15 and 18, the LCDR2 has an amino acid sequence selected from any one of SEQ ID NOs: 13, 16 and 19, and the LCDR3 has an amino acid sequence selected from any one of SEQ ID NOs: 14, 17 and 20; or

[0361] (ii)

[0362] (a) the VH comprises three complementary determining regions HCDR1, HCDR2, and HCDR3 in sequence, wherein the HCDR1 has an amino acid sequence selected from any one of SEQ ID NOs: 39, 42, and 45, the HCDR2 has an amino acid sequence selected from any one of SEQ ID NOs: 40, 43, and 46, and the HCDR3 has an amino acid sequence selected from any one of SEQ ID NOs: 41, 44, and 47; and

[0363] (b) the VL comprises three complementarity determining regions LCDR1, LCDR2 and LCDR3 in sequence, wherein the LCDR1 has an amino acid sequence selected from any one of SEQ ID NOs: 50, 53 and 56, the LCDR2 has an amino acid sequence selected from any one of SEQ ID NOs: 51, 54 and 57, and the LCDR3 has an amino acid sequence selected from any one of SEQ ID NOs: 52, 55 and 58.

[0364] In particular, the present disclosure provides multispecific antibodies comprising a first domain that specifically binds to IL-17A and a second domain that specifically binds to TNFα, wherein the first domain comprises (i) (a) HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs: 1, 2, and 3, respectively, and (b) LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs: 12, 13, and 14, respectively; or (ii) (a) HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs: 39, 40, and 41, respectively, and (b) LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs: 50, 51, and 52, respectively.

[0365] Other domains of the present disclosure that specifically bind IL-17A include amino acid sequences that have been mutated but still have at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identity in the CDR regions to the CDR regions described in the sequences described in Table 1. Suitably, other domains of the present disclosure that specifically bind IL-17A include mutant amino acid sequences in which no more than 1, 2, 3, 4, 5, or 10 amino acids have been mutated by deletion, insertion, or substitution of amino acids in the CDR regions when compared to the CDR regions described in the sequences described in Table 1. Mutations, such as substitutions, may be made at any residue within the CDR group and may be within CDR1, CDR2, and / or CDR3.

[0366] The term "amino acid" refers to naturally occurring amino acids and synthetic amino acids, as well as amino acid analogs and amino acid mimetics that function in a manner similar to the naturally occurring amino acids. Naturally occurring amino acids are the amino acids encoded by the genetic code, as well as amino acids that are later modified, such as hydroxyproline, γ-carboxyglutamate, and O-phosphoserine. The terms "polypeptide" and "protein" are used interchangeably herein to refer to a polymer of amino acid residues. The terms apply to amino acid polymers in which one or more amino acid residues is an artificial chemical mimetic of the corresponding naturally occurring amino acid, as well as to naturally occurring amino acid polymers and non-naturally occurring amino acid polymers. Unless otherwise indicated, a particular polypeptide sequence also implicitly encompasses conservatively modified variants thereof.

[0367] Techniques for making mutations, such as substitutions, within the CDRs, antibody VH or VL domains, and antibody amino acid sequences are generally available in the art. Variant sequences can be prepared in which the mutations, such as substitutions, may or may not be predicted to have minimal or beneficial effects on activity and tested for their ability to bind and / or neutralize IL-17A, TNFα, or human serum albumin, and / or any other desired properties. Appropriate mutations, such as substitutions, within the CDRs do not result in loss of function, so that an antibody or binding domain thereof comprising such a mutated amino acid sequence retains the ability to bind and / or neutralize IL-17A, TNFα, or human serum albumin. For example, it may retain the same quantitative binding and / or neutralizing ability as an unaltered domain thereof, for example, as measured in the assays described herein. Suitably, a domain or antigen-binding fragment thereof comprising such a mutated amino acid sequence may have improved binding and / or neutralization ability to IL-17A, TNFα, or human serum albumin. Suitably, the first domain of the present disclosure that specifically binds to IL-17A and comprises an amino acid sequence so mutated has the ability to neutralize IL-17A with a potency relative to secukinumab (relative potency) of greater than 2, such as greater than 5, greater than 10, greater than 15, greater than 20, greater than 25, greater than 30, greater than 35, greater than 40, greater than 45, preferably greater than 50, as determined by measuring Gro-α secretion in the HT-29 assay, and wherein the relative potency is the IC of secukinumab in ng / mL as measured in the HT-29 assay. 50 The values ​​are comparable to the IC values ​​in ng / mL of the multispecific antibodies measured in the HT-29 assay. 50 The ratio of values.

[0368] Furthermore, appropriate mutations, such as substitutions, within the CDRs do not result in a loss of solubility, stability, or high-yield manufacturability of the multispecific antibodies of the present disclosure, and thus the multispecific antibodies of the present disclosure, or their binding domains, comprising such mutated amino acid sequences retain biophysical properties. For example, they may retain the same high-yield manufacturability and / or stability as the unmodified multispecific antibodies of the present disclosure, for example, as measured in the assays described herein. Suitably, the multispecific antibodies of the present disclosure, or their binding domains, comprising such mutated amino acid sequences may have improved biophysical properties.

[0369] In the context of two or more nucleic acid or polypeptide sequences, the term "identical" or percent "identity" refers to two or more sequences or subsequences that are identical. "Percent amino acid sequence identity (%)" and "homology" with respect to nucleic acid, peptide, polypeptide or antibody sequences are defined as the percentage of amino acid residues in a candidate sequence that are identical to the amino acid residues in a particular peptide or polypeptide sequence, after aligning the sequences and introducing gaps, if necessary, to achieve maximum percent sequence identity, and not considering any conservative substitutions as part of the sequence identity. Alignments for determining percent amino acid sequence identity can be achieved in various ways within the skill of the art, for example, using disclosed computer software such as BLAST, BLAST-2 or ALIGN software. One skilled in the art can determine appropriate parameters for measuring alignment, including any algorithm required to achieve maximum alignment over the full length of the compared sequences.

[0370] To perform a sequence comparison, one sequence is typically used as a reference sequence to which a test sequence is compared. When using a sequence comparison algorithm, the test and reference sequences are input into a computer, subsequence coordinates are specified, if necessary, and sequence algorithm program parameters are specified. Default program parameters can be used, or alternative parameters can be specified. The sequence comparison algorithm then calculates the percent sequence identity of the test sequence relative to the reference sequence based on the program parameters.

[0371] Two examples of algorithms suitable for determining percent sequence identity and sequence similarity are the BLAST and BLAST 2.0 algorithms, which are described in Altschul et al., Nuc Acids Res. 25:3389-3402, 1977; and Altschul et al., J. Mol. Biol. 215:403-410, 1990, respectively. Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information.

[0372] The percent identity between two amino acid sequences can also be determined using the algorithm of E. Meyers and W. Miller (Comput. Appl. Biosci., 4: 11-17, 1988), which has been incorporated into the ALIGN program (version 2.0), using the PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4. Additionally, the percent identity between two amino acid sequences can be determined using the Needleman and Wunsch (J. Mol, Biol. 48: 444-453, 1970) algorithm, which has been incorporated into the GAP program in the GCG software package (available at www.gcg.com), using either a Blossom 62 matrix or a PAM250 matrix, a gap weight of 16, 14, 12, 10, 8, 6, or 4, and a length weight of 1, 2, 3, 4, 5, or 6.

[0373] Suitably, the first domain of the multispecific antibody of the present disclosure that specifically binds to IL-17A comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein:

[0374] (i)

[0375] (a) the VH comprises three complementary determining regions HCDR1, HCDR2 and HCDR3 in sequence,

[0376] the HCDR1 having an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to any one of SEQ ID NOs: 1, 4 and 7, preferably SEQ ID NO: 1;

[0377] the HCDR2 having an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to any one of SEQ ID NOs: 2, 5 and 8, preferably SEQ ID NO: 2;

[0378] The HCDR3 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to any one of SEQ ID NOs: 3, 6 and 9, preferably SEQ ID NO: 3; and / or

[0379] (b) the VL comprises three complementary determining regions LCDR1, LCDR2 and LCDR3 in sequence,

[0380] The LCDR1 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to any one of SEQ ID NOs: 12, 15 and 18, preferably SEQ ID NO: 12;

[0381] The LCDR2 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to any one of SEQ ID NOs: 13, 16 and 19, preferably SEQ ID NO: 13;

[0382] The LCDR3 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to any one of SEQ ID NOs: 14, 17 and 20, preferably SEQ ID NO: 14; or

[0383] (ii)

[0384] (a) the VH comprises three complementary determining regions HCDR1, HCDR2 and HCDR3 in sequence,

[0385] the HCDR1 having an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to any one of SEQ ID NOs: 39, 42 and 45, preferably SEQ ID NO: 39;

[0386] the HCDR2 having an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to any one of SEQ ID NOs: 40, 43 and 46, preferably SEQ ID NO: 40;

[0387] The HCDR3 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to any one of SEQ ID NO: 41, 44 and 47, preferably SEQ ID NO: 41; and / or

[0388] (b) the VL comprises three complementary determining regions LCDR1, LCDR2 and LCDR3 in sequence,

[0389] The LCDR1 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to any one of SEQ ID NOs: 50, 53 and 56, preferably SEQ ID NO: 50;

[0390] The LCDR2 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to any one of SEQ ID NOs: 51, 54 and 57, preferably SEQ ID NO: 51;

[0391] The LCDR3 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to any one of SEQ ID NO: 52, 55 and 58, preferably SEQ ID NO: 52.

[0392] Suitably, the first domain that specifically binds to IL-17A comprises: (i) a HCDR1, HCDR2 and HCDR3 that are at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to the sequences of SEQ ID NOs: 1, 2 and 3, respectively, and / or a LCDR1, LCDR2 and LCDR3 that are at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to the sequences of SEQ ID NOs: 12, 13 and 14, respectively; or (ii) a HCDR1, HCDR2 and HCDR3 that are at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to the sequences of SEQ ID NOs: 39, 40 and 41, respectively, and / or a LCDR1, LCDR2 and LCDR3 that are at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to the sequences of SEQ ID NOs: The sequences of NOs: 50, 51 and 52 have LCDR1, LCDR2 and LCDR3 that are at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical.

[0393] In another embodiment, the first domain that specifically binds to IL-17A comprises a heavy chain variable region VHA and a light chain variable region VLA.

[0394] In the context of the present disclosure, the terms "VH" (variable heavy chain or heavy chain variable region), "VL" (variable light chain or light chain variable region), "Vκ" and "Vλ" refer to families of antibody heavy and light chain sequences, grouped according to sequence identity and homology. Methods for determining sequence homology, for example, by using a homology search matrix such as BLOSUM (Henikoff, S. & Henikoff, JG, Proc. Natl. Acad. Sci. USA 89 (1992) 10915-10919), and methods for grouping sequences according to homology are well known to those of ordinary skill in the art. For VH, Vκ and Vλ, different subfamilies can be defined, for example, as shown in Knappik et al., J. Mol. Biol. 296 (2000) 57-86, which group VH into VH1A, VH1B and VH2 to VH6, Vκ into Vκ1 to Vκ4, and Vλ into Vλ1 to Vλ3. In vivo, antibody Vκ chains, Vλ chains, and VH chains are the result of random rearrangement of germline κ chain V and J segments, germline λ chain V and J segments, and heavy chain V, D, and J segments, respectively. The subfamily to which a given antibody variable chain belongs is determined by the corresponding V segment, in particular, by the framework regions FR1 to FR3. Therefore, any VH sequence characterized in this application by only a specific set of framework regions HFR1 to HFR3 can be combined with any HFR4 sequence, for example, an HFR4 sequence obtained from one of the heavy chain germline J segments, or an HFR4 sequence obtained from a rearranged VH sequence.

[0395] Suitably, the first domain of the multispecific antibody of the present disclosure that specifically binds to IL-17A comprises a heavy chain variable region VHA, wherein the VHA is VH1A, VH1B, VH3, or VH4. In one embodiment, the first domain of the multispecific antibody of the present disclosure that specifically binds to IL-17A comprises a heavy chain variable region VHA, wherein the VHA is VH4. In a preferred embodiment, the first domain of the multispecific antibody of the present disclosure that specifically binds to IL-17A comprises a heavy chain variable region VHA, wherein the VHA is VH3.

[0396] Suitably, the first domain of the multispecific antibody of the present disclosure that specifically binds to IL-17A comprises a light chain variable region, VLA, wherein the VLA comprises a Vκ framework FR1, FR2, and FR3, particularly Vκ1 or Vκ3 FR1 to FR3, preferably a Vκ1 framework FR1 to FR3, and a framework FR4 selected from the group consisting of a VκFR4, particularly a Vκ1 FR4, a Vκ3 FR4, and a VλFR4. Suitable VλFR4s are shown in SEQ ID NOs: 26 to 32. In one embodiment, the first domain that specifically binds to IL-17A comprises a VλFR4 comprising an amino acid sequence that is at least 60, 70, 80, or 90% identical to an amino acid sequence selected from any one of SEQ ID NOs: 26 to 32, preferably SEQ ID NO: 26 or SEQ ID NO: 27, and more preferably SEQ ID NO: 27. Suitably, the first domain that specifically binds to IL-17A comprises VλFR4, which comprises an amino acid sequence selected from any one of SEQ ID NO: 26 to SEQ ID NO: 32, preferably, VλFR4 is as shown in SEQ ID NO: 26 or 27, more preferably, VλFR4 is as shown in SEQ ID NO: 27.

[0397] Thus, in one embodiment, the present disclosure provides a multispecific antibody comprising a first domain that specifically binds IL-17A and a second domain that specifically binds TNFα, wherein the first domain comprises:

[0398] (i)(a) the HCDR1, HCDR2 and HCDR3 sequences of SEQ ID NOs: 1, 2 and 3, respectively, and the LCDR1, LCDR2 and LCDR3 sequences of SEQ ID NOs: 12, 13 and 14, respectively; or (i)(b) the HCDR1, HCDR2 and HCDR3 sequences of SEQ ID NOs: 39, 40 and 41, respectively, and the LCDR1, LCDR2 and LCDR3 sequences of SEQ ID NOs: 50, 51 and 52, respectively;

[0399] (ii) a VH3 or VH4 domain, preferably a VH3 domain; and

[0400] (iii) a VL domain comprising a VL framework comprising a Vκ framework FR1, FR2 and FR3, in particular Vκ1 or Vκ3 FR1 to FR3, preferably Vκ1 FR1 to FR3, and a framework FR4 selected from the group consisting of VκFR4, in particular Vκ1FR4, Vκ3 FR4, and VλFR4, in particular VλFR4 comprising an amino acid sequence that is at least 60, 70, 80, 90% identical to an amino acid sequence selected from any one of SEQ ID NO: 26 to SEQ ID NO: 32, preferably a VλFR4 as shown in an amino acid sequence selected from any one of SEQ ID NO: 26 to SEQ ID NO: 32, more preferably a VλFR4 as shown in SEQ ID NO: 27.

[0401] Suitably, the present disclosure provides a multispecific antibody comprising a first domain that specifically binds to IL-17A and a second domain that specifically binds to TNFα, wherein the first domain comprises a VH listed in Table 1. Suitably, the present disclosure also provides a multispecific antibody comprising a first domain that specifically binds to IL-17A and a second domain that specifically binds to HSA, wherein the first domain comprises (or alternatively consists of) a VH amino acid sequence listed in Table 1, wherein no more than about 20 amino acids in the framework sequence (e.g., a sequence that is not a CDR) have been mutated (wherein, as various non-limiting examples, the mutation is an addition, substitution, or deletion), preferably no more than about 10. Other domains of the present disclosure that specifically bind to IL-17A include amino acid sequences that have been mutated but still have at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identity in the VH region to the VH region described in the sequences described in Table 1.

[0402] Suitably, the present disclosure provides a multispecific antibody comprising a first domain that specifically binds to IL-17A and a second domain that specifically binds to TNFα, wherein the first domain comprises a VL domain listed in Table 1. Suitably, the present disclosure also provides a multispecific antibody comprising a first domain that specifically binds to IL-17A and a second domain that specifically binds to HSA, wherein the first domain comprises (or alternatively consists of) a VL amino acid sequence listed in Table 1, wherein no more than about 20 amino acids in the framework sequence (e.g., a sequence that is not a CDR) have been mutated (wherein, as various non-limiting examples, the mutation is an addition, substitution, or deletion), preferably no more than about 10 amino acids. Other domains of the present disclosure that specifically bind to IL-17A include amino acid sequences that have been mutated but still have at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identity in the VL region to the VL region described in the sequences described in Table 1.

[0403] In one embodiment, the present disclosure provides a multispecific antibody comprising a first domain that specifically binds to IL-17A and a second domain that specifically binds to TNFα, wherein the first domain comprises a heavy chain variable region comprising an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90%, identical to the amino acid sequence of SEQ ID NO: 10 or SEQ ID NO: 11, preferably SEQ ID NO: 10, and in particular, wherein the domain comprises the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs: 1, 2, and 3, respectively. In another embodiment, the first domain that specifically binds to IL-17A comprises a heavy chain variable region comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 11, and wherein the heavy chain variable region comprises Q14K, G16E, and G56A (AHo numbering).

[0404] In another embodiment, the present disclosure provides a multispecific antibody comprising a first domain that specifically binds to IL-17A and a second domain that specifically binds to TNFα, wherein the first domain comprises a heavy chain variable region comprising an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90%, identical to the amino acid sequence of SEQ ID NO: 48 or SEQ ID NO: 49, preferably SEQ ID NO: 48, and in particular, wherein the antibody comprises the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs: 39, 40, and 41, respectively. In another embodiment, the present disclosure provides an isolated antibody that specifically binds to human IL-17A, wherein the antibody comprises a heavy chain variable region comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 49, and wherein the heavy chain variable region comprises R20T and Q141P (AHo numbering).

[0405] In another embodiment, the present disclosure provides a multispecific antibody comprising a first domain that specifically binds IL-17A and a second domain that specifically binds TNFα, wherein the first domain (i) comprises a light chain variable region comprising an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99%, preferably at least 90%, identical to the amino acid sequence of SEQ ID NO: 21 or SEQ ID NO: 22, preferably SEQ ID NO: 21, and in particular, wherein the antibody comprises LCDR1, LCDR2 and LCDR3 of SEQ ID NO: 12, 13 and 14, respectively; or (ii) comprises a light chain variable region comprising an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99%, preferably at least 90%, identical to the amino acid sequence of SEQ ID NO: 12, 13 and 14, respectively. NO:59, at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99%, preferably at least 90% identical amino acid sequence, and in particular, wherein the antibody comprises LCDR1, LCDR2 and LCDR3 sequences of SEQ ID NO:50, 51 and 52, respectively.

