Multispecific antibodies having specificity for TNFα and IL-17A, antibodies targeting IL-17A, and methods for using them.

Abstract

To provide improved therapeutic antibodies that effectively neutralize the activity of both IL-17A and TNFα, with beneficial affinity and efficacy, and an improved safety profile, e.g., less immunogenicity.SOLUTION: The present invention relates to an isolated multispecific antibody comprising a first domain specifically binding to TNFα and a second domain specifically binding to IL-17A and, optionally, a third domain specifically binding to human serum albumin. The present invention further relates to an isolated antibody which specifically binds to human IL-17A. Additionally, the present invention further relates to methods of use of the antibodies, pharmaceutical compositions containing the antibodies and methods of use and kits thereof. The present invention also relates to a nucleic acid comprising a nucleotide sequence encoding the antibodies, a vector comprising the nucleic acid, a host cell comprising the nucleic acid or the vector, and a method of producing the antibodies.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to an isolated multispecific antibody comprising a first domain that specifically binds to TNFα, a second domain that specifically binds to IL-17A, and a third domain that selectively binds to human serum albumin. The present invention further relates to a method of using the antibody, a pharmaceutical composition comprising the antibody, a method of using the same, and a kit. The present invention also relates to a nucleic acid comprising a nucleotide sequence encoding the antibody, a vector comprising the nucleic acid, a host cell comprising the nucleic acid or the vector, and a method for producing the antibody.

[0002] The present invention further relates to an isolated antibody that specifically binds to human IL-17A, a multispecific molecule containing the isolated antibody of the present invention, a pharmaceutical composition, and a method of using the same. The present invention further relates to a kit containing the antibody, a nucleic acid containing a nucleotide sequence encoding the antibody, a vector containing the nucleic acid, a host cell containing the nucleic acid or the vector, and a method of producing the antibody. [Background technology]

[0003] TNFα is a homotrimeric pro-inflammatory cytokine released by immune system cells and interacting with those cells. TNFα is found as a soluble protein and a precursor called transmembrane TNFα, and is 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), releasing 157 amino acid residues of soluble TNFα. Soluble TNFα is a homotrimer of a 17 kDa cleaved monomer. Transmembrane TNFα also exists as a homotrimer of a 26 kDa uncleaved monomer. Both transmembrane TNFα and soluble TNFα are biologically active. TNFα can bind to two receptors, TNF receptor 1 (TNFR1) and 2 (TNFR2), where transmembrane TNFα primarily acts via TNFR2. TNFR1 is widely expressed in various cells, and its involvement triggers pro-inflammatory responses. TNFR2 expression is almost exclusively limited to immune cells, and its binding promotes cell survival and proliferation (Bazzoni F, Beutler B, N Engl J Med (1996) 334(26):1717-25; Locksley RM, et al., Cell (2001) 104(4):487-501; Cabal-Hierro L, Lazo PS, Cell Signal (2012) 24(6):1297-305; Brenner D, et al., Nat Rev Immunol (2015) 15(6):362-74).

[0004] TNFα has been shown to be upregulated in many human diseases, including inflammatory and autoimmune diseases, as well as 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) have investigated the therapeutic potential of anti-TNFα antibodies in the treatment of septic shock. The use of anti-TNFα antibodies in the treatment of septic shock has also been discussed by Kirschenbaum et al. (Critical Care Medicine, 26, 1625-1626, 1998). Collagen-induced arthritis can be effectively treated with anti-TNFα monoclonal antibodies (Williams et al. (Proc. Natl. Acad. Sci. USA 89, 9784-9788, 1992)). The use of anti-TNFα antibodies in the treatment of rheumatoid arthritis and Crohn's disease has been discussed by 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). Antibodies against TNFα previously used in such treatments are generally chimeric antibodies, as described in U.S. Patent No. 5,919,452.

[0005] Monoclonal antibodies against TNFα are described in the prior art. Meager et al. (Hybridoma, 6, 305-311, 1987) describe a mouse monoclonal antibody against recombinant TNFα. Fendly et al. (Hybridoma, 6, 359-370, 1987) describe the use of a mouse monoclonal antibody against recombinant TNFα in defining a neutralizing epitope on TNFα. Furthermore, international patent application WO92 / 11383 discloses recombinant antibodies, including a CDR-transplant antibody specific to TNFα. Rankin et al. (British J. Rheumatology, 34, 334-342, 1995) describe the use of such a CDR-transplant antibody 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 medical conditions associated with the presence of TNFα. Further, anti-TNFα antibodies are described in Stephens et al. (Immunology, 85, 668-674, 1995), GB-A-2 246570, GB-A-2 297145, US8,673,310, US2014 / 0193400, EP2390267B1, US8,293,235, US8,697,074, WO2009 / 155723A2, and WO2006 / 131013A2.

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

[0007] However, anti-TNFα therapy has been shown to have certain limitations. Not all patients necessarily achieve a sufficient clinical response or maintain a clinical response to anti-TNFα over the long term, necessitating a switch to new therapies to control the disease. For example, in rheumatoid arthritis (RA) patients, approximately 40% do not respond to anti-TNFα therapy, and only 20% experience a significant reduction in disease activity. Therefore, there remains a great unmet clinical need for therapies that more effectively suppress the progression of disorders such as inflammation and autoimmune disorders.

[0008] Activation of complement biological pathways in patients after anti-TNFα therapy may be one reason why many patients do not respond to or only partially respond to anti-TNFα therapy. A recent series of pieces of evidence have revealed the role of IL-17A in the pathogenesis of inflammatory and autoimmune diseases, such as rheumatoid arthritis.

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

[0010] Interleukin-17A (IL-17A or IL17A, which is synonymous with IL-17 or cytotoxic T lymphocyte-associated antigen-8 (CTLA-8)) is a homodimeric pro-inflammatory cytokine. IL-17A is produced by memory CD4+ T cells (called Th17), CD8+ T cells (Tc17), invariant NKT cells, γδT cells, non-T non-B lymphocytes (called type 3 innate lymphocytes), and a subset of neutrophils. IL-17A and IL-17F form distinct subgroups within the IL-17 family. They share the greatest sequence homology and identity within the IL-17 family, but other members of the IL-17 family have significantly lower sequence identity with respect to 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 transmit signals via a heterodimer 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-bonded homodimers and IL-17A / F disulfide-bonded 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 pro-inflammatory cytokines and antimicrobial peptides.

[0011] Human IL-17A (CTLA-8, Swiss Prot Q16552, also known as IL-17 or IL17; Sequence ID No. 33) is involved in various inflammatory conditions, 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; For a review, see Gu C., et al., Cytokine. 2013 Nov 64(2). IL-17A plays a role in inducing other pro-inflammatory cytokines and chemokines, acute phase proteins, antimicrobial peptides, mucins, matrix metalloproteinases, and adhesion molecules for neutrophil mobilization. IL-17A also synergistically induces chemokine expression in conjunction with other cytokines, including TNFα and IL-1 beta (Chabaud M., et al., J. Immunol. 161(1):409-14 (1998)).

