Multi-specific antibodies specific for TNF alpha and IL-17A, antibodies targeting IL-17A, and methods of use thereof

Abstract

The present invention relates to an isolated multispecific antibody comprising a first domain that specifically binds to TNF [alpha], a second domain that specifically binds to IL-17A, and optionally a third domain that specifically binds to human serum albumin. The invention also relates to isolated antibodies that specifically bind to human IL-17A. In addition, the invention also relates to a use method of the antibody, a pharmaceutical composition and a use method thereof, and a kit comprising the antibody. The 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 of producing the antibody.

Description

[0001] The present invention is a divisional application of Chinese invention patent application No. 202080025103.7, filed on January 31, 2020, entitled "Multispecific antibodies specific to TNFα and IL-17A, antibodies targeting IL-17A and methods of use thereof". Technical Field

[0002] 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 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 nucleotide sequences encoding the antibody, vectors comprising the nucleic acids, host cells comprising the nucleic acids or the vectors, and methods of producing the antibodies.

[0003] The present invention also relates to an isolated antibody that specifically binds to human IL-17A, a multispecific molecule comprising the isolated antibody of the present invention, a pharmaceutical composition and a method of using the same. The present invention also relates to a kit comprising the antibody, 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 of producing the antibody. Background Art

[0004] TNFα is a homotrimeric proinflammatory cytokine that is released by and interacts with cells of the immune system. 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 the 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. Both membrane TNFα and soluble TNFα are biologically active. TNFα can bind to two receptors, TNF receptor 1 (TNFR1) and 2 (TNFR2), of which transmembrane TNFα mainly acts through TNFR2. TNFR1 is widely expressed on various cells, and its engagement triggers a proinflammatory response. The expression of TNFR2 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).

[0005] 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. The TNFα antibodies previously used in such treatments are generally chimeric antibodies, such as those described in U.S. Pat. No. 5,919,452.

[0006] 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) describes 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 B1, US 8,293,235, US 8,697,074, WO 2009 / 155723 A2 and WO 2006 / 131013 A2.

[0007] 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).

[0008] However, anti-TNFα therapy has been shown to have certain limitations. Not all patients are able to achieve an adequate clinical response to anti-TNFα or maintain a clinical response long-term, resulting in the need to switch to new therapies to control their disease. For example, when anti-TNFα therapy is used in patients with rheumatoid arthritis (RA), approximately 40% of patients never respond and only 20% of patients 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 conditions.

[0009] 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 therapy. Recently, a large amount of evidence has emerged that IL-17A plays a role in the pathogenesis of inflammatory and autoimmune disorders, such as RA.

[0010] The interleukin 17 (IL-17) family in humans and mice consists of six cytokines, namely 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, namely IL-17RA, IL-17RB, IL-17RC, IL-17RD, and IL-17RE.

[0011] 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 memory CD4+ T cells (known as Th17), CD8+ T cells (Tc17), invariant NKT cells, γδ T cells, non-T non-B lymphocytes (known as type 3 innate lymphocytes), and a subset of neutrophils. IL-17A and IL-17F form a unique subset within the IL-17 family. They have the greatest sequence homology and identity in 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.

[0012] 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)).

[0013] Pathological production of IL-17A leads to excessive inflammation and tissue damage (see for review Gu et al., Cytokine. 2013 November; 64(2)). High levels of IL-17A have been found in patients with multiple sclerosis (MS), psoriasis, asthma, Crohn's disease, and rheumatoid arthritis. Treatment of animals with an IL-17A neutralizing antibody reduces the disease incidence and severity of autoimmune encephalomyelitis (Komiyama, Y. et al., J. Immunol. 177 (2006) 566-573). In addition, IL-17A neutralizing antibodies reduce the severity and incidence of the rheumatoid arthritis model in mice, and high levels of IL-17A can be detected in the inflamed synovial fluid of patients with rheumatoid arthritis (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).

[0014] Experimental evidence from a large number of human and animal models supports the development of IL-17A targeted therapies. A number of 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), RG4943 (Roche), etc. WO 2006 / 013107, WO 2006 / 054059, WO2007 / 070750, WO2007 / 149032, WO 2008 / 001063, WO 2008 / 021156, WO 2010 / 034443, WO2010 / 102251, WO 2012 / 018767, WO 2014 / 161570, WO 2014 / 001368, WO2014 / 122613, WO2015 / 070697, WO 2015 / 137843, WO 2016 / 048188, WO2016 / 113557, WO 2016 / 138842, WO2017 / 068472 disclose examples of anti-IL-17A antibodies.

[0015] A number of clinical trials have been conducted or are still ongoing for various molecules that block IL-17A signaling. Biological agents targeting IL-17A or its receptor and their efficacy are being evaluated in inflammatory or autoimmune disorders such as rheumatoid arthritis, ankylosing spondyloarthropathy, Crohn's disease, psoriasis, multiple sclerosis, and ozone-induced neutrophilia.

[0016] For example, secukinumab is a fully human IgG1κ anti-IL-17A monoclonal antibody (U.S. Patent Nos. 7,807,155 and WO 2006 / 013107) and 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 signs and symptoms of psoriatic arthritis (Mease PJ. et al. N Engl J Med 2015;373:1329-39; McInnes IB. et al. The Lancet;386:1137-46).

[0017] Ixekizumab is a humanized anti-IL-17A monoclonal antibody (U.S. Patent Nos. 7,838,638 and 8,110,191 and WO 2007 / 070750) and was studied in a 24-week phase III trial (SPIRIT-P1) in patients with active psoriatic arthritis who had not received biotherapy (Mease PJ. et al., Ann Rheum Dis. 2017 Jan;76(1):79-87). The study showed that in patients with active psoriatic arthritis who had not received biotherapy, ixekizumab treatment led to improvement in disease activity and physical function, as well as inhibition of the progression of structural damage. Ixekizumab is also effective in treating patients with moderate to severe plaque psoriasis (Griffiths CEM et al., The Lancet;386:541-51).

[0018] 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).

[0019] Therefore, IL-17A is a promising target for the treatment of inflammatory and autoimmune disorders. 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 method that simultaneously blocks the biological pathways of TNFα and IL-17A has the potential to significantly increase the response rate and address the unmet needs in the treatment of disorders 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, WO2014 / 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 bivalent ("2+2" construct) or monovalent ("2+2" construct) to each of the two antigens. According to WO 2015 / 014979, bivalent constructs are superior to monovalent alternatives.

[0020] However, there is still 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 is still a need to develop improved therapeutic antibodies that effectively neutralize the activity of IL-17A and TNFα, have beneficial affinity and efficacy, and improved safety, such as lower immunogenicity. In addition, therapeutic antibodies should have improved biophysical properties, thereby having better developability, high yield manufacturability, and excellent antibody stability. Summary of the invention

[0021] It is an object of the present invention to provide a medicament for improving the treatment of inflammatory and autoimmune diseases.

[0022] 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 inhibit IL-17A and TNFa simultaneously and has further improved properties that are beneficial to treatment, such as higher affinity, improved efficacy, selectivity, safety (e.g., low immunogenicity) and improved biophysical properties (e.g., developability and stability).

[0023] In one aspect, the disclosure relates to an isolated multispecific antibody comprising 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.

[0024] In one aspect, the present disclosure relates to a pharmaceutical composition comprising a multispecific antibody of the present invention and a pharmaceutically acceptable carrier.

[0025] 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.

[0026] In another aspect, the 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.

[0027] In one aspect, the present disclosure provides the use of the multispecific antibody of the invention or the pharmaceutical composition of the invention for 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.

[0028] In one aspect, the present disclosure provides the 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.

[0029] In yet another aspect, the present disclosure provides a method of treating a disorder mediated by IL-17A and / or TNFα, the method 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.

[0030] In one aspect, the present disclosure provides a kit comprising the multispecific antibody of the present invention, or the pharmaceutical composition of the present invention.

[0031] In another aspect, the present disclosure provides a nucleic acid comprising a nucleotide sequence encoding a multispecific antibody of the present invention. In another aspect, the present disclosure provides a vector comprising the nucleic acid. In another aspect, the present disclosure provides a host cell comprising the nucleic acid or the vector.

[0032] 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 comprising a nucleic acid or vector encoding the multispecific antibody or its binding domain or fragment thereof of the present invention.

[0033] The aspects, advantageous features and preferred embodiments of the present disclosure outlined in the following, respectively alone or in combination, further contribute to solving the objects of the present invention:

[0034] 1. An isolated multispecific antibody comprising a first domain that specifically binds to IL-17A and a second domain that specifically binds to TNFα.

[0035] 2. The multispecific antibody according to item 1, wherein the antibody comprises only one domain that specifically binds to IL-17A and / or only one domain that specifically binds to TNFα.

[0036] 3. The multispecific antibody according to item 1 or item 2, wherein the antibody is capable of neutralizing the biological activity of human TNFα and human IL-17A.

[0037] 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.

[0038] 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.

[0039] 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.

[0040] 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.

[0041] 8. The multispecific antibody according to any one 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:

[0042] 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 include but are 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.

[0043] 9. The multispecific antibody according to any one 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-Fv 2 , Morrison (IgGCH3-scFv fusion (Morrison L) or IgG CL-scFv fusion (Morrison H)), triabodies, scDb-scFv, bispecific Fab 2, diabodies, tetrabodies, scFv-Fc-scFv fusion, scFv-HSA-scFv fusion, diabodies, DVD-Ig, COVD, IgG-scFab, scFab-dsscFv, Fv 2 -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), 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) fused to the N- and / or C-terminus of either chain of a heterodimeric Fc domain or any other heterodimeric domain 2 , Fab-(scFv) 1 , Fab, Fab-Fv 2 , COVD, MATCH and DuoBody, preferably a trio or scDb-scFv.

[0044] 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.

[0045] 11. The multispecific antibody of any of the preceding items, wherein the antibody is a triabody.

[0046] 12. A multispecific antibody according to item 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.

[0047] 13. A multispecific antibody according to item 9, wherein the antibody is scDb-scFv, preferably wherein the scFv part 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.

[0048] 14. The multispecific antibody according to item 13, wherein the antibody is represented by the following formula:

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

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

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

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

[0053] Preferably VLB-L1-VHA-L2-VLA-L3-VHB-L4-VLC-L5–VHC or VLA-L1-VHB-L2-VLB-L3-VHA-L4-VLC-L5–VHC,

[0054] More preferably VLB-L1-VHA-L2-VLA-L3-VHB-L4-VLC-L5–VHC,

[0055] 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.

[0056] 15. The multispecific antibody according to item 14, wherein L1 and L3 are as shown in SEQ ID NO:132.

[0057] 16. A multispecific antibody according to any one of items 14 to 15, wherein L2, L4 and L5 are as shown in SEQ ID NO: 23.

[0058] 17. The multispecific antibody according to any of the preceding items, wherein the antibody has the following characteristics

[0059] (a) having 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 the values; and

[0060] (b) having the ability to neutralize TNFα with a potency (relative potency) relative to the scDb (A13) according to SEQ ID NO: 149 as determined by measuring Gro-α secretion in the HT-29 assay 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, 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 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

[0061] (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

[0062] (d) optionally, having the ability to neutralize TNFα with a potency (relative potency) relative to the scDb (A13) according to SEQ ID NO: 149 of at least 0.4, e.g. at least 0.5, preferably at least 1 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 Values ​​are comparable to the IC values ​​in nM of the multispecific antibodies measured in the L929 assay. 50 value ratio; and / or

[0063] (e) Binding to human IL-17A, dissociation constant (K D ) is less than 5 nM, such as 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;

[0064] (f) Binding 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

[0065] (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.

[0066] 18. The multispecific antibody according to any one of the preceding items, wherein the antibody has the following characteristics

[0067] (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;

[0068] (b) when the multispecific antibody is present at an initial concentration of 10 mg / ml in phosphate buffered saline (PBS) at pH 7.4, the monomer content loss is less than 5% after five consecutive freeze-thaw cycles,

[0069] For example, less than 4%, less than 3%, less than 2%, preferably 1% or less;

[0070] (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

[0071] (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, in particular at least four weeks, the loss of monomer content is less than 20%, preferably less than 15%.

[0072] 19. The multispecific antibody of any of the preceding items, wherein each domain comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein:

[0073] (a) the VH comprises three complementary determining regions HCDR1, HCDR2 and HCDR3 in sequence, and

[0074] (b) The VL comprises three complementary determining regions LCDR1, LCDR2 and LCDR3 in sequence.

[0075] 20. A multispecific antibody according to 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 such a set of CDRs: (i) HCDR1' is shown in SEQ ID NO:1; HCDR2' is shown in SEQ ID NO:2; HCDR3' is shown in SEQ ID NO:3; LCDR1' is shown in SEQ ID NO:12; LCDR2' is shown in SEQ ID NO:13; LCDR3' is shown in SEQ ID NO:14, or (ii) HCDR1' is shown in SEQ ID NO:39; HCDR2' is shown in SEQ ID NO:40; HCDR3' is shown in SEQ ID NO:41; LCDR1' is shown in SEQ ID NO:50; LCDR2' is shown in SEQ ID NO:51; LCDR3' is shown in SEQ ID NO:52.

[0076] 21. The multispecific antibody according to claim 20, wherein

[0077] (i)

[0078] (a) the HCDR1 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 1, 4 and 7;

[0079] (b) the HCDR2 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 2, 5 and 8;

[0080] (c) the HCDR3 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 3, 6 and 9;

[0081] (d) the LCDR1 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 12, 15 and 18;

[0082] (e) the LCDR2 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 13, 16 and 19; and

[0083] (f) the LCDR3 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 14, 17 and 20; or

[0084] (ii)

[0085] (a) the HCDR1 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 39, 42 and 45;

[0086] (b) the HCDR2 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 40, 43 and 46;

[0087] (c) the HCDR3 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 41, 44 and 47;

[0088] (d) the LCDR1 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 50, 53 and 56;

[0089] (e) the LCDR2 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 51, 54 and 57; and

[0090] (f) the LCDR3 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 52, 55 and 58.

[0091] 22. A multispecific antibody according to 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.

[0092] 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.

[0093] 24. A 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 Vκ1FR1 to FR3, and a framework FR4, wherein the framework FR4 is selected from VκFR4, in particular Vκ1FR4, 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.

[0094] 25. A multispecific antibody according to 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.

[0095] 26. A multispecific antibody according to 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.

[0096] 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.

[0097] 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.

[0098] 29. The multispecific antibody according to item 28, wherein

[0099] (a) the HCDR1 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 63, 66 and 69;

[0100] (b) the HCDR2 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 64, 67 and 70;

[0101] (c) the HCDR3 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 65, 68 and 71;

[0102] (d) the LCDR1 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 76, 79 and 82;

[0103] (e) the LCDR2 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 77, 80 and 83; and

[0104] (f) The LCDR3 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 78, 81 and 84.

[0105] 30. The multispecific antibody according to claim 29, comprising 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.