[0406] In another embodiment, the present disclosure provides a multispecific antibody comprising a first domain that specifically binds IL-17A and a second domain that specifically binds TNFα, wherein the first domain comprises a light chain variable region comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 22, and wherein the light chain variable region comprises A51P (AHo numbering).

[0407] In another embodiment, the present disclosure provides a multispecific antibody comprising a first domain that specifically binds to IL-17A and a second domain that specifically binds to TNFα, wherein the first domain (i) comprises a heavy chain variable region comprising an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90%, identical to the amino acid sequence of SEQ ID NO: 10; and the light chain variable region comprises an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90% identical to the amino acid sequence of SEQ ID NO: 21; or (ii) comprises a heavy chain variable region comprising an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90% identical to the amino acid sequence of SEQ ID NO: NO:48 is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99%, preferably at least 90% identical to an amino acid sequence; the light chain variable region comprises an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99%, preferably at least 90% identical to the amino acid sequence of SEQ ID NO:59.

[0408] Preferably, the first domain that specifically binds to IL-17A of the multispecific antibody of the present disclosure comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprising (i) an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90%, identical to the amino acid sequence of SEQ ID NO: 10; and a light chain variable region comprising an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90% identical to the amino acid sequence of SEQ ID NO: 21, and wherein the antibody comprises the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs: 1, 2, and 3, respectively, and / or the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs: 12, 13, and 14, respectively; in particular, wherein the antibody comprises the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs: 12, 13, and 14, respectively. NO: 1, 2 and 3, and LCDR1, LCDR2 and LCDR3 sequences of SEQ ID NO: 12, 13 and 14, respectively; or the heavy chain variable region comprises (ii) an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99%, preferably at least 90% identical to the amino acid sequence of SEQ ID NO: 48; the light chain variable region comprises an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99%, preferably at least 90% identical to the amino acid sequence of SEQ ID NO: 59, and wherein the antibody comprises HCDR1, HCDR2 and HCDR3 sequences of SEQ ID NO: 39, 40 and 41, respectively, and / or SEQ ID NO: 42, respectively. ID NOs: 50, 51 and 52; in particular, wherein the antibody comprises the HCDR1, HCDR2 and HCDR3 sequences of SEQ ID NOs: 39, 40 and 41, respectively, and the LCDR1, LCDR2 and LCDR3 sequences of SEQ ID NOs: 39, 40 and 41, respectively.

[0409] In a specific embodiment, the present disclosure provides a multispecific antibody comprising a first domain that specifically binds to IL-17A and a second domain that specifically binds to TNFα, wherein the first domain comprises (i) a VH comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 10 and 11; and / or a VL thereof comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 21 and 22, or (ii) a VH comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 48 and 19; and / or a VL thereof comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 59 and 60.

[0410] In one specific embodiment, the first domain that specifically binds to IL-17A comprises (i) a VH sequence of SEQ ID NO: 10 and a VL sequence of SEQ ID NO: 21, or (ii) a VH sequence of SEQ ID NO: 48 and a VL sequence of SEQ ID NO: 59. In yet another specific embodiment, the first domain that specifically binds to IL-17A comprises (i) a VH sequence of SEQ ID NO: 11 and a VL sequence of SEQ ID NO: 22, or (ii) a VH sequence of SEQ ID NO: 49 and a VL sequence of SEQ ID NO: 60.

[0411] In one embodiment, the domain that specifically binds to human IL-17A is a domain described in Table 1. In one embodiment, the domain that specifically binds to human IL-17A comprises at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99%, preferably at least 90%, of an amino acid sequence selected from the group consisting of (i) SEQ ID NOs: 24 and 25, preferably SEQ ID NO: 24, or (ii) SEQ ID NOs: 61 and 62, preferably SEQ ID NO: 61. In one embodiment, the domain that specifically binds to human IL-17A is as shown in (i) SEQ ID NO: 24 or SEQ ID NO: 25, preferably SEQ ID NO: 24, or (ii) SEQ ID NO: 61 or SEQ ID NO: 62, preferably SEQ ID NO: 61.

[0412] Other domains of the present disclosure that have binding specificity for human IL-17A include those in which the amino acids or nucleic acids encoding the amino acids have been mutated, but have at least 60, 70, 80, 90, or 95% identity with the sequences set forth in Table 1. In one embodiment, the amino acid sequences are comprised in which no more than 1, 2, 3, 4, or 5 amino acids in the variable region have been mutated when compared to the variable region set forth in the sequences set forth in Table 1, while retaining substantially the same activity. As used herein, the term "substantially the same activity" means that the activity represented by substantially the same activity is at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 98% or even at least 100% or at least 110%, or at least 120%, or at least 130%, or at least 140%, or at least 150%, or at least 160%, or at least 170%, or at least 180%, or at least 190%, for example up to 200% of the activity determined for the parent antibody (e.g., a multispecific antibody of the disclosure, in particular a multispecific antibody of the disclosure comprising a first domain that specifically binds to human IL-17A as described in Table 1 and / or a second domain that specifically binds to human TNFα as described in Table 1).

[0413] In yet another embodiment, the present disclosure provides a multispecific antibody comprising a first domain that specifically binds to IL-17A and a second domain that specifically binds to TNFα, wherein the first domain comprises an amino acid sequence homologous to the sequences described in Table 1, and the first domain binds to human IL-17A and retains the desired functional properties of those domains described in Table 1.

[0414] In one embodiment, a domain of the invention that specifically binds to IL-17A has a heavy chain variable region comprising a HCDR1, HCDR2, and HCDR3 sequence and a light chain variable region comprising a LCDR1, LCDR2, and LCDR3 sequence, wherein one or more of these CDR sequences has a specified amino acid sequence based on the domain described herein or conservative modifications thereof, wherein the domain retains the desired functional properties of the antibodies of the present disclosure.

[0415] The terms "conservatively modified variants" or "conservative variants" apply to both amino acid and nucleic acid sequences. With respect to a particular nucleic acid sequence, conservatively modified variants refer to those nucleic acids that encode identical or substantially identical amino acid sequences, or, when the nucleic acid does not encode an amino acid sequence, to substantially identical sequences. Due to the degeneracy of the genetic code, a large number of functionally identical nucleic acids encode any given protein. For example, the codons GCA, GCC, GCG, and GCU all encode the amino acid alanine. Thus, at every position where alanine is explicitly specified by a codon, that codon can be changed to any of the described corresponding codons without altering the encoded polypeptide. Such nucleic acid variations are "silent variations," a type of conservatively modified variation. Each nucleic acid sequence encoding a polypeptide herein also describes every possible silent variation of the nucleic acid. Those skilled in the art will recognize that every codon in a nucleic acid (except AUG, which is typically the only codon for methionine, and TGG, which is typically the only codon for tryptophan) can be modified to produce functionally identical molecules. Thus, every silent variation of a nucleic acid encoding a polypeptide is implicit in each recited sequence.

[0416] For polypeptide sequences, "conservatively modified variants" or "conservative variants" include single substitutions, deletions, or additions to a polypeptide sequence that result in the replacement of an amino acid with a chemically similar amino acid. Conservative substitution tables providing functionally similar amino acids are well known in the art. These conservatively modified variants (i.e., having one or more "conservative modifications") are in addition to and do not exclude the following: polymorphic variants, interspecies homologs, and alleles of the present disclosure. The following eight groups contain amino acids that are conservative substitutions for each other: 1) alanine (A), glycine (G); 2) aspartic acid (D), glutamic acid (E); 3) asparagine (N), glutamine (Q); 4) arginine (R), lysine (K); 5) isoleucine (I), leucine (L), methionine (M), valine (V); 6) phenylalanine (F), tyrosine (Y), tryptophan (W); 7) serine (S), threonine (T); and 8) cysteine ​​(C), methionine (M) (see, e.g., Creighton, Proteins (1984)). In one embodiment, the term "conservative sequence modification" is used to refer to amino acid modifications that do not significantly affect or alter the binding properties of the antibody containing the amino acid sequence.

[0417] Thus, the present disclosure provides a multispecific antibody comprising a first domain that specifically binds to IL-17A and a second domain that specifically binds to TNFα, wherein the first domain comprises (or consists of):

[0418] (i)

[0419] A heavy chain variable region (VH), which comprises three complementarity determining regions HCDR1, HCDR2 and HCDR3 in sequence, wherein the HCDR1 is the amino acid sequence of SEQ ID NO: 1, or a conservative variant thereof; the HCDR2 is the amino acid sequence of SEQ ID NO: 2, or a conservative variant thereof; the HCDR3 is an amino acid sequence selected from any one of SEQ ID NO: 3, or a conservative variant thereof; and

[0420] A light chain variable region (VL) comprising three complementarity determining regions LCDR1, LCDR2, and LCDR3, wherein the LCDR1 has the amino acid sequence of SEQ ID NO: 12, or a conservative variant thereof; the LCDR2 has the amino acid sequence of SEQ ID NO: 13, or a conservative variant thereof; the LCDR3 has the amino acid sequence of SEQ ID NO: 14, or a conservative variant thereof; or

[0421] (ii)

[0422] A heavy chain variable region (VH), which comprises three complementarity determining regions HCDR1, HCDR2 and HCDR3 in sequence, wherein the HCDR1 is the amino acid sequence of SEQ ID NO: 39, or a conservative variant thereof; the HCDR2 is the amino acid sequence of SEQ ID NO: 40, or a conservative variant thereof; the HCDR3 is an amino acid sequence selected from any one of SEQ ID NO: 41, or a conservative variant thereof; and

[0423] A light chain variable region (VL), which comprises three complementarity determining regions LCDR1, LCDR2 and LCDR3 in sequence, wherein the LCDR1 has the amino acid sequence of SEQ ID NO: 50, or a conservative variant thereof; the LCDR2 has the amino acid sequence of SEQ ID NO: 51, or a conservative variant thereof; the LCDR3 has the amino acid sequence of SEQ ID NO: 52, or a conservative variant thereof,

[0424] The antibody specifically binds to human IL-17A and / or neutralizes IL-17A.

[0425] Anti-IL-17A antibodies of the present disclosure

[0426] The present disclosure is based on the discovery of antibody molecules that specifically bind to human IL-17A with improved affinity, efficacy, and selectivity. In addition, the antibodies of the present disclosure have improved biophysical properties, such as improved solubility, developability, and high manufacturability, relatively low impurities (>98%, particularly >99% monomeric, as determined by SE-HPLC), and stability.

[0427] In one aspect, the present disclosure provides an isolated antibody with binding specificity to human IL-17A, comprising a set of CDRs: HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3, wherein the set of CDRs has 10 or fewer amino acid substitutions, e.g., 9 or fewer amino acid substitutions, 8 or fewer amino acid substitutions, 7 or fewer amino acid substitutions, 6 or fewer amino acid substitutions, 5 or fewer amino acid substitutions, 4 or fewer amino acid substitutions, 3 or fewer amino acid substitutions, 2 or fewer amino acid substitutions, 1 or 0 amino acid substitutions, preferably 0 amino acid substitutions, relative to such a set of CDRs: (i) HCDR1′ is an amino acid sequence selected from any one of SEQ ID NOs: 1, 4, and 7, preferably SEQ ID NO: 1; HCDR2′ is an amino acid sequence selected from any one of SEQ ID NOs: 2, 5, and 8, preferably SEQ ID NO: 2; HCDR3′ is an amino acid sequence selected from any one of SEQ ID NOs: 3, 6, and 9, preferably SEQ ID NO: NO:3; LCDR1' is an amino acid sequence selected from any one of SEQ ID NOs:12, 15 and 18, preferably SEQ ID NO:12; LCDR2' is an amino acid sequence selected from any one of SEQ ID NOs:13, 16 and 19, preferably SEQ ID NO:13; and LCDR3' has an amino acid sequence selected from any one of SEQ ID NOs:14, 17 and 20, preferably SEQ ID NO:14; or (ii) wherein HCDR1' is an amino acid sequence selected from any one of SEQ ID NOs:39, 42 and 45, preferably SEQ ID NO:39; HCDR2' is an amino acid sequence selected from any one of SEQ ID NOs:40, 43 and 46, preferably SEQ ID NO:40; HCDR3' is an amino acid sequence selected from any one of SEQ ID NOs:41, 44 and 47, preferably SEQ ID NO:41; LCDR1' is an amino acid sequence selected from any one of SEQ ID NOs:50, 53 and 56, preferably SEQ ID NO:50; LCDR2' is an amino acid sequence selected from any one of SEQ ID NOs:51, 54 and 57, preferably SEQ ID NO:51; and LCDR3' has an amino acid sequence selected from any one of SEQ ID NOs:52, 55 and 58, preferably SEQ ID NO:52.

[0428] The term "IL-17A" or "IL17A" refers specifically to human IL-17A with UniProt ID No. Q16552, reproduced herein as SEQ ID NO: 33. The term "cynomolgus IL-17A" or "cynomolgus monkey IL-17A" refers to Macaca fascicularis IL-17A with UniProt ID No. G1QUS7.

[0429] The term "IL-17B" refers in particular to human IL-17B having UniProt ID No. Q9UHF5, reproduced herein as SEQ ID NO: 34. The term "IL-17C" refers in particular to human IL-17C having UniProt ID No. Q9P0M4, reproduced herein as SEQ ID NO: 35. The term "IL-17D" refers in particular to human IL-17D having UniProt ID No. Q8TAD2, reproduced herein as SEQ ID NO: 36. The term "IL-17E" refers in particular to human IL-17E having UniProt ID No. Q9H293, reproduced herein as SEQ ID NO: 37. The term "IL-17F" refers in particular to human IL-17F having UniProt ID No. Q96PD4, reproduced herein as SEQ ID NO: 38.

[0430] The term "epitope" refers to a local region of an antigen to which an antibody can specifically bind. For example, an epitope can be a continuous amino acid of a polypeptide, or, for example, an epitope can be derived from two or more non-contiguous regions of a polypeptide or polypeptides.

[0431] As used herein, the term "antibody" and the like include: complete antibodies; any antigen-binding fragment (i.e., "antigen-binding portion") or single chain of a complete antibody; and molecules (including but not limited to multispecific antibodies) comprising antibody CDRs, VH regions, or VL regions. A naturally occurring "complete antibody" is a glycoprotein comprising at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds. Each heavy chain consists of a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region. The heavy chain constant region consists of three domains, CH1, CH2, and CH3. Each light chain consists of a light chain variable region (abbreviated herein as VL) and a light chain constant region. The light chain constant region consists of one domain, CL. The VH and VL regions can be further subdivided into hypervariable regions, called complementarity determining regions (CDRs), interspersed with more conserved regions, called framework regions (FRs). Each VH and VL is composed of three CDRs and four FRs, which are arranged in the following order from amino terminus to carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavy and light chains contain binding domains that interact with antigens. The constant region of an antibody can mediate the binding of immunoglobulins to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (Clq) of the classical complement system.

[0432] As used herein, the term "isotype" refers to the antibody class (e.g., IgM, IgE, IgD, IgA, IgY, and IgG, such as IgG1 or IgG4) provided by the heavy chain constant region gene. Isotypes also include modified forms of one of these classes, wherein the modification is to alter Fc function, e.g., to enhance or reduce effector function or binding to Fc receptors. Suitably, the antibodies of the present disclosure are IgGs selected from the group consisting of IgG1, IgG2, IgG3, and IgG4. More suitably, the antibodies of the present disclosure are IgG1 or IgG4.

[0433] As used herein, the terms "antigen-binding fragment," "antigen-binding portion," and the like refer to one or more fragments of an intact, whole antibody that retain the ability to specifically bind to a given antigen (e.g., IL-17A). Examples of binding fragments encompassed within the term "antigen-binding portion" of an antibody include a Fab fragment, a monovalent fragment consisting of the VL, VH, CL, and CH1 domains; a F(ab)2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bond at the hinge region; an Fd fragment consisting of the VH and CH1 domains; an Fv fragment consisting of the VL and VH domains of a single arm of an antibody; a single-domain antibody (dAb) fragment (Ward et al., 1989 Nature 5576). 341:544-546), which consists of a VH domain; isolated complementarity determining regions (CDRs), dsFv, scAb, STAB, single domain antibodies (sdAb or dAb), single domain heavy chain antibodies, and single domain light chain antibodies, VHH, VNAR, single domain antibodies based on shark VNAR structures, and binding domains based on alternative scaffolds including but not limited to ankyrin-based domains, fynomers, avimers, anticalins, fibronectin, and binding sites built into antibody constant regions (e.g., Modular Antibody Technology of F-star). TM ).

[0434] The term "complementarity determining region" ("CDR") refers to an amino acid sequence whose boundaries are defined using any of a number of well-known schemes, including those described by Kabat et al. (1991), "Sequences of Proteins of Immunological Interest," 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD ("Kabat" numbering scheme); Al-Lazikani et al., (1997) JMB 273, 927-948 ("Chothia" numbering scheme); ImMunoGenTics (IMGT) numbering (Lefranc, M.-P., The Immunologist, 7, 132-136 (1999); Lefranc, M.-P. et al., (1997) JMB 273, 927-948 ("Chothia" numbering scheme); ImMunoGenTics (IMGT) numbering (Lefranc, M.-P., The Immunologist, 7, 132-136 (1999); Lefranc, M.-P. et al., (1997) JMB 273, 927-948 ("Chothia" numbering scheme); al., Dev. Comp. Immunol., 27, 55-77 (2003) ("IMGT" numbering scheme), and the numbering scheme described in Honegger & Plückthun, J. Mol. Biol. 309 (2001) 657-670 ("AHo" numbering scheme). For example, for the classical form, under Kabat, the CDR amino acid residues in the heavy chain variable domain (VH) are numbered 31-35 (HCDR1), 50-65 (HCDR2), and 95-102 (HCDR3); the CDR amino acid residues in the light chain variable domain (VL) are numbered 24-34 (LCDR1), 50-56 (LCDR2), and 89-97 (LCDR3). Under Chothia, the CDR amino acid residues in VH are numbered 26-32 (HCDR1), 52-56 (HCDR2), and 95-102 (HCDR3). The amino acid residues in the VL are numbered as 24-34 (LCDR1), 50-56 (LCDR2), and 89-97 (LCDR3). By combining the CDR definitions of Kabat and Chothia, the CDRs consist of amino acid residues 26-35 (HCDR1), 50-65 (HCDR2), and 95-102 (HCDR3) in human VH, and amino acid residues 24-34 (LCDR1), 50-56 (LCDR2), and 89-97 (LCDR3) in human VL. Under IMGT, the CDR amino acid residues in VH are numbered as approximately 26-35 (HCDR1), 51-57 (HCDR2), and 93-102 (HCDR3), and the CDR amino acid residues in VL are numbered as approximately 27-32 (LCDR1), 50-52 (LCDR2), and 89-97 (LCDR3) (according to "Kabat" numbering).Under IMGT, the CDRs of an antibody can be determined using the program IMGT / DomainGap Align.