[0012] Pathological production of IL-17A causes excessive inflammation and tissue damage (see review article Gu et al., Cytokine. 2013 November; 64(2)). High levels of IL-17A have been observed in patients with multiple sclerosis (MS), psoriasis, asthma, Crohn's disease, and rheumatoid arthritis. Treatment of animals with IL-17A neutralizing antibodies reduces the incidence and severity of autoimmune encephalomyelitis (Komiyama, Y. et al, J. Immunol. 177 (2006) 566-573). Furthermore, IL-17A neutralizing antibodies reduce the severity and incidence of collagen-induced arthritis in a mouse model of rheumatoid arthritis, and high levels of IL-17A can be detected in the synovial fluid of inflamed joints in 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 PW et al, Am. J. Resp. Cell Mol. Biol. 28 (2003) 42-50).

[0013] The majority of experimental evidence in human and animal models has supported the development of IL-17A-targeted therapies. Several anti-IL-17 antibodies have been developed, including AIN457 (secukinumab; see U.S. Patent No. 7,807,155 and WO2006 / 013107), LY2439821 (ixekizumab; see U.S. Patent Nos. 7,838,638 and 8,110,191 and WO2007 / 070750), SCH900117 (Merck), and RG4943 (Roche). Examples of anti-IL-17A antibodies include WO2006 / 013107, WO2006 / 054059, WO2007 / 070750, WO2007 / 149032, WO2008 / 001063, WO2008 / 021156, WO2010 / 034443, WO2010 / 102251, and WO2012 / 0187. It is disclosed in 67, WO2014 / 161570, WO2014 / 001368, WO2014 / 122613, WO2015 / 070697, WO2015 / 137843, WO2016 / 048188, WO2016 / 113557, WO2016 / 138842, and WO2017 / 068472.

[0014] Several clinical trials using various molecules that block IL-17A signaling are underway or currently ongoing. Biologics targeting IL-17A or its receptor, along with their efficacy, are being evaluated in the context of inflammatory or autoimmune diseases such as rheumatoid arthritis, ankylosing spondyloarthritis, Crohn's disease, psoriasis, multiple sclerosis, and ozone-induced neutropenia.

[0015] For example, secukinumab (US Patent No. 7,807,155 and WO2006 / 013107), a fully human IgG1κ anti-IL-17A monoclonal antibody, is currently approved for the treatment of psoriasis, psoriatic arthritis, and ankylosing spondylitis (see Wang et al., Eur J Rheumatol 2017 (4) 272-7 for a review). Phase III trials (FUTURE I and FUTURE II) evaluating the long-term efficacy and safety of secukinumab in subjects with psoriatic arthritis showed that 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).

[0016] Ixekizumab (U.S. Patent Nos. 7,838,638 and 8,110,191 and WO2007 / 070750), a humanized anti-IL-17A monoclonal antibody, was tested in a 24-week Phase III trial (SPIRIT-P1) in biologic-naive patients with active psoriatic arthritis (Mease PJ. Et al., Ann Rheum Dis. 2017 Jan;76(1):79-87). In biologic-naive patients with active psoriatic arthritis, ixekizumab treatment demonstrated improvement in disease activity and physical function, as well as inhibition of structural damage progression. Ixekizumab has also been shown to be effective in treating patients with moderate to severe plaque psoriasis (Griffiths CEM, et al., The Lancet; 386: 541-51).

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

[0018] Therefore, IL-17A is a promising target in the treatment of inflammatory and autoimmune diseases. While many anti-IL-17A antibodies have been identified to date, there is still a need for improved therapeutic antibodies that can effectively reduce or eliminate IL-17A activity in inflammatory responses and autoimmune diseases, while simultaneously possessing an improved safety profile and being suitable for development. Therapeutic antibodies need to have improved biophysical properties, in addition to beneficial affinity, efficacy, and immunogenicity, resulting in improved developability, high-yield productivity, and protein stability. Therapies that simultaneously block both the TNFα and IL-17A biological pathways could significantly improve response rates and address unmet needs in the treatment of disorders mediated by TNFα and IL-17A. Several biotherapeutic agents that specifically bind to IL-17 and TNFα have been proposed. WO2010 / 102251 (Abbvie Inc.) discloses a bispecific tetravalent antibody that binds to both TNFα and IL-17. WO2013 / 063110 (Abbvie Inc.) discloses a polyvalent DVD-Ig binding protein capable of binding to TNF and IL-17. WO2014 / 044758 (Covagen ACJ) discloses a fusion construct capable of inhibiting glycosylated IL-17A and binding to TNFα. WO2014 / 137961 (Eli Lilly and Company) and WO2017 / 132457 (Janssen Biotech) also disclose anti-TNF and anti-IL-17A bispecific antibodies. WO2017 / 102830 (UCB Biopharma) discloses a multispecific antibody capable of inhibiting TNFα, IL-17A, and IL-17F, and in particular, comprising a binding domain specific to human TNFα and a binding domain specific to human IL-17A and human IL-17F. Xu et al., Oncotarget, 8 (2017) 81860-81872 generated an IgG-like bispecific antibody (bsAb) in which two arms of an IgG-like molecule are directed towards TNFα and IL-17A, respectively.Interestingly, WO2015 / 014979 (see also Roche, Fischer et al., Arthritis & Rheumatology 67 (2015) 51-62) discloses bispecific tetravalent IL-17AxTNF antibody constructs, which are either bivalent ("2+2" construct) or monovalent ("2+2" construct) for each of the two antigens. According to WO2015 / 014979, the bivalent construct is preferred over the monovalent substitute. [Overview of the Initiative] [Problems that the invention aims to solve]

[0019] However, there remains a need for improved anti-inflammatory drugs that can effectively neutralize the activity of both IL-17A and TNFα for the treatment of disorders such as inflammatory and autoimmune diseases, for example, rheumatoid arthritis, in which the majority of patients still do not respond adequately to treatment. There is still a need for the development of improved therapeutic antibodies that effectively neutralize the activity of both IL-17A and TNFα, with beneficial affinity and efficacy, and an improved safety profile, such as lower immunogenicity. Furthermore, therapeutic antibodies should possess improved biophysical properties that lead to better developability, higher yield productivity, and superior antibody stability. [Means for solving the problem]

[0020] The object of the present invention is to provide agents for improving the treatment of inflammatory and autoimmune disorders.

[0021] The antibody of the present invention provides a new treatment option for patients with unmet medical needs. The present invention provides a novel multispecific antibody that can simultaneously inhibit IL-17A and TNFα and has further improved properties beneficial for therapeutic use, such as higher affinity, improved efficacy, selectivity, safety, such as lower immunogenicity, and improved biophysical properties, such as scalability and stability.

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

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

[0024] In a further embodiment, this disclosure provides multispecific antibodies or pharmaceutical compositions of the present invention for use as pharmaceutical agents.