[0106] 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.

[0107] 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 Vκ3FR1 to FR3, preferably Vκ1FR1 to FR3, and a framework FR4, wherein the framework FR4 is selected from VκFR4, in particular Vκ1FR4, 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.

[0108] 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 NOs: 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 NOs: 85, 86, 87, 88 and 89, preferably an amino acid sequence that is at least 90% identical to SEQ ID NO: 85.

[0109] 34. A multispecific antibody according to 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.

[0110] 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.

[0111] 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:

[0112] (a) HCDR1' is shown in SEQ ID NO:90; HCDR2' is shown in SEQ ID NO:91;

[0113] HCDR3' is shown in SEQ ID NO:92; LCDR1' is shown in SEQ ID NO:100;

[0114] LCDR2' is shown in SEQ ID NO: 101; LCDR3' is shown in SEQ ID NO: 102;

[0115] or

[0116] (b) HCDR1' is shown in SEQ ID NO:111; HCDR2' is shown in SEQ ID NO:112;

[0117] HCDR3' is shown in SEQ ID NO:113; LCDR1' is shown in SEQ ID NO:120;

[0118] LCDR2' is shown in SEQ ID NO:121; LCDR3' is shown in SEQ ID NO:122.

[0119] 37. The multispecific antibody according to item 36, wherein

[0120] (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

[0121] and 106; the LCDR2 is selected from SEQ ID NO:

[0122] The amino acid sequence of any one of SEQ ID NOs: 101, 104 and 107; and the LCDR3 is shown as an amino acid sequence selected from any one of SEQ ID NOs: 102, 105 and 108; or

[0123] (b) the HCDR1 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 111, 114 and 117; the HCDR2 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 112, 115 and 118; the HCDR3 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 113, 116

[0124] and 119; the LCDR1 is selected from SEQ ID NO:

[0125] The LCDR2 is shown as an amino acid sequence selected from any one of SEQ ID NOs: 122, 125 and 128; and the LCDR3 is shown as an amino acid sequence selected from any one of SEQ ID NOs: 123, 126 and 129.

[0126] 38. The multispecific antibody according to item 37, comprising

[0127] (a) HCDR1, HCDR2 and HCDR3 sequences of SEQ ID NOs: 90, 91, and 92, respectively, and LCDR1, LCDR2 sequences of SEQ ID NOs: 100, 101, and 102, respectively

[0128] and LCDR3 sequence; or

[0129] (b) HCDR1, HCDR2 and HCDR3 of SEQ ID NOs: 111, 112 and 113, respectively

[0130] Sequence, and LCDR1, LCDR2 of SEQ ID NO: 121, 122, and 123, respectively

[0131] and LCDR3 sequences.

[0132] 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.

[0133] 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 Vκ3FR1 to FR3, preferably Vκ1FR1 to FR3, and a framework FR4, wherein the framework FR4 is selected from 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 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.

[0134] 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.

[0135] 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.

[0136] 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.

[0137] 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.

[0138] 45. The multispecific antibody according to any of the preceding items, wherein the antibody is humanized.

[0139] 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 CDR has a sequence according to items 22, 30 and 38(a).

[0140] 47. A multispecific antibody according to any of the preceding items, wherein the antibody comprises an amino acid sequence selected from any one of SEQ ID NO: 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, and 148, preferably 143.

[0141] 48. A pharmaceutical composition comprising the multispecific antibody according to any one of the preceding items and a pharmaceutically acceptable carrier.

[0142] 49. The multispecific antibody of any one of items 1 to 47, or the pharmaceutical composition of item 48, for use as a medicament.

[0143] 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 that can be treated by inhibiting Gro-α secretion.

[0144] 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.

[0145] 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, allogeneic transplant rejection, graft-versus-host disease, systemic sclerosis, urological inflammatory disorders, cardiovascular disease, vasculitis, periodic fever, 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.

[0146] 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 that can be treated by inhibiting Gro-α secretion.

[0147] 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.

[0148] 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, extraurological inflammatory disorders, cardiovascular disease, vasculitis, periodic fever, 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.

[0149] 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 condition is alleviated.

[0150] 57. A method according to item 56, wherein the condition mediated by IL-17A and / or TNFα is an inflammatory condition or an autoimmune disease.

[0151] 58. A method according to claim 56, wherein the condition 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, periodic fever, 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.

[0152] 59. A nucleic acid encoding the multispecific antibody or fragment thereof according to any one of items 1 to 47.

[0153] 60. A vector comprising the nucleic acid described in item 59.

[0154] 61. A host cell comprising the nucleic acid described in item 59 or the vector described in item 60.

[0155] 62. A method for producing the multispecific antibody described in any one of items 1 to 47, the method comprising the following steps: culturing a host cell, the host cell comprising a nucleic acid or vector encoding the multispecific antibody or fragment thereof described in any one of items 1 to 47.

[0156] 63. A kit comprising the multispecific antibody of any one of items 1 to 47, or the pharmaceutical composition of item 48.

[0157] 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.

[0158] 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.

[0159] Therefore, in one aspect, the present disclosure provides an isolated antibody having binding specificity to human IL-17A, in particular, it comprises a set of CDRs: HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, wherein the set of CDRs has 10 or less, 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; 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.

[0160] On the other hand, the present disclosure provides an isolated antibody having binding specificity to human IL-17A, in particular, it includes a set of CDRs: HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, wherein the set of CDRs has 10 or less, 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; 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.

[0161] 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.

[0162] 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.

[0163] In one aspect, the disclosure relates to an antibody, or a multispecific molecule comprising the isolated antibody, or a pharmaceutical composition of the disclosure for use in treating a disorder mediated by IL-17A or a disorder treatable by inhibiting GRO-α secretion.

[0164] 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, 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.

[0165] In another aspect, the present disclosure relates to a method for treating a condition mediated by IL-17A, the method comprising administering an effective amount of an antibody of the present disclosure, or a multispecific molecule of the present disclosure, or a pharmaceutical composition of the present disclosure to a subject in need thereof. In another aspect, the present disclosure relates to a nucleic acid encoding an antibody of the present disclosure. In another aspect, the present disclosure relates to a vector comprising the nucleic acid. In another aspect, the present disclosure relates to a host cell comprising the nucleic acid or the vector.

[0166] In another aspect, the present disclosure relates to a method for producing the antibody of the present disclosure, the method comprising the steps of: culturing a host cell comprising a nucleic acid or a vector encoding the antibody of the present disclosure.

[0167] The aspects, advantageous features and preferred embodiments of the present disclosure outlined in the following, respectively alone or in combination, further contribute to solving the objects of the present invention:

[0168] 1. An isolated antibody having binding specificity for human IL-17A, comprising a set of CDRs:

[0169] 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:

[0170] (i) HCDR1′ is an amino acid sequence selected from any one of SEQ ID NOs: 1, 4 and 7;

[0171] HCDR2' is an amino acid sequence selected from any one of SEQ ID NOs: 2, 5 and 8;

[0172] HCDR3' is an amino acid sequence selected from any one of SEQ ID NOs: 3, 6 and 9;

[0173] LCDR1′ is an amino acid sequence selected from any one of SEQ ID NOs: 12, 15 and 18;

[0174] LCDR2' is an amino acid sequence selected from any one of SEQ ID NOs: 13, 16 and 19;

[0175] LCDR3' has an amino acid selected from any one of SEQ ID NOs: 14, 17 and 20; or

[0176] (ii) HCDR1′ is an amino acid sequence selected from any one of SEQ ID NOs: 39, 42 and 45;

[0177] HCDR2' is an amino acid sequence selected from any one of SEQ ID NOs: 40, 43 and 46;

[0178] HCDR3′ is an amino acid sequence selected from any one of SEQ ID NOs: 41, 44 and 47;

[0179] LCDR1′ is an amino acid sequence selected from any one of SEQ ID NOs: 50, 53 and 56;

[0180] LCDR2' is an amino acid sequence selected from any one of SEQ ID NOs: 51, 54 and 57;

[0181] LCDR3′ has an amino acid sequence selected from any one of SEQ ID NOs: 52, 55 and 58.

[0182] 2. An antibody according to item 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:

[0183] (i) HCDR1′ is as shown in SEQ ID NO: 1;

[0184] HCDR2' is shown in SEQ ID NO:1;

[0185] HCDR3' is shown in SEQ ID NO:3;

[0186] LCDR1' is shown in SEQ ID NO:12;

[0187] LCDR2' is shown in SEQ ID NO:13;

[0188] LCDR3' is shown in SEQ ID NO:14; or

[0189] (ii) HCDR1′ is as shown in SEQ ID NO:39;

[0190] HCDR2' is shown in SEQ ID NO:40;

[0191] HCDR3' is shown in SEQ ID NO:41;

[0192] LCDR1' is shown in SEQ ID NO:50;

[0193] LCDR2' is shown in SEQ ID NO:51;

[0194] LCDR3' is shown in SEQ ID NO:52.

[0195] 3. The antibody according to item 1 or 2, comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein:

[0196] (c) the VH comprises three complementary determining regions HCDR1, HCDR2 and HCDR3 in sequence, and

[0197] (d) The VL comprises three complementary determining regions LCDR1, LCDR2 and LCDR3 in sequence.

[0198] 4. The antibody according to item 3, wherein

[0199] (i)

[0200] (a) the HCDR1 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 1, 4 and 7;

[0201] (b) the HCDR2 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 2, 5 and 8;

[0202] (c) the HCDR3 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 3, 6 and 9;

[0203] (d) the LCDR1 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 12, 15 and 18;

[0204] (e) the LCDR2 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 13, 16 and 19; and

[0205] (f) the LCDR3 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 14, 17 and 20; or (ii)

[0206] (a) the HCDR1 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 39, 42 and 45;

[0207] (b) The HCDR2 is as shown by the amino acid sequence selected from any one of SEQ ID NO: 40, 43, and 46;

[0208] (c) The HCDR3 is as shown by the amino acid sequence selected from any one of SEQ ID NO: 41, 44, and 47;

[0209] (d) The LCDR1 is as shown by the amino acid sequence selected from any one of SEQ ID NO: 50, 53, and 56;

[0210] (e) The LCDR2 is as shown by the amino acid sequence selected from any one of SEQ ID NO: 51, 54, and 57; and

[0211] (f) The LCDR3 is as shown by the amino acid sequence selected from any one of SEQ ID NO: 52, 55, and 58.

[0212] 5. The antibody according to item 4, wherein the antibody comprises (i) the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NO: 1, 2, and 3 respectively, and the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NO: 12, 13, and 14 respectively; or (ii) 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.

[0213] 6. The antibody according to any one of items 3 to 5, wherein the VH is VH3 or VH4, preferably VH3.

[0214] 7. The antibody according to any one of items 3 to 6, wherein the VL comprises Vκ framework FR1, FR2, and FR3, particularly Vκ1 or Vκ3 FR1 to FR3, preferably Vκ1 FR1 to FR3, and framework FR4, and the framework FR4 is selected from Vκ FR4, particularly Vκ1 FR4, Vκ3 FR4, and Vλ FR4, particularly Vλ FR4 comprising an amino acid sequence having at least 60%, 70%, 80%, 90% identity with the amino acid sequence selected from any one of SEQ ID NO: 26 to SEQ ID NO: 32, preferably Vλ FR4 as shown by any one of SEQ ID NO: 26 to SEQ ID NO: 32, preferably Vλ FR4 as shown by SEQ ID NO: 26 or 27, more preferably Vλ FR4 as shown by SEQ ID NO: 27.

[0215] 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.

[0216] 9. An antibody according to 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.

[0217] 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.

[0218] 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.

[0219] 12. The antibody according to any of the preceding items, wherein the antibody has binding specificity for cynomolgus monkey IL-17A.

[0220] 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.

[0221] 14. The antibody according to any one of the preceding items, wherein it binds to IL-17A

[0222] (c) inhibiting or blocking the binding between IL-17A and its receptor (IL-17RA), and

[0223] (d) reducing or neutralizing IL-17A activity.

[0224] 15. The antibody of claim 14, wherein the antibody is capable of inhibiting GRO-α secretion when evaluated in vitro in the HT-29 assay.

[0225] 16. The antibody according to any of the preceding items, wherein the antibody:

[0226] (k) 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, and wherein the relative potency is the IC of secukinumab in ng / mL as measured by ELISA 50 The values ​​are compared with the IC values ​​in ng / mL of the scFv form of the antibody of the present invention measured by ELISA. 50 value ratio; and / or

[0227] (l) 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 value ratio; and / or

[0228] (m) 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, said inhibitory activity being determined by measuring the GRO-α secretion induced by human IL-17A in the presence of 50 pg / ml TNFα in a HT-29 assay.

[0229] 17. The antibody according to any of the preceding items, wherein the antibody:

[0230] (c) 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 setup; and

[0231] (d) 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.

[0232] 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.

[0233] 19. The antibody according to any of the preceding items, wherein the antibody:

[0234] (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 at pH 6.4, 150 mM NaCl;

[0235] (f) when in scFv format, when the starting concentration of the antibody of the invention is 10 mg / ml, after 5 consecutive freeze-thaw cycles, the monomer content loss is less than 5%, particularly less than 3%, more particularly less than 1%, particularly wherein the antibody is in phosphate buffered saline (PBS) at pH 7.4;

[0236] (g) When in scFv form, when the initial concentration of the antibody of the present invention is 10 mg / ml, at 4°C

[0237] The monomer content loss is 5% or less, particularly less than 4%, less than 3%, less than 2%, more particularly less than 1% after storage for at least two weeks, particularly at least 4 weeks, particularly wherein the antibody is in phosphate buffered saline (PBS) at pH 7.4; and / or

[0238] (h) When the initial concentration of the antibody of the present invention is 10 mg / ml, the loss of monomer content is less than 5% after storage at 37°C for at least two weeks, especially at least four weeks.

[0239] 20. The antibody of any of the preceding items, wherein the antibody is 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 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 scFv.

[0240] 21. An 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.

[0241] 22. An 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.

[0242] 23. The isolated antibody according to any of the preceding items, wherein the antibody is humanized.

[0243] 24. The antibody of any one of items 1 to 23, which is a multispecific molecule.

[0244] 25. The antibody according to item 24, wherein the form of the 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, triabody (Fab-(scFv) 2 ) or diabody (Fab-(scFv) 1 ), Fab, Fab-Fv 2 Morrison (IgG CH 3-scFv fusion (Morrison L) or IgG CL-scFv fusion (Morrison H)), triabodies, scDb-scFv, bispecific Fab 2 , diabodies, tetrabodies, scFv-Fc-scFv fusion, scFv-HSA-scFv fusion, diabodies, DVD-Ig, COVD, IgG-scFab, scFab-dsscFv, Fv 2 -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), 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) fused to the N- and / or C-terminus of either chain of a heterodimeric Fc domain or any other heterodimeric domain 2 , Fab-(scFv)1, Fab, Fab-Fv 2 , COVD, MATCH and DuoBody.