[0435] In the context of the present disclosure, unless otherwise specifically indicated, the numbering system suggested by Honegger & Plückthun ("AHo") is used (Honegger & Plückthun, J. Mol. Biol. 309 (2001) 657-670). In addition, according to the AHo numbering scheme, the following residues are defined as CDRs: LCDR1 (also known as CDR-L1): L24-L42; LCDR2 (also known as CDR-L2): L58-L72; LCDR3 (also known as CDR-L3): L107-L138; HCDR1 (also known as CDR-H1): H27-H42; HCDR2 (also known as CDR-H2): H57-H76; HCDR3 (also known as CDR-H3): H108-H138. For the sake of clarity, the numbering system according to Honegger & Plückthun takes into account the length diversity found in naturally occurring antibodies, in both different VH and VL subfamilies, in particular in the CDRs, and provides for gaps in the sequence. Thus, in a given antibody variable domain, not all positions 1 to 149 are usually occupied by amino acid residues.

[0436] Preferably, the "antigen-binding region" comprises at least amino acid residues 4 to 138 of the variable light (VL) chain and amino acid residues 5 to 138 of the variable heavy (VH) chain (in each case numbered according to Honegger & Plückthun), more preferably amino acid residues 3 to 144 of VL and amino acid residues 4 to 144 of VH, and particularly preferably complete VL and VH chains (amino acids 1 to 149 of VL and amino acids 1 to 149 of VH). The antigen-binding portion can also be incorporated into single-domain antibodies, maxibodies, minibodies, intrabodies, diabodies, triabodies, tetrabodies, v-NARs, and bis-scFvs (see, e.g., Holliger and Hudson, 2005, Nature Biotechnology, 23, 9, 1 He1 36). The antigen-binding portion of the antibody can be grafted onto a scaffold based on a polypeptide, such as type III fibronectin (Fn3) (see U.S. Pat. No. 6,703,199, which describes a fibronectin polypeptide monoclonal antibody). The antigen-binding portion can be incorporated into a single-chain molecule comprising a pair of tandem Fv segments (VH-CH1-VH-CH1), which together with a complementary light chain polypeptide form a pair of antigen-binding regions (Zapata et al., 1995 Protein Eng. 8(10):1057-1062; and U.S. Pat. No. 5,641,870).

[0437] As used herein, the terms "domain" or "domain that specifically binds to X" or "binding domain" of an antibody, "antigen-binding fragment thereof," "antigen-binding portion thereof," and the like refer to one or more fragments of an intact, complete antibody that retain the ability to specifically bind to a given antigen (e.g., IL-17A, TNFα, HSA). The antigen-binding function of an antibody can be performed by fragments of an intact antibody. In some embodiments, the binding domain of the multispecific antibodies of the present disclosure is selected from the group consisting of: a Fab fragment, a monovalent fragment consisting of the VL, VH, CL, and CH1 domains; a F(ab)2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bond at the hinge region; an Fd fragment consisting of the VH and CH1 domains; an Fv fragment consisting of the VL and VH domains of a single arm of an antibody; a single-domain antibody (dAb) fragment (Ward et al., 2001). al., 1989 Nature 341: 544-546), which consists of a VH domain; isolated complementarity determining regions (CDRs), dsFv, scAb, STAB, single domain antibodies (sdAb or dAb), single domain heavy chain antibodies, and single domain light chain antibodies, VHH, VNAR, single domain antibodies based on shark VNAR structures, and binding domains based on alternative scaffolds including but not limited to ankyrin-based domains, fynomers, avimers, anticalins, fibronectin, and binding sites constructed into antibody constant regions (e.g., Modular Antibody Technology of F-star) TM Suitably, the binding domain of the present disclosure is an Fv fragment (Fv). Suitably, the binding domain of the present disclosure is a single-chain Fv fragment (scFv). Suitably, the binding domain of the present disclosure is a Fab fragment.

[0438] Preferably, the "domain" or "domain that specifically binds to X" or "binding domain", "antigen-binding fragment thereof", or "antigen-binding portion" comprises at least amino acid residues 4 to 138 of the variable light (VL) chain and amino acid residues 5 to 138 of the variable heavy (VH) chain (in each case, numbered according to Honegger & Plückthun), more preferably amino acid residues 3 to 144 of VL and amino acid residues 4 to 144 of VH, and particularly preferably complete VL and VH chains (amino acids 1 to 149 of VL and amino acids 1 to 149 of VH). The antigen-binding portion can also be incorporated into single-domain antibodies, maxibodies, minibodies, inobodies, diabodies, triabodies, tetrabodies, v-NARs, and bis-scFvs (see, e.g., Holliger and Hudson, 2005, Nature Biotechnology, 23, 9, 1 He1 1 36). The antigen-binding portion of the antibody can be grafted onto a scaffold based on a polypeptide, such as type III fibronectin (Fn3) (see U.S. Pat. No. 6,703,199, which describes a fibronectin polypeptide monoclonal antibody). The antigen-binding portion can be incorporated into a single-chain molecule comprising a pair of tandem Fv segments (VH-CH1-VH-CH1), which together with a complementary light chain polypeptide form a pair of antigen-binding regions (Zapata et al., 1995 Protein Eng. 8(10):1057-1062; and U.S. Pat. No. 5,641,870).

[0439] As used herein, the term "binding specificity" or "specific binding" refers to the ability of a single antibody or antibody domain to react with a kind of antigenic determinant and not with different antigenic determinants. As used herein, the term "with ... specifically binds" or "specific for" refers to a measurable and reproducible interaction, such as the combination between a target and an antibody or antibody domain, which is in the presence of a heterogeneous population of molecules (including biomolecules), and there is a determinant of the target. For example, an antibody or antibody domain that specifically binds to a target (which can be an epitope) is an antibody or antibody domain that has a greater affinity, avidity, easier and / or longer duration than that of the target than that of other targets. In its most general form (and when no specified standard is mentioned), "specific binding" refers to the ability of an antibody or antibody domain to distinguish between a target of interest and an unrelated molecule, such as determined according to specific assay methods known in the art. Such methods include, but are not limited to, Western blotting, ELISA, RIA, ECL, IRMA, SPR (surface plasmon resonance) tests and peptide scanning. For example, standard ELISA assays can be performed. Scoring can be performed by standard color development (e.g., a secondary antibody with horseradish peroxide and tetramethylbenzidine with hydrogen peroxide). The reactions in certain wells are scored by optical density (e.g., at 450 nm). A typical background (= negative reaction) may be about 0.1 OD; a typical positive reaction may be about 1 OD. This means that the ratio between the positive and negative scores can be 10 times or more. In another example, an SPR assay can be performed, in which a difference between the background and the signal of at least 10 times, preferably at least 100 times, indicates specific binding. Typically, rather than using a single reference molecule, a panel of about three to five unrelated molecules, such as milk powder, transferrin, etc., is used to determine binding specificity. The specific antibodies or antibody domains disclosed herein have binding specificity for human IL-17A or human TNFα.

[0440] The multispecific antibodies of the present disclosure comprise a first domain that specifically binds to IL-17A and a second domain that specifically binds to TNFα, and thus have binding specificity for IL-17A and TNFα, in particular for human IL-17A and human TNFα. In one embodiment, the antibodies of the present disclosure have binding specificity for human IL-17A and cynomolgus monkey (Macaca fascicularis) (also known as Cynomolgus monkey or "Cynomolgus") IL-17A. In one embodiment, the antibodies of the present disclosure have binding specificity for human TNFα and cynomolgus monkey (Macaca fascicularis) (also known as Cynomolgus monkey or "Cynomolgus") TNFα.

[0441] In another aspect, the disclosure relates to antibodies or antibody domains that have binding specificity for human IL-17A and Macaca fascicularis (also known as Cynomolgus monkey or "Cynomolgus") IL-17A.

[0442] Suitably, the anti-IL-17A antibodies of the present disclosure are isolated antibodies.

[0443] Suitably, the anti-IL-17A antibodies of the present disclosure are monoclonal antibodies.

[0444] The anti-IL-17A antibodies of the present disclosure include, but are not limited to, chimeric antibodies and humanized antibodies.

[0445] Suitably, the anti-IL-17A antibodies of the present disclosure are humanized. Suitably, the anti-IL-17A antibodies of the present disclosure are humanized and comprise rabbit-derived CDRs.

[0446] Antibodies disclosed herein include, but are not limited to, humanized monoclonal antibodies isolated as described herein, including in the Examples. Examples of such anti-human IL-17A antibodies are the antibodies whose sequences are listed in Table 1. Additional details regarding the generation and characterization of the antibodies described herein are provided in the Examples.

[0447] The isolated antibody of the present invention that has binding specificity for human IL-17A comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein: (a) the VH comprises three complementarity determining regions HCDR1, HCDR2, and HCDR3 in sequence, and (b) the VL comprises three complementarity determining regions LCDR1, LCDR2, and LCDR3 in sequence.

[0448] The present disclosure provides antibodies that specifically bind to an IL-17A protein, the antibodies comprising a VH CDR having the amino acid sequence of any one of the VH CDRs listed in Table 1. In particular, the present disclosure provides antibodies that specifically bind to an IL-17A protein VH CDR, the antibodies comprising one, two, three or more VH CDRs having the amino acid sequence of any one of the VH CDRs listed in Table 1.

[0449] The present disclosure provides an antibody with binding specificity to human IL-17A, comprising a heavy chain variable region (VH), wherein the VH comprises (i) three complementarity determining regions HCDR1, HCDR2, and HCDR3, in sequence, wherein the HCDR1 has an amino acid sequence selected from any one of SEQ ID NOs: 1, 4, and 7, the HCDR2 has an amino acid sequence selected from any one of SEQ ID NOs: 2, 5, and 8, and the HCDR3 has an amino acid sequence selected from any one of SEQ ID NOs: 3, 6, and 9. In particular, the present disclosure provides antibodies that have binding specificity to human IL-17A and comprise the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs: 1, 2, and 3, respectively; or (ii) three complementarity determining regions HCDR1, HCDR2, and HCDR3, wherein the HCDR1 has an amino acid sequence selected from any one of SEQ ID NOs: 39, 42, and 45, the HCDR2 has an amino acid sequence selected from any one of SEQ ID NOs: 40, 43, and 46, and the HCDR3 has an amino acid sequence selected from any one of SEQ ID NOs: 41, 44, and 47. In particular, the present disclosure provides antibodies that have binding specificity to human IL-17A and comprise the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs: 39, 40, and 41, respectively.

[0450] The present disclosure also provides an antibody that specifically binds to an IL-17A protein, the antibody comprising a VL CDR having the amino acid sequence of any one of the VL CDRs listed in Table 1. In particular, the present disclosure provides an antibody that specifically binds to an IL-17A protein, the antibody comprising one, two, three or more VL CDRs having the amino acid sequence of any one of the VL CDRs listed in Table 1.

[0451] The present disclosure provides an antibody with binding specificity to human IL-17A, comprising a light chain variable region (VL), wherein the VL comprises (i) three complementarity determining regions LCDR1, LCDR2, and LCDR3, in sequence, wherein the LCDR1 has an amino acid sequence selected from any one of SEQ ID NOs: 12, 15, and 18, the LCDR2 has an amino acid sequence selected from any one of SEQ ID NOs: 13, 16, and 19, and the LCDR3 has an amino acid sequence selected from any one of SEQ ID NOs: 14, 17, and 20. In particular, the present disclosure provides antibodies that have binding specificity to human IL-17A and comprise the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs: 12, 13, and 14, respectively; or (ii) three complementarity determining regions, LCDR1, LCDR2, and LCDR3, wherein LCDR1 has an amino acid sequence selected from any one of SEQ ID NOs: 50, 53, and 56, LCDR2 has an amino acid sequence selected from any one of SEQ ID NOs: 51, 54, and 57, and LCDR3 has an amino acid sequence selected from any one of SEQ ID NOs: 52, 55, and 58. In particular, the present disclosure provides antibodies that have binding specificity to human IL-17A and comprise the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs: 50, 51, and 52, respectively.

[0452] Suitably, the present disclosure provides an isolated antibody having binding specificity for human IL-17A, comprising a heavy chain variable region (VH) and a light chain variable region (VL),

[0453] (i)

[0454] (a) the VH comprises three complementary determining regions HCDR1, HCDR2, and HCDR3 in sequence, wherein the HCDR1 has an amino acid sequence selected from any one of SEQ ID NOs: 1, 4, and 7, the HCDR2 has an amino acid sequence selected from any one of SEQ ID NOs: 2, 5, and 8, and the HCDR3 has an amino acid sequence selected from any one of SEQ ID NOs: 3, 6, and 9; and

[0455] (b) the VL comprises three complementary determining regions, LCDR1, LCDR2 and LCDR3, in sequence, wherein the LCDR1 has an amino acid sequence selected from any one of SEQ ID NOs: 12, 15 and 18, the LCDR2 has an amino acid sequence selected from any one of SEQ ID NOs: 13, 16 and 19, and the LCDR3 has an amino acid sequence selected from any one of SEQ ID NOs: 14, 17 and 20; or

[0456] (ii)

[0457] (a) the VH comprises three complementary determining regions HCDR1, HCDR2, and HCDR3 in sequence, wherein the HCDR1 has an amino acid sequence selected from any one of SEQ ID NOs: 39, 42, and 45, the HCDR2 has an amino acid sequence selected from any one of SEQ ID NOs: 40, 43, and 46, and the HCDR3 has an amino acid sequence selected from any one of SEQ ID NOs: 41, 44, and 47; and

[0458] (b) the VL includes three complementarity determining regions LCDR1, LCDR2 and LCDR3 in sequence,

[0459] The LCDR1 has an amino acid sequence selected from any one of SEQ ID NOs: 50, 53 and 56, the LCDR2 has an amino acid sequence selected from any one of SEQ ID NOs: 51, 54 and 57, and the LCDR3 has an amino acid sequence selected from any one of SEQ ID NOs: 52, 55 and 58.

[0460] In particular, the present disclosure provides antibodies that have binding specificity to human IL-17A and comprise (i) (a) HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs: 1, 2, and 3, respectively, and (b) LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs: 12, 13, and 14, respectively; or (ii) (a) HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs: 39, 40, and 41, respectively, and (b) LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs: 50, 51, and 52, respectively.

[0461] Other antibodies of the present disclosure include amino acid sequences that have been mutated but still have at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identity in the CDR regions to the CDR regions described in the sequences described in Table 1. Suitably, other antibodies of the present disclosure include mutant amino acid sequences in which no more than 1, 2, 3, 4, 5 or 10 amino acids have been mutated by deletion, insertion or substitution of amino acids in the CDR regions when compared to the CDR regions described in the sequences described in Table 1. Mutations, such as substitutions, may be made at any residue within the CDR group and may be within CDR1, CDR2 and / or CDR3.

[0462] Suitably, the antibodies of the present disclosure comprising the mutated amino acid sequence are capable of inhibiting the activity of 1 ng of human IL-17A by 50% when the antibody concentration is 50 ng / ml, preferably 20 ng / ml, preferably 10 ng / ml, preferably 5 ng / ml, more preferably 1 ng / ml, more preferably 0.5 ng / ml, even more preferably 0.2 ng / ml or less, as determined by measuring human IL-17A-induced GRO-α secretion in the HT-29 assay in the presence of 50 pg / ml TNF.

[0463] Suitably, the isolated antibodies of the present disclosure having binding specificity for human IL-17A comprise: a heavy chain variable region (VH) and a light chain variable region (VL), wherein:

[0464] (i)

[0465] (a) the VH comprises three complementary determining regions HCDR1, HCDR2 and HCDR3 in sequence,

[0466] the HCDR1 having an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to any one of SEQ ID NOs: 1, 4 and 7, preferably SEQ ID NO: 1;

[0467] the HCDR2 having an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to any one of SEQ ID NOs: 2, 5 and 8, preferably SEQ ID NO: 2;

[0468] The HCDR3 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to any one of SEQ ID NOs: 3, 6 and 9, preferably SEQ ID NO: 3; and / or

[0469] (b) the VL comprises three complementary determining regions LCDR1, LCDR2 and LCDR3 in sequence,

[0470] The LCDR1 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to any one of SEQ ID NOs: 12, 15 and 18, preferably SEQ ID NO: 12;

[0471] The LCDR2 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to any one of SEQ ID NOs: 13, 16 and 19, preferably SEQ ID NO: 13;

[0472] The LCDR3 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to any one of SEQ ID NOs: 14, 17 and 20, preferably SEQ ID NO: 14; or

[0473] (ii)

[0474] (a) the VH comprises three complementary determining regions HCDR1, HCDR2 and HCDR3 in sequence,

[0475] the HCDR1 having an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to any one of SEQ ID NOs: 39, 42 and 45, preferably SEQ ID NO: 39;

[0476] the HCDR2 having an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to any one of SEQ ID NOs: 40, 43 and 46, preferably SEQ ID NO: 40;

[0477] The HCDR3 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to any one of SEQ ID NO: 41, 44 and 47, preferably SEQ ID NO: 41; and / or

[0478] (b) the VL comprises three complementary determining regions LCDR1, LCDR2 and LCDR3 in sequence,

[0479] The LCDR1 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to any one of SEQ ID NOs: 50, 53 and 56, preferably SEQ ID NO: 50;

[0480] The LCDR2 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to any one of SEQ ID NOs: 51, 54 and 57, preferably SEQ ID NO: 51;

[0481] The LCDR3 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to any one of SEQ ID NO: 52, 55 and 58, preferably SEQ ID NO: 52.