[0025] In a further embodiment, the present disclosure provides multispecific antibodies or pharmaceutical compositions of the present invention for use in the treatment of disorders mediated by IL-17A and / or TNFα, or disorders that can be treated by inhibiting Gro-α secretion, particularly inflammatory conditions or autoimmune diseases.

[0026] In one embodiment, the present disclosure provides the use of the multispecific antibodies or pharmaceutical compositions of the present invention for use in the treatment of disorders mediated by IL-17A and / or TNFα, or disorders that can be treated by inhibiting Gro-α secretion, particularly in the treatment of inflammatory conditions or autoimmune diseases.

[0027] In one embodiment, the present disclosure provides the use of the multispecific antibodies or pharmaceutical compositions of the present invention in the manufacture of agents for the treatment of disorders mediated by IL-17A and / or TNFα, or disorders that can be treated by inhibiting Gro-α secretion, particularly for the treatment of inflammatory conditions or autoimmune diseases.

[0028] In yet another embodiment, the present disclosure provides a method for treating IL-17A and / or TNFα-mediated disorders, wherein the method comprises administering an effective amount of the multispecific antibody or pharmaceutical composition of the present invention to a subject of interest.

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

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

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

[0032] The aspects, advantageous features, and preferred embodiments of this disclosure, summarized in the following sections, each individually or in combination, further contribute to solving the objectives of the present invention:

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

[0034] 2. The multispecific antibody described in item 1, wherein the antibody comprises a single domain that specifically binds to IL-17A and / or a single domain that specifically binds to TNFα.

[0035] 3. The multispecific antibody described in item 1 or item 2, which can neutralize the biological activity of human TNFα and human IL-17A.

[0036] 4. A multispecific antibody as described in any one of the above items, wherein, when measured by ELISA, the antibody selectively binds to human IL-17A more than human IL-17B, IL-17C, IL-17D, IL-17E, and IL-17F.

[0037] 5. A multispecific antibody according to any one of the above items, further comprising a third domain that has specificity for antigens other than IL-17A or TNFα.

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

[0039] 7. A multispecific antibody according to any one of the above items, wherein the domains can simultaneously bind to each of those antigens or receptors.

[0040] 8. A 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 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, but not limited to, ankyrin-based domains, finomers, avimers, anticarin, binding domains based on alternative scaffolds including fibronectin, and binding sites incorporated into the constant region of the antibody (e.g., F-star's Modular Antibody Technology®), preferably selected from the group consisting of Fab, Fv, and scFv, more preferably, where the first domain and / or the second domain and / or the third domain is Fv or scFv.

[0041] 9. A multispecific antibody described in any one of the above items, wherein the multispecific antibody is a single-chain diabody (scDb), tandem scDb (Tandab), linear dimer scDb (LD-scDb), cyclic dimer scDb (CD-scDb), bispecific T cell engager (BiTE; tandem di-scFv), tandem tri-scFv, tribody (Fab-(scFv)2) or vibody (Fab-(scFv)1), Fab, Fab-Fv2, Morrison (IgG CH3-scFv fusion (Morrison L) or IgG) CL-scFv fusion (Morrison H), triabody, scDb-scFv, bispecific Fab2, dimini antibody, tetrabody, scFv-Fc-scFv fusion, scFv-HSA-scFv fusion, didiabody, DVD-Ig, COVD, IgG-scFab, scFab-dsscFv, Fv2-Fc, IgG-scFv fusion, e.g., 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) The multispecific antibody is in a form selected from the group consisting of cFv), Ts2Ab (dsscFv linked to the C-terminus of the heavy chain), bispecific antibodies based on a heterodimeric Fc domain, such as Knob-into-Hole antibody (KiH); Fv, scFv, scDb, tandem-di-scFv, tandem-tri-scFv, Fab-(scFv)2, Fab-(scFv)1, Fab, Fab-Fv2, COVD, MATCH, and duobody, preferably tribody, or scDb-scFv fused to the N-terminus and / or C-terminus of any chain of a heterodimeric Fc domain or any other heterodimerized domain.

[0042] 10. A multispecific antibody according to any one of the above items, wherein the antibody does not contain an immunoglobulin Fc region polypeptide and optionally does not contain the CH1 and / or CL region.

[0043] 11. A multispecific antibody as described in any one of the above items, wherein the antibody is a tribody.

[0044] 12. The 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 the second domain is Fab and the first and third domains are scFv.

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

[0046] 14. A multispecific antibody as described in item 13, wherein the antibody has the formula: VLA-L1-VHC-L2-VLC-L3-VHA-L4-VLB-L5-VHB, or VLB-L1-VHA-L2-VLA-L3-VHB-L4-VLC-L5-VHC or is VLC-L1-VHB-L2-VLB-L3-VHC-L4-VLA-L5-VHA or VLA-L1-VHB-L2-VLB-L3-VHA-L4-VLC-L5-VHC, Preferably, VLB-L1-VHA-L2-VLA-L3-VHB-L4-VLC-L5-VHC, or VLA-L1-VHB-L2-VLB-L3-VHA-L4-VLC-L5-VHC, More preferably, represented as VLB-L1-VHA-L2-VLA-L3-VHB-L4-VLC-L5-VHC, Herein, VLA and VHA are the light chain variable region and heavy chain variable region of the first domain, respectively; VLB and VHB are the light chain variable region and heavy chain variable region of the second domain, respectively; VLC and VHC are the light chain variable region and heavy chain variable region of the third domain, respectively; and herein, L1, L2, L3, L4, and L5 are polypeptide linkers, the multispecific antibody.

[0047] 15. A multispecific antibody as described in item 14, wherein L1 and L3 are those described in Sequence ID No. 132.

[0048] 16. A multispecific antibody as described in any one of items 14-15, wherein L2, L4, and L5 are those described in Sequence ID No. 23.

[0049] 17. A multispecific antibody according to any one of the above items, wherein the antibody has the following characteristics:

[0050] (a) Having the ability to neutralize IL-17A at a titer (relative titer) greater than 2, e.g., greater than 5, greater than 10, greater than 15, greater than 20, greater than 25, greater than 30, greater than 35, greater than 40, greater than 45, preferably greater than 50, compared to the titer of secukinumab determined by measuring Gro-α secretion in the HT-29 assay, where the relative titer is the IC of secukinumab measured by the HT-29 assay. 50 The value (ng / mL) and the IC50 of the multispecific antibody measured by the HT-29 assay. 50 It is the ratio to the value (ng / mL); and

[0051] (b) Having the ability to neutralize TNFα at a titer (relative titer) of at least 1, for example, 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, compared to the titer of scDb of SEQ ID NO: 149(A13) as determined by measuring Gro-α secretion in the HT-29 assay, where the relative titer is the IC2C of scDb of SEQ ID NO: 149(A13) as measured by the HT-29 assay. 50 The nM value and the IC50 of the multispecific antibody measured by the HT-29 assay. 50 It is the ratio to the value (nM); and