[0245] 26. A pharmaceutical composition comprising the antibody according to any one of items 1 to 25 and a pharmaceutically acceptable carrier.

[0246] 27. The antibody of any one of items 1 to 25, or the pharmaceutical composition of item 26, for use as a medicament.

[0247] 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.

[0248] 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.

[0249] 30. An antibody as described in any one of items 1 to 25, or a pharmaceutical composition as described in item 26, for use in the treatment of 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, extraurological inflammatory disorders, cardiovascular disease, vasculitis, periodic fever, 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.

[0250] 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.

[0251] 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.

[0252] 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, allogeneic transplant rejection, graft-versus-host disease, systemic sclerosis, urinary inflammatory disorders, cardiovascular disease, vasculitis, periodic fever, 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.

[0253] 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 condition is alleviated.

[0254] 35. The method of claim 34, wherein the disorder mediated by IL-17A is an inflammatory condition or an autoimmune disease.

[0255] 36. A method according to 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, allogeneic transplant rejection, graft-versus-host disease, systemic sclerosis, urological inflammatory disorders, cardiovascular disease, vasculitis, periodic fever, 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.

[0256] 37. A nucleic acid encoding the antibody according to items 1-25.

[0257] 38. A vector comprising the nucleic acid described in item 37.

[0258] 39. A host cell comprising the nucleic acid described in item 37 or the vector described in item 38.

[0259] 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.

[0260] 41. A kit comprising the antibody of any one of items 1 to 25, or the pharmaceutical composition of item 26.

[0261] 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.

[0262] 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

[0263] Figure 1Shown that labeled IL-17A is suitable for the sorting process. Biological activity of labeled IL-17A in HT-29 assay. Serial 3-fold 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 50 values of labeled IL-17A (IL-17A-RPE) and unlabeled IL-17A (IL-17A) were 82.8 ng / ml and 55 ng / ml, respectively.

[0264] Figure 2 Shown the potency of 27-07-G02 rabbit IgG (A) and 27-31-C04 IgG (B) to neutralize IL-17A in HT-29 assay.

[0265] Figure 3 Shown the potency of 27-07-G02 rabbit IgG (A) and 27-31-C04 IgG (B) to inhibit the interaction between IL-17A and IL-17RA.

[0266] Figure 4 Shown the potency of humanized scFv A1 and A2 (A) and PRO571 and PRO592 (B) to neutralize IL-17A in HT-29 assay.

[0267] Figure 5 Shown the potency of anti-IL-17A scFv A1 and A2 (A) and PRO571 and PRO592 (B) to inhibit the interaction between human IL-17A and IL-17RA (ELISA).

[0268] Figure 6 Shown the target specificity of scFv A1 (A) and scFv PRO571 and PRO592 (B). The potential of IL-17B to IL-17F to inhibit the interaction of biotinylated IL-17A with scFv was analyzed by competitive ELISA. Dose-dependent effects of IL-17A and IL-17B to IL-17F were shown.

[0269] Figure 7 Shown the thermal unfolding curves of scFv A1 and A2 (A) and scFv PRO571 and PRO592 (B) measured by DSF. The resulting Tm values were determined by fitting the data to the Boltzmann equation to obtain the midpoint of the transition.

[0270] Figure 8Storage stability studies of scFv A1 (A) and scFv PRO571 and PRO592 (B) were performed 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.

[0271] Fig. 9 Shown are the monitoring of monomer content of scFv A1 (A) and scFv PRO571 and PRO592 (B) during 5 repeated freeze and thaw cycles.

[0272] Fig.10 A trispecific format is shown. A, Fab-(scFv) was designed 2 A, Three domain arrangements of the molecule. ScFv fusions at the CL and CH1 positions of the trimeric form were considered equivalent, resulting in three variants of this form. B, Three scDb-scFv domain arrangement variants 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.

[0273] Fig.11 The general process of lead manufacturing is shown. A, Overlay of SE-HPLC traces of final A3-A5 (left) and A6-A8 (right) samples. Peaks with retention times of 7-8 min correspond to the apparent molecular weight of the monomer of the respective molecule; peaks with retention times >10 min are buffer- and salt-related artifacts. 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 For antibody forms (A3-A5), the band shifted to ~50 kDa under reducing conditions, while for A6-A8, a band with a molecular weight of ~75 kDa could also be observed under reducing conditions.

[0274] Fig.12 The potency to neutralize 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.

[0275] Fig.13 Comparison of potency in neutralizing human and cynomolgus monkey TNF-α is shown. Absorbance measured using the Cell Counting Kit-8 in the presence of human or cynomolgus monkey TNF-α is expressed as a function of A5 and A7 concentration.

[0276] Fig.14 Simultaneous blockade of TNF-α and IL-17A in the HT-29 assay in the presence of HSA is shown. The simultaneous in vitro blockade of TNFα and IL-17A by six trispecific molecules A3-A8 was analyzed using an HT-29 cell-based assay in the presence of 1 mg / ml HSA. Secukinumab (IL-17A blocker) and A13 (parental bispecific, HSA binder and TNFα blocker) were used as references. The GRO-α secretion data obtained are expressed as a function of the molecular concentration in nM (A, C and E) and ng / ml (B, D and F). "Without TNF" represents GRO-α secretion when only IL-17A is added, which corresponds to the maximum effect of TNFα blockade. "Without IL17a" represents GRO-α secretion after only TNFα is added, which corresponds to the maximum effect of IL-17A blockade. "No IL17a, no TNFa" represents the background GRO-α secretion without the addition of IL-17A and TNFα, which corresponds to the maximal effect of simultaneous blockade of TNFα and IL-17A.

[0277] Fig.15 Neutralization of IL-17A binding to IL-17RA in a competitive ELISA is shown. The 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.

[0278] Fig.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 molecules were 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.

[0279] Fig.17Storage stability studies at 10 mg / mL protein concentration for 4 weeks at 37° C., 4° C., and -80° C. are shown. The % monomer content and % monomer loss over time were recorded at d0, d2, d7, d14, d21, and d28.

[0280] Fig.18 Shown are overlays of SE-HPLC traces of d0 (black, shaded) and d28 (grey) stability samples of A5 (left), A7 (middle) and A8 (right).

[0281] Fig.19 Shown are the pharmacokinetic profiles and ADA analysis of A7 following intravenous (n=3) and subcutaneous (n=3) injections in cynomolgus monkeys.

[0282] Fig. 20 Schematic representation of the structures of Morrison L constructs A14 and A15 is shown.

[0283] Fig.21 Shown are the average sizes of soluble complexes of scDb-scFvA7 determined by dynamic light scattering.

[0284] Fig. 22 Shown are the average sizes of soluble complexes of A14 determined by dynamic light scattering.

[0285] Fig.23 Shown are the average sizes of soluble complexes of A15 determined by dynamic light scattering.

[0286] Fig.24 The protein concentration recovery of the soluble complex of scDb-scFv A7 is shown.

[0287] Fig.25 The protein concentration recovery of the soluble complex of A14 is shown.

[0288] Fig.26 The protein concentration recovery of the soluble complex of A15 is shown. DETAILED DESCRIPTION

[0289] 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 disclosed in 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.), immune complexes are likely to form, which may lead to immunogenicity or other adverse reactions. In contrast, the monovalent bispecific and trispecific constructs disclosed in 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 disclosed in the present disclosure have better biophysical properties, such as developability and high-yield producibility, relatively few impurities, and superior stability.

[0290] The present disclosure also provides antibodies that specifically bind to human IL-17A protein, as well as pharmaceutical compositions, methods of production, and methods of using such antibodies and pharmaceutical compositions.

[0291] 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.

[0292] 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."

[0293] 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" and "an" and "the" and similar references should be interpreted as covering 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 plural form, this is also considered to refer to a single compound, salt, etc.

[0294] Multispecific Antibodies of the Disclosure

[0295] In one aspect, the disclosure provides an isolated multispecific antibody comprising a first domain that specifically binds IL-17A and a second domain that specifically binds TNFα.

[0296] The term "TNFα" or "TNF-α" or "tumor necrosis factor" refers in particular to human TNFα. TNFα exists in the form of a soluble protein and a precursor, which is called transmembrane TNFα, expressed as a cell surface type II polypeptide. 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 an uncleaved monomer of 26-kD. As used herein, the term "TNFα" includes soluble and transmembrane forms. The term "TNFa" refers in particular to human transmembrane TNFa with UniProt ID number P01375, reproduced herein as SEQ ID NO:134. The term "TNFa" refers in particular to the soluble, transmembrane TNFa having UniProt ID No. P01375, reproduced herein as SEQ ID NO: 135.

[0297] 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, an isolated antibody that specifically binds 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 materials and / or chemicals.

[0298] 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.

[0299] Antibodies of the present disclosure include, but are not limited to, chimeric and humanized.

[0300] The term "chimeric antibody" is an antibody molecule in which (a) the constant region or a portion thereof is altered, replaced or exchanged so that the antigen binding site (variable region) is linked to a constant region of a different or altered class, effector function and / or type, or to a completely different molecule that confers new properties to the chimeric antibody, such as an enzyme, toxin, hormone, growth factor, drug, etc.; or (b) the variable region or a portion thereof is altered, replaced or exchanged for a variable region with a different or altered antigen specificity. For example, a mouse antibody can be modified by replacing the constant region of a human immunoglobulin with that of a human immunoglobulin. Due to the replacement by the human constant region, the chimeric antibody can retain its specificity for recognizing an antigen while having reduced human antigenicity compared to the original mouse antibody.

[0301] As used herein, a "humanized" antibody is an antibody that retains the reactivity of a non-human antibody while being less immunogenic to humans. For example, this can be achieved by retaining the non-human CDRs and replacing the remainder of the antibody with a human counterpart (i.e., the framework portion of the constant region and the variable region). 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 include 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, e.g., 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 techniques include, but are not limited to, the Xoma technique disclosed in U.S. Pat. No. 5,766,886.

[0302] As used herein, the term "recombinant humanized antibody" includes all humanized antibodies of the present disclosure 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 recombinant combinatorial human antibody libraries; 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 regions and constant regions (if present) from human germline immunoglobulin sequences. However, such antibodies can be subjected to in vitro mutagenesis (or, when transgenic animals of human Ig sequences are used, in vivo somatic cell 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 exist in the human antibody germline.

[0303] Suitably, the antibodies or binding domains thereof of the disclosure are humanized. Suitably, the antibodies or binding domains thereof of the disclosure are humanized and comprise CDRs derived from rabbit.

[0304] 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.

[0305] 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 an antigen in a molecule. Therefore, the terms "monovalent", "bivalent", "tetravalent" and "hexavalent" refer to the presence of one, two, four and six binding domains that are specific for an antigen in a molecule, respectively. As used herein, the term "monovalent antibody" refers to an antibody with a single antigen binding moiety 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.

[0306] The multispecific antibodies disclosed herein can be monovalent or multivalent, such as bivalent, trivalent or tetravalent, with monovalent being preferred, for binding to IL-17A.

[0307] The multispecific antibodies disclosed herein may be monovalent or multivalent, such as bivalent, trivalent or tetravalent, preferably monovalent, for binding to TNFα. Since TNFα forms a trimer, it may be trivalent and may form a three-dimensional immune complex in which the antibody has several domains that specifically bind to TNFα, such as a bivalent, trivalent or multivalent antibody for binding to TNFα. To illustrate, at different antigen / antibody ratios, the size of the immune complex formed between TNF and infliximab (chimeric TNFα IgG antibody) and etanercept (TNFR2 dimer fusion protein with IgG1Fc) showed that each antibody produced an immune complex with unique size characteristics (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.

[0308] 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, there is an optional polypeptide linker and consists of a polypeptide having 4 to 25 amino acid residues.

[0309] Suitably, the multispecific antibodies of the present disclosure are advantageously capable of neutralizing the biological activity of human TNFα and human IL-17A. It will be appreciated that the term "neutralization" as used herein refers to a reduction in biological signal activity, which reduction may be partial or complete. Suitable methods for determining neutralization are known in the art, and certain such detection methods are provided in the Examples herein.

[0310] In one embodiment, the antibody or first domain thereof of the present disclosure selectively binds 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" means that an antibody, composition, formulation, etc., does not significantly bind to IL-17B / C / D / E / F, but binds to IL-17A. Selective binding is characterized by high affinity (or low K D ) and low to moderate IC 50 , which is distinguished from nonspecific binding, which usually has low affinity (or high K D ) and medium to high IC 50 Typically, when the antibody binds to the K D Less than 10 -7 Suitably, the antibody or first domain thereof of the present disclosure binds to human IL-17A with a higher affinity or K than it binds 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 of the present disclosure for IL-17B, IL-17C, IL-17D, IL-17E and IL-17F are 50 Values ​​compared to the IC of IL-17A 50At least 100 times higher, such as at least 200 times higher, at least 300 times higher, at least 400 times higher, as measured by ELISA. In one embodiment, the antibody or the first domain thereof of the present disclosure 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.

[0311] In another embodiment, the multispecific antibody of the present disclosure further comprises a third domain having specificity for an antigen different from IL-17A and TNFα. Suitably, the multispecific antibody of the present disclosure is a trispecific antibody. As used herein, a "trispecific antibody" refers to an antibody molecule having three antigen binding domains, for example, one of which binds to human TNFα, another binds to human IL-17A, and another binds to an antigen that can extend the half-life of the antibody molecule, such as human serum albumin.

[0312] In particular, the multispecific antibody of the present disclosure further comprises a third domain that specifically binds to human serum albumin (HSA).

[0313] 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:

[0314] (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

[0315] (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.

[0316] The term "HSA" refers in particular to human serum albumin with UniProt ID number P02768. Human serum albumin (HSA) is an abundant protein of 66.4 kDa in human serum (50% of total protein) and is composed of 585 amino acids (Sugio, Protein Eng, Vol. 12, 1999, 439-446). The multifunctional HSA protein is associated with its structure, which allows the binding and transport of many metabolites, such as fatty acids, metal ions, bilirubin and certain drugs (Fanali, Molecular Aspects of Medicine, Vol. 33, 2012, 209-290). The HSA concentration in serum is about 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.

[0317] Suitably, in one embodiment, the multispecific antibody of the present disclosure comprises a first domain that specifically binds to IL-17A and a second domain that specifically binds to TNFα, and a third domain that specifically binds to human serum albumin. The multispecific antibody of the present disclosure may be monovalent or multivalent, such as bivalent, trivalent or tetravalent, preferably monovalent, for binding to human serum albumin. Suitably, the multispecific antibody of the present disclosure comprises only one domain that specifically binds to IL-17A and 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 antibody of the present disclosure consists 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, there is an optional polypeptide linker and consists of a polypeptide having 4 to 25 amino acid residues.

[0318] 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.