[0482] Suitably, the isolated antibody of the invention having binding specificity for human IL-17A comprises: (i) a HCDR1, HCDR2 and HCDR3 that are at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to the sequences of SEQ ID NOs: 1, 2 and 3, respectively, and / or a LCDR1, LCDR2 and LCDR3 that are at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to the sequences of SEQ ID NOs: 12, 13 and 14, respectively; or (ii) a HCDR2, HCDR3 that are at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to the sequences of SEQ ID NOs: The sequences of SEQ ID NOs: 39, 40 and 41 have HCDR1, HCDR2 and HCDR3 that are at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical, and / or LCDR1, LCDR2 and LCDR3 that are at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to the sequences of SEQ ID NOs: 50, 51 and 52, respectively.

[0483] In a further embodiment, the present disclosure provides an antibody that specifically binds to human IL-17A, wherein the antibody comprises a VH domain and a VL domain.

[0484] Suitably, the present disclosure provides antibodies that specifically bind to human IL-17A, wherein the antibody comprises VH1A, VH1B, VH3 or VH4. In one embodiment, the isolated antibody of the present disclosure comprises a VH4 domain. In a preferred embodiment, the isolated antibody of the present disclosure comprises a VH3 domain.

[0485] Suitably, the present disclosure provides an isolated antibody that specifically binds to human IL-17A, wherein the antibody comprises a Vκ framework FR1, FR2 and FR3, particularly a Vκ1 or Vκ3 FR1 to FR3, preferably a Vκ1 framework FR1 to FR3, and a framework FR4 selected from the group consisting of a VκFR4, particularly a Vκ1FR4, a Vκ3FR4, and a VλFR4. Suitable VλFR4s are shown in SEQ ID NO: 26 to SEQ ID NO: 32. In one embodiment, the present disclosure provides an isolated antibody that specifically binds to human IL-17A, wherein the antibody comprises a VλFR4 comprising an amino acid sequence selected from any one of SEQ ID NO: 26 to SEQ ID NO: 32, preferably SEQ ID NO: 26 or SEQ ID NO: 27, more preferably SEQ ID NO: 27, having at least 60, 70, 80, 90% identity. Suitably, the present disclosure provides an isolated antibody that specifically binds to human IL-17A, wherein the antibody comprises VλFR4 comprising an amino acid sequence selected from any one of SEQ ID NO: 26 to SEQ ID NO: 32, preferably VλFR4 as shown in SEQ ID NO: 26 or SEQ ID NO: 27, more preferably VλFR4 as shown in SEQ ID NO: 27.

[0486] Thus, in one embodiment, the present disclosure provides an antibody comprising:

[0487] (i)(a) the HCDR1, HCDR2 and HCDR3 sequences of SEQ ID NOs: 1, 2 and 3, respectively, and the LCDR1, LCDR2 and LCDR3 sequences of SEQ ID NOs: 12, 13 and 14, respectively; or (i)(b) the HCDR1, HCDR2 and HCDR3 sequences of SEQ ID NOs: 39, 40 and 41, respectively, and the LCDR1, LCDR2 and LCDR3 sequences of SEQ ID NOs: 50, 51 and 52, respectively;

[0488] (ii) a VH3 or VH4 domain, preferably a VH3 domain; and

[0489] (iii) a VL domain comprising a VL framework comprising Vκ framework FR1, FR2 and FR3, in particular FR1 to FR3 of Vκ1 or Vκ3, preferably Vκ1FR1 to FR3, and a framework FR4 selected from the group consisting of VκFR4, in particular Vκ1FR4, Vκ3FR4, and VλFR4, in particular VλFR4 comprising an amino acid sequence that is at least 60, 70, 80, 90% identical to an amino acid sequence selected from any one of SEQ ID NO: 26 to SEQ ID NO: 32, preferably a VλFR4 as shown in an amino acid sequence selected from any one of SEQ ID NO: 26 to SEQ ID NO: 32, more preferably a VλFR4 as shown in SEQ ID NO: 27.

[0490] Suitably, the present disclosure provides an isolated antibody that specifically binds to human IL-17A, wherein the antibody comprises a VH domain listed in Table 1.

[0491] Suitably, the present disclosure also provides an isolated antibody that specifically binds to human IL-17A, wherein the antibody comprises a VH amino acid sequence listed in Table 1, wherein no more than about 10 amino acids in the framework sequence (e.g., sequences that are not CDRs) have been mutated (wherein, by way of various non-limiting examples, the mutation is an addition, substitution, or deletion).

[0492] Suitably, the present disclosure also provides an isolated antibody that specifically binds to human IL-17A, wherein the antibody comprises a VH amino acid sequence listed in Table 1, wherein no more than about 20 amino acids in the framework sequence (e.g., sequences that are not CDRs) have been mutated (wherein, by way of various non-limiting examples, the mutation is an addition, substitution, or deletion).

[0493] Other antibodies of the present disclosure include amino acid sequences that have been mutated but still have at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identity in the VH region to the VH region depicted in the sequences depicted in Table 1.

[0494] Suitably, the present disclosure provides an isolated antibody that specifically binds to human IL-17A, wherein the antibody comprises a VL domain listed in Table 1. Suitably, the present disclosure also provides an isolated antibody that specifically binds to human IL-17A, wherein the antibody comprises a VL amino acid sequence listed in Table 1, wherein no more than about 10 amino acids in the framework sequence (e.g., sequences that are not CDRs) have been mutated (wherein, by way of various non-limiting examples, the mutation is an addition, substitution or deletion).

[0495] Suitably, the present disclosure also provides an isolated antibody that specifically binds to human IL-17A, wherein the antibody comprises a VL amino acid sequence listed in Table 1, wherein no more than about 20 amino acids in the framework sequence (e.g., sequences that are not CDRs) have been mutated (wherein, by way of various non-limiting examples, the mutation is an addition, substitution, or deletion).

[0496] Other antibodies of the present disclosure include amino acid sequences that have been mutated but still have at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identity in the VL region to the VL region depicted in the sequences depicted in Table 1.

[0497] In one embodiment, the present disclosure provides an isolated antibody that specifically binds to human IL-17A, wherein the antibody comprises a heavy chain variable region comprising an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90%, identical to the amino acid sequence of SEQ ID NO: 10 or SEQ ID NO: 11, preferably SEQ ID NO: 10, and in particular, wherein the antibody comprises the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NO: 1, 2, and 3, respectively. In another embodiment, the present disclosure provides an isolated antibody that specifically binds to human IL-17A, wherein the antibody comprises a heavy chain variable region comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 11, and wherein the heavy chain variable region comprises Q14K, G16E, and G56A (AHo numbering).

[0498] In one embodiment, the present disclosure provides an isolated antibody that specifically binds to human IL-17A, wherein the antibody comprises a heavy chain variable region comprising an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90%, identical to the amino acid sequence of SEQ ID NO: 48 or SEQ ID NO: 49, preferably SEQ ID NO: 48, and in particular, wherein the antibody comprises the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs: 39, 40, and 41, respectively. In another embodiment, the present disclosure provides an isolated antibody that specifically binds to human IL-17A, wherein the antibody comprises a heavy chain variable region comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 49, and wherein the heavy chain variable region comprises R20T and Q141P (AHo numbering).

[0499] In another embodiment, the present disclosure provides an isolated antibody that specifically binds to human IL-17A, wherein the antibody (i) comprises a light chain variable region comprising an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90%, identical to the amino acid sequence of SEQ ID NO: 21 or SEQ ID NO: 22, preferably SEQ ID NO: 21, and in particular, wherein the antibody comprises LCDR1, LCDR2, and LCDR3 of SEQ ID NO: 12, 13, and 14, respectively; or (ii) comprises a light chain variable region comprising an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90%, identical to the amino acid sequence of SEQ ID NO: 59 or SEQ ID NO: 60, preferably SEQ ID NO: 59, and in particular, wherein the antibody comprises SEQ ID NO: 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85 LCDR1, LCDR2 and LCDR3 sequences of NO:50, 51 and 52.

[0500] In yet another embodiment, the present disclosure provides an isolated antibody that specifically binds human IL-17A, wherein the antibody comprises a light chain variable region comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 22, and wherein the light chain variable region comprises A51P (AHo numbering).

[0501] In another embodiment, the present disclosure provides an antibody that specifically binds to human IL-17A, wherein (i) the antibody comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprising an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90%, identical to the amino acid sequence of SEQ ID NO: 10; the light chain variable region comprises an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90% identical to the amino acid sequence of SEQ ID NO: 21; or (ii) the antibody comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprising an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90% identical to the amino acid sequence of SEQ ID NO: NO:48 is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99%, preferably at least 90% identical to an amino acid sequence; the light chain variable region comprises an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99%, preferably at least 90% identical to the amino acid sequence of SEQID NO:59.

[0502] Thus, the present disclosure provides an isolated antibody that specifically binds to human IL-17A, wherein (i) the antibody comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprising an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99%, preferably at least 90%, identical to the amino acid sequence of SEQ ID NO: 10; the light chain variable region comprising an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99%, preferably at least 90% identical to the amino acid sequence of SEQ ID NO: 21, and wherein the antibody comprises the HCDR1, HCDR2 and HCDR3 sequences of SEQ ID NOs: 1, 2 and 3, respectively, and / or the LCDR1, LCDR2 and LCDR3 sequences of SEQ ID NOs: 12, 13 and 14, respectively, preferably wherein the antibody comprises the HCDR1, HCDR2 and HCDR3 sequences of SEQ ID NOs: 13, 14 and 15, respectively. NO: 1, 2 and 3, and LCDR1, LCDR2 and LCDR3 sequences of SEQ ID NO: 12, 13 and 14, respectively; or (ii) the antibody comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprising an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99%, preferably at least 90% identical to the amino acid sequence of SEQ ID NO: 48; the light chain variable region comprising an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99%, preferably at least 90% identical to the amino acid sequence of SEQ ID NO: 59, and wherein the antibody comprises a HCDR1, HCDR2 and HCDR3 sequence of SEQ ID NO: 39, 40 and 41, respectively, and / or a HCDR1, HCDR2 and LCDR3 sequence of SEQ ID NO: 41, respectively. NO: 50, 51 and 52, preferably, wherein the antibody comprises the HCDR1, HCDR2 and HCDR3 sequences of SEQ ID NO: 39, 40 and 41, respectively, and the LCDR1, LCDR2 and LCDR3 sequences of SEQ ID NO: 39, 40 and 41, respectively.

[0503] In another embodiment, the present disclosure provides an isolated antibody that specifically binds to human IL-17A, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90%, identical to the amino acid sequence of SEQ ID NO: 49; and the light chain variable region comprises an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90%, identical to the amino acid sequence of SEQ ID NO: 60, particularly, wherein the heavy chain variable region comprises R20T and Q141P (AHo numbering).

[0504] Thus, the present disclosure provides an isolated antibody that specifically binds to human IL-17A, wherein the antibody comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprising an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99%, preferably at least 90%, identical to the amino acid sequence of SEQ ID NO: 49; the light chain variable region comprising an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99%, preferably at least 90% identical to the amino acid sequence of SEQ ID NO: 60, in particular, wherein the heavy chain variable region comprises R20T and Q141P (AHo numbering), and wherein the antibody comprises the HCDR1, HCDR2 and HCDR3 sequences of SEQ ID NOs: 39, 40 and 41, respectively, and / or the HCDR2 and HCDR3 sequences of SEQ ID NOs: 39, 40 and 41, respectively. NO:50, 51 and 52 LCDR1, LCDR2 and LCDR3 sequences; in particular, wherein the antibody comprises the HCDR1, HCDR2 and HCDR3 sequences of SEQ ID NO:39, 40 and 41, respectively, and the LCDR1, LCDR2 and LCDR3 sequences of SEQ ID NO:50, 51 and 52, respectively.

[0505] In a specific embodiment, the present disclosure provides an isolated antibody that specifically binds to human IL-17A and comprises (i) a VH comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 10 and 11; and / or a VL comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 21 and 22; or (ii) a VH comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 48 and 49; and / or a VL comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 59 and 60. In specific embodiments, the antibodies of the present disclosure comprise (i) the VH sequence of SEQ ID NO: 10 and the VL sequence of SEQ ID NO: 21; or (ii) the VH sequence of SEQ ID NO: 48 and the VL sequence of SEQ ID NO: 59. In yet another specific embodiment, an antibody of the present disclosure comprises (i) the VH sequence of SEQ ID NO: 11 and the VL sequence of SEQ ID NO: 22; or (ii) the VH sequence of SEQ ID NO: 49 and the VL sequence of SEQ ID NO: 60.

[0506] In one embodiment, the antibody that specifically binds to human IL-17A is an antibody described in Table 1. In one embodiment, the antibody that specifically binds to human IL-17A comprises at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90%, of an amino acid sequence selected from the group consisting of (i) SEQ ID NOs: 24 and 25, preferably SEQ ID NO: 24, or (ii) SEQ ID NOs: 61 and 62, preferably SEQ ID NO: 61. In one embodiment, the antibody that specifically binds to human IL-17A is as shown in (i) SEQ ID NO: 24 or SEQ ID NO: 25, preferably SEQ ID NO: 24; or (ii) SEQ ID NO: 61 or SEQ ID NO: 62, preferably SEQ ID NO: 61.

[0507] Other antibodies of the present disclosure that have binding specificity for human IL-17A include those in which the amino acids or nucleic acids encoding the amino acids have been mutated, but have at least 60, 70, 80, 90, or 95% identity with the sequences described in Table 1. In one embodiment, the amino acid sequences include those in which no more than 1, 2, 3, 4, or 5 amino acids have been mutated in the variable region when compared to the variable region described in the sequences described in Table 1, while retaining substantially the same activity. As used herein, the term "substantially the same activity" means that the activity represented by substantially the same activity is at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 98%, or even at least 100%, or at least 110%, or at least 120%, or at least 130%, or at least 140%, or at least 150%, or at least 160%, or at least 170%, or at least 180%, or at least 190%, for example, up to 200% of the activity determined for the parent antibody (e.g., an antibody of the invention, particularly an antibody of the invention described in Table 1).

[0508] In yet another embodiment, the present disclosure provides an antibody comprising an amino acid sequence homologous to the sequence described in Table 1, and said antibody binds to human IL-17A and retains the desired functional properties of those antibodies described in Table 1.

[0509] In one embodiment, an antibody of the present disclosure has a heavy chain variable region comprising CDR1, CDR2, and CDR3 sequences and a light chain variable region comprising CDR1, CDR2, and CDR3 sequences, wherein one or more of these CDR sequences has a specified amino acid sequence based on an antibody described herein or conservative modifications thereof, wherein the antibody retains the desired functional properties of an antibody of the present disclosure.

[0510] Therefore, the present disclosure provides a monoclonal antibody comprising:

[0511] (i)

[0512] A heavy chain variable region (VH), which comprises three complementarity determining regions HCDR1, HCDR2 and HCDR3 in sequence, wherein the HCDR1 is the amino acid sequence of SEQ ID NO: 1, or a conservative variant thereof; the HCDR2 is the amino acid sequence of SEQ ID NO: 2, or a conservative variant thereof; the HCDR3 is an amino acid sequence selected from any one of SEQ ID NO: 3, or a conservative variant thereof; and

[0513] A light chain variable region (VL) comprising three complementarity determining regions LCDR1, LCDR2, and LCDR3, wherein the LCDR1 has the amino acid sequence of SEQ ID NO: 12, or a conservative variant thereof; the LCDR2 has the amino acid sequence of SEQ ID NO: 13, or a conservative variant thereof; the LCDR3 has the amino acid sequence of SEQ ID NO: 14, or a conservative variant thereof; or

[0514] (ii)

[0515] a heavy chain variable region (VH), which comprises three complementarity determining regions HCDR1, HCDR2, and HCDR3, wherein the HCDR1 is the amino acid sequence of SEQ ID NO: 39, or a conservative variant thereof; the HCDR2 is the amino acid sequence of SEQ ID NO: 40, or a conservative variant thereof; and the HCDR3 is an amino acid sequence selected from any one of SEQ ID NO: 41, or a conservative variant thereof; and

[0516] A light chain variable region (VL), which comprises three complementarity determining regions LCDR1, LCDR2 and LCDR3 in sequence, wherein the LCDR1 has the amino acid sequence of SEQ ID NO: 50, or a conservative variant thereof; the LCDR2 has the amino acid sequence of SEQ ID NO: 51, or a conservative variant thereof; the LCDR3 has the amino acid sequence of SEQ ID NO: 52, or a conservative variant thereof,

[0517] The antibody specifically binds to human IL-17A and / or neutralizes IL-17A.

[0518] In one embodiment, an antibody of the present disclosure is optimized for expression in mammalian cells, comprising a heavy chain variable region and a light chain variable region, wherein one or more of these sequences has a specified amino acid sequence based on an antibody described herein or a conservative modification thereof, wherein the antibody retains the desired functional properties of the antibody of the present disclosure. Accordingly, the present invention provides a monoclonal antibody optimized for expression in mammalian cells, comprising a heavy chain variable region and a light chain variable region, wherein: the heavy chain variable region comprises an amino acid sequence selected from the group consisting of: (i) any one of SEQ ID NOs: 10 and 11, and conservative modifications thereof; or (ii) any one of SEQ ID NOs: 48 and 49, and conservative modifications thereof; and the light chain variable region comprises an amino acid sequence selected from the group consisting of: (i) any one of SEQ ID NOs: 21 and 22, and conservative modifications thereof; or (ii) any one of SEQ ID NOs: 59 and 60, and conservative modifications thereof; wherein the antibody specifically binds to human IL-17A and / or neutralizes IL-17A.

[0519] In one embodiment, an antibody of the present disclosure is optimized for expression in mammalian cells, having a full-length heavy chain sequence and a full-length light chain sequence, wherein one or more of these sequences has a specified amino acid sequence based on an antibody described herein or conservative modifications thereof, wherein the antibody retains the desired functional properties of the antibody of the present disclosure.