[0052] (c) Optionally, having the ability to block the interaction between IL-17A and IL-17RA with a titer (relative titer) greater than 2, such as 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, compared to the titer of secukinumab determined by ELISA, wherein the relative titer is the ratio of the IC 50 value (ng / mL) of secukinumab measured by ELISA to the IC 50 value (ng / mL) of the multispecific antibody measured by ELISA; and

[0053] (d) Optionally, having the ability to neutralize TNFα with a titer (relative titer) of at least 0.4, such as at least 0.5, preferably at least 1, compared to the titer of the scDb of SEQ ID NO: 149 (A13) determined by the L929 assay, wherein the relative titer is the ratio of the IC 50 value (nM) of the scDb of SEQ ID NO: 149 measured by the L929 assay to the IC 50 value (nM) of the multispecific antibody measured by the L929 assay; and / or

[0054] (e) Binding to human IL-17A with a dissociation constant (K D ) of 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, and optionally binding to cynomolgus IL-17A with a K D of 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; and

[0055] (f) Binding to human TNFα with a dissociation constant (K D ) of 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, more preferably less than 0.25 nM; and

[0056] (g) When measured by surface plasmon resonance, the dissociation constant (K) is less than 5 nM, for example less than 4 nM, less than 3 nM, preferably less than 2 nM. D ) Binds to human serum albumin, and when optionally measured by surface plasmon resonance, the dissociation constant (K) is less than 5 nM, for example less than 4 nM, less than 3 nM, preferably less than 2 nM. D ) binds to cynomolgus monkey serum albumin.

[0057] 18. A multispecific antibody as described in any one of the above items, wherein the antibody has the following characteristics: (a) In a pH 6.4, 150 mM NaCl phosphate-citrate buffer, the melting temperature (Tm) determined by differential scanning fluorescence assay is at least 55°C, preferably at least 58°C, and more preferably at least 60°C; (b) When the multispecific antibody is at an initial concentration of 10 mg / ml in phosphate-buffered saline (PBS), pH 7.4, it has a monomer content loss of less than 5%, for example less than 4%, less than 3%, less than 2%, preferably less than 1%, after 5 consecutive freeze-thaw cycles; (c) When the multispecific antibody is at an initial concentration of 10 mg / ml in phosphate-buffered saline (PBS), pH 7.4, it has a loss of monomer content of less than 10%, preferably less than 5%, after being stored at 4°C for at least 2 weeks, more specifically at least 4 weeks; and / or (d) When the multispecific antibody is at an initial concentration of 10 mg / ml in phosphate-buffered saline (PBS), pH 7.4, it has a loss of monomer content of less than 20%, preferably less than 15%, after being stored at 37°C for at least 2 weeks, more specifically at least 4 weeks.

[0058] 19. A multispecific antibody as described in any one of the above items, wherein each domain includes a heavy chain variable region (VH) and a light chain variable region (VL), where, (a) The VH includes, in order, three complementarity determination regions HCDR1, HCDR2, and HCDR3, and (b) The multispecific antibody wherein the VL comprises, in order, three complementarity-determining regions LCDR1, LCDR2, and LCDR3.

[0059] 20. A multispecific antibody according to any one of the above items, wherein the first domain that specifically binds to IL-17A comprises a set of CDRs (HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3), and the set of CDRs has 10 or fewer amino acid substitutions from a set of CDRs [(i) HCDR1' is the one described in SEQ ID NO: 1; HCDR2' is the one described in SEQ ID NO: 2; HCDR3' is the one described in SEQ ID NO: 3; LCDR1' is the one described in SEQ ID NO: 12; LCDR2' is the one described in SEQ ID NO: 13; LCDR3' is the one described in SEQ ID NO: 14; or (ii) HCDR1' is the one described in SEQ ID NO: 39; HCDR2' is the one described in SEQ ID NO: 40; HCDR3' is the one described in SEQ ID NO: 41; LCDR1' is the one described in SEQ ID NO: 50; LCDR2' is the one described in SEQ ID NO: 51; LCDR3' is the one described in SEQ ID NO: 52].

[0060] 21. A multispecific antibody as described in item 20, (i) (a) The HCDR1 is described in an amino acid sequence selected from any one of Sequence IDs 1, 4, and 7; (b) The HCDR2 is described in an amino acid sequence selected from any one of Sequence IDs 2, 5, and 8; (c) The HCDR3 is described in the amino acid sequence selected from any one of Sequence ID Nos. 3, 6, and 9; (d) The LCDR1 is an amino acid sequence selected from any one of sequence numbers 12, 15, and 18; (e) The LCDR2 is described in an amino acid sequence selected from any one of sequence numbers 13, 16, and 19; and (f) The LCDR3 is an amino acid sequence selected from any one of sequence numbers 14, 17, and 20; or (ii) (a) The HCDR1 is described in an amino acid sequence selected from any one of Sequence IDs 39, 42, and 45; (b) The HCDR2 is described in an amino acid sequence selected from any one of Sequence IDs 40, 43, and 46; (c) The HCDR3 is described in an amino acid sequence selected from any one of Sequence IDs 41, 44, and 47; (d) The LCDR1 is described in an amino acid sequence selected from any one of sequence numbers 50, 53, and 56; (e) The LCDR2 is described in an amino acid sequence selected from any one of sequence numbers 51, 54, and 57; and (f) The multispecific antibody wherein the LCDR3 is described in an amino acid sequence selected from any one of SEQ ID NOs. 52, 55, and 58.

[0061] 22. A multispecific antibody as described in item 21, comprising (i) 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 (ii) (i) 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.

[0062] 23. A multispecific antibody according to any one of the above items, wherein the first domain that specifically binds to IL-17A includes a heavy chain variable region VHA, and the VHA is VH3 or VH4, preferably VH3.

[0063] 24. A multispecific antibody according to any one of the above items, wherein the first domain that specifically binds to IL-17A includes a light chain variable region VLA, the VLA includes Vκ framework FR1, FR2, and FR3, more specifically Vκ1 or Vκ3 FR1~FR3, preferably Vκ1 FR1~FR3, and framework FR4, wherein framework FR4 includes Vλ FR4 having at least 60, 70, 80, or 90% identity with an amino acid sequence selected from any of SEQ ID NOs: 26 to 32, preferably Vλ FR4 selected from any of SEQ ID NOs: 26 to 32, preferably Vλ FR4 described in SEQ ID NOs: 26 or 27, more preferably Vλ FR4 described in SEQ ID NOs: 27.

[0064] 25. A multispecific antibody described in any one of items 23-24, wherein the VHA is (i) The VLA contains an amino acid sequence that is at least 90% identical to amino acid sequence SEQ ID NO: 10, and / or the VLA contains an amino acid sequence that is at least 90% identical to amino acid sequence SEQ ID NO: 21, or (ii) The multispecific antibody comprising an amino acid sequence that is at least 90% identical to amino acid sequence SEQ ID NO: 48, and / or the VLA comprising an amino acid sequence that is at least 90% identical to amino acid sequence SEQ ID NO: 59.