[0319] The domains of the multispecific antibodies of the present disclosure, for example, the first domain, the second domain, and 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 domains based on ankyrin, fynomers, avimers, anticalins, fibronectin, and binding sites constructed in 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.

[0320] The multispecific antibodies of the present disclosure may be in any suitable form. In one embodiment, the form 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, tri-Fab (Fab-(scFv) 2 ) or diabody (Fab-(scFv) 1 ), Fab, Fab-Fv 2 , Morrison (IgG CH3-scFv fusion (Morrison L) or IgG CL-scFv fusion (Morrison H)), triabodies, scDb-scFv, bispecific Fab 2 , diabodies, tetrabodies, scFv-Fc-scFv fusion, scFv-HSA-scFv fusion, diabodies, DVD-Ig, COVD, IgG-scFab, scFab-dsscFv, Fv 2-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), 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) fused to the N- and / or C-terminus of either chain of a heterodimeric Fc domain or any other heterodimeric domain 2 , Fab-(scFv) 1 , Fab, Fab-Fv 2 , COVD, MATCH and DuoBody, preferably a trio or scDb-scFv.

[0321] The term "diabody" refers to an antibody fragment with two antigen binding sites, which fragments include VH (VH-VL) connected to VL in the same polypeptide chain. By using a linker that is too short to allow pairing between 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 the first antigen, and VLB and VHB together form an antigen binding site for the second antigen. The linker L1 is particularly a peptide of 2-10 amino acids, more particularly 3-7 amino acids, and most particularly 5 amino acids, and the 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.

[0322] 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 those that bind IgG antibodies (γ receptors), and include receptors of the FcγRI, FcγRII, and FcγRIII subclasses, including allelic variants and splice forms of these receptors. The FcγRII receptor includes FcγRIIA ("activating receptor") and FcγRIIB ("inhibitory receptor"), which have similar amino acid sequences, with the main difference being 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 inhibitory motif (ITIM) in its cytoplasmic domain (see M. Daeron, Annu. Rev. Immunol. 5:203-234 (1997)). An overview of FcRs is provided in 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" 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 with variant Fc regions. WO 2004 / 42072 (Presta) describes antibody variants that improve or reduce binding to FcR. See also, for example, Shields et al., J. Biol. Chem. 9 (2): 6591-6604 (2001).

[0323] In order to increase the number of specificities / functionalities 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 the complete antibody and also show improved tissue permeability and pharmacokinetic properties compared to the complete immunoglobulin molecule. Although such fragments seem to show many advantages relative to complete immunoglobulins, they are also plagued by high serum clearance rates because they lack the Fc domain with 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), so there is hope for improved efficacy at the same or lower doses. Suitably, the antibodies of the present disclosure do not include immunoglobulin Fc region polypeptides, and, optionally, do not include CH1 and CL regions, particularly when the multispecific antibody comprises a third domain that specifically binds to human serum albumin.

[0324] 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 when in a trimeric format, 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, the multispecific antibodies of the present disclosure have favorable properties.

[0325] Preferably, the antibody of the present disclosure is 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 antibody of the present disclosure, when in the form of scDb-scFv, has an scFv domain that is fused at the C-terminus to an scDb consisting of two other domains. The multispecific antibody of the present disclosure, when in the form of scDb-Fv, can be represented by the following formula:

[0326] VLA-L1-VHC-L2-VLC-L3-VHA-L4-VLB-L5–VHB or

[0327] VLB-L1-VHA-L2-VLA-L3-VHB-L4-VLC-L5–VHC or

[0328] VLC-L1-VHB-L2-VLB-L3-VHC-L4-VLA-L5–VHA or

[0329] VLA-L1-VHB-L2-VLB-L3-VHA-L4-VLC-L5–VHC,

[0330] Preferred VLB-L1-VHA-L2-VLA-L3-VHB-L4-VLC-L5–VHC or VLA-

[0331] L1-VHB-L2-VLB-L3-VHA-L4-VLC-L5–VHC,

[0332] More preferably VLB-L1-VHA-L2-VLA-L3-VHB-L4-VLC-L5–VHC,

[0333] 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, wherein L1, L2, L3, L4 and L5 are polypeptide linkers.

[0334] In the context of the present disclosure, the term "polypeptide linker" refers to a linker 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 linker. The length of a polypeptide linker should be enough to connect two molecules so that they have the correct conformation to each other, thereby maintaining the desired activity. In a specific embodiment, a polypeptide linker has a continuous chain of 2 to 30 amino acid residues (e.g., 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 the activity of the polypeptide. Therefore, the linker peptide should not show an inconsistency with the 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 other interacting charges that will seriously hinder the combination of the monomer domain. 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 materials that may be globular glycine chains. Thirdly, similar chains have been shown to effectively connect subunits of recombinant proteins such as single-chain antibodies.

[0335] Suitably, the L1 and L3 of the present disclosure are as shown in SEQ ID NO: 132. Suitably, the L2, L4 and L5 of the present disclosure are as shown in SEQ ID NO: 23.

[0336] The present 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 when in the scDb-scFv format, when the first domain and the second domain that specifically bind to IL-17A and TNFα, respectively, form scDb, and the third domain that specifically binds to HSA is scFv, the multispecific antibody of the present disclosure has favorable properties.

[0337] The multispecific antibodies of the present disclosure have the following advantageous properties:

[0338] (a) having 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 the values; and

[0339] (b) having the ability to neutralize TNFα with a potency (relative potency) relative to the scDb (A13) according to SEQ ID NO: 149 as determined by measuring Gro-α secretion in the HT-29 assay 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, 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 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

[0340] (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 Value and

[0341] IC of the multispecific antibody in ng / mL measured by ELISA 50 The ratio of the values; and

[0342] (d) optionally, having the ability to neutralize TNFα with a potency (relative potency) relative to the scDb (A13) according to SEQ ID NO: 149 of at least 0.4, e.g. at least 0.5, preferably at least 1 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 Values ​​are comparable to the IC values ​​in nM of the multispecific antibodies measured in the L929 assay. 50 value ratio; and / or

[0343] (e) Dissociation constant (K) for binding to human IL-17A D ) is less than 5 nM, such as less than 4 nM, less than 3 nM,

[0344] 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 of 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;

[0345] (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

[0346] (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

[0347] Optionally, the 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.

[0348] 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 region of the antibody "arm" interacts with the antigen at many sites through weak non-covalent forces; the greater the interaction, the stronger the affinity.

[0349] "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 reflecting 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) is indicated. 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 readily, while high-affinity antibodies generally bind antigen more quickly and tend to remain bound for longer periods. A variety of methods for measuring binding affinity are known in the art, and any of these methods 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.

[0350] As used herein, the term "K assoc ", "Ka" or "K on " refers to the association rate of a specific antibody-antigen interaction, and the term "K dis ", "Kd" or "K off " refers to the dissociation rate of a specific antibody-antigen interaction. In one embodiment, the term "K D " as used herein refers to the dissociation constant, which is obtained as the ratio of Kd to Ka (i.e., Kd / Ka) and is expressed as a molar concentration (M). In one embodiment, the "K D " or "K D value" or "KD" or "KD value" according to the present disclosure is measured by surface plasmon resonance assay using a MASS-1SPR instrument (Sierra Sensors), as described in the examples. The binding affinity of an antibody can be determined, for example, by the dissociation constant (K D ). A lower K D indicates a stronger affinity, while a higher K D indicates a weaker affinity.

[0351] 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 ) 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 ) 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 (KD ) 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).

[0352] Suitably, the dissociation constant (K) of the antibodies of the present disclosure for binding to human TNFα 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, 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 dissociation constant (K) of an antibody of the present disclosure for binding to human TNFα is 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.

[0353] In suitable embodiments, the dissociation constant (K) of an antibody of the present disclosure for binding to human serum albumin is D ) is 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 dissociation constant (K) of an antibody of the present disclosure for binding to human serum albumin is 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).

[0354] Suitably, the antibodies of the present disclosure have beneficial biophysical characteristics.

[0355] (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;

[0356] (b) when the multispecific antibody is at an initial 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;

[0357] (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

[0358] (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, in particular at least four weeks, the loss of monomer content is less than 20%, preferably less than 15%.

[0359] 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 transition midpoint of thermal unfolding was determined by differential scanning fluorimetry (DSF) using fluorescent dyes Orange determined (see Wong & Raleigh, Protein Science 25 (2016) 1834-1840). Samples were prepared at a final protein concentration of 50 μg / mL in citrate phosphate buffer at pH 6.4 and contained 5x The final concentration of Orange, with a total volume of 100 μl. Twenty-five microliters of the prepared sample were added in triplicate to a white-wall AB gene PCR plate. The assay was performed in a qPCR machine used as a thermal cycler, and fluorescence emission was detected using the custom dye calibration program of the software. The PCR plate containing the test samples was subjected to a temperature change from 25 °C to 96 °C in 1 °C increments, and after each temperature increment, a pause of 30 seconds was made. The total assay time was approximately two hours. The Tm was calculated by using the mathematical second derivative method with GraphPad Prism software to calculate the inflection point of the curve. The reported Tm is the average of three measurements.

[0360] 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. This 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 permeation volume (VT) of a specific column. The measurement by SE-HPLC was carried out on a Chromaster HPLC system (Hitachi High-Technologies Corporation) equipped with an automatic sample injector and a UV detector with the detection wavelength set at 280 nm. The device was controlled by the software EZChrom Elite (Agilent Technologies, Version 3.3.2SP2), which also supported the analysis of the resulting chromatogram. The protein sample was cleared by centrifugation before injection, and the temperature of the autosampler was maintained at 6 °C. For the analysis of scFv samples, a Shodex KW403-4F column (Showa Denko K.K., #F6989202) was used, with a standard buffered saline mobile phase (50 mM sodium phosphate pH 6.5, 300 mM sodium chloride), and the recommended flow rate was 0.35 mL / min. The target sample loading amount for each injection was 5 μg. The samples were detected by a UV detector at a wavelength of 280 nm, and the data were recorded by a suitable software suite. The resulting chromatogram was analyzed in the range from V0 to VT, thus excluding matrix-related peaks with an elution time > 10 minutes.

[0361] Exemplary domain that specifically binds to IL-17A

[0362] The multispecific antibody of the present disclosure comprises 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 sequentially comprises three complementarity-determining regions HCDR1, HCDR2, and HCDR3, and (b) the VL sequentially includes three complementarity-determining regions LCDR1, LCDR2, and LCDR3.

[0363] Suitable domains for use in the multispecific antibodies of the present disclosure that specifically bind to IL-17A include, but are not limited to:

[0364] 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;

[0365] 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;

[0366] LY2439821 (also known as ixekizumab; disclosed in U.S. Pat. Nos. 7,838,638 and 8,110,191 and WO2007 / 070750, the entire contents of which are incorporated herein by reference) or an antigen-binding fragment thereof;

[0367] SCH900117 or its antigen-binding fragment (Merck);

[0368] RG4943 or its antigen-binding fragment (Roche);

[0369] ·WO 2006 / 013107,WO 2006 / 054059,WO 2007 / 070750,WO 2007 / 149032,

[0370] WO 2008 / 001063,WO 2008 / 021156,WO 2010 / 034443,WO 2010 / 102251,

[0371] The anti-IL-17A antibodies or antigen-binding fragments thereof disclosed in WO 2012 / 018767, WO 2014 / 161570, WO 2014 / 001368, WO 2014 / 122613, WO 2015 / 070697, WO 2015 / 137843, WO 2016 / 048188, WO 2016 / 113557, WO 2016 / 138842, and WO 2017 / 068472, the entire contents of which are incorporated herein by reference.

[0372] 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, and their sequences are listed in Table 1.

[0373] 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 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: 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:44 and 47, preferably SEQ ID NO:41; 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.

[0374] 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 of the VH CDRs listed in Table 1.

[0375] Suitably, the first domain that specifically binds to IL-17A comprises a heavy chain variable region (VH), wherein the VH comprises, in sequence, (i) three complementarity determining regions HCDR1, HCDR2 and HCDR3, the HCDR1 having an amino acid sequence selected from any one of SEQ ID NOs: 1, 4 and 7, the HCDR2 having an amino acid sequence selected from any one of SEQ ID NOs: 2, 5 and 8, and the HCDR3 having an amino acid sequence selected from any one of SEQ ID NOs: 3, 6 and 9. In particular, the present disclosure provides an antibody that has binding specificity to human IL-17A and comprises the HCDR1, HCDR2 and HCDR3 sequences of SEQ ID NOs: 1, 2 and 3, respectively; or (ii) three complementary 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 an antibody that has binding specificity to human IL-17A and comprises the HCDR1, HCDR2 and HCDR3 sequences of SEQ ID NOs: 39, 40 and 41, respectively.

[0376] 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.

[0377] 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, the LCDR1 having an amino acid sequence selected from any one of SEQ ID NOs: 12, 15 and 18, the LCDR2 having an amino acid sequence selected from any one of SEQ ID NOs: 13, 16 and 19, and the LCDR3 having an amino acid sequence selected from any one of SEQ ID NOs: 14, 17 and 20. In particular, the present disclosure provides an antibody having binding specificity to human IL-17A and comprising LCDR1, LCDR2 and LCDR3 sequences of SEQ ID NOs: 12, 13 and 14, respectively; or (ii) three complementary determining regions LCDR1, LCDR2 and LCDR3, 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. In particular, the present disclosure provides an antibody having binding specificity to human IL-17A and comprising LCDR1, LCDR2 and LCDR3 sequences of SEQ ID NOs: 50, 51 and 52, respectively.

[0378] 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),

[0379] (i)

[0380] (a) the VH comprises three complementary determining regions HCDR1, HCDR2 and HCDR3 in sequence, the HCDR1 has an amino acid sequence selected from any one of SEQ ID NOs: 1, 4 and 7, the

[0381] HCDR2 has an amino acid sequence selected from any one of SEQ ID NOs: 2, 5 and 8, and HCDR3 has an amino acid sequence selected from any one of SEQ ID NOs: 3, 6 and 9; and

[0382] (b) the VL includes three complementary determination regions LCDR1, LCDR2 and LCDR3 in sequence, the

[0383] LCDR1 has an amino acid sequence selected from any one of SEQ ID NOs: 12, 15 and 18,

[0384] LCDR2 has an amino acid sequence selected from any one of SEQ ID NOs: 13, 16 and 19,

[0385] LCDR3 has an amino acid sequence selected from any one of SEQ ID NOs: 14, 17 and 20; or

[0386] (ii)

[0387] (a) the VH comprises three complementary determining regions HCDR1, HCDR2 and HCDR3 in sequence, the HCDR1 having an amino acid sequence selected from any one of SEQ ID NOs: 39, 42 and 45, the HCDR2 having an amino acid sequence selected from any one of SEQ ID NOs: 40, 43 and 46, and the HCDR3 having an amino acid sequence selected from any one of SEQ ID NOs: 41, 44 and 47; and

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

[0389] LCDR1 has an amino acid sequence selected from any one of SEQ ID NOs: 50, 53 and 56,

[0390] LCDR2 has an amino acid sequence selected from any one of SEQ ID NOs: 51, 54 and 57,

[0391] LCDR3 has an amino acid sequence selected from any one of SEQ ID NOs: 52, 55 and 58.