[0520] As used herein, the term "optimized" refers to a nucleotide sequence that is changed to a codon-encoded amino acid sequence that is preferably used in a production cell or organism (typically a eukaryotic cell, such as a Pichia cell, a Chinese hamster ovary cell (CHO) or a human cell). The optimized nucleotide sequence is engineered to retain the amino acid sequence originally encoded by the starting nucleotide sequence (also referred to as the "parent" sequence) completely or as much as possible. The optimized sequence herein has been modified to have a codon preferred in mammalian cells. However, the optimized expression of these sequences in other eukaryotic cells or prokaryotic cells is also contemplated herein. The amino acid sequence encoded by the optimized nucleotide sequence is also referred to as optimized.

[0521] Another type of variable region modification is to mutate amino acid residues within the VH and / or VL CDR1, CDR2 and / or CDR3 regions to improve one or more binding properties (e.g., affinity) of the target antibody, referred to as "affinity maturation." Site-directed mutagenesis or PCR-mediated mutagenesis can be performed to introduce mutations, and the effects on antibody binding or other desired functions can be assessed in in vitro or in vivo assays as described herein and provided in the Examples. Conservative modifications (as described above) can be introduced. Mutations can be amino acid substitutions, additions, or deletions. In addition, typically no more than 1, 2, 3, 4, or 5 residues are changed in the CDR regions.

[0522] An "affinity matured" antibody is an antibody that has one or more alterations in one or more of its variable domains that result in improved affinity for the antigen compared to a parent antibody that does not have those alterations. In one embodiment, affinity matured antibodies have nanomolar or even picomolar affinities for the target antigen. Affinity matured antibodies are produced by protocols known in the art. For example, Marks et al., Bio / Technology 10:779-783 (1992) describes affinity maturation by shuffling of VH and VL domains. Random mutagenesis of HVR and / or framework residues is described in, for example, Barbas et al. Proc Natl. Acad. Sci. USA 91:3809-3813 (1994); Schier et al. Gene 169:147-155 (1995); Jackson et al., J. Immunol. 154(7):3310-9 (1995); and Hawkins et al., J. Mol. Biol. 226:889-896 (1992). Thus, the present disclosure provides antibodies, wherein the antibodies are affinity matured.

[0523] The antibodies of the present disclosure can be further prepared by the following method: using an antibody having one or more VH and / or VL sequences as described herein as a starting material to engineer a modified antibody that may have different properties from the starting antibody. The antibody can be engineered by modifying one or more residues within one or both variable regions (i.e., VH and / or VL), such as within one or more CDR regions and / or within one or more framework regions. Additionally or alternatively, the antibody can be engineered by modifying residues within the constant region, for example, to alter the effector function of the antibody.

[0524] One type of variable region engineering that can be performed is CDR grafting. Antibodies interact with target antigens primarily through amino acid residues located in the six heavy and light chain complementary determining regions (CDRs). Therefore, the amino acid sequences in the CDRs are more diverse between individual antibodies than the sequences outside the CDRs. Because CDR sequences are responsible for most antibody-antigen interactions, recombinant antibodies that mimic the properties of a specific naturally occurring antibody can be expressed by constructing an expression vector that contains the CDR sequences from that specific naturally occurring antibody and grafted onto framework sequences from a different antibody with different properties (see, e.g., Riechmann, L. et al., 1998 Nature 332:323-327; Jones, P. et al., 1986 Nature 321:522-525; Queen, C. et al., 1989 Proc. Natl. Acad. Sci. USA 86:10029-10033; U.S. Pat. No. 5,225,539 to Winter, and U.S. Pat. Nos. 5,530,101; 5,585,089; 5,693,762 and 6,180,370 to Queen et al.).

[0525] Such framework sequences can be obtained from public DNA databases or published references that include germline antibody gene sequences or rearranged antibody sequences. For example, germline DNA sequences of human heavy and light chain variable region genes can be found in the "VBase" human germline sequence database (available online at www.mrc-cpe.cam.ac.uk / vbase) and in Kabat, EA et al., 1991 Sequences of Proteins of Immunological Interest, Fifth Edition, US Department of Health and Human Services, NIH Publication No. 91-3242; Tomlinson, IM et al., 1992 J. fol. Biol. 227:776-798; and Cox, JPL et al., 1994 Eur. J Immunol. 24:827-836; the contents of each of which are expressly incorporated herein by reference. For example, germline DNA sequences of human heavy and light chain variable region genes and rearranged antibody sequences can be found in the "IMGT" database (available online at www.imgt.org; see Lefranc, MP et al., 1999 Nucleic Acids Res. 27:209-212; the contents of each of which are expressly incorporated herein by reference).

[0526] Examples of framework sequences for use with the antibodies of the present invention are those that are structurally similar to the framework sequences used with the selected antibodies of the present invention, for example, the consensus sequences and / or framework sequences used with the monoclonal antibodies of the present invention. The VH CDR1, 2, and 3 sequences and the VL CDR1, 2, and 3 sequences can be grafted onto framework regions having sequences identical to those found in the germline immunoglobulin genes from which the framework sequences are derived, or the CDR sequences can be grafted onto framework regions that contain one or more mutations compared to the germline sequences. For example, it has been found that, in certain cases, it is beneficial to mutate residues within the framework regions to maintain or enhance the antigen-binding ability of the antibody (see, e.g., U.S. Patent Nos. 5,530,101; 5,585,089; 5,693,762 and 6,180,370 to Queen et al.).

[0527] A variety of antibody / immunoglobulin frameworks or scaffolds can be used, so long as the resulting polypeptide includes at least one binding region that specifically binds to IL-17A. Such frameworks or scaffolds include the five major idiotypes of human immunoglobulins, antigen-binding fragments thereof, and immunoglobulins from other animal species, preferably with humanized characteristics.

[0528] In one aspect, the present disclosure relates to a method for producing non-immunoglobulin-based antibodies using a non-immunoglobulin scaffold onto which the CDRs of the present disclosure can be grafted. Known or future non-immunoglobulin frameworks and scaffolds can be used, as long as they contain binding regions specific for the target IL-17A protein. Known non-immunoglobulin frameworks or scaffolds include, but are not limited to, fibronectin (Compound Therapeutics, Inc., Waltham, Mass.), ankyrin (Molecular Partners AG, Zurich, Switzerland), domain antibodies (Domantis, Ltd., Cambridge, Mass. and Ablynx nv, Zwijnaarde, Belgium), lipocalin (Pieris Proteolab AG, Freising, Germany), small modular immuno-pharmaceuticals (Trubion Pharmaceuticals Inc., Seattle, Wash.), maxybodies (Avidia, Inc., Mountain View, Calif), protein A (Affibody AG, Sweden), and affilin (γ-crystallin or ubiquitin) (Scil Proteins GmbH, Halle, Germany).

[0529] The antibodies according to the present disclosure have valuable properties that are predicted to be beneficial to human patients in need of human IL-17A targeted therapy. The antibodies according to the present disclosure are characterized by having one or more of the following properties (as determined in Examples 1 and 2):

[0530] This antibody specifically binds to human IL-17A and:

[0531] (a) has binding specificity for cynomolgus monkey IL-17A;

[0532] (b) selective binding to human IL-17A compared to human IL-17B, IL-17C, IL-17D, IL-17E, and IL-17F, as measured by ELISA;

[0533] (c) inhibiting or blocking the binding between IL-17A and its receptor (IL-17RA);

[0534] (d) reducing or neutralizing IL-17A activity;

[0535] (e) able to inhibit GRO-α secretion when assessed in an in vitro HT-29 assay;

[0536] (f) having the ability to block the interaction between IL-17A and IL-17RA, with a potency relative to secukinumab (relative potency) of greater than 5, particularly greater than 10, greater than 15, more particularly greater than 20 as determined in an ELISA assay, wherein the relative potency is the IC of secukinumab in ng / mL as measured by ELISA 50 The values ​​are similar to the IC values ​​in ng / mL of the scFv form of the present invention measured by ELISA. 50 The ratio of values;

[0537] (g) having the ability to neutralize IL-17A with a potency relative to secukinumab (relative potency) of greater than 50, particularly greater than 100, more particularly greater than 150 as determined by measuring GRO-α secretion in the HT-29 assay, and wherein the relative potency is the IC of secukinumab in ng / mL as measured in the HT-29 assay 50 The values ​​are comparable to the IC values ​​in ng / mL of the scFv forms of the antibodies of the invention measured in the HT-29 assay. 50 The ratio of values;

[0538] (h) capable of inhibiting the activity of 1 ng of human IL-17A by 50% at a concentration of 1 ng / ml or less, particularly 0.5 ng / ml or less, more particularly 0.2 ng / ml or less, as determined by measuring GRO-α secretion induced by human IL-17A in the presence of 50 pg / ml TNFα in a HT-29 assay;

[0539] (i) Binding to human IL-17A, dissociation constant (K D ) less than 5 nM, in particular less than 1 nM, less than 0.5 nM, less than 0.2 nM, in particular less than 100 pM, more in particular less than 50 pM, as measured by surface plasmon resonance, preferably by surface plasmon resonance in a direct setting;

[0540] (j) Binds to cynomolgus monkey IL-17A, K D less than 10 nM, such as less than 7 nM, less than 5 nM, less than 2 nM, less than 1 nM, in particular less than 0.5 nM, as measured by surface plasmon resonance, in particular as measured by surface plasmon resonance in a trapping setup;

[0541] (k) when in scFv format, a melting temperature (Tm) of at least 60°C, particularly at least 62°C, more particularly at least 65°C, even more particularly at least 70°C, as determined by differential scanning fluorimetry, particularly wherein the antibody is in phosphate-citrate buffer, pH 6.4, 150 mM NaCl;

[0542] (1) when in scFv format, the antibody of the invention loses less than 5%, particularly less than 3%, more particularly less than 1% of its monomer content after 5 consecutive freeze-thaw cycles when the starting concentration is 10 mg / ml, particularly wherein the antibody is in phosphate buffered saline (PBS) at pH 7.4; and / or

[0543] (m) when in scFv format, the antibody of the invention has a monomer content loss of 5% or less, particularly less than 4%, less than 3%, less than 2%, more particularly less than 1%, after storage at 4°C for at least two weeks, particularly at least 4 weeks, when the initial concentration is 10 mg / ml, particularly wherein the antibody is in phosphate buffered saline (PBS) at pH 7.4; and / or

[0544] (n) When the initial concentration of the antibody of the present invention is 10 mg / ml, the monomer content loss is less than 5% after storage at 37°C for at least two weeks, particularly at least four weeks.

[0545] In one embodiment, the antibodies of the present disclosure selectively bind to human IL-17A relative to human IL-17B, IL-17C, IL-17D, IL-17E, and IL-17F, as measured by ELISA. As used herein, the term "selectively binds" means that the antibody, composition, formulation, etc. does not significantly bind to IL-17B / C / D / E / F, but does bind to IL-17A. Selective binding is characterized by high affinity (or low K D ) and low to moderate IC 50 , as distinguished from nonspecific binding, which typically has low affinity and moderate to high IC 50 Typically, when the antibody binds to the K D Less than 10 -7 Suitably, the antibodies of the present disclosure bind to human IL-17A with a higher affinity or K than they bind to human IL-17B, IL-17C, IL-17D, IL-17E and IL-17F. D Suitably, the IC values ​​of the antibodies of the present disclosure for IL-17B, IL-17C, IL-17D, IL-17E and IL-17F are lower than those for IL-17B, IL-17C, IL-17D, IL-17E and IL-17F. 50 Comparison of IC of IL-17A 50More than 100 times greater, particularly more than 200 times greater, more than 300 times greater, more than 400 times greater, as determined by ELISA.

[0546] The antibodies of the present disclosure specifically bind to IL-17A, wherein binding to IL-17A (a) inhibits or blocks the binding between IL-17A and its receptor (IL-17RA), and (b) reduces or neutralizes the activity of IL-17A.

[0547] As used herein, the term "neutralizing antibody" describes an antibody that is capable of neutralizing the biosignaling activity of IL-17A, for example, by blocking the binding of IL-17A to one or more of its receptors, particularly by blocking the binding of IL-17A to IL-17RA. The antibodies disclosed herein are IL-17A neutralizing antibodies. It will be understood that the term "neutralizing" as used herein refers to a reduction in biosignaling activity, which may be partial or complete. Neutralization of IL-17A can be determined using various assays, examples of which are described elsewhere herein.

[0548] Thus, the antibodies of the present disclosure are capable of inhibiting the secretion of GRO-α when assessed in vitro in the HT-29 assay (described in Examples 1 and 2). In one embodiment, the antibodies of the present disclosure have the ability to neutralize IL-17A with a potency relative to secukinumab (relative potency) as determined by measuring GRO-α secretion in the HT-29 assay of greater than 50, preferably greater than 100, more preferably greater than 150, and wherein the relative potency is the IC of secukinumab in ng / mL as measured in the HT-29 assay. 50 The values ​​are comparable to the IC values ​​in ng / mL for the scFv forms of the disclosed antibodies measured in the HT-29 assay. 50 In another embodiment, the antibodies of the present disclosure are capable of inhibiting the activity of 1 ng of human IL-17A by 50% at a concentration of 1 ng / ml or less, preferably 0.5 ng / ml or less, more preferably 0.2 ng / ml or less, as determined by measuring GRO-α secretion induced by human IL-17A in the HT-29 assay in the presence of 50 pg / ml TNFα.

[0549] In one embodiment, the antibodies of the present disclosure have the ability to block the interaction between IL-17A and IL-17RA with a potency relative to secukinumab (relative potency) of greater than 5, preferably greater than 10, greater than 15, more particularly greater than 20 as determined in an ELISA assay, and wherein the relative potency is the IC of secukinumab in ng / mL as measured by ELISA. 50 The values ​​are comparable to the IC values ​​in ng / mL of the disclosed antibodies in scFv form as measured by ELISA. 50The ratio of values.

[0550] As used herein, the term "affinity" refers to the strength of the interaction between an antibody and an antigen at a single antigenic site. Within each antigenic site, the variable regions of the antibody "arms" interact with the antigen at many sites through weak non-covalent forces; the greater the interaction, the stronger the affinity.

[0551] "Binding affinity" generally refers to the strength of the sum of non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless otherwise indicated, as used herein, "binding affinity" or "binding to" refers to intrinsic binding affinity that reflects a 1:1 interaction between members of a binding pair (e.g., an antibody fragment and an antigen). The affinity of a molecule X for its partner Y can generally be expressed in terms of the dissociation constant (K D ) represents. Affinity can be measured by conventional methods known in the art, including the methods described herein. Low-affinity antibodies generally bind antigen slowly and tend to dissociate easily, while high-affinity antibodies generally bind antigen faster and tend to remain bound longer. Various methods for measuring binding affinity are known in the art, any of which can be used for the purposes of the present disclosure. Specific illustrative and exemplary embodiments for measuring binding affinity, i.e., binding strength, are described below.

[0552] The term "K assoc ”, “Ka” or “K on ” refers to the on-rate of a particular antibody-antigen interaction, and the term “K dis ”, “Kd” or “K off " refers to the off-rate of a particular antibody-antigen interaction. In one embodiment, the term "K" as used herein D " refers to the dissociation constant, which is obtained from the ratio of Kd to Ka (i.e., Kd / Ka) and is expressed as a molar concentration (M). In one embodiment, the "Kd" according to the present disclosure is D ” or “K D"KD" or "KD value" was measured using a T200 device (Biacore, GE Healthcare). To measure the affinity of humanized scFv to human IL-17A, biotinylated human IL-17A was captured using Biacore's Biotin-CAPture kit. After each analyte injection cycle, the CAP sensor chip was regenerated and the neoantigen was captured. scFv was injected as analyte using a dose-response multi-cycle kinetic assay with an analyte concentration range of 0.35 to 90 nM diluted in running buffer. The obtained sensorgrams were fitted using a 1:1 binding model. Alternatively, the affinity of the humanized scFv can be measured and analyzed using an alternative SPR detection setup: IL-17A was immobilized on a CM5 sensor chip (GE Healthcare) by amine coupling, and serial dilutions of the scFv were injected over the immobilized IL-17A at concentrations ranging from 0.35 to 90 nM.

[0553] Suitably, the antibodies of the present disclosure may have a higher affinity for IL-17A than the affinity of IL-17A for IL-17RA. It will be appreciated that the higher affinity of the antibodies of the present disclosure compared to the affinity of IL-17A for IL-17RA may be particularly useful for dissociating or neutralizing pre-formed IL-17RA / IL-17A complexes. In one embodiment, the antibodies of the present disclosure neutralize the IL-17RA / IL-17A interaction. In another embodiment, the antibodies of the present disclosure inhibit or block the binding between IL-17A and its receptor (IL-17RA). In one embodiment, the antibodies of the present disclosure neutralize the activity of IL-17A.

[0554] Suitably, the affinity of the antibodies of the present disclosure for IL-17A can be comparable to or greater than, and preferably greater than, the affinity of secukinumab for IL-17A. In one embodiment, the antibodies of the present disclosure neutralize IL-17A activity with potency equal to or greater than, and preferably greater than, secukinumab. In another embodiment, the antibodies of the present disclosure neutralize IL-17RA / IL-17A interaction with potency equal to or greater than, and preferably greater than, secukinumab.

[0555] The binding affinity of an antibody can be measured, for example, by the dissociation constant (K D ) OK. Lower K D indicates stronger affinity, while higher K D Indicates weaker affinity.

[0556] Thus, in one suitable embodiment, the K of an antibody of the present disclosure is D1 to 10000 pM, 1 to 7000 pM, 1 to 5000 pM, 1 to 2500 pM, 1 to 2000 pM, 1 to 1000 pM, 1 to 750 pM, 1 to 500 pM, 1 to 400 pM, 1 to 300 pM, 1 to 200 pM, 1 to 100 pM, 1 to 50 pM, preferably as measured by surface plasmon, more preferably by surface plasmon resonance in a direct setting. In a suitable embodiment, the K of an antibody of the present disclosure is D In a specific embodiment, the K of an antibody of the present disclosure is 1 to 200 pM, in particular 1 to 100 pM, as measured by surface plasmon resonance in a direct setting. D In a suitable embodiment, the K of an antibody of the present disclosure is 1 to 50 pM, as measured by surface plasmon resonance in a direct setting. D The K of an antibody of the present disclosure may be less than 5 nM, less than 4 nM, less than 3 nM, less than 2 nM, less than 1 nM, less than 0.5 nM, less than 0.4 nM, less than 0.3 nM, less than 0.25 nM, less than 0.2 nM, less than 150 pM, less than 100 pM, or less than 50 pM, preferably as measured by surface plasmon resonance, more preferably as measured by surface plasmon resonance in a direct setting. Suitably, the K of an antibody of the present disclosure may be less than 5 nM, less than 4 nM, less than 3 nM, less than 2 nM, less than 1 nM, less than 0.5 nM, less than 0.4 nM, less than 0.3 nM, less than 0.25 nM, less than 0.2 nM, less than 150 pM, less than 100 pM, or less than 50 pM, preferably as measured by surface plasmon resonance, more preferably as measured by surface plasmon resonance in a direct setting. D less than 1 nM, particularly less than 100 pM. Suitably, the K D Less than 0.5 nM, particularly less than 50 pM. More suitably, the K D Less than 0.2 nM, in particular less than 50 pM, as measured by surface plasmon resonance in a direct setting.