[0065] 26. A multispecific antibody as described in item 25, wherein the VHA is (i) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 10 and 11, and / or the VLA comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 21 and 22, or (ii) The multispecific antibody comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 48 and 49, and / or the VLA comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 59 and 60.

[0066] 27. A multispecific antibody as described in 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.

[0067] 28. A multispecific antibody according to any one of the above 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 from a set of CDRs [wherein HCDR1' is the one described in SEQ ID NO: 63; HCDR2' is the one described in SEQ ID NO: 64; HCDR3' is the one described in SEQ ID NO: 65; LCDR1' is the one described in SEQ ID NO: 76; LCDR2' is the one described in SEQ ID NO: 77; and LCDR3' is the one described in SEQ ID NO: 78].

[0068] 29. (a) The HCDR1 is described in an amino acid sequence selected from any one of sequence numbers 63, 66, and 69; (b) The HCDR2 is described in an amino acid sequence selected from any one of Sequence IDs 64, 67, and 70; (c) The HCDR3 is described in an amino acid sequence selected from any one of Sequence IDs 65, 68, and 71; (d) The LCDR1 is an amino acid sequence selected from any one of sequence numbers 76, 79, and 82; (e) The LCDR2 is described in an amino acid sequence selected from any one of sequence numbers 77, 80, and 83; and (f) The multispecific antibody described in item 28, wherein the LCDR3 is described in an amino acid sequence selected from any one of sequence numbers 78, 81, and 84.

[0069] 30. A multispecific antibody as described in item 29, comprising the HCDR1, HCDR2, and HCDR3 sequences of sequence numbers 63, 64, and 65, respectively, and the LCDR1, LCDR2, and LCDR3 sequences of sequence numbers 76, 77, and 78, respectively.

[0070] 31. A multispecific antibody according to any one of the above items, wherein the second domain that specifically binds to TNFα includes a heavy chain variable region VHB, and the VHB is VH3 or VH4, preferably VH3.

[0071] 32. A multispecific antibody according to any one of the preceding items, wherein the second domain that specifically binds to TNFα comprises a light chain variable region VLB, and the VLB comprises Vκ framework FR1, FR2, and FR3, more specifically Vκ1 or Vκ3 FR1~FR3, preferably Vκ1 FR1~FR3, and framework FR4, wherein framework FR4 comprises Vλ FR4 having at least 60, 70, 80, or 90% identity with an amino acid sequence selected from any of SEQ ID NOs: 26 to 32, preferably Vλ FR4 described in any of SEQ ID NOs: 26 to 32, preferably Vλ FR4 described in SEQ ID NOs: 26 or 27, more preferably Vλ FR4 described in SEQ ID NOs: 27.

[0072] 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 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 at least 90% identical to SEQ ID NO. 85.

[0073] 34. The multispecific antibody according to item 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.

[0074] 35. (i) the VHB sequence of sequence number 72 and / or the VLB sequence of sequence number 85; or (ii) the VHB sequence of sequence number 75 and / or the VLB sequence of sequence number 88; or (iii) A multispecific antibody as described in item 34, comprising the VHB sequence of SEQ ID NO: 75 and / or the VLB sequence of SEQ ID NO: 89.

[0075] 36. A 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), where the set of CDRs is a set of CDRs [where, (a) HCDR1' is the one described in SEQ ID NO: 90; HCDR2' is the one described in SEQ ID NO: 91; HCDR3' is the one described in SEQ ID NO: 92; LCDR1' is the one described in SEQ ID NO: 100; LCDR2' is the one described in SEQ ID NO: 101; LCDR3' is the one described in SEQ ID NO: 102; or (b) The multispecific antibody having 10 or fewer amino acid substitutions from HCDR1', which is described in SEQ ID NO: 111; HCDR2', which is described in SEQ ID NO: 112; HCDR3', which is described in SEQ ID NO: 113; LCDR1', which is described in SEQ ID NO: 120; LCDR2', which is described in SEQ ID NO: 121; and LCDR3', which is described in SEQ ID NO: 122.

[0076] 37. A multispecific antibody as described in item 36, (a) The HCDR1 is an amino acid sequence selected from any one of SEQ ID NOs: 90, 93, and 96; the HCDR2 is an amino acid sequence selected from any one of SEQ ID NOs: 91, 94, and 97; the HCDR3 is an amino acid sequence selected from any one of SEQ ID NOs: 92, 95, and 98; the LCDR1 is an amino acid sequence selected from any one of SEQ ID NOs: 100, 103, and 106; the LCDR2 is an amino acid sequence selected from any one of SEQ ID NOs: 101, 104, and 107; the LCDR3 is an amino acid sequence selected from any one of SEQ ID NOs: 102, 105, and 108; or (b) The multispecific antibody wherein HCDR1 is described in an amino acid sequence selected from any one of SEQ ID NOs: 111, 114, and 117; HCDR2 is described in an amino acid sequence selected from any one of SEQ ID NOs: 112, 115, and 118; HCDR3 is described in an amino acid sequence selected from any one of SEQ ID NOs: 113, 116, and 119; LCDR1 is described in an amino acid sequence selected from any one of SEQ ID NOs: 121, 124, and 127; LCDR2 is described in an amino acid sequence selected from any one of SEQ ID NOs: 122, 125, and 128; and LCDR3 is described in an amino acid sequence selected from any one of SEQ ID NOs: 123, 126, and 129.

[0077] 38. Multispecific antibodies as described in item 37, including the following: (a) the HCDR1, HCDR2, and HCDR3 sequences of sequence numbers 90, 91, and 92, respectively, and the LCDR1, LCDR2, and LCDR3 sequences of sequence numbers 100, 101, and 102, respectively; or (b) The HCDR1, HCDR2, and HCDR3 sequences of sequence numbers 111, 112, and 113, respectively, and the LCDR1, LCDR2, and LCDR3 sequences of sequence numbers 121, 122, and 123, respectively.

[0078] 39. A multispecific antibody according to any one of items 6 to 38, wherein the third domain that specifically binds to HSA includes a heavy chain variable region VHC, and the VHC is VH3 or VH4, preferably VH3.

[0079] 40. A multispecific antibody according to any one of items 6 to 39, wherein the third domain that specifically binds to HSA comprises a light chain variable region VLC, the VLC comprising Vκ framework FR1, FR2, and FR3, more particularly Vκ1 or Vκ3 FR1 to FR3, preferably Vκ1 FR1 to FR3, and framework FR4, wherein framework FR4 comprises Vλ FR4 having at least 60, 70, 80, or 90% identity with an amino acid sequence selected from any of SEQ ID NOs: 26 to 32, preferably Vλ FR4 described in any of SEQ ID NOs: 26 to 32, preferably Vλ FR4 of SEQ ID NO: 26 or 27, more preferably Vλ FR4 described in SEQ ID NO: 27.

[0080] 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 amino acid sequence sequence number 99; and / or the VLC comprises an amino acid sequence that is at least 90% identical to amino acid sequence sequence number 109.