[0392] 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 (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.

[0393] 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 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 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.

[0394] 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 term applies to amino acid polymers in which one or more amino acid residues are artificial chemical mimetics 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.

[0395] Mutations, such as substitutions, are made within the CDRs, antibody VH or VL domains, and the amino acid sequences of antibodies, and the techniques required are generally available in the art. Variant sequences can be made in which mutations, such as substitutions, may or may not be predicted to have minimal or beneficial effects on activity and tested for the ability to bind and / or neutralize IL-17A, or, 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 an antibody or binding domain thereof of the present disclosure comprising such a mutated amino acid sequence retains the ability to bind and / or neutralize IL-17A, or, TNFα or human serum albumin. For example, it may retain the same quantitative binding and / or neutralizing ability as an unchanged domain of the present disclosure, for example, measured in an assay described herein. Suitably, a domain or antigen-binding fragment of the present disclosure comprising such a mutated amino acid sequence may have an improved ability to bind and / or neutralize IL-17A, or, TNFα or human serum albumin. Suitably, the first domain of the present disclosure that specifically binds to IL-17A and comprises the 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 value.

[0396] In addition, appropriate mutations, such as substitutions, within the CDRs do not result in a loss of solubility, stability, and 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, it may retain the same high-yield manufacturability and / or stability as the unaltered multispecific antibodies of the present disclosure, for example, as measured in the assays described herein. Suitably, the multispecific antibodies or their binding domains comprising such mutated amino acid sequences may have improved biophysical properties.

[0397] In the context of two or more nucleic acid or polypeptide sequences, the term "identical" or percentage "identity" refers to two or more sequences or subsequences that are identical. "Amino acid sequence identity percentage (%)" 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 percentage sequence identity, and without considering any conservative substitutions as part of the sequence identity. Alignment for determining amino acid sequence identity percentage 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 suitable parameters for measuring alignment, including any algorithm required for achieving maximum alignment over the full length of the compared sequence.

[0398] To perform sequence comparison, one sequence is typically used as a reference sequence to which a test sequence is compared. When a sequence comparison algorithm is used, the test sequence and the reference sequence are entered into a computer, subsequence coordinates are specified if necessary, and sequence algorithm program parameters are specified. Default program parameters may be used, or other parameters may 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.

[0399] Two examples of algorithms that are 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.

[0400] 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. In addition, 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 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.

[0401] 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:

[0402] (i)

[0403] (a) the VH comprises three complementary determining regions HCDR1, HCDR2 and HCDR3 in sequence,

[0404] The HCDR1 has the same sequence as any one of SEQ ID NO: 1, 4 and 7, preferably SEQ ID NO:

[0405] 1, with at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99%

[0406] amino acid sequence of identity;

[0407] The HCDR2 has the same sequence as any one of SEQ ID NO: 2, 5 and 8, preferably SEQ ID NO:

[0408] 2, with at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99%

[0409] amino acid sequence of identity;

[0410] The HCDR3 has the same sequence as any one of SEQ ID NO: 3, 6 and 9, preferably SEQ ID NO:

[0411] 3. Having at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99%

[0412] amino acid sequence; and / or

[0413] (b) the VL comprises three complementary determining regions LCDR1, LCDR2 and LCDR3 in sequence,

[0414] 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;

[0415] 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;

[0416] The LCDR3 has the same structure as any one of SEQ ID NOs: 14, 17 and 20, preferably SEQ ID NO: 14, and has at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98

[0417] or 99% identical amino acid sequence; or (ii)

[0418] (a) the VH comprises three complementary determining regions HCDR1, HCDR2 and HCDR3 in sequence,

[0419] 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: 39, 42 and 45, preferably SEQ ID NO: 39;

[0420] 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: 40, 43 and 46, preferably SEQ ID NO: 40;

[0421] The HCDR3 has the same structure as any one of SEQ ID NO:41, 44 and 47, preferably SEQ ID NO:41, and has at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98

[0422] or an amino acid sequence with 99% identity; and / or

[0423] (b) The VL sequentially includes three complementarity determining regions LCDR1, LCDR2, and LCDR3,

[0424] The LCDR1 has an amino acid sequence having at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identity with any one of SEQ ID NO: 50, 53, and 56, preferably SEQ ID NO: 50;

[0425] The LCDR2 has an amino acid sequence having at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identity with any one of SEQ ID NO: 51, 54, and 57, preferably SEQ ID NO: 51;

[0426] The LCDR3 has an amino acid sequence having at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identity with any one of SEQ ID NO: 52, 55, and 58, preferably SEQ ID NO: 52.

[0427] Suitably, the first domain that specifically binds to IL-17A comprises: (i) HCDR1, HCDR2, and HCDR3 having at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identity with the sequences of SEQ ID NO: 1, 2, and 3, respectively, and / or LCDR1, LCDR2, and LCDR3 having at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identity with the sequences of SEQ ID NO: 12, 13, and 14, respectively; or (ii) HCDR1, HCDR2, and HCDR3 having at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identity with the sequences of SEQ ID NO: 39, 40, and 41, respectively, and / or LCDR1, LCDR2, and LCDR3 having at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identity with the sequences of SEQ ID NO: 50, 51, and 52, respectively.

[0428] 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.

[0429] 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, such as 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 determined, for example, as shown in Knappik et al., J. Mol. Biol. 296 (2000) 57-86, which groups 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 rearrangements of germline κ chain V and J segments, germline λ chain V and J segments, and heavy chain V, D and J segments, respectively. Which subfamily a given antibody variable chain belongs to is determined by the corresponding V segment, in particular by the framework regions FR1 to FR3. Therefore, any VH sequence characterized only by a specific set of framework regions HFR1 to HFR3 in the present application can be combined with any HFR4 sequence, such as an HFR4 sequence obtained from one of the heavy chain germline J segments, or an HFR4 sequence obtained from a rearranged VH sequence.

[0430] 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 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 present disclosure that specifically binds to IL-17A comprises a heavy chain variable region VHA, wherein the VHA is VH3.

[0431] 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, and wherein the VLA comprises Vκ framework FR1, FR2 and FR3, in particular Vκ1 or Vκ3FR1 to FR3, preferably Vκ1 framework FR1 to FR3, and framework FR4, the framework FR4 is selected from VκFR4, in particular Vκ1FR4, Vκ3FR4, and VλFR4. Suitable VλFR4 is shown in SEQ ID NO:26 to SEQ ID NO:32. In one embodiment, the first domain that specifically binds to IL-17A comprises VλFR4, the 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 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.

[0432] 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:

[0433] (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;

[0434] (ii) a VH3 or VH4 domain, preferably a VH3 domain; and

[0435] (iii) a VL domain comprising a VL framework comprising Vκ framework FR1, FR2 and FR3, in particular Vκ1 or Vκ3FR1 to FR3, preferably Vκ1FR1 to FR3, and a framework FR4, wherein the framework FR4 is selected from 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 VλFR4 as shown in an amino acid sequence selected from any one of SEQ ID NO:26 to SEQ ID NO:32, more preferably VλFR4 as shown in SEQ ID NO:27.

[0436] 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 TNFa, 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 a number of 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 with the VH region described in the sequence described in Table 1.

[0437] Suitably, the present disclosure provides multispecific antibodies comprising a first domain that specifically binds IL-17A and a second domain that specifically binds TNFα, wherein the first domain comprises the VL domain listed in Table 1. Suitably, the present disclosure also provides multispecific antibodies comprising a first domain that specifically binds IL-17A and a second domain that specifically binds HSA, wherein the first domain comprises (alternatively, consists of) the VL amino acid sequence listed in Table 1, wherein no more than about 20 amino acids, preferably no more than about 10 amino acids, have been mutated in the framework sequences (e.g., sequences that are not CDRs) (wherein, as a plurality of non-limiting examples, the mutations are additions, substitutions, or deletions). Other domains that specifically bind IL-17A 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 to the VL domain described in the sequences described in Table 1 in the VL region.

[0438] In one embodiment, the present disclosure provides multispecific antibodies comprising a first domain that specifically binds IL-17A and a second domain that specifically binds 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 HCDR1, HCDR2, and HCDR3 sequences that are SEQ ID NOs:1, 2, and 3, respectively. In another embodiment, the first domain that specifically binds 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 number).

[0439] In another embodiment, the 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 NO: 39, 40 and 41, respectively. In another embodiment, the 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).

[0440] 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 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.

[0441] 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 light chain variable region, the light chain variable region comprises 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).

[0442] 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 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 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 SEQ ID NO: 21; or (ii) 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 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 SEQ ID NO: 59.

[0443] Preferably, the first domain of the multispecific antibody of the present disclosure that specifically binds to IL-17A 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 amino acid sequence of SEQ ID NOs: 1, 2 and 3, 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 the HCDR1, HCDR2 and HCDR3 sequences of SEQ ID NO:39, 40 and 41, respectively, and / or the 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: ID 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: 39, 40 and 41, respectively.

[0444] 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.

[0445] In a 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.

[0446] 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 the amino acid sequence of (i) an amino acid sequence selected from the group consisting of SEQ ID NOs: 24 and 25, preferably SEQ ID NO: 24, or (ii) an amino acid sequence selected from the group consisting of 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.

[0447] Other domains of the present disclosure having binding specificity for human IL-17A 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, the amino acid sequences are comprised of no more than 1, 2, 3, 4 or 5 amino acids in the variable region that have been mutated when compared to the variable region described in the sequence 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., a multispecific antibody of the present disclosure, in particular a multispecific antibody of the present disclosure comprising a first domain that specifically binds to human IL-17A described in Table 1 and / or a second domain that specifically binds to human TNFα described in Table 1).

[0448] 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 sequence 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.

[0449] In one embodiment, a domain of the invention that specifically binds to IL-17A has a heavy chain variable region comprising HCDR1, HCDR2, and HCDR3 sequences and a light chain variable region comprising LCDR1, LCDR2, and LCDR3 sequences, wherein one or more of these CDR sequences has a specified amino acid sequence based on the domains described herein or conservative modifications thereof, wherein the domains retain the desired functional properties of the antibodies of the present disclosure.

[0450] The term "conservatively modified variant" or "conservative variant" applies to amino acid and nucleic acid sequences. With respect to a particular nucleic acid sequence, conservatively modified variants refer to those nucleic acids encoding the same or substantially the same amino acid sequence, or when the nucleic acid does not encode an amino acid sequence, it refers to substantially the same sequence. 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. Therefore, at each position where alanine is clearly specified by a codon, the codon can be changed to any of the corresponding codons described without changing the encoded polypeptide. Such nucleic acid variations are "silent variations", which are a type of conservatively modified variations. Each nucleic acid sequence encoding a polypeptide herein also describes each possible silent variation of the nucleic acid. Those skilled in the art will recognize that each codon in a nucleic acid (except AUG and TGG, AUG is usually the only codon for methionine, and TGG is usually the only codon for tryptophan) can be modified to produce functionally identical molecules. Therefore, each silent variation of a nucleic acid encoding a polypeptide is implicit in each recorded sequence.

[0451] For polypeptide sequences, "conservatively modified variants" or "conservative variants" include single substitutions, deletions or additions to a polypeptide sequence which 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 conservatively substituted 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 change the binding properties of the antibody containing the amino acid sequence.

[0452] 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):

[0453] (i)

[0454] A heavy chain variable region (VH), which comprises three complementary 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

[0455] A light chain variable region (VL), which comprises three complementary determining regions LCDR1, LCDR2 and LCDR3 in sequence, 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

[0456] (ii)

[0457] A heavy chain variable region (VH), which comprises three complementary 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

[0458] A light chain variable region (VL), which comprises three complementary 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,

[0459] The antibody specifically binds to human IL-17A and / or neutralizes IL-17A.

[0460] Anti-IL-17A Antibodies of the Disclosure

[0461] The present disclosure is based on the discovery of antibody molecules that specifically bind to human IL-17A and have 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 few impurities (>98%, especially>99% monomers, as detected by SE-HPLC), and stability.

[0462] In one aspect, the present disclosure provides an isolated antibody having 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.

[0463] The term "IL-17A" or "IL17A" refers in particular to human IL-17A with UniProt ID number Q16552, reproduced herein as SEQ ID NO: 33. The term "cynomolgus IL-17A" or "cynomolgus monkey IL-17A" refers to Macacafascicularis IL-17A with UniProt ID number G1QUS7.

[0464] The term "IL-17B" particularly refers to human IL-17B with UniProt ID number Q9UHF5, reproduced herein as SEQ ID NO: 34. The term "IL-17C" particularly refers to human IL-17C with UniProt ID number Q9P0M4, reproduced herein as SEQ ID NO: 35. The term "IL-17D" particularly refers to human IL-17D with UniProt ID number Q8TAD2, reproduced herein as SEQ ID NO: 36. The term "IL-17E" particularly refers to human IL-17E with UniProt ID number Q9H293, reproduced herein as SEQ ID NO: 37. The term "IL-17F" particularly refers to human IL-17F with UniProt ID number Q96PD4, reproduced herein as SEQ ID NO: 38.

[0465] 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 come from two or more non-contiguous regions of a polypeptide or polypeptides together.

[0466] 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 consists of three CDRs and four FRs, which are arranged in the following order from the amino terminus to the 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.

[0467] 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 change Fc function, for example, to enhance or reduce effector function or binding to Fc receptors. Suitably, the antibody of the present disclosure is an IgG selected from the group consisting of IgG1, IgG2, IgG3 and IgG4. More suitably, the antibody of the present disclosure is IgG1 or IgG4.

[0468] 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 Fab fragments, a monovalent fragment consisting of the VL, VH, CL, and CH1 domains; F(ab) 2Fragments, a bivalent fragment comprising two Fab fragments connected by a disulfide bond at the hinge region; an Fd fragment consisting of a VH and CH1 domain; 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 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 the VNAR structure of sharks, and binding domains based on alternative scaffolds include but are not limited to domains based on ankyrin, fynomers, avimers, anticalins, fibronectin, and binding sites constructed in antibody constant regions (e.g., Modular Antibody Technology of F-star). TM ).

[0469] 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); 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 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.

[0470] 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 referred to as CDR-L1): L24-L42; LCDR2 (also referred to as CDR-L2): L58-L72; LCDR3 (also referred to as CDR-L3): L107-L138; HCDR1 (also referred to as CDR-H1): H27-H42; HCDR2 (also referred to as CDR-H2): H57-H76; HCDR3 (also referred to 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, typically not all positions 1 to 149 are occupied by amino acid residues.

[0471] 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 may 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 Hel l 36). The antigen binding portion of the antibody can be grafted into a framework based on a polypeptide (e.g., 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).