[0557] In another embodiment, the antibodies of the present disclosure bind to cynomolgus monkey IL-17A, K D Less than 10 nM, such as less than 7 nM, less than 5 nM, less than 2 nM, less than 1 nM, in particular less than 0.5 nM as measured by surface plasmon resonance (SPR) in a trapping setup.

[0558] Suitably, the antibodies of the present disclosure bind to human IL-17A, K on The rate is at least 10 3 M -1 s -1 or higher by at least 10 4 M - 1 s -1 or higher at least 5x10 4 M -1 s -1 or higher by at least 10 5 M -1 s-1 or higher at least 5x10 5 M -1 s -1 or higher by at least 10 6 M -1 s -1 or higher, as measured by surface plasmon resonance (SPR), preferably as measured by surface plasmon resonance in a direct setting. Preferably, the K of the antibodies of the present disclosure is on The rate is at least 10 5 M -1 s -1 or higher, especially at least 5x10 5 M -1 s -1 or higher, especially at least 10 6 M -1 s -1 or higher, as measured by SPR, preferably as measured by surface plasmon resonance in a direct setup.

[0559] Suitably, the antibodies of the present disclosure bind to human IL-17A, K off The rate is 5x10 -3 s -1 or lower 3x10 -3 s -1 or lower 10 -3 s -1 or lower 5x10 -4 s -1 or lower 3x10 -4 s -1 or lower 10 -4 s -1 or lower 5x10 -5 s -1 or lower, as measured by surface plasmon resonance (SPR), preferably as measured by surface plasmon resonance in a direct setup. Preferably, the K of the antibodies of the present disclosure is off Rate is 10 -4 s -1 or lower, especially 5x10 -5 s -1 or lower, as measured by SPR, preferably as measured by surface plasmon resonance in a direct setup.

[0560] Suitably, the antibodies of the present disclosure have favorable biophysical properties.

[0561] Suitably, when expressed in the form of a scFv (single chain variable fragment) antibody, the melting temperature (Tm) of the antibodies of the present disclosure is at least 60°C, preferably at least 62°C, more preferably at least 65°C, even more preferably at least 70°C, as determined by differential scanning fluorimetry (DSF), as previously described (Egan et al., MAbs, 9(1)(2017), 68-84; Niesen et al., Nature Protocols, 2(9)(2007) 2212-2221). The transition midpoint of the thermal unfolding of the scFv construct is determined by differential scanning fluorimetry using a fluorescent dye Orange determined (see Wong & Raleigh, Protein Science 25 (2016) 1834-1840). Samples were prepared in citrate phosphate buffer at pH 6.4 at a final protein concentration of 50 μg / mL and contained 5x The final concentration of Orange was 100 μl in a total volume of 100 μl. 25 microliters of the prepared sample was added in triplicate to a white-walled AB gene PCR plate. The analysis was performed in a qPCR machine used as a thermal cycler, and fluorescence emission was detected using the software's custom dye calibration program. The PCR plate containing the test samples was temperature-changed from 25°C to 96°C in increments of 1°C, with a 30s pause after each temperature increase. The total assay time was approximately two hours. Tm was calculated by calculating the inflection point of the curve using the mathematical second derivative method using GraphPad Prism software. The reported Tm is the average of the three measurements.

[0562] The antibodies of the present disclosure, particularly when expressed as scFv (single-chain variable fragment) antibodies, are characterized by a loss of monomer content of less than 5%, particularly less than 3%, and more particularly less than 1%, after five consecutive freeze-thaw cycles when the starting concentration of the antibodies of the present disclosure is 10 mg / ml.

[0563] After storage at 4°C for at least two weeks, and in particular for at least four weeks, the antibodies of the present disclosure, in particular when expressed in the form of scFv (single-chain variable fragment) antibodies, are characterized by a monomer content loss of 5% or less, in particular less than 4%, less than 3%, less than 2%, and preferably less than 1%, when the starting concentration of the antibodies of the present invention is 10 mg / ml. The loss of monomer content is determined by calculating the area under the curve of the SE-HPLC chromatogram. SE-HPLC is a separation technique based on a solid stationary phase and a liquid mobile phase, as described in USP Chapter 621. The method utilizes a hydrophobic stationary phase and an aqueous mobile phase to separate molecules based on their size and shape. The separation of molecules occurs between the void volume (V0) and the total permeate volume (VT) of a specific column. SE-HPLC measurements were performed on a Chromaster HPLC system (Hitachi High-Technologies Corporation) equipped with automatic sample injection and a UV detector with a detection wavelength set to 280 nm. The device is controlled by the software EZChrom Elite (Agilent Technologies, Version 3.3.2SP2), which also supports the analysis of the obtained chromatograms. Protein samples were cleared by centrifugation before injection, and the temperature of the autosampler was maintained at 4-6°C. For the analysis of scFv samples, a Shodex KW403-4F column (Showa Denko Co., Ltd., #F6989202) was used, with a standard buffered saline mobile phase (50mM sodium phosphate pH 6.5, 300mM sodium chloride) and a recommended flow rate of 0.35mL / min. The target sample loading amount for each injection was 5μg. The samples were detected by a UV detector at a wavelength of 280nm, and the data were recorded by a suitable software suite. The obtained chromatograms were analyzed in the range of V0 to VT, thereby excluding matrix-related peaks with elution times>10 minutes.

[0564] Suitably, the isolated antibodies of the present disclosure are selected from the group consisting of monoclonal antibodies, chimeric antibodies, Fab, Fv, scFv, dsFv, scAb, STAB, single domain antibodies (sdAb or dAb), single domain heavy chain antibodies, and single domain light chain antibodies, VHH, VNAR, single domain antibodies based on the VNAR structure of sharks, and binding domains based on alternative scaffolds including but not limited to ankyrin-based domains, fynomers, avimers, anticalins, fibronectin, and binding sites built into antibody constant regions (e.g., Modular Antibody Technology of F-star). TM ), preferably scFv.

[0565] Suitably, the antibodies of the present disclosure are Fv. Suitably, the antibodies of the present disclosure are scFv antibody fragments. A "single-chain Fv" or "scFv" or "sFv" antibody fragment comprises the VH and VL domains of an antibody, wherein these domains are present in a single polypeptide chain. Typically, the Fv polypeptide further comprises a polypeptide linker between the VH and VL domains that enables the sFv to form a desired target binding structure. A "single-chain Fv" or "scFv" antibody fragment comprises the VH and VL domains of an antibody, wherein these domains are present in a single polypeptide chain. Typically, the scFv polypeptide further comprises a polypeptide linker between the VH and VL domains that enables the scFv to form a desired antigen binding structure (see, e.g., Plückthun, The Pharmacology of Monoclonal Antibodies, Vol. 113, Rosenburg and Moore, eds., (Springer-Verlag, New York, 1994), pp. 269-315). In a specific embodiment, the functional fragment is in the form of an scFv comprising a linker according to SEQ ID NO: 23. In one embodiment, the antibody that specifically binds to human IL-17A is an antibody described in Table 1. In one embodiment, the antibody that specifically binds to human IL-17A comprises an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99%, preferably at least 90%, identical to an amino acid sequence selected from (i) the group consisting of SEQ ID NO: 24 and SEQ ID NO: 25; or (ii) the group consisting of SEQ ID NO: 61 and SEQ ID NO: 62. In another embodiment, the antibody of the present disclosure is (i) a single chain variable fragment (scFv) as shown in SEQ ID NO: 24 or SEQ ID NO: 25; or (ii) a single chain variable fragment (scFv) as shown in SEQ ID NO: 61 or SEQ ID NO: 62.

[0566] Suitably, the antibodies of the present disclosure are IgG antibodies. In one embodiment, the antibodies of the present disclosure are IgG selected from the group consisting of IgG1, IgG2, IgG3 and IgG4, preferably IgG1.

[0567] Exemplary domains that specifically bind TNFα

[0568] The multispecific antibodies of the present disclosure comprise a second domain that specifically binds to TNFα, wherein the domain comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein: (a) the VH comprises three complementarity determining regions HCDR1, HCDR2 and HCDR3 in sequence, and (b) the VL comprises three complementarity determining regions LCDR1, LCDR2 and LCDR3 in sequence.

[0569] Suitable domains that specifically bind to TNFα for use in the multispecific antibodies of the present disclosure include, but are not limited to:

[0570] A humanized monoclonal antibody or a binding domain thereof, the sequences of which are listed in Table 1 (described in WO2017 / 158101, which is incorporated herein by reference in its entirety);

[0571] ·incixImab( U.S. Patent Nos. 6,277,969, 6,284,471, 6,790,444, and 6,835,823, all of which are incorporated herein by reference);

[0572] adalimumab / D2E7( described in U.S. Patent No. 6,090,382, which is incorporated herein by reference in its entirety);

[0573] certolizumab, a PEGylated Fab fragment ( described in US 7,012,135 and US 7,186,820, all of which are incorporated herein by reference);

[0574] ·golimumab( Disclosed in U.S. Application 2009 / 214528, which is incorporated herein by reference in its entirety).

[0575] Preferred domains that specifically bind to TNFα for use in the multispecific antibodies of the present disclosure include, but are not limited to, humanized monoclonal antibodies or their binding domains, the sequences of which are listed in Table 1 (described in WO 2017 / 158101, which is incorporated herein by reference in its entirety).

[0576] Thus, in one embodiment, the present disclosure provides a multispecific antibody comprising a first domain that specifically binds to IL-17A and a second domain that specifically binds to TNFα, wherein the second domain comprises a set of CDRs: HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3,

[0577] wherein the set of CDRs has 10 or fewer amino acid substitutions, e.g., 9 or fewer amino acid substitutions, 8 or fewer amino acid substitutions, 7 or fewer amino acid substitutions, 6 or fewer amino acid substitutions, 5 or fewer amino acid substitutions, 4 or fewer amino acid substitutions, 3 or fewer amino acid substitutions, 2 or fewer amino acid substitutions, 1 or 0 amino acid substitutions, preferably 0 amino acid substitutions, relative to a set of CDRs: HCDR1′ is an amino acid sequence selected from any one of SEQ ID NOs: 25, 28, and 31, preferably SEQ ID NO: 25; HCDR2′ is an amino acid sequence selected from any one of SEQ ID NOs: 26, 29, and 32, preferably SEQ ID NO: 26; HCDR3′ is an amino acid sequence selected from any one of SEQ ID NOs: 27, 30, and 33, preferably SEQ ID NO: 27; LCDR1′ is an amino acid sequence selected from any one of SEQ ID NOs: 38, 41, and 44, preferably SEQ ID NO: 38; LCDR2′ is an amino acid sequence selected from any one of SEQ ID NOs: 39, 41, and 45, preferably SEQ ID NO: 46; NO: 39, 42 and 45, preferably SEQ ID NO: 39; and LCDR3' has an amino acid sequence selected from any one of SEQ ID NO: 40, 43 and 46, preferably SEQ ID NO: 40.

[0578] In particular, the present disclosure provides a multispecific antibody comprising a first domain that specifically binds to IL-17A and a second domain that specifically binds to TNFα, wherein the second domain comprises a VH CDR having the amino acid sequence of any one of the VH CDRs listed in Table 1. In particular, the second domain that specifically binds to TNFα comprises (or alternatively consists of) one, two, three or more VH CDRs having the amino acid sequence of any one of the VH CDRs listed in Table 1.

[0579] Suitably, the present disclosure provides a second domain that specifically binds to TNFα, wherein the second domain comprises a heavy chain variable region (VH), wherein the VH comprises three complementarity determining regions HCDR1, HCDR2 and HCDR3 in sequence, the HCDR1 having an amino acid sequence selected from any one of SEQ ID NOs: 25, 28 and 31, the HCDR2 having an amino acid sequence selected from any one of SEQ ID NOs: 26, 29 and 32, and the HCDR3 having an amino acid sequence selected from any one of SEQ ID NOs: 27, 30 and 33. In particular, the second domain comprises the HCDR1, HCDR2 and HCDR3 sequences of SEQ ID NOs: 25, 26 and 27, respectively.

[0580] The present disclosure also provides a multispecific antibody comprising a first domain that specifically binds to IL-17A and a second domain that specifically binds to TNFα, wherein the second domain comprises a VL CDR having the amino acid sequence of any one of the VL CDRs listed in Table 1. In particular, the second domain that specifically binds to TNFα comprises (or alternatively consists of) one, two, three or more VL CDRs having the amino acid sequence of any one of the VL CDRs listed in Table 1.

[0581] Suitably, the second domain that specifically binds to TNFα comprises a light chain variable region (VL), wherein the VL comprises three complementary determining regions LCDR1, LCDR2 and LCDR3 in sequence, the LCDR1 having an amino acid sequence selected from any one of SEQ ID NOs: 38, 41 and 44, the LCDR2 having an amino acid sequence selected from any one of SEQ ID NOs: 39, 42 and 45, and the LCDR3 having an amino acid sequence selected from any one of SEQ ID NOs: 40, 43 and 46. In particular, the second domain comprises the LCDR1, LCDR2 and LCDR3 sequences of SEQ ID NOs: 38, 39 and 40, respectively.

[0582] Suitably, the present disclosure provides a multispecific antibody comprising a first domain that specifically binds to IL-17A and a second domain that specifically binds to TNFα, wherein the second domain comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein:

[0583] (a) the VH comprises three complementary determining regions HCDR1, HCDR2, and HCDR3 in sequence, wherein the HCDR1 has an amino acid sequence selected from any one of SEQ ID NOs: 25, 28, and 31, the HCDR2 has an amino acid sequence selected from any one of SEQ ID NOs: 26, 29, and 32, and the HCDR3 has an amino acid sequence selected from any one of SEQ ID NOs: 27, 30, and 33; and

[0584] (b) the VL comprises three complementarity determining regions LCDR1, LCDR2 and LCDR3 in sequence, wherein the LCDR1 has an amino acid sequence selected from any one of SEQ ID NOs: 38, 41 and 44, the LCDR2 has an amino acid sequence selected from any one of SEQ ID NOs: 39, 42 and 45, and the LCDR3 has an amino acid sequence selected from any one of SEQ ID NOs: 40, 43 and 46.

[0585] In particular, the present disclosure provides a multispecific antibody comprising a first domain that specifically binds to IL-17A and a second domain that specifically binds to TNFα, wherein the second domain comprises (a) HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs: 25, 26, and 27, respectively, and (b) LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs: 38, 39, and 40, respectively.

[0586] Other domains of the present disclosure that specifically bind to TNFα include amino acid sequences that have been mutated but still have at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identity in the CDR regions to the CDR regions described in the sequences described in Table 1. Suitably, other domains of the present disclosure that specifically bind to TNFα include mutant amino acid sequences in which no more than 1, 2, 3, 4, 5, or 10 amino acids have been mutated by deletion, insertion, or substitution of amino acids in the CDR regions when compared to the CDR regions described in the sequences described in Table 1. Mutations, such as substitutions, may be made at any residue within the CDR group and may be within CDR1, CDR2, and / or CDR3.

[0587] Suitably, the second domain that specifically binds to TNFα of the multispecific antibody of the present disclosure comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein:

[0588] (a) the VH comprises, in order, three complementarity determining regions: HCDR1, HCDR2, and HCDR3, wherein the HCDR1 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to any one of SEQ ID NOs: 25, 28, and 31; the HCDR2 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to any one of SEQ ID NOs: 26, 29, and 32; and the HCDR3 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to any one of SEQ ID NOs: 27, 30, and 33; and / or

[0589] (b) the VL comprises three complementary determining regions, LCDR1, LCDR2 and LCDR3, in sequence, wherein the LCDR1 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to any one of SEQ ID NOs: 38, 41 and 44; the LCDR2 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to any one of SEQ ID NOs: 39, 42 and 45; and the LCDR3 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to any one of SEQ ID NOs: 40, 43 and 46.

[0590] Suitably, the second domain that specifically binds to TNFα comprises: a HCDR1 , a HCDR2 and a HCDR3 that are at least 60, 70, 80, 90, 91 , 92, 93, 94, 95, 96, 97, 98 or 99% identical to the sequences of SEQ ID NOs: 25, 26 and 27, respectively, and / or a LCDR1 , a LCDR2 and a LCDR3 that are at least 60, 70, 80, 90, 91 , 92, 93, 94, 95, 96, 97, 98 or 99% identical to the sequences of SEQ ID NOs: 38, 39 and 40, respectively.

[0591] In a further embodiment, the second domain that specifically binds to TNFα comprises a heavy chain variable region VHB and a light chain variable region VLB.

[0592] Suitably, the present disclosure provides a multispecific antibody comprising a first domain that specifically binds to IL-17A and a second domain that specifically binds to TNFα, wherein the second domain comprises a heavy chain variable region VHB, wherein the VHB is VH1A, VH1B, VH3, or VH4. In one embodiment, the second domain that specifically binds to TNFα of the present disclosure comprises a heavy chain variable region VHB, wherein the VHB is VH4. In a preferred embodiment, the second domain that specifically binds to TNFα of the present disclosure comprises a heavy chain variable region VHB, wherein the VHB is VH3.