[0081] 42. A multispecific antibody as described in item 41, comprising the VHC sequence of SEQ ID NO: 99 and / or the VLC sequence of SEQ ID NO: 109.

[0082] 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 amino acid sequence sequence number 110; and / or the VLC comprises an amino acid sequence that is at least 90% identical to amino acid sequence sequence number 120.

[0083] 44. A multispecific antibody as described in item 43, comprising the VHC sequence of SEQ ID NO: 110 and / or the VLC sequence of SEQ ID NO: 120.

[0084] 45. A multispecific antibody described in any one of the above items, wherein the antibody is humanized.

[0085] 46. ​​A multispecific antibody according to any one of the above items, wherein the antibody comprises an amino acid sequence having at least 80% identity, preferably at least 90% identity, with a sequence selected from any of SEQ ID NOs: 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, and 148, preferably 143, and the CDR has the sequences of items 22, 30, and 38(a).

[0086] 47. A multispecific antibody according to any one of the above items, wherein the antibody comprises an amino acid sequence selected from any of SEQ ID NOs: 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, and 148, preferably 143.

[0087] 48. A pharmaceutical composition comprising a multispecific antibody described in any one of the preceding items and a pharmaceutically acceptable carrier.

[0088] 49. A multispecific antibody as described in any one of items 1 to 47, or a pharmaceutical composition as described in item 48, for use as a pharmaceutical agent.

[0089] 50. A multispecific antibody according to any one of items 1 to 47, or a pharmaceutical composition according to item 48, for use in the treatment of disorders mediated by L-17A and / or TNFα, or disorders that can be treated by inhibiting Gro-α secretion.

[0090] 51. A multispecific antibody as described in any one of items 1 to 47, or a pharmaceutical composition as described in item 48, for use in the treatment of an inflammatory condition or autoimmune disease.

[0091] 52. A multispecific antibody as described in any one of items 1 to 47, or a pharmaceutical composition as described in item 48, for use in the treatment of the following diseases: cancer, arthritis, rheumatoid arthritis, osteoarthritis of the elbow, reactive arthritis, psoriasis, chronic obstructive pulmonary disease, systemic lupus erythematosus (SLE), lupus nephritis, autoimmune inflammatory bowel disease, asthma, multiple sclerosis, cystic fibrosis, bone loss, airway hypersensitivity, demyelinating disorders, skin hypersensitivity, acute transplant rejection, allograft rejection, graft-versus-host disease, systemic sclerosis, urinary tract inflammatory diseases, cardiovascular diseases, vasculitis, periodic fever, glucose metabolism disorders, lung diseases, periodontitis, hepatic interstitial keratitis, allergies, inflammatory pain, spondyloarthropathy, sepsis, septic or endotoxin shock, meningitis, surgical trauma, autoimmune hematological disorders, Alzheimer's disease, sarcoidosis, cirrhosis, hepatitis, glomerulonephritis, or dyslipidemia.

[0092] 53. Use of a multispecific antibody described in any one of items 1 to 47, or a pharmaceutical composition described in item 48, in the manufacture of a drug for use in the treatment of disorders mediated by L-17A and / or TNFα, or disorders that can be treated by inhibiting Gro-α secretion.

[0093] 54. Use of any one of items 1 to 47, or a pharmaceutical composition described in item 48, in the manufacture of a drug for use in the treatment of an inflammatory condition or autoimmune disease.

[0094] 55. Use of any multispecific antibody described in item 1 to 47, or any pharmaceutical composition described in item 48, in the manufacture of a drug for use in the treatment of the following diseases: cancer, arthritis, rheumatoid arthritis, osteoarthritis of the elbow, 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, and skin hypersensitivity. Acute transplant rejection, allograft rejection, graft-versus-host disease, systemic sclerosis, urinary tract inflammatory diseases, cardiovascular diseases, vasculitis, periodic fever, glucose metabolism disorders, lung diseases, periodontitis, hepatic interstitial keratitis, allergies, inflammatory pain, spondyloarthropathy, sepsis, septic or endotoxin shock, meningitis, surgical trauma, autoimmune hematological disorders, Alzheimer's disease, sarcoidosis, cirrhosis, hepatitis, glomerulonephritis, or dyslipidemia.

[0095] 56. A method for treating a disorder mediated by IL-17A and / or TNFα, comprising administering an effective amount of a multispecific antibody described in any one of items 1 to 47 or a pharmaceutical composition described in item 48, such that the condition is alleviated.

[0096] 57. The method according to item 56, wherein the disorder mediated by IL-17A and / or TNFα is an inflammatory condition or an autoimmune disease.

[0097] 58. The method according to item 56, wherein the disorder mediated by IL-17A and / or TNFα is cancer, arthritis, rheumatoid arthritis, osteoarthritis of the elbow, reactive arthritis, psoriasis, chronic obstructive pulmonary disease, systemic lupus erythematosus (SLE), lupus nephritis, autoimmune inflammatory bowel disease, asthma, multiple sclerosis, cystic fibrosis, osteoporosis, airway hyperresponsiveness, demyelinating disorders, skin hypersensitivity, acute transplant rejection, allograft rejection, graft-versus-host disease, systemic sclerosis, urinary tract inflammatory diseases, cardiovascular diseases, vasculitis, periodic fever, glucose metabolism disorders, lung diseases, periodontitis, hepatic interstitial keratitis, allergies, inflammatory pain, spondyloarthropathy, sepsis, septic or endotoxin shock, meningitis, surgical trauma, autoimmune hematological disorders, Alzheimer's disease, sarcoidosis, cirrhosis, hepatitis, glomerulonephritis, or dyslipidemia.

[0098] 59. A nucleic acid encoding a multispecific antibody or a fragment thereof as described in any one of items 1 to 47.

[0099] 60. A vector containing the nucleic acid described in item 59.

[0100] 61. Host cells containing nucleic acids as described in item 59 or vectors as described in item 60.

[0101] 62. A method for producing a multispecific antibody according to any one of items 1 to 47, comprising the step of culturing a host cell containing a nucleic acid or vector encoding a multispecific antibody or a fragment thereof according to any one of items 1 to 47.

[0102] 63. A kit comprising a multispecific antibody described in any one of items 1 to 47, or a pharmaceutical composition described in item 48.

[0103] Another object of the present invention is to provide an anti-IL-17A antibody having improved affinity, efficacy, and improved biophysical properties, such as improved solubility, developability, and stability.

[0104] The anti-IL-17A antibody of the present invention has improved properties beneficial for therapeutic use, such as higher affinity, improved efficacy, selectivity, and improved biophysical properties, such as solubility, developability, and stability.