[0472] As used herein, the term "domain" or "domain that specifically binds to X" or "binding domain" of an antibody, "antigen binding fragment thereof", "antigen binding portion" and the like refers 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 a fragment of an intact antibody. In some embodiments, the binding domain of the multispecific antibody of the present disclosure is selected from the group consisting of: a Fab fragment, a monovalent fragment consisting of a VL, VH, CL and CH1 domain; a F(ab) fragment; 2Fragments, a bivalent fragment comprising two Fab fragments connected by a disulfide bond at the hinge region; an Fd fragment consisting of a VH and CH1 domain; 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 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 the VNAR structure of sharks, and binding domains based on alternative scaffolds include but are not limited to domains based on ankyrin, fynomers, avimers, anticalins, fibronectin, and binding sites constructed in 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.

[0473] Preferably, the "domain" or "domain that specifically binds to X" or "binding domain", "antigen-binding fragment thereof", "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 may also be incorporated into single domain antibodies, large antibodies, minibodies, intrabodies, 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 into a framework based on a polypeptide (e.g., 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).

[0474] As used herein, the term "binding specificity" or "specific binding" refers to the ability of a single antibody or antibody domain to react with an antigenic determinant but not with a different antigenic determinant. As used herein, the term "specific binding with ... " or "specific for " refers to a measurable and reproducible interaction, such as the binding between a target and an antibody or antibody domain, which is a determinant of the presence of a heterogeneous population of molecules (including biomolecules). For example, an antibody or antibody domain that specifically binds to a target (which may be an epitope) is an antibody or antibody domain that has a greater affinity, greater avidity, easier and / or longer duration than that of binding to the target than to 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 and an unrelated molecule, such as determined by a specific determination method known in the art. Such methods include, but are not limited to, Western blot, 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., secondary antibodies with horseradish peroxide and tetramethylbenzidine with hydrogen peroxide). The reaction in certain wells is scored by optical density (e.g., at 450nm). A typical background (=negative reaction) may be about 0.1OD; a typical positive reaction may be about 1OD. This means that the ratio between positive and negative scores can be 10 times or higher. In another example, an SPR assay can be performed, in which a difference of at least 10 times, preferably at least 100 times, between background and signal indicates specific binding. Typically, a group of about three to five unrelated molecules, such as milk powder, transferrin, etc., is used, rather than a single reference molecule, to determine binding specificity. The specific antibodies or antibody domains disclosed herein have binding specificity for human IL-17A or human TNFα.

[0475] The multispecific antibodies of the present disclosure include a first domain that specifically binds to IL-17A and a second domain that specifically binds to TNFα, and thus have binding specificity to IL-17A and TNFα, in particular, have binding specificity to human IL-17A and human TNFα. In one embodiment, the antibodies of the present disclosure have binding specificity to 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 to human TNFα and cynomolgus monkey (Macaca fascicularis) (also known as Cynomolgus monkey or "Cynomolgus") TNFα.

[0476] On the other hand, the present disclosure relates to antibodies or antibody domains that have binding specificity for human IL-17A and cynomolgus monkey (Macaca fascicularis) (also known as cynomolgus monkey or "cynomolgus") IL-17A.

[0477] Suitably, the anti-IL-17A antibodies of the present disclosure are isolated antibodies.

[0478] Suitably, the anti-IL-17A antibodies of the present disclosure are monoclonal antibodies.

[0479] The anti-IL-17A antibodies of the present disclosure include, but are not limited to, chimeric antibodies and humanized antibodies.

[0480] 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 contain rabbit CDRs.

[0481] The antibodies of the present disclosure include, but are not limited to, isolated humanized monoclonal antibodies 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.

[0482] The isolated antibodies of the present invention that have binding specificity for human IL-17A comprise a heavy chain variable region (VH) and a light chain variable region (VL), wherein: (a) the VH sequentially comprises three complementarity determining regions HCDR1, HCDR2, and HCDR3, and (b) the VL sequentially comprises three complementarity determining regions LCDR1, LCDR2, and LCDR3.

[0483] The present disclosure provides antibodies that specifically bind to the IL-17A protein, the antibodies comprising VH CDRs having the amino acid sequence of any one of the VH CDRs listed in Table 1. In particular, the present disclosure provides antibodies to the VH CDRs of the IL-17A protein, 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.

[0484] 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, the HCDR1 having an amino acid sequence selected from any one of SEQ ID NOs: 1, 4 and 7, the HCDR2 having an amino acid sequence selected from any one of SEQ ID NOs: 2, 5 and 8, and the HCDR3 having an amino acid sequence selected from any one of SEQ ID NOs: 3, 6 and 9. In particular, the present disclosure provides an antibody that has binding specificity to human IL-17A and comprises the HCDR1, HCDR2 and HCDR3 sequences of SEQ ID NOs: 1, 2 and 3, respectively; or (ii) three complementary 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 an antibody that has binding specificity to human IL-17A and comprises the HCDR1, HCDR2 and HCDR3 sequences of SEQ ID NOs: 39, 40 and 41, respectively.

[0485] The present disclosure also provides an antibody that specifically binds to an IL-17A protein, the antibody comprising a VL CDR, the VL CDR having an 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, the VL CDR having an amino acid sequence of any one of the VL CDRs listed in Table 1.

[0486] 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, the LCDR1 having an amino acid sequence selected from any one of SEQ ID NOs: 12, 15 and 18, the LCDR2 having an amino acid sequence selected from any one of SEQ ID NOs: 13, 16 and 19, and the LCDR3 having an amino acid sequence selected from any one of SEQ ID NOs: 14, 17 and 20. In particular, the present disclosure provides an antibody having binding specificity to human IL-17A and comprising LCDR1, LCDR2 and LCDR3 sequences of SEQ ID NOs: 12, 13 and 14, respectively; or (ii) three complementary 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 an antibody having binding specificity to human IL-17A and comprising LCDR1, LCDR2 and LCDR3 sequences of SEQ ID NOs: 50, 51 and 52, respectively.

[0487] Suitably, the present disclosure provides an isolated antibody having binding specificity to human IL-17A, comprising a heavy chain variable region (VH) and a light chain variable region (VL),

[0488] (i)

[0489] (a) the VH comprises three complementary determining regions HCDR1, HCDR2 and HCDR3 in sequence, the HCDR1 having an amino acid sequence selected from any one of SEQ ID NOs: 1, 4 and 7, the HCDR2 having an amino acid sequence selected from any one of SEQ ID NOs: 2, 5 and 8, and the HCDR3 having an amino acid sequence selected from any one of SEQ ID NOs: 3, 6 and 9; and

[0490] (b) 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: 12, 15 and 18, the LCDR2 having an amino acid sequence selected from any one of SEQ ID NOs: 13, 16 and 19, and the LCDR3 having an amino acid sequence selected from any one of SEQ ID NOs: 14, 17 and 20; or

[0491] (ii)

[0492] (a) the VH comprises three complementary determining regions HCDR1, HCDR2 and HCDR3 in sequence, the HCDR1 having an amino acid sequence selected from any one of SEQ ID NOs: 39, 42 and 45, the HCDR2 having an amino acid sequence selected from any one of SEQ ID NOs: 40, 43 and 46, and the HCDR3 having an amino acid sequence selected from any one of SEQ ID NOs: 41, 44 and 47; and

[0493] (b) the VL comprises three complementary determining regions LCDR1, LCDR2 and LCDR3 in sequence,

[0494] 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.

[0495] In particular, the present disclosure provides antibodies having binding specificity to human IL-17A and comprising (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.

[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 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.

[0497] 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, and the inhibitory activity is determined by measuring human IL-17A-induced GRO-α secretion in the HT-29 assay in the presence of 50 pg / ml TNF.

[0498] 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:

[0499] (i)

[0500] (a) the VH comprises three complementary determining regions HCDR1, HCDR2 and HCDR3 in sequence,

[0501] The HCDR1 has the same sequence as any one of SEQ ID NO: 1, 4 and 7, preferably SEQ ID NO:

[0502] 1, with at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99%

[0503] amino acid sequence of identity;

[0504] The HCDR2 has the same sequence as any one of SEQ ID NO: 2, 5 and 8, preferably SEQ ID NO:

[0505] 2. an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical;

[0506] The HCDR3 has the same sequence as any one of SEQ ID NO: 3, 6 and 9, preferably SEQ ID NO:

[0507] 3. an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identical to the amino acid sequence; and / or

[0508] (b) the VL comprises three complementary determining regions LCDR1, LCDR2 and LCDR3 in sequence,

[0509] 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;

[0510] 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;

[0511] The LCDR3 has the same structure as any one of SEQ ID NOs: 14, 17 and 20, preferably SEQ ID NO: 14, and has at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98

[0512] or 99% identical amino acid sequence; or (ii)

[0513] (a) the VH comprises three complementary determining regions HCDR1, HCDR2 and HCDR3 in sequence,

[0514] 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: 39, 42 and 45, preferably SEQ ID NO: 39;

[0515] 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: 40, 43 and 46, preferably SEQ ID NO: 40;

[0516] The HCDR3 has the same structure as any one of SEQ ID NO:41, 44 and 47, preferably SEQ ID NO:41, and has at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98

[0517] or an amino acid sequence with 99% identity; and / or

[0518] (b) the VL comprises three complementary determining regions LCDR1, LCDR2 and LCDR3 in sequence,

[0519] 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;

[0520] 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;

[0521] 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.

[0522] Suitably, the isolated antibody of the invention having binding specificity for human IL-17A comprises: (i) HCDR1, HCDR2 and HCDR3 having at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identity to the sequences of SEQ ID NOs: 1, 2 and 3, respectively, and / or LCDR1, LCDR2 and LCDR3 having at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identity to the sequences of SEQ ID NOs: 12, 13 and 14, respectively; or (ii) 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.

[0523] 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.

[0524] Suitably, the present disclosure provides an antibody that specifically binds 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.

[0525] Suitably, the present disclosure provides an isolated antibody that specifically binds to human IL-17A, wherein the antibody comprises Vκ framework FR1, FR2 and FR3, in particular Vκ1 or Vκ3FR1 to FR3, preferably Vκ1 framework FR1 to FR3, and framework FR4, the framework FR4 is selected from VκFR4, in particular Vκ1FR4, Vκ3FR4, and VλFR4. Suitable VλFR4 is shown as 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 VλFR4, the 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, and the VλFR4 comprises 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.

[0526] Therefore, in one embodiment, the present disclosure provides an antibody comprising:

[0527] (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;

[0528] (ii) a VH3 or VH4 domain, preferably a VH3 domain; and

[0529] (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, wherein the framework FR4 is selected from 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 VλFR4 as shown in an amino acid sequence selected from any one of SEQ ID NO:26 to SEQ ID NO:32, more preferably VλFR4 as shown in SEQ ID NO:27.

[0530] 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.

[0531] 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, as a number of non-limiting examples, the mutation is an addition, substitution or deletion).

[0532] 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, as a number of non-limiting examples, the mutation is an addition, substitution or deletion).

[0533] 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 described in the sequences described in Table 1.

[0534] 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., a sequence that is not a CDR) have been mutated (wherein, as a number of non-limiting examples, the mutation is an addition, substitution or deletion).

[0535] 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, as a number of non-limiting examples, the mutation is an addition, substitution or deletion).

[0536] 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 described in the sequences described in Table 1.

[0537] 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).

[0538] 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 NO: 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).

[0539] 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 includes 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 includes SEQ ID NO: 12, 13 and 14, respectively. LCDR1, LCDR2 and LCDR3 sequences of NO:50, 51 and 52.

[0540] In yet another embodiment, the present disclosure provides an isolated antibody that specifically binds to human IL-17A, wherein the antibody comprises a light chain variable region, the 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).

[0541] 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: 21. 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 the amino acid sequence of SEQID NO:59; 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.

[0542] Therefore, 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 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, 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 amino acid sequence 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 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 the HCDR1, HCDR2 and HCDR3 sequences of SEQ ID NO:39, 40 and 41, respectively, and / or the LCDR1, LCDR2 and LCDR3 sequences of SEQ ID NO:12, 13 and 14, 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.

[0543] 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, 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 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, in particular, wherein the heavy chain variable region comprises R20T and Q141P (AHo numbering).

[0544] Therefore, 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 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, 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, 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.

[0545] 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 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 49; and / or a VL thereof comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 59 and 60. In a specific embodiment, the antibody of the present disclosure 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 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.

[0546] 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 SEQ ID NOs: 24 and 25, preferably SEQ ID NO: 24, or (ii) an amino acid sequence selected from the group consisting of 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.

[0547] Other antibodies of the present disclosure having binding specificity to 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, 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. 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%, such as up to 200% of the activity determined for the parent antibody (e.g., an antibody of the present invention, particularly an antibody of the present invention described in Table 1).

[0548] 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.

[0549] 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 have 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 an antibody of the present disclosure.

[0550] Therefore, the present disclosure provides a monoclonal antibody comprising:

[0551] (i)

[0552] A heavy chain variable region (VH), which comprises three complementary 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

[0553] A light chain variable region (VL), which comprises three complementary determining regions LCDR1, LCDR2 and LCDR3 in sequence, 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

[0554] (ii)

[0555] A heavy chain variable region (VH), which comprises three complementary 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

[0556] A light chain variable region (VL), which comprises three complementary 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,

[0557] The antibody specifically binds to human IL-17A and / or neutralizes IL-17A.

[0558] In one embodiment, an antibody of the present disclosure is optimized for expression in a mammalian cell, having a heavy chain variable region and a light chain variable region, wherein one or more of these sequences have 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 an antibody of the present disclosure. Accordingly, the present invention provides a monoclonal antibody optimized for expression in a mammalian cell, 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.

[0559] 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 a conservative modification thereof, wherein the antibody retains the desired functional properties of an antibody of the present disclosure.

[0560] As used herein, the term "optimized" refers to a nucleotide sequence that is changed to use codons that are preferred 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) to encode an amino acid. 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 engineered to have preferred codons 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.

[0561] 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 intended 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.

[0562] An "affinity matured" antibody is an antibody that has one or more changes in one or more of its variable domains that result in improved affinity of the antibody for the antigen compared to a parent antibody that does not have those changes. 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, for example, in: 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.

[0563] The antibodies disclosed herein can be further prepared by the following method: using antibodies having one or more VH and / or VL sequences as described herein as starting materials to engineer modified antibodies, which modified antibodies may have different properties from the starting antibodies. Antibodies 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, antibodies can be engineered by modifying residues within the constant region, for example to alter the effector function of the antibody.

[0564] One type of variable region engineering that can be performed is CDR grafting. Antibodies interact with target antigens mainly through amino acid residues located in six heavy and light chain complementarity determining regions (CDRs). Thus, the amino acid sequences in the CDRs are more diverse among individual antibodies than the sequences outside the CDRs. Since 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 that CDR sequences are 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. U.S.A. 86:10029-10033; U.S. Patent No. 5,225,539 to Winter, and U.S. Patent Nos. 5,530,101; 5,585,089; 5,693,762 and 6,180,370 to Queen et al.).