[0593] Suitably, the present disclosure provides a multispecific antibody comprising a first domain that specifically binds to IL-17A and a second domain that specifically binds to TNFα, wherein the second domain comprises a light chain variable region, VLB, and wherein the VLB comprises a Vκ framework FR1, FR2, and FR3, particularly FR1 to FR3 of Vκ1 or Vκ3, preferably Vκ1 framework FR1 to FR3, and a framework FR4 selected from the group consisting of VκFR4, particularly Vκ1FR4, Vκ3FR4, and VλFR4. Suitable VλFR4s are shown in SEQ ID NO: 97 to SEQ ID NO: 103. In one embodiment, the second domain that specifically binds to TNFα comprises a VλFR4 comprising an amino acid sequence selected from any one of SEQ ID NO: 97 to SEQ ID NO: 103, preferably SEQ ID NO: 97 or SEQ ID NO: 98, more preferably SEQ ID NO: 97, having at least 60, 70, 80, 90% identity. Suitably, the second domain that specifically binds to TNFα comprises VλFR4, which comprises an amino acid sequence selected from any one of SEQ ID NO: 97 to SEQ ID NO: 103, preferably, VλFR4 is as shown in SEQ ID NO: 97 or 98, more preferably, VλFR4 is as shown in SEQ ID NO: 98.

[0594] Thus, in one embodiment, the present disclosure provides a multispecific antibody comprising a first domain that specifically binds IL-17A and a second domain that specifically binds TNFα, wherein the second domain comprises:

[0595] (a) the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs: 25, 26, and 27, respectively, and the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs: 38, 39, and 40, respectively;

[0596] (b) VHB, wherein the VHB is VH3 or VH4, preferably VH3; and

[0597] (c) VLB, which comprises a VL framework, said VL framework comprising Vκ framework FR1, FR2 and FR3, in particular FR1 to FR3 of Vκ1 or Vκ3, preferably Vκ1FR1 to FR3, and a framework FR4, said framework FR4 being selected from VκFR4, in particular Vκ1FR4, Vκ3FR4, and VλFR4, in particular VλFR4 comprising an amino acid sequence having at least 60, 70, 80, 90% identity with an amino acid sequence selected from any one of SEQ ID NO: 97 to SEQ ID NO: 103, preferably VλFR4 as shown in any one of SEQ ID NO: 97 to SEQ ID NO: 103, preferably VλFR4 as shown in SEQ ID NO: 97 or 98, more preferably VλFR4 as shown in SEQ ID NO: 98.

[0598] Suitably, the present disclosure provides a multispecific antibody comprising a first domain that specifically binds to IL-17A and a second domain that specifically binds to TNFα, wherein the second domain comprises a VH listed in Table 1. Suitably, the present disclosure also provides a multispecific antibody comprising a first domain that specifically binds to IL-17A and a second domain that specifically binds to HSA, wherein the second domain comprises (or alternatively consists of) a VH amino acid sequence listed in Table 1, wherein no more than about 20 amino acids in the framework sequence (e.g., a sequence that is not a CDR) have been mutated (wherein, as various non-limiting examples, the mutation is an addition, substitution, or deletion), preferably no more than about 10. Other domains of the present disclosure that specifically bind to TNFα include amino acid sequences that have been mutated but still have at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identity in the VH region to the VH region described in the sequences described in Table 1.

[0599] Suitably, the present disclosure provides a multispecific antibody comprising a first domain that specifically binds to IL-17A and a second domain that specifically binds to TNFα, wherein the second domain comprises a VL domain listed in Table 1. Suitably, the present disclosure also provides a multispecific antibody comprising a first domain that specifically binds to IL-17A and a second domain that specifically binds to TNFα, wherein the second domain comprises (or alternatively consists of) a VL amino acid sequence listed in Table 1, wherein no more than about 20 amino acids in the framework sequence (e.g., a sequence that is not a CDR) have been mutated (wherein, as various non-limiting examples, the mutation is an addition, substitution, or deletion), preferably no more than about 10 amino acids. Other domains of the present disclosure that specifically bind to TNFα include amino acid sequences that have been mutated but still have at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identity in the VL region to the VL region described in the sequences described in Table 1.

[0600] In one embodiment, the present disclosure provides a multispecific antibody comprising a first domain that specifically binds IL-17A and a second domain that specifically binds TNFα, wherein the second domain comprises a heavy chain variable region comprising an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99%, preferably at least 90%, identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 34, 35, 36 and 37, preferably SEQ ID NO: 34, and in particular, wherein the domains include the HCDR1, HCDR2 and HCDR3 sequences of SEQ ID NOs: 25, 26 and 27, respectively. In a preferred embodiment, the second domain that specifically binds to TNFα comprises a heavy chain variable region comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 34, and wherein the heavy chain variable region comprises G56A, R82L, S85A, K86Q (AHo numbering), and in particular, wherein the domain comprises the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs: 25, 26, and 27, respectively. Suitably, the second domain that specifically binds to TNFα may comprise a heavy chain variable region comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 36, and wherein the heavy chain variable region comprises A24K, G56A, R82L (AHO numbering), in particular, wherein the domain comprises the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs: 25, 26, and 27, respectively. Suitably, the second domain that specifically binds to TNFα may comprise a heavy chain variable region comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 37, and wherein the heavy chain variable region comprises G56A, R82L, S85A, K86Q (AHo numbering), and in particular, wherein the domain includes the HCDR1, HCDR2 and HCDR3 sequences of SEQ ID NOs: 25, 26 and 27, respectively.

[0601] In another embodiment, the present disclosure provides a multispecific antibody comprising a first domain that specifically binds IL-17A and a second domain that specifically binds TNFα, wherein the second domain comprises a light chain variable region comprising an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99%, preferably at least 90%, identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 47, 48, 49, 50 and 51, preferably SEQ ID NO: 47, and in particular, wherein the domains include LCDR1, LCDR2 and LCDR3 sequences of SEQ ID NOs: 38, 39 and 40, respectively.

[0602] In a preferred embodiment, the second domain that specifically binds to TNFα may comprise a light chain variable region comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 47, and wherein the light chain variable region comprises T22N, A51R, F89Y (AHO numbering), in particular, wherein the domain comprises the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs: 38, 39, and 40, respectively. Suitably, the second domain that specifically binds to TNFα may comprise a light chain variable region comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 49, and wherein the light chain variable region comprises T22N, D88E, S95G, E99A (AHO numbering), in particular, wherein the domain LCDR1, LCDR2, and LCDR3 sequences are SEQ ID NOs: 38, 39, and 40, respectively. Suitably, the second domain that specifically binds to TNFα may comprise a light chain variable region comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 50, and wherein the light chain variable region comprises T22N, A51R, F89Y, S95G, G141T, L145V (AHO numbering), in particular, wherein the domains LCDR1, LCDR2, and LCDR3 are sequences of SEQ ID NOs: 38, 39, and 40, respectively. Suitably, the second domain that specifically binds to TNFα may comprise a light chain variable region comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 51, and wherein the light chain variable region comprises T22N, K50Q, A51R, F89Y, G141T, L145V (AHO numbering), in particular, wherein the domains LCDR1, LCDR2, and LCDR3 are sequences of SEQ ID NOs: 38, 39, and 40, respectively.

[0603] In another embodiment, the present disclosure provides a multispecific antibody comprising a first domain that specifically binds IL-17A and a second domain that specifically binds TNFα, wherein the second domain comprises a heavy chain variable region comprising an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99%, preferably at least 90% identical to the amino acid sequence of SEQ ID NO: 34; and a light chain variable region comprising an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99%, preferably at least 90% identical to the amino acid sequence of SEQ ID NO: 47.

[0604] Preferably, the second domain that specifically binds to TNFα comprises a heavy chain variable region comprising an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99%, preferably at least 90%, identical to the amino acid sequence of SEQ ID NO: 34; and a light chain variable region comprising an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99%, preferably at least 90% identical to the amino acid sequence of SEQ ID NO: 47, and wherein the domain comprises the HCDR1, HCDR2 and HCDR3 sequences of SEQ ID NOs: 25, 26 and 27, respectively, and / or the LCDR1, LCDR2 and LCDR3 sequences of SEQ ID NOs: 38, 39 and 40, respectively, preferably wherein the domain comprises the HCDR1, HCDR2 and HCDR3 sequences of SEQ ID NOs: 39, 40, respectively. The HCDR1, HCDR2 and HCDR3 sequences are SEQ ID NOs: 25, 26 and 27, and the LCDR1, LCDR2 and LCDR3 sequences are SEQ ID NOs: 38, 39 and 40, respectively.

[0605] Suitably, the second domain that specifically binds to TNFα comprises a heavy chain variable region comprising an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99%, preferably at least 90%, identical to the amino acid sequence of SEQ ID NO: 37; and a light chain variable region comprising an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99%, preferably at least 90% identical to the amino acid sequence of SEQ ID NO: 50 or 51, and wherein the domain comprises the HCDR1, HCDR2 and HCDR3 sequences of SEQ ID NO: 25, 26 and 27, respectively, and / or the LCDR1, LCDR2 and LCDR3 sequences of SEQ ID NO: 38, 39 and 40, respectively, preferably wherein the domain comprises the HCDR1, HCDR2 and LCDR3 sequences of SEQ ID NO: 39, 40, respectively. The HCDR1, HCDR2 and HCDR3 sequences are SEQ ID NOs: 25, 26 and 27, and the LCDR1, LCDR2 and LCDR3 sequences are SEQ ID NOs: 38, 39 and 40, respectively.

[0606] In a specific embodiment, the present disclosure provides a multispecific antibody comprising a first domain that specifically binds IL-17A and a second domain that specifically binds TNFα, wherein the second domain comprises a VH comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 34, 35, 36, and 37; and / or a VL comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 47, 48, 49, 50, and 51. In a specific embodiment, the second domain that specifically binds TNFα comprises the VH sequence of SEQ ID NO: 34 and the VL sequence of SEQ ID NO: 47. In yet another specific embodiment, the second domain that specifically binds TNFα comprises the VH sequence of SEQ ID NO: 37 and the VL sequence of SEQ ID NO: 50 or SEQ ID NO: 51.

[0607] In one embodiment, the domain that specifically binds to human TNFα is a domain described in Table 1. Other domains of the present disclosure that have binding specificity for human TNFα include those antibodies in which the amino acids or nucleic acids encoding the amino acids have been mutated, but have at least 60, 70, 80, 90, or 95% identity with the sequences described in Table 1. In one embodiment, an amino acid sequence is included in which no more than 1, 2, 3, 4, or 5 amino acids have been mutated in the variable region when compared to the variable region described in the sequence described in Table 1 while retaining substantially the same activity.

[0608] In yet another embodiment, the present disclosure provides a multispecific antibody comprising a first domain that specifically binds to IL-17A and a second domain that specifically binds to TNFα, wherein the second domain comprises an amino acid sequence homologous to the sequence described in Table 1, and the domains bind to human TNFα and retain the desired functional properties of those domains described in Table 1.

[0609] In one embodiment, a domain of the invention that specifically binds to TNFα has a heavy chain variable region comprising a HCDR1, HCDR2, and HCDR3 sequence and a light chain variable region comprising a LCDR1, LCDR2, and LCDR3 sequence, wherein one or more of these CDR sequences has a specified amino acid sequence based on the domain described herein or conservative modifications thereof, wherein the domain retains the desired functional properties of the antibodies of the present disclosure.

[0610] Thus, the present disclosure provides a multispecific antibody comprising a first domain that specifically binds IL-17A and a second domain that specifically binds TNFα, wherein the second domain comprises (or consists of):

[0611] A heavy chain variable region (VH), which comprises three complementarity determining regions HCDR1, HCDR2 and HCDR3 in sequence, wherein the HCDR1 is the amino acid sequence of SEQ ID NO: 25, or a conservative variant thereof; the HCDR2 is the amino acid sequence of SEQ ID NO: 26, or a conservative variant thereof; the HCDR3 is an amino acid sequence selected from any one of SEQ ID NO: 27, or a conservative variant thereof; and

[0612] A light chain variable region (VL), which comprises three complementarity determining regions LCDR1, LCDR2, and LCDR3, wherein LCDR1 has the amino acid sequence of SEQ ID NO: 38, or a conservative variant thereof; LCDR2 has the amino acid sequence of SEQ ID NO: 39, or a conservative variant thereof; and LCDR3 has the amino acid sequence of SEQ ID NO: 40, or a conservative variant thereof;

[0613] The domain specifically binds to human TNFα and / or neutralizes TNFα.

[0614] Exemplary domains that specifically bind to human serum albumin (HSA)

[0615] The multispecific antibodies of the present disclosure comprise a third domain that specifically binds to human serum albumin, wherein the domain comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein: (a) the VH comprises three complementarity determining regions HCDR1, HCDR2 and HCDR3 in sequence, and (b) the VL comprises three complementarity determining regions LCDR1, LCDR2 and LCDR3 in sequence.

[0616] Suitably, the multispecific antibodies of the present disclosure may comprise a third binding domain having a third specificity different from the specificity of the first and second domains. Suitably, the multispecific antibodies of the present disclosure may comprise a third domain that specifically binds to human serum albumin (HSA). Thus, in one embodiment, the multispecific antibodies of the present disclosure comprise a first domain that specifically binds to IL-17A, a second domain that specifically binds to TNFα, and a third domain that specifically binds to human serum albumin.

[0617] Suitable domains that specifically bind to human serum albumin for use in the multispecific antibodies of the present disclosure include, but are not limited to, the humanized monoclonal antibodies or their binding domains whose sequences are listed in Table 1.

[0618] In one embodiment, the domain that specifically binds to human serum albumin comprises a set of CDRs: HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3, wherein the set of CDRs has 10 or fewer amino acid substitutions, e.g., 9 or fewer amino acid substitutions, 8 or fewer amino acid substitutions, 7 or fewer amino acid substitutions, 6 or fewer amino acid substitutions, 5 or fewer amino acid substitutions, 4 or fewer amino acid substitutions, 3 or fewer amino acid substitutions, 2 or fewer amino acid substitutions, 1 or 0 amino acid substitutions, preferably 0 amino acid substitutions, relative to such a set of CDRs:

[0619] (a) HCDR1′ is set forth in SEQ ID NO: 52; HCDR2′ is set forth in SEQ ID NO: 53; HCDR3′ is set forth in SEQ ID NO: 54; LCDR1′ is set forth in SEQ ID NO: 62; LCDR2′ is set forth in SEQ ID NO: 63; and LCDR3′ is set forth in SEQ ID NO: 64; or

[0620] (b) HCDR1' is shown in SEQ ID NO: 73; HCDR2' is shown in SEQ ID NO: 74; HCDR3' is shown in SEQ ID NO: 75; LCDR1' is shown in SEQ ID NO: 83; LCDR2' is shown in SEQ ID NO: 84; LCDR3' is shown in SEQ ID NO: 85.

[0621] In particular, the domain that specifically binds to human serum albumin comprises a VH CDR having the amino acid sequence of any one of the VH CDRs listed in Table 1. In particular, the present disclosure provides multispecific antibodies comprising a third domain that specifically binds to HSA, wherein the domain comprises (or consists of) one, two, three or more VH CDRs having the amino acid sequence of any one of the VH CDRs listed in Table 1.

[0622] Suitably, the domain that specifically binds to human serum albumin comprises a heavy chain variable region (VH), wherein said VH comprises three complementarity determining regions HCDR1, HCDR2 and HCDR3 in sequence, wherein

[0623] (a) the HCDR1 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 52, 55, and 58; the HCDR2 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 53, 56, and 59; and the HCDR3 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 54, 57, and 60; or

[0624] (b) the HCDR1 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 73, 76 and 79; the HCDR2 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 74, 77 and 80; and the HCDR3 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 75, 78 and 81.

[0625] In a preferred embodiment, the domain that specifically binds to human serum albumin comprises the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs: 52, 53, and 54, respectively. In another embodiment, the domain that specifically binds to human serum albumin comprises the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs: 73, 74, and 75, respectively.

[0626] Suitably, the domain that specifically binds to human serum albumin comprises a VL CDR having the amino acid sequence of any one of the VL CDRs listed in Table 1. In particular, the present disclosure provides multispecific antibodies comprising a third domain that specifically binds to HSA, wherein said domain comprises (or alternatively consists of) one, two, three or more VL CDRs having the amino acid sequence of any of the VL CDRs listed in Table 1.

[0627] Suitably, the domain that specifically binds to human serum albumin comprises a light chain variable region (VL), wherein said VL comprises three complementarity determining regions LCDR1, LCDR2 and LCDR3 in sequence, wherein

[0628] (a) the LCDR1 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 62, 65, and 68; the LCDR2 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 63, 66, and 69; and the LCDR3 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 64, 67, and 70; or

[0629] (b) the LCDR1 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 83, 86 and 89; the LCDR2 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 84, 87 and 90; and the LCDR3 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 85, 88 and 91.

[0630] In a preferred embodiment, the domain that specifically binds to human serum albumin comprises the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs: 62, 63, and 64, respectively. In another embodiment, the domain that specifically binds to human serum albumin comprises the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs: 83, 84, and 85, respectively.

[0631] Suitably, the present disclosure provides a multispecific antibody comprising a third domain that specifically binds to HSA, wherein the third domain comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein:

[0632] (a) the VH comprises, in sequence, three complementarity determining regions HCDR1, HCDR2, and HCDR3, the HCDR1 having an amino acid sequence selected from any one of SEQ ID NOs: 52, 55, and 58, the HCDR2 having an amino acid sequence selected from any one of SEQ ID NOs: 53, 56, and 59, and the HCDR3 having an amino acid sequence selected from any one of SEQ ID NOs: 54, 57, and 60; and the VL comprises, in sequence, three complementarity determining regions LCDR1, LCDR2, and LCDR3, the LCDR1 having an amino acid sequence selected from any one of SEQ ID NOs: 62, 65, and 68, the LCDR2 having an amino acid sequence selected from any one of SEQ ID NOs: 63, 66, and 69, and the LCDR3 having an amino acid sequence selected from any one of SEQ ID NOs: 64, 67, and 70, or

[0633] (b) the VH comprises, in sequence, three complementarity determining regions HCDR1, HCDR2 and HCDR3, the HCDR1 having an amino acid sequence selected from any one of SEQ ID NOs: 73, 76 and 79, the HCDR2 having an amino acid sequence selected from any one of SEQ ID NOs: 74, 77 and 80, and the HCDR3 having an amino acid sequence selected from any one of SEQ ID NOs: 75, 78 and 81; and the VL comprises, in sequence, three complementarity determining regions LCDR1, LCDR2 and LCDR3, the LCDR1 having an amino acid sequence selected from any one of SEQ ID NOs: 83, 86 and 89, the LCDR2 having an amino acid sequence selected from any one of SEQ ID NOs: 84, 87 and 90, and the LCDR3 having an amino acid sequence selected from any one of SEQ ID NOs: 85, 88 and 91.