[0105] Accordingly, in one embodiment, the present disclosure provides an isolated antibody having binding specificity to human IL-17A, in particular, comprising a set of CDRs (HCDR1, HCDR2, HCDR3, LDCR1, LDCR2, and LDCR3), wherein the set of CDRs has 10 or fewer, preferably 0, amino acid substitutions from a given set of CDRs [wherein HCDR1' is an amino acid sequence selected from any one of SEQ ID NOs: 1, 4, and 7; HCDR2' is an amino acid sequence selected from any one of SEQ ID NOs: 2, 5, and 8; HCDR3' is an amino acid sequence selected from any one of SEQ ID NOs: 3, 6, and 9; LCDR1' is an amino acid sequence selected from any one of SEQ ID NOs: 12, 15, and 18; LCDR2' is an amino acid sequence selected from any one of SEQ ID NOs: 13, 16, and 19; and LCDR3' is an amino acid sequence selected from any one of SEQ ID NOs: 14, 17, and 20]. In one embodiment, this disclosure relates to a multispecific molecule comprising an isolated antibody of the present disclosure.

[0106] In another embodiment, the present disclosure provides isolated antibodies having binding specificity to human IL-17A, in particular comprising a set of CDRs (HCDR1, HCDR2, HCDR3, LDCR1, LDCR2, and LDCR3), wherein the set of CDRs has 10 or fewer, preferably 0, amino acid substitutions from a given set of CDRs [wherein HCDR1' is an amino acid sequence selected from any one of SEQ ID NOs. 39, 42, and 45; HCDR2' is an amino acid sequence selected from any one of SEQ ID NOs. 40, 43, and 46; HCDR3' is an amino acid sequence selected from any one of SEQ ID NOs. 41, 44, and 47; LCDR1' is an amino acid sequence selected from any one of SEQ ID NOs. 50, 53, and 56; LCDR2' is an amino acid sequence selected from any one of SEQ ID NOs. 51, 54, and 57; and LCDR3' is an amino acid sequence selected from any one of SEQ ID NOs. 52, 55, and 58]. In one embodiment, this disclosure relates to a multispecific molecule comprising an isolated antibody of the present disclosure.

[0107] In one embodiment, the present disclosure relates to a pharmaceutical composition comprising an isolated antibody of the present disclosure, or a multispecific molecule containing an isolated antibody of the present disclosure, and a pharmaceutically acceptable carrier.

[0108] In another embodiment, the present disclosure relates to an antibody of the present disclosure, or a multispecific molecule containing the isolated antibody, or a pharmaceutical composition of the present disclosure, for use as a pharmaceutical agent.

[0109] In one embodiment, the present disclosure relates to an antibody of the present disclosure, or a multispecific molecule containing the isolated antibody, or a pharmaceutical composition for use in the treatment of a disorder mediated by IL-17A or a disorder that can be treated by inhibiting GRO-α secretion.

[0110] In one embodiment, the Disclosure relates to the use of the Antibody of the Disclosure, or a multispecific molecule containing the Isolated Antibody, or a Pharmaceutical Composition of the Disclosure, in the manufacture of a drug for use in the treatment of a disorder mediated by IL-17A or a disorder that can be treated by inhibiting GRO-α secretion.

[0111] In another embodiment, the Disclosure relates to a method for treating IL-17A-mediated disorders, the method comprising administering an effective amount of the antibody of the Disclosure, or a multispecific molecule of the Disclosure, or a pharmaceutical composition of the Disclosure, to a subject in need thereof. In yet another embodiment, the Disclosure relates to a nucleic acid encoding the antibody of the Disclosure. In a further embodiment, the Disclosure relates to a vector comprising the nucleic acid. In a further embodiment, the Disclosure relates to a host cell comprising the nucleic acid or the vector.

[0112] In another embodiment, the Disclosure relates to a method for producing the antibody of the Disclosure, the method comprising the step of culturing a host cell containing a nucleic acid or vector encoding the antibody of the Disclosure.

[0113] Individually or in combination, the aspects, advantageous features, and preferred embodiments of this disclosure, summarized in the following sections, further contribute to solving the objectives of the present invention.

[0114] 1. An antibody having binding specificity to human IL-17A, comprising a set of CDRs (HCDR1, HCDR2, HCDR3, LDCR1, LDCR2, and LDCR3), wherein the set of CDRs is a set of CDRs [where, (i) HCDR1' is an amino acid sequence selected from one of sequence numbers 1, 4, and 7, HCDR2' is an amino acid sequence selected from one of sequence numbers 2, 5, and 8. HCDR3' is an amino acid sequence selected from one of sequence numbers 3, 6, and 9. LCDR1' is an amino acid sequence selected from one of sequence numbers 12, 15, and 18. LCDR2' is an amino acid sequence selected from one of sequence numbers 13, 16, and 19. LCDR3' has an amino acid sequence selected from any one of SEQ ID NOs: 14, 17, and 20; or (ii) HCDR1' is an amino acid sequence selected from one of sequence numbers 39, 42, and 45, HCDR2' is an amino acid sequence selected from one of sequence numbers 40, 43, and 46. HCDR3' is an amino acid sequence selected from one of sequence numbers 41, 44, and 47. LCDR1' is an amino acid sequence selected from one of sequence numbers 50, 53, and 56. LCDR2' is an amino acid sequence selected from one of sequence numbers 51, 54, and 57. The antibody having 10 or fewer amino acid substitutions from the above antibody, wherein LCDR3' has an amino acid sequence selected from any one of SEQ ID NOs. 52, 55, and 58.

[0115] 2. An antibody of item 1 comprising a set of CDRs (HCDR1, HCDR2, HCDR3, LDCR1, LDCR2, and LDCR3), wherein the set of CDRs is a set of CDRs [where, (i) HCDR1' is the one described in Sequence ID No. 1, HCDR2' is the one described in Sequence ID No. 2. HCDR3' is the one described in Sequence ID No. 3, LCDR1' is the one described in Sequence ID No. 12. LCDR2' is the one described in Sequence ID No. 13. LCDR3' is the one described in Sequence ID No. 14; or (ii) HCDR1' is the one described in Sequence ID No. 39, HCDR2' is the one described in Sequence ID No. 40. HCDR3' is the one described in Sequence ID No. 41. LCDR1' is the one described in Sequence ID No. 50. LCDR2' is the one described in Sequence ID No. 51. The above antibody having 10 or fewer amino acid substitutions from LCDR3', which is described in SEQ ID NO: 52.

[0116] 3. An antibody of item 1 or item 2, comprising a heavy chain variable region (VH) and a light chain variable region (VL), (c) The VH includes, in order, three complementarity determination regions HCDR1, HCDR2, and HCDR3, and (d) The antibody wherein the VL comprises, in order, three complementarity-determining regions LCDR1, LCDR2, and LCDR3.