[0565] 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) as well as Kabat, E.A. et al., 1991 Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242; Tomlinson, I.M. et al., 1992 J. Mol. Biol. 227:776-798; and Cox, J.P.L. et al., 1994 Eur. J Immunol. 24:827-836; the respective contents of which are hereby expressly incorporated by reference. For example, germline DNA sequences and rearranged antibody sequences of human heavy and light chain variable region genes can be found in the "IMGT" database (available online at www.imgt.org; see Lefranc, M.P. et al., 1999 Nucleic Acids Res. 27:209-212; the respective contents of which are hereby expressly incorporated by reference).

[0566] Examples of framework sequences for antibodies of the present disclosure are those that are structurally similar to the framework sequences used by the selected antibodies of the present disclosure, for example, the consensus sequences and / or framework sequences used by the monoclonal antibodies of the present disclosure. The VH CDR1, 2 and 3 sequences and the VL CDR1, 2 and 3 sequences can be transplanted onto a framework region having the same sequence as that found in the germline immunoglobulin gene from which the framework sequence is derived, or the CDR sequences can be transplanted onto a framework region comprising one or more mutations compared to the germline sequence. For example, it has been found that, in some cases, it is beneficial to mutate residues within the framework region to maintain or enhance the antigen binding ability of the antibody (see, e.g., U.S. Pat. Nos. 5,530,101; 5,585,089; 5,693,762 and 6,180,370 of Queen et al.).

[0567] 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 features.

[0568] In one aspect, the present disclosure relates to a method for producing non-immunoglobulin-based antibodies using a non-immunoglobulin backbone to which the CDRs of the present disclosure can be grafted. Known or future non-immunoglobulin frameworks and backbones can be used as long as they contain binding regions that are specific for the target IL-17A protein. Known non-immunoglobulin frameworks or backbones 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).

[0569] 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):

[0570] The antibody specifically binds to human IL-17A and:

[0571] (a) has binding specificity for cynomolgus monkey IL-17A;

[0572] (b) selectively binds to human IL-17A compared to human IL-17B, IL-17C, IL-17D, IL-17E and IL-17F,

[0573] As measured by ELISA;

[0574] (c) inhibiting or blocking the binding between IL-17A and its receptor (IL-17RA);

[0575] (d) reducing or neutralizing IL-17A activity;

[0576] (e) was able to inhibit GRO-α secretion when assessed in an in vitro HT-29 assay;

[0577] (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, and 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 antibody of the present invention measured by ELISA. 50 The ratio of values;

[0578] (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;

[0579] (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; (i) binding to human IL-17A with a 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, in particular less than 100 pM, more particularly less than 50 pM, such as by surface

[0580] measured by plasmon resonance, preferably by surface plasmon resonance in a direct setting; (j) binding 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;

[0581] (k) when in scFv form, a melting temperature (Tm) of at least 60° C. as determined by differential scanning fluorimetry,

[0582] particularly at least 62°C, more particularly at least 65°C, even more particularly at least 70°C, particularly wherein the antibody is in phosphate-citrate buffer, pH 6.4, 150 mM NaCl;

[0583] (l) when in scFv format, when the initial concentration of the antibody of the invention is 10 mg / ml, after 5 consecutive freeze-thaw cycles, the monomer content is lost by less than 5%, particularly less than 3%, more particularly less than 1%, particularly wherein the antibody is in phosphate buffered saline (PBS) at pH 7.4; and / or

[0584] (m) when in scFv form, when the antibody of the invention is stored at a starting concentration of 10 mg / ml at 4°C for at least two weeks, in particular at least 4 weeks, the loss in monomer content is 5% or less, in particular less than 4%, less than 3%, less than 2%, more particularly less than 1%, in particular, wherein the antibody is in phosphate buffered saline (PBS) at pH 7.4; and / or

[0585] (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, especially at least four weeks.

[0586] 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 an 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 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 50 Comparison of IC of IL-17A 50 More than 100 times greater, particularly more than 200 times greater, more than 300 times greater, more than 400 times greater, as determined by ELISA.

[0587] The antibodies of the present disclosure specifically bind to IL-17A, wherein the 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.

[0588] As used herein, the term "neutralizing antibody" describes an antibody that is capable of neutralizing the biological signaling 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 of the present disclosure are IL-17A neutralizing antibodies. It is understood that the term "neutralization" as used herein refers to a reduction in biological signaling activity, which reduction may be partial or complete. Neutralization of IL-17A can be determined by various assays, examples of which are described elsewhere herein.

[0589] Thus, the antibodies of the present disclosure are capable of inhibiting the secretion of GRO-α when evaluated 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) 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 Values ​​are comparable to the IC values ​​in ng / mL of the disclosed antibodies in scFv form measured in the HT-29 assay. 50 In another embodiment, the antibody of the present disclosure is 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 the secretion of GRO-α induced by human IL-17A in the HT-29 assay in the presence of 50 pg / ml TNFα.

[0590] 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 value of secukinumab in ng / mL as measured by ELISA. 50 The values ​​are similar to the IC values ​​in ng / mL of the disclosed antibodies in scFv form measured by ELISA. 50 The ratio of value.

[0591] 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 region of the antibody "arm" interacts with the antigen at many sites through weak non-covalent forces; the greater the interaction, the stronger the affinity.

[0592] "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 reflecting 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 ) is represented by. Affinity can be measured by conventional methods known in the art, including the methods described herein. Low-affinity antibodies will generally bind antigen slowly and tend to dissociate easily, while high-affinity antibodies will generally bind antigen faster and tend to remain bound longer. A variety of 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.

[0593] The term "K assoc ”, “Ka” or “K on " refers to the association rate of a specific antibody-antigen interaction, and the term "K" as used herein dis ”, “Kd” or “K off " refers to the dissociation 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 by 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 the Biotin-CAPture kit from Biacore. 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 a concentration range of analyte from 0.35 to 90 nM diluted in running buffer. The obtained sensorgrams were fit using a 1:1 binding model. Alternatively, the affinity of humanized scFv can be measured and analyzed with another SPR detection setup: IL-17A was immobilized on a CM5 sensor chip (GE Healthcare) by amine coupling and serial dilutions of scFv from 0.35 to 90 nM were injected on the immobilized IL-17A.

[0594] Suitably, the affinity of the antibodies of the present disclosure for IL-17A may be higher than the affinity of IL-17A for IL-17RA. It is understood 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 preformed IL-17RA / IL-17A complexes. In one embodiment, the antibodies of the present disclosure can neutralize the interaction of IL-17RA / IL-17A. In another embodiment, the antibodies of the present disclosure can inhibit or block the binding between IL-17A and its receptor (IL-17RA). In one embodiment, the antibodies of the present disclosure can neutralize the activity of IL-17A.

[0595] Suitably, the affinity of the antibodies of the present disclosure for IL-17A may be comparable to or higher than the affinity of secukinumab for IL-17A, preferably higher than the latter. In one embodiment, the potency of the antibodies of the present disclosure in neutralizing IL-17A activity is equal to or higher than, preferably higher than, secukinumab. In another embodiment, the potency of the antibodies of the present disclosure in neutralizing IL-17RA / IL-17A interaction is equal to or higher than, preferably higher than, secukinumab.

[0596] The binding affinity of an antibody can be measured, for example, by the dissociation constant (K D ) is determined. The lower K D indicates stronger affinity, while higher K D Indicates weaker affinity.

[0597] 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 of the antibodies of the present disclosure is D Less than 0.5 nM, particularly less than 50 pM. More suitably, the K of the antibodies of the present disclosure is D Less than 0.2 nM, in particular less than 50 pM, as measured by surface plasmon resonance in a direct setting.

[0598] 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.

[0599] 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.

[0600] 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 less than 10 -3 s -1 or less 5x10 -4 s -1 or lower 3x10 -4 s -1 or less than 10 -4 s -1 or less 5x10 -5 s -1 or lower, 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 off The 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.

[0601] Suitably, the antibodies of the present disclosure have beneficial biophysical properties.

[0602] Suitably, when expressed in the form of scFv (single chain variable fragment) antibodies, 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 at a final protein concentration of 50 μg / mL in citrate phosphate buffer at pH 6.4 and contained 5x The final concentration of Orange was 100 μl in a total volume of 100 μl. 25 microliters of the prepared sample were 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 the fluorescence emission was detected using the custom dye calibration program of the software. The PCR plate containing the test samples was subjected to a temperature change from 25°C to 96°C in increments of 1°C, with a pause of 30s 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.

[0603] The antibodies of the present disclosure, particularly when expressed in the form of 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 initial concentration of the antibodies of the present disclosure is 10 mg / ml.

[0604] After storage at 4°C for at least two weeks, 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 initial 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. The 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 for each injection was 5μg. Samples were detected by a UV detector at a wavelength of 280nm, and 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.

[0605] Suitably, the isolated antibody of the present disclosure is 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 constructed in antibody constant regions (e.g., Modular Antibody Technology of F-star). TM ), preferably scFv.

[0606] Suitably, the antibody of the present disclosure is an Fv. Suitably, the antibody of the present disclosure is an scFv antibody fragment. 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 a 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.

[0607] Suitably, the antibody of the present disclosure is an IgG antibody. In one embodiment, the antibody of the present disclosure is an IgG selected from the group consisting of IgG1, IgG2, IgG3 and IgG4, preferably IgG1.

[0608] Exemplary domains that specifically bind to TNFα

[0609] The multispecific antibodies disclosed herein 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.

[0610] Suitable domains that specifically bind to TNFα for use in the multispecific antibodies of the present disclosure include, but are not limited to:

[0611] 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);

[0612] ·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);

[0613] adalimumab / D2E7( described in U.S. Pat. No. 6,090,382, which is incorporated herein by reference in its entirety);

[0614] certolizumab, a PEGylated Fab fragment ( described in US7,012,135 and US7,186,820, all of which are incorporated herein by reference);

[0615] ·golimumab Disclosed in U.S. Application 2009 / 214528, which is incorporated herein by reference in its entirety).

[0616] 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 WO2017 / 158101, which is incorporated herein by reference in its entirety).

[0617] Therefore, 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,

[0618] 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: 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 SEQ ID NOs: 39, 40 and 45, preferably SEQ ID NO: 39; 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.

[0619] 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 of the VH CDRs listed in Table 1.

[0620] 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 complementary 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.

[0621] 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 of the VL CDRs listed in Table 1.

[0622] 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.

[0623] 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:

[0624] (a) the VH comprises three complementary 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; and

[0625] (b) the VL comprises three complementarity 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.

[0626] 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.

[0627] 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.

[0628] Suitably, the second domain of the multispecific antibody of the present disclosure that specifically binds to TNFα comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein:

[0629] (a) the VH comprises three complementary determining regions HCDR1, HCDR2 and HCDR3 in sequence, 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: 25, 28 and 31; 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: 26, 29 and 32; the HCDR3 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: 27, 30 and 33

[0630] amino acid sequence; and / or

[0631] (b) the VL comprises three complementary determining regions, LCDR1, LCDR2 and LCDR3, in sequence, the LCDR1 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: 38, 41 and 44; the LCDR2 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; the LCDR3 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

[0632] Amino acid sequence of similarity.

[0633] Suitably, the second domain that specifically binds to TNFα comprises: 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: 25, 26 and 27, respectively, 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: 38, 39 and 40, respectively.

[0634] 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.

[0635] 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.

[0636] 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 Vκ framework FR1, FR2 and FR3, in particular FR1 to FR3 of Vκ1 or Vκ3, preferably Vκ1 framework FR1 to FR3, and framework FR4, the framework FR4 is selected from VκFR4, in particular Vκ1FR4, Vκ3FR4, and VλFR4. Suitable VλFR4 is shown in SEQ ID NO:97 to SEQ ID NO:103. In one embodiment, the second domain that specifically binds to TNFα comprises VλFR4, the 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.

[0637] 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:

[0638] (a) HCDR1, HCDR2 and HCDR3 sequences of SEQ ID NOs: 25, 26 and 27, respectively, and LCDR1, LCDR2 and LCDR3 sequences of SEQ ID NOs: 38, 39 and 40, respectively;

[0639] (b) VHB, wherein the VHB is VH3 or VH4, preferably VH3; and

[0640] (c) a VLB comprising a VL framework comprising Vκ frameworks FR1, FR2 and FR3,

[0641] In particular, FR1 to FR3 of Vκ1 or Vκ3, preferably Vκ1FR1 to FR3, and a framework FR4, wherein the framework FR4 is selected from 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:97 to SEQ ID NO:103, preferably VλFR4 as shown in any one of SEQID 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.

[0642] Suitably, the 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 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 a number of non-limiting examples, the mutation is an addition, substitution or deletion), preferably no more than about 10. Other domains of the 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 with the VH region described in the sequence described in Table 1.

[0643] Suitably, the 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 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 have been mutated (wherein, as a number of non-limiting examples, the mutation is an addition, substitution or deletion) in the framework sequence (e.g., a sequence that is not a CDR), preferably no more than about 10. Other domains of the 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 with the VL region described in the sequence described in Table 1.

[0644] 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 the 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 domain includes 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, the 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 NO: 25, 26 and 27, respectively. Suitably, the second domain that specifically binds to TNFα may comprise a heavy chain variable region, the 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 NO: 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.

[0645] 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, 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 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.

[0646] In a preferred embodiment, the second domain that specifically binds to TNFα may include a light chain variable region, the 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 LCDR1, LCDR2 and LCDR3 sequences of SEQ ID NOs: 38, 39 and 40, respectively. Suitably, the second domain that specifically binds to TNFα may include a light chain variable region, the 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 include a light chain variable region, the 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 sequences are SEQ ID NOs: 38, 39 and 40, respectively. Suitably, the second domain that specifically binds to TNFα may include a light chain variable region, the 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 sequences are SEQ ID NOs: 38, 39 and 40, respectively.

[0647] 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 second domain 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: 34; 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: 47.

[0648] Preferably, the second domain that specifically binds to TNFα comprises a heavy 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: 34; and a light chain variable region, 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: 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 sequences of SEQ ID NOs: The HCDR1, HCDR2 and HCDR3 sequences of SEQ ID NOs: 25, 26 and 27, and the LCDR1, LCDR2 and LCDR3 sequences of SEQ ID NOs: 38, 39 and 40, respectively.

[0649] 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 sequences of SEQ ID NO: 41, 42 and 43, respectively. The HCDR1, HCDR2 and HCDR3 sequences of SEQ ID NOs: 25, 26 and 27, and the LCDR1, LCDR2 and LCDR3 sequences of SEQ ID NOs: 38, 39 and 40, respectively.