[0634] In a preferred embodiment, the present disclosure provides a multispecific antibody comprising a third domain that specifically binds to HSA, wherein the third domain comprises (a) HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs: 52, 53, and 54, respectively, and (b) LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs: 62, 63, and 64, respectively. In another embodiment, the present disclosure provides a multispecific antibody comprising a third domain that specifically binds to HSA, wherein the third domain comprises (a) HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs: 73, 74, and 75, respectively, and (b) LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs: 83, 84, and 85, respectively.

[0635] Other domains of the present disclosure that specifically bind to HSA include amino acid sequences that have been mutated but still have at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identity in the CDR regions to the CDR regions described in the sequences described in Table 1. Suitably, other domains of the present disclosure that specifically bind to HSA include mutant amino acid sequences in which no more than 1, 2, 3, 4, 5, or 10 amino acids have been mutated by deletion, insertion, or substitution of amino acids in the CDR regions when compared to the CDR regions described in the sequences described in Table 1. Mutations, such as substitutions, may be made at any residue within the CDR group and may be within CDR1, CDR2, and / or CDR3.

[0636] Suitably, the third domain of the multispecific antibody of the present disclosure that specifically binds to HSA comprises: a heavy chain variable region (VH) and a light chain variable region (VL), wherein:

[0637] (a) the VH comprises, in order, three complementarity determining regions: HCDR1, HCDR2, and HCDR3, wherein the HCDR1 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to any one of SEQ ID NOs: 52, 55, and 58; the HCDR2 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to any one of SEQ ID NOs: 53, 56, and 59; the HCDR3 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to any one of SEQ ID NOs: 54, 57, and 60; and / or

[0638] (b) the VL comprises three complementarity determining regions, LCDR1, LCDR2 and LCDR3, in sequence, wherein the LCDR1 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to any one of SEQ ID NOs: 62, 65 and 68; the LCDR2 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to any one of SEQ ID NOs: 63, 66 and 69; the LCDR3 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to any one of SEQ ID NOs: 64, 67 and 70.

[0639] Suitably, the multispecific antibody comprises a third domain that specifically binds to HSA, wherein the third domain comprises: a HCDR1 , a HCDR2, and a HCDR3 that are at least 60, 70, 80, 90, 91 , 92, 93, 94, 95, 96, 97, 98, or 99% identical to the sequences of SEQ ID NOs: 52, 53, and 54, respectively, and / or a LCDR1 , a LCDR2, and a LCDR3 that are at least 60, 70, 80, 90, 91 , 92, 93, 94, 95, 96, 97, 98, or 99% identical to the sequences of SEQ ID NOs: 62, 63, and 64, respectively.

[0640] Suitably, the third domain of the multispecific antibody of the present disclosure that specifically binds to HSA comprises: a heavy chain variable region (VH) and a light chain variable region (VL), wherein:

[0641] (a) the VH comprises, in order, three complementarity determining regions: HCDR1, HCDR2, and HCDR3, wherein the HCDR1 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to any one of SEQ ID NOs: 73, 76, and 79; the HCDR2 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to any one of SEQ ID NOs: 74, 77, and 80; the HCDR3 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to any one of SEQ ID NOs: 75, 78, and 81; and / or

[0642] (b) the VL comprises three complementarity determining regions, LCDR1, LCDR2 and LCDR3, in sequence, wherein the LCDR1 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to any one of SEQ ID NOs: 83, 86 and 89; the LCDR2 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to any one of SEQ ID NOs: 84, 87 and 90; the LCDR3 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to any one of SEQ ID NOs: 85, 88 and 91.

[0643] Suitably, the multispecific antibody comprises a third domain that specifically binds to HSA, wherein the third domain comprises: a HCDR1 , a HCDR2, and a HCDR3 that are at least 60, 70, 80, 90, 91 , 92, 93, 94, 95, 96, 97, 98, or 99% identical to the sequences of SEQ ID NOs: 73, 74, and 75, respectively, and / or a LCDR1 , a LCDR2, and a LCDR3 that are at least 60, 70, 80, 90, 91 , 92, 93, 94, 95, 96, 97, 98, or 99% identical to the sequences of SEQ ID NOs: 83, 84, and 85, respectively.

[0644] In a further embodiment, the present disclosure provides a multispecific antibody comprising a third domain that specifically binds to HSA, wherein the third domain comprises a heavy chain variable region VHC and a light chain variable region VLC.

[0645] Suitably, the present disclosure provides a multispecific antibody comprising a third domain that specifically binds to HSA, wherein the third domain comprises a heavy chain variable region VHC, wherein the VHC is VH1A, VH1B, VH3, or VH4. In one embodiment, the third domain that specifically binds to HSA of the present disclosure comprises a heavy chain variable region VHC, wherein the VHC is VH4. In a preferred embodiment, the third domain that specifically binds to HSA of the present disclosure comprises a heavy chain variable region VHC, wherein the VHC is VH3.

[0646] Suitably, the present disclosure provides a multispecific antibody comprising a third domain that specifically binds to HSA, wherein the third domain comprises a light chain variable region, VLC, and wherein the VLC comprises Vκ framework FR1, FR2, and FR3, particularly Vκ1 or Vκ3 FR1 to FR3, preferably Vκ1 framework FR1 to FR3, and a framework FR4 selected from the group consisting of VκFR4, particularly Vκ1FR4, Vκ3FR4, and VλFR4. Suitable VλFR4s are shown in SEQ ID NO: 97 to SEQ ID NO: 103. In one embodiment, the third domain comprises VλFR4, comprising an amino acid sequence that is at least 60, 70, 80, 90% identical to an amino acid sequence selected from any one of SEQ ID NO: 97 to SEQ ID NO: 103, preferably to SEQ ID NO: 97 or SEQ ID NO: 98, more preferably to SEQ ID NO: 97. Suitably, the third domain comprises VλFR4 comprising an amino acid sequence selected from any one of SEQ ID NO: 97 to SEQ ID NO: 103, preferably VλFR4 as shown in SEQ ID NO: 97 or 98, more preferably VλFR4 as shown in SEQ ID NO: 98.

[0647] Therefore, in a preferred embodiment, the present disclosure provides a multispecific antibody comprising a third domain that specifically binds to HSA, wherein the third domain comprises:

[0648] (a) the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs: 52, 53, and 54, respectively, and the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs: 62, 63, and 64, respectively;

[0649] (b) VHC, wherein the VHC is VH3 or VH4, preferably VH3; and

[0650] (c) VLC, which comprises a VL framework, said VL framework comprising Vκ framework FR1, FR2 and FR3, in particular Vκ1 or Vκ3FR1 to FR3, preferably Vκ1FR1 to FR3, and a framework FR4, said framework FR4 being selected from VκFR4, in particular Vκ1FR4, Vκ3FR4, and VλFR4, in particular VλFR4 comprising an amino acid sequence having at least 60, 70, 80, 90% identity with an amino acid sequence selected from any one of SEQ ID NO: 97 to SEQ ID NO: 103, preferably VλFR4 as shown in an amino acid sequence selected from any one of SEQ ID NO: 97 to SEQ ID NO: 103, preferably VλFR4 as shown in SEQ ID NO: 97 or 98, more preferably VλFR4 as shown in SEQ ID NO: 98.

[0651] In a further embodiment, the present disclosure provides a multispecific antibody comprising a third domain that specifically binds to HSA, wherein the third domain comprises:

[0652] (i) the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs: 73, 74, and 75, respectively, and the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs: 83, 84, and 85, respectively;

[0653] (ii) VHC, wherein the VHC is VH3 or VH4, preferably VH3; and

[0654] (iii) VLC, which comprises a VL framework, said VL framework comprising Vκ framework FR1, FR2 and FR3, in particular FR1 to FR3 of Vκ1 or Vκ3, preferably Vκ1FR1 to FR3, and a framework FR4, said framework FR4 being selected from VκFR4, in particular Vκ1FR4, Vκ3FR4, and VλFR4, in particular VλFR4 comprising an amino acid sequence having at least 60, 70, 80, 90% identity with an amino acid sequence selected from any one of SEQ ID NO: 97 to SEQ ID NO: 103, preferably VλFR4 as shown in an amino acid sequence selected from any one of SEQ ID NO: 97 to SEQ ID NO: 103, preferably VλFR4 as shown in SEQ ID NO: 97 or 98, more preferably VλFR4 as shown in SEQ ID NO: 98.

[0655] Suitably, the present disclosure provides a multispecific antibody comprising a third domain that specifically binds to HSA, wherein the third domain comprises a VH listed in Table 1. Suitably, the present disclosure also provides a multispecific antibody comprising a third domain that specifically binds to HSA, wherein the third domain comprises (or alternatively consists of) a VH amino acid sequence listed in Table 1, wherein no more than about 20 amino acids in the framework sequence (e.g., a sequence that is not a CDR) have been mutated (wherein, as various non-limiting examples, the mutation is an addition, substitution, or deletion), preferably no more than about 10. Other domains of the present disclosure that specifically bind to HSA include amino acid sequences that have been mutated but still have at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identity in the VH region to the VH region described in the sequences described in Table 1.

[0656] Suitably, the present disclosure provides multispecific antibodies comprising a third domain that specifically binds to HSA, wherein the third domain comprises a VL domain listed in Table 1. Suitably, the present disclosure also provides multispecific antibodies comprising a third domain that specifically binds to HSA, wherein the third domain comprises (or alternatively consists of) a VL amino acid sequence listed in Table 1, wherein no more than about 20 amino acids in the framework sequence (e.g., a sequence that is not a CDR) have been mutated (wherein, as various non-limiting examples, the mutation is an addition, substitution, or deletion), preferably no more than about 10. Other domains of the present disclosure that specifically bind to HSA include amino acid sequences that have been mutated but still have at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identity in the VL region to the VL region described in the sequences described in Table 1.

[0657] In one embodiment, the present disclosure provides a multispecific antibody comprising a third domain that specifically binds to HSA, wherein the third domain comprises a heavy chain variable region comprising an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99%, preferably at least 90%, identical to the amino acid sequence of SEQ ID NO: 61, and in particular, wherein the domain includes the HCDR1, HCDR2 and HCDR3 sequences of SEQ ID NOs: 52, 53 and 54, respectively.

[0658] In another embodiment, the present disclosure provides a multispecific antibody comprising a third domain that specifically binds to HSA, wherein the third domain comprises a light chain variable region comprising an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99%, preferably at least 90%, identical to the amino acid sequence of SEQ ID NO: 71, and in particular, wherein the domain includes the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs: 62, 63, and 64, respectively.

[0659] In a further embodiment, the present disclosure provides a multispecific antibody comprising a third domain that specifically binds to HSA, wherein the third domain comprises a heavy chain variable region comprising an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90%, identical to the amino acid sequence of SEQ ID NO: 61; and a light chain variable region comprising an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90% identical to the amino acid sequence of SEQ ID NO: 71.

[0660] In a preferred embodiment, the present disclosure provides a multispecific antibody comprising a third domain that specifically binds to HSA, wherein the third domain comprises a heavy chain variable region comprising an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90%, identical to the amino acid sequence of SEQ ID NO: 61; and a light chain variable region comprising an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90% identical to the amino acid sequence of SEQ ID NO: 71, and wherein the domain comprises the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs: 52, 53, and 54, respectively, and / or the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs: 62, 63, and 64, respectively. Preferably, the domain comprises the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs: 52, 53, and 54, respectively. The HCDR1, HCDR2 and HCDR3 sequences are SEQ ID NOs: 52, 53 and 54, and the LCDR1, LCDR2 and LCDR3 sequences are SEQ ID NOs: 62, 63 and 64, respectively.

[0661] In certain embodiments, the present disclosure provides a multispecific antibody comprising a third domain that specifically binds to HSA, wherein the third domain comprises a VH comprising the amino acid sequence of SEQ ID NO: 61, and / or its VL comprises the amino acid sequence of SEQ ID NO: 71. In a preferred embodiment, the third domain that specifically binds to HSA comprises the VH sequence of SEQ ID NO: 61 and the VL sequence of SEQ ID NO: 71.

[0662] In another embodiment, the present disclosure provides a multispecific antibody comprising a third ...

Claims

1. An isolated multispecific antibody comprising a first domain that specifically binds to IL-17A, a second domain that specifically binds to TNFα, and a third domain that specifically binds to human serum albumin, wherein (a) the first domain comprises a set of CDRs: HCDR1, HCDR2 and HCDR3 sequences of SEQ ID NOs: 1, 2 and 3, respectively, and LCDR1, LCDR2 and LCDR3 sequences of SEQ ID NOs: 12, 13 and 14, respectively, wherein the first domain comprises a combination of a VH domain having at least 95% sequence identity with the VH domain of SEQ ID NO: 10 and a VL domain having at least 95% sequence identity with the VL domain of SEQ ID NO: 21; and (b) the second domain comprises a set of CDRs: HCDR1, HCDR2 and HCDR3 sequences of SEQ ID NOs: 63, 64 and 65, respectively, and LCDR1, LCDR2 and LCDR3 sequences of SEQ ID NOs: 76, 77 and 78, respectively, wherein the second domain comprises a combination of a VH domain having at least 95% sequence identity to the VH domain of SEQ ID NO: 72 and a VL domain having at least 95% sequence identity to the VL domain of SEQ ID NO: 85; and (c) the third domain comprises a set of CDRs: HCDR1, HCDR2 and HCDR3 sequences of SEQ ID NOs: 90, 91 and 92, respectively, and LCDR1, LCDR2 and LCDR3 sequences of SEQ ID NOs: 100, 101 and 102, respectively, wherein the third domain comprises a combination of a VH domain having at least 95% sequence identity with the VH domain of SEQ ID NO: 99 and a VL domain having at least 95% sequence identity with the VL domain of SEQ ID NO: 109; and The multispecific antibody has one or more of the following characteristics: (a) having the ability to neutralize IL-17A with a potency relative to secukinumab as determined by measuring Gro-α secretion in the HT-29 assay of greater than 50, wherein the relative potency is the IC of secukinumab in ng / mL as measured in the HT-29 assay 50 The values ​​are comparable to the IC values ​​in ng / mL of the multispecific antibodies measured in the HT-29 assay. 50 The ratio of values; (b) having the ability to neutralize TNFα with a potency of at least 1 relative to the scDb according to SEQ ID NO: 149 as determined by measuring Gro-α secretion in the HT-29 assay, wherein the relative potency is the IC in nM of the scDb according to SEQ ID NO: 149 as measured in the HT-29 assay. 50 Values ​​are comparable to the IC values ​​in nM of the multispecific antibodies measured in the HT-29 assay. 50 The ratio of values; (c) having the ability to block the interaction between IL-17A and IL-17RA, with a potency relative to secukinumab measured in an ELISA assay of greater than 4, wherein the relative potency is the IC value of secukinumab in ng / mL as measured by ELISA 50 The values ​​were compared with the IC values ​​of the multispecific antibodies in ng / mL measured by ELISA. 50 The ratio of values; (d) having the ability to neutralize TNFα with a potency of at least 0.4 relative to the scDb according to SEQ ID NO: 149 as determined in the L929 assay, wherein said relative potency is the IC in nM of said scDb according to SEQ ID NO: 149 as measured in the L929 assay. 50 Values ​​are comparable to the IC values ​​in nM of the multispecific antibodies measured in the L929 assay. 50 The ratio of values; (e) Dissociation constant K for binding to human IL-17A measured by surface plasmon resonance D less than 2 nM; and the K for binding to cynomolgus monkey IL-17A as measured by surface plasmon resonance D Less than 1nM; (f) Dissociation constant K for binding to human TNFα measured by surface plasmon resonance D Less than 0.25nM; (g) Dissociation constant K for binding to human serum albumin measured by surface plasmon resonance D Less than 5 nM, and / or a dissociation constant K for binding to cynomolgus monkey serum albumin as measured by surface plasmon resonance D Less than 5 nM; and / or (h) the multispecific antibody selectively binds human IL-17A relative to human IL-17B, IL-17C, IL-17D, IL-17E and IL-17F as measured by ELISA. 2 . The multispecific antibody according to claim 1 , wherein the antibody is monovalent to IL-17A and / or monovalent to TNFα and / or monovalent to human serum albumin.

3. The multispecific antibody of claim 1 or claim 2, wherein the domains are capable of binding to their respective antigens or receptors simultaneously.

4. The multispecific antibody of claim 1, wherein the antibody has the following properties: (a) a melting temperature, Tm, of at least 60° C. as measured by differential scanning fluorimetry in phosphate-citrate buffer, pH 6.4, 150 mM NaCl; (b) when the multispecific antibody is at an initial concentration of 10 mg / ml in phosphate buffered saline at pH 7.4, the monomer content loss is less than 1% after five consecutive freeze-thaw cycles; (c) when the multispecific antibody is stored at an initial concentration of 10 mg / ml in phosphate buffered saline at pH 7.4, the monomer content loss is less than 5% after 4 weeks at 4°C; and / or (d) when the multispecific antibody is stored at a starting concentration of 10 mg / ml in phosphate buffered saline at pH 7.4, the loss of monomer content is less than 15% after storage at 37°C for 4 weeks. The multispecific antibody according to claim 1 , wherein the format of the multispecific antibody is selected from the group consisting of: Fab-(scFv) 2 and scDb-scFv.

6. The multispecific antibody of claim 1, wherein the antibody comprises an amino acid sequence that is at least 95% identical to a sequence selected from any one of SEQ ID NOs: 136, 137, 138, 139, 140, 141, 142, 143, and 144.

7. The multispecific antibody of claim 6, wherein the multispecific antibody comprises an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO:

143.

8. The multispecific antibody of claim 7, wherein the antibody comprises an amino acid sequence selected from any one of SEQ ID NOs: 136, 137, 138, 139, 140, 141, 142, 143 and 144.

9. The multispecific antibody of claim 8, wherein the antibody comprises the amino acid sequence of SEQ ID NO:

143. 10 . A pharmaceutical composition comprising the multispecific antibody according to claim 1 and a pharmaceutically acceptable carrier.

11. Use of the multispecific antibody according to any one of claims 1 to 9 in the preparation of a medicament for treating an inflammatory condition, an autoimmune disease or cancer.

12. Use of the pharmaceutical composition according to claim 10 in the preparation of a medicament for treating inflammatory conditions, autoimmune diseases or cancer.

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