[0117] 4. (i) (g) The HCDR1 is described in an amino acid sequence selected from any one of Sequence ID Nos. 1, 4, and 7, (h) The HCDR2 is described in an amino acid sequence selected from any one of Sequence IDs 2, 5, and 8, (i) The HCDR3 is described in an amino acid sequence selected from any one of Sequence ID Nos. 3, 6, and 9, (j) The LCDR1 is described in an amino acid sequence selected from any one of sequence numbers 12, 15, and 18, (k) The LCDR2 is described in an amino acid sequence selected from any one of sequence numbers 13, 16, and 19, and (l) The LCDR3 is an amino acid sequence selected from any one of sequence numbers 14, 17, and 20; or (ii) (a) The HCDR1 is described in an amino acid sequence selected from any one of Sequence IDs 39, 42, and 45, (b) The HCDR2 is described in an amino acid sequence selected from any one of Sequence ID Nos. 40, 43, and 46, (c) The HCDR3 is described in the amino acid sequence selected from any one of Sequence IDs 41, 44, and 47, (d) The LCDR1 is described in an amino acid sequence selected from any one of sequence numbers 50, 53, and 56, (e) The LCDR2 is described in an amino acid sequence selected from any one of sequence numbers 51, 54, and 57, and (f) The antibody of item 3, wherein the LCDR3 is described in an amino acid sequence selected from any one of sequence numbers 52, 55, and 58.

[0118] 5. (i) 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 (ii) The antibody of item 4, comprising 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.

[0119] 6. One antibody from item 3 to 5, wherein the VH is VH3 or VH4, preferably VH3.

[0120] 7. The VL comprises Vκ framework FR1, FR2, and FR3, more specifically Vκ1 or Vκ3 FR1-FR3, preferably Vκ1 FR1-FR3, and framework FR4, wherein framework FR4 comprises Vλ FR4 having at least 60, 70, 80, or 90% identity with Vκ FR4, more specifically Vκ1 FR4, Vκ3 FR4, and Vλ FR4, more specifically Vλ FR4 having at least 60, 70, 80, or 90% identity with an amino acid sequence selected from any of SEQ ID NOs. 26-32, preferably Vλ FR4 as described in any of SEQ ID NOs. 26-32, preferably Vλ FR4 as described in SEQ ID NOs. 26 or 27, more preferably Vλ FR4 as described in SEQ ID NOs. 7. The antibody comprising any one of items 3-6.

[0121] 8. Any one antibody from items 3 to 7, wherein (i) the VH contains an amino acid sequence that is at least 90% identical to amino acid sequence sequence number 10, and / or the VL contains an amino acid sequence that is at least 90% identical to amino acid sequence sequence number 21; or (ii) the VH contains an amino acid sequence that is at least 90% identical to amino acid sequence sequence number 48, and / or the VL contains an amino acid sequence that is at least 90% identical to amino acid sequence sequence number 59.

[0122] 9. The antibody of item 8, wherein (i) VH comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 10 and 11, and / or VL comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 21 and 22; or (ii) VH comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 48 and 49, and / or VL comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 59 and 60.

[0123] 10. The antibody of 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.

[0124] 11. (i) The VH sequence of SEQ ID NO: 11 and / or the VL sequence of SEQ ID NO: 49; or (ii) The antibody of item 9, comprising the VH sequence of SEQ ID NO: 11 and / or the VL sequence of SEQ ID NO: 60.

[0125] 12. One of the antibodies listed above, wherein the antibody has binding specificity to cynomolgus monkey IL-17A.

[0126] 13. One of the aforementioned antibodies that, when measured by ELISA, selectively binds to human IL-17A rather than human IL-17B, IL-17C, IL-17D, IL-17E, and IL-17F.

[0127] 14. The binding to IL-17A (a) Inhibit or block the binding of IL-17A to its receptor (IL-17RA), (b) One of the antibodies listed above that reduces or neutralizes IL-17A activity.

[0128] 15. The antibody of item 14, which, when evaluated in vitro using the HT-29 assay, can inhibit GRO-α secretion.

[0129] 16. An antibody of any of the items mentioned above, (h) Having the ability to inhibit the interaction between IL-17A and IL-17RA at a titer (relative titer) greater than 5, preferably greater than 10, more preferably greater than 15, and even more preferably greater than 20 compared to the titer of secukinumab determined by ELISA assay, where the relative titer is the IC of secukinumab measured by ELISA. 50 The IC50 value (ng / mL) and the IC50 value of the antibody of the present invention in scFv format as measured by ELISA. 50 It is the ratio to the value (ng / mL); and / or (i) Having the ability to neutralize IL-17A at a titer (relative titer) greater than 50, preferably greater than 100, more preferably greater than 150, compared to the titer of secukinumab determined by measuring Gro-α secretion in the HT-29 assay, where the relative titer is the IC of secukinumab as measured by the HT-29 assay. 50 The IC50 value (ng / mL) and the IC50 value of the antibody of the present invention in scFv format as measured by the HT-29 assay. 50 It is the ratio to the value (ng / mL); and / or (j) The antibody can inhibit 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, and more preferably 0.2 ng / mL or less, wherein the inhibitory activity is determined by measuring the GRO-α secretion induced by human IL-17A in an HT-29 assay in the presence of 50 pg / ml TNFα.

[0130] 17. An antibody of any of the items mentioned above, (a) When measured by surface plasmon resonance, more specifically when measured directly by surface plasmon resonance, the dissociation constant (K) for human IL-17A is less than 5 nM, more specifically less than 1 nM, less than 0.5 nM, less than 0.2 nM, more specifically less than 100 pM, and more specifically less than 50 pM. D ) and (b) When measured by surface plasmon resonance, specifically in the capture setting, the cynomolgus monkey IL-17A had a K content of less than 10 nM, specifically less than 7 nM, less than 5 nM, less than 2 nM, less than 1 nM, and more specifically less than 0.5 nM. D The above antibody binds to it.

[0131] 18. When the antibody is measured by surface plasmon resonance, more specifically by direct measurement using surface plasmon resonance, the dissociation constant (K) for human IL-17A is less than 0.5 nM, less than 0.2 nM, less than 100 pM, and more specifically less than 50 pM. D ) The antibody of item 17 that binds to it.

[0132] 19. An antibody of any of the items mentioned above, (e) If in scFv format, more specifically, when the antibody is in pH 6.4, 150 mM NaCl phosphate-citrate buffer, the melting temperature (Tm) determined by differential scanning fluorescence assay is at least 60°C, more specifically at least 62°C, at least 65°C, and more specifically at least 70°C; (f) In the case of scFv format, if the antibody of the present invention is at an initial concentration of 10 mg / ml, more specifically, if the antibody is in phosphate-buffered saline (PBS), pH 7.4, after 5 consecutive freeze-thaw cycles, the loss of monomer content is less than 5%, more specifically less than 3%, and more specifically less than 1%; (g) When in scFv format, when the antibody of the present invention is at an initial concentration of 10 mg / ml, more specifically when the antibody is in phosphate-buffered saline (PBS), pH 7.4, after storage at 4°C for at least 2 weeks, more specifically at least 4 weeks, the loss of monomer content is 5% or less, more specifically less than 4%, less than 3%, less than 2%, more specifically less than 1%, and / or (h) The antibody of the present invention, when the initial concentration is 10 mg / ml, exhibits a loss of monomer content of less than 5% after storage at 37°C for at least 2 weeks, and more specifically, at least 4 weeks.

[0133] 20. 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 the VNAR structure of