[0650] 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 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 its VL comprises 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 to TNFα comprises a VH sequence of SEQ ID NO: 34 and a VL sequence of SEQ ID NO: 47. In yet another specific embodiment, the second domain that specifically binds to TNFα comprises a VH sequence of SEQ ID NO: 37 and a VL sequence of SEQ ID NO: 50 or SEQ ID NO: 51.

[0651] 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.

[0652] 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.

[0653] In one embodiment, a domain of the invention that specifically binds to TNFα has a heavy chain variable region comprising HCDR1, HCDR2, and HCDR3 sequences and a light chain variable region comprising LCDR1, LCDR2, and LCDR3 sequences, wherein one or more of these CDR sequences has a specified amino acid sequence based on the domains described herein or conservative modifications thereof, wherein the domains retain the desired functional properties of the antibodies of the present disclosure.

[0654] 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 second domain comprises (or consists of):

[0655] A heavy chain variable region (VH), which comprises three complementary 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

[0656] A light chain variable region (VL), which comprises three complementary determining regions LCDR1, LCDR2 and LCDR3 in sequence, wherein the LCDR1 has the amino acid sequence of SEQ ID NO: 38, or a conservative variant thereof; the LCDR2 has the amino acid sequence of SEQ ID NO: 39, or a conservative variant thereof; the LCDR3 has the amino acid sequence of SEQ ID NO: 40, or a conservative variant thereof;

[0657] The domain specifically binds to human TNFα and / or neutralizes TNFα.

[0658] Exemplary domains that specifically bind to human serum albumin (HSA)

[0659] The multispecific antibodies disclosed herein 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.

[0660] 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 include 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.

[0661] 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.

[0662] 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:

[0663] (a) HCDR1' is as shown in SEQ ID NO:52; HCDR2' is as shown in SEQ ID NO:53; HCDR3'

[0664] as shown in SEQ ID NO:54; LCDR1' as shown in SEQ ID NO:62; LCDR2' as shown in SEQ ID NO:63; LCDR3' as shown in SEQ ID NO:64; or

[0665] (b) HCDR1' is shown in SEQ ID NO: 73; HCDR2' is shown in SEQ ID NO: 74; HCDR3'

[0666] As 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.

[0667] 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 a multispecific antibody 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.

[0668] Suitably, the domain that specifically binds to human serum albumin comprises a heavy chain variable region (VH), wherein said VH comprises three complementary determining regions HCDR1, HCDR2 and HCDR3 in sequence, wherein

[0669] (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; the HCDR3 is represented by an amino acid sequence selected from any one of SEQ ID NOs: 54, 57 and 60; or

[0670] (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.

[0671] 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.

[0672] 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 the 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.

[0673] Suitably, the domain that specifically binds to human serum albumin comprises a light chain variable region (VL), wherein said VL comprises three complementary determining regions LCDR1, LCDR2 and LCDR3 in sequence, wherein

[0674] (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

[0675] (b) the LCDR1 is shown as an amino acid sequence selected from any one of SEQ ID NOs: 83, 86 and 89; the LCDR2 is shown as an amino acid sequence selected from any one of SEQ ID NOs: 84, 87 and 90; and the LCDR3 is shown as an amino acid sequence selected from any one of SEQ ID NOs: 85, 88 and 91.

[0676] In a preferred embodiment, the domain that specifically binds to human serum albumin comprises 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 LCDR1, LCDR2 and LCDR3 sequences of SEQ ID NOs: 83, 84 and 85, respectively.

[0677] 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:

[0678] (a) the VH comprises three complementary determining regions HCDR1, HCDR2 and HCDR3 in sequence, 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 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: 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

[0679] (b) 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: 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 three complementarity determining regions LCDR1, LCDR2 and LCDR3 in sequence, 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.

[0680] 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.

[0681] 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.

[0682] 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:

[0683] (a) the VH comprises three complementary determining regions, HCDR1, HCDR2 and HCDR3, in sequence, 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: 52, 55 and 58; 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: 53, 56 and 59; the HCDR3 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: 54, 57 and 60; and / or

[0684] (b) the VL comprises three complementary determining regions, LCDR1, LCDR2 and LCDR3, in sequence, the LCDR1 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: 62, 65 and 68; the LCDR2 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: 63, 66 and 69; the LCDR3 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: 64, 67 and 70.

[0685] 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.

[0686] 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:

[0687] (a) the VH comprises three complementary determining regions, HCDR1, HCDR2 and HCDR3, in sequence, 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: 73, 76 and 79; 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: 74, 77 and 80; the HCDR3 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: 75, 78 and 81; and / or

[0688] (b) the VL comprises three complementary determining regions, LCDR1, LCDR2 and LCDR3, in sequence, the LCDR1 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: 83, 86 and 89; the LCDR2 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: 84, 87 and 90; the LCDR3 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: 85, 88 and 91.

[0689] 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.

[0690] 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.

[0691] 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.

[0692] Suitably, the present disclosure provides multispecific antibodies that comprise a third domain that specifically binds HSA, wherein the third domain comprises a light chain variable region VLC, and wherein the VLC comprises Vκ framework regions FR1, FR2, and FR3, particularly Vκ1 or Vκ3 FR1 to FR3, preferably Vκ1 framework regions FR1 to FR3, and a framework region FR4, wherein the framework region FR4 is selected from Vκ FR4, particularly Vκ1 FR4, Vκ3 FR4, and Vλ FR4. Suitable Vλ FR4s are shown in SEQ ID NOs: 97 to 103. In one embodiment, the third domain comprises Vλ FR4, and the Vλ FR4 comprises an amino acid sequence having at least 60%, 70%, 80%, 90% identity to an amino acid sequence selected from any one of SEQ ID NOs: 97 to 103, preferably to SEQ ID NO: 97 or SEQ ID NO: 98, more preferably to SEQ ID NO: 97. Suitably, the third domain comprises Vλ FR4, and the Vλ FR4 comprises an amino acid sequence selected from any one of SEQ ID NOs: 97 to 103, preferably Vλ FR4 as shown in SEQ ID NO: 97 or 98, more preferably Vλ FR4 as shown in SEQ ID NO: 98.

[0693] Accordingly, in a preferred embodiment, the present disclosure provides multispecific antibodies that comprise a third domain that specifically binds HSA, wherein the third domain comprises:

[0694] (a) HCDR1, HCDR2, and HCDR3 sequences that are SEQ ID NOs: 52, 53, and 54, respectively, and LCDR1, LCDR2, and LCDR3 sequences that are SEQ ID NOs: 62, 63, and 64, respectively;

[0695] (b) VHC, wherein the VHC is VH3 or VH4, preferably VH3; and

[0696] (c) VLC, which comprises a VL framework that comprises Vκ framework regions FR1, FR2, and FR3,

[0697] In particular, Vκ1 or Vκ3FR1 to FR3, preferably Vκ1FR1 to FR3, and a framework FR4, wherein the framework FR4 is selected from 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: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.

[0698] 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:

[0699] (i) HCDR1, HCDR2 and HCDR3 sequences of SEQ ID NOs: 73, 74 and 75, respectively, and LCDR1, LCDR2 and LCDR3 sequences of SEQ ID NOs: 83, 84 and 85, respectively;

[0700] (ii) VHC, wherein the VHC is VH3 or VH4, preferably VH3; and

[0701] (iii) VLC, which comprises a VL framework, wherein the VL framework comprises Vκ framework FR1, FR2 and FR3, in particular FR1 to FR3 of Vκ1 or Vκ3, preferably Vκ1FR1 to FR3, and a framework FR4, wherein the framework FR4 is 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.

[0702] 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 a number of 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 sequence described in Table 1.

[0703] Suitably, the disclosure provides a multispecific antibody comprising a third domain that specifically binds to HSA, wherein the third domain comprises a VL domain listed in Table 1. Suitably, the 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 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 a number of non-limiting examples, the mutation is an addition, substitution or deletion), preferably no more than about 10. Other domains of the 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 sequence described in Table 1.

[0704] 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 NO: 52, 53 and 54, respectively.

[0705] 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 LCDR1, LCDR2 and LCDR3 sequences of SEQ ID NO: 62, 63 and 64, respectively.

[0706] 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 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: 61; 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: 71.

[0707] 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 and a light chain variable region, the heavy cha...

Claims

1. An antibody with binding specificity to human IL-17A, 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, and wherein VH comprises an amino acid sequence in the framework region that is at least 90% identical to the amino acid sequence of SEQ ID NO: 10; and VL comprises an amino acid sequence in the framework region that is at least 90% identical to the amino acid sequence of SEQ ID NO: 21; 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, and wherein VH comprises an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 48; and VL comprises an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:

59.

2. The antibody of claim 1, wherein (i) the VH comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 10 and 11; and 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 the VL comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 59 and 60.

3. The antibody according to claim 2, comprising (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. 4 . The antibody according to claim 2 , comprising (i) the VH sequence of SEQ ID NO: 11 and the VL sequence of SEQ ID NO: 49; or (ii) the VH sequence of SEQ ID NO: 11 and the VL sequence of SEQ ID NO:

60.

5. The antibody of any one of claims 1 to 4, wherein the antibody has binding specificity for cynomolgus monkey IL-17A.

6. The antibody according to any one of claims 1 to 5, 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.

7. The antibody according to any one of claims 1 to 6, which binds to IL-17A (a) inhibiting or blocking the binding between IL-17A and its receptor (IL-17RA), and (b) Reducing or neutralizing IL-17A activity.

8. The antibody according to claim 7, wherein said antibody is capable of inhibiting GRO-α secretion when assessed in vitro in the HT-29 assay.

9. The antibody according to any one of claims 1 to 8, wherein the antibody: (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, and 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 antibody of the present invention measured by ELISA. 50 value ratio; and / or (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 value ratio; and / or (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, said inhibitory activity being determined by measuring the secretion of GRO-α induced by human IL-17A in the HT-29 assay in the presence of 50 pg / ml TNFα.

10. The antibody according to any one of claims 1 to 9, wherein the antibody: (a) The dissociation constant (K) for binding to human IL-17A measured by surface plasmon resonance, in particular by surface plasmon resonance in a direct setup D ) is less than 5 nM, in particular less than 1 nM, less than 0.5 nM, less than 0.2 nM, more in particular less than 100 pM, more in particular less than 50 pM; and (b) optionally, measuring the K of binding to cynomolgus monkey IL-17A by surface plasmon resonance, in particular by surface plasmon resonance in a trapping setup D Less than 10 nM, particularly less than 7 nM, less than 5 nM, less than 2 nM, less than 1 nM, more particularly less than 0.5 nM.

11. The antibody according to claim 10, wherein the dissociation constant (K) of the antibody for binding to human IL-17A is measured by surface plasmon resonance, in particular by surface plasmon resonance in a direct setting. D ) is less than 0.5 nM, less than 0.2 nM, less than 100 pM, and in particular less than 50 pM.

12. The antibody according to any one of claims 1 to 11, wherein the antibody: (a) 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 at pH 6.4, 150 mM NaCl; (b) when in scFv format, when the initial concentration of the antibody of the invention is 10 mg / ml, after 5 consecutive freeze-thaw cycles, the monomer content loss is less than 5%, particularly less than 3%, more particularly less than 1%, particularly wherein the antibody is in phosphate buffered saline (PBS) at pH 7.4; (c) when in scFv form, when the initial concentration of the antibody of the invention is 10 mg / ml, the loss of monomer content is 5% or less after storage at 4°C for at least two weeks, in particular at least four weeks, In particular less than 4%, less than 3%, less than 2%, more particularly less than 1%, in particular, wherein the antibody is in phosphate buffered saline (PBS) at pH 7.4; and / or (d) When the initial concentration of the antibody of the present invention is 10 mg / ml, the loss of monomer content is less than 5% after storage at 37°C for at least two weeks, especially at least four weeks.

13. according to the antibody described in any one of claims 1 to 12, wherein said antibody is selected from the group consisting of: monoclonal antibody, chimeric antibody, Fab, Fv, scFv, dsFv, scAb, STAB, single domain antibody (sdAb or dAb), single domain heavy chain antibody, and single domain light chain antibody, VHH, VNAR, single domain antibody based on shark VNAR structure, and binding domain based on alternative scaffold including but not limited to ankyrin-based domain, fynomer, avimer, anticalin, fibronectin, and binding site constructed in antibody constant region (e.g., Modular Antibody Technology of F-star). TM ), preferably scFv.

14. The antibody according to claim 13, wherein the scFv has (i) an amino acid sequence selected from 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 (ii) an amino acid sequence selected from 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.

15. The antibody according to claim 13, 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.

16. The antibody according to any one of claims 1 to 15, which is a multispecific molecule.

17. The antibody according to claim 16, 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-Fv 2 Morrison (IgG CH 3 -scFv fusion (Morrison L) or IgG CL-scFv fusion (Morrison H)), triabodies, scDb-scFv, bispecific Fab 2 , diabodies, tetrabodies, scFv-Fc-scFv fusions, scFv-HSA-scFv fusions, diabodies, DVD-Ig, COVD, IgG-scFab, scFab-dsscFv, Fv 2 -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 both the heavy and light chains), Ts2Ab (dsscFv linked to the C-terminus of the heavy chain), 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) fused to the N- and / or C-terminus of any one chain of a heterodimeric Fc domain or any other heterodimeric domain 2 , Fab-(scFv)1, Fab, Fab-Fv 2 , COVD, MATCH and DuoBody.

18. A pharmaceutical composition comprising the antibody of any one of claims 1 to 17 and a pharmaceutically acceptable carrier.

19. The antibody of any one of claims 1 to 17, or the pharmaceutical composition of claim 18, for use as a medicament.

20. The antibody of any one of claims 1 to 17, or the pharmaceutical composition of claim 18, for use in treating an inflammatory condition or an autoimmune disease.

21. The antibody of any one of claims 1 to 17, or the pharmaceutical composition of claim 18, for use in the treatment of 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 fever, glucose metabolism disorders, lung disease, periodontitis, 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 dyslipidemia.

22. Use of the antibody of any one of claims 1 to 17 or the pharmaceutical composition of claim 18 in the preparation of a medicament for treating an inflammatory condition or an autoimmune disease.

23. Use of the antibody of any one of claims 1 to 17 or the pharmaceutical composition of claim 18 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, cystic fibrosis, bone loss, airway hypersensitivity, demyelinating disorders, skin hypersensitivity, acute transplant rejection, allogeneic transplant rejection, graft-versus-host disease, systemic sclerosis, urinary inflammatory disorders, cardiovascular disease, vasculitis, periodic fever, glucose metabolism disorders, lung disease, periodontitis, 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 dyslipidemia.

24. A nucleic acid encoding the antibody of any one of claims 1 to 17. A vector comprising the nucleic acid of claim 24 .

26. A host cell comprising the nucleic acid of claim 24 or the vector of claim 25.

27. A method for producing the antibody of any one of claims 1 to 17, said method The following steps are involved: Cultivating a host cell comprising a nucleic acid or a vector encoding the antibody of any one of claims 1 to 17.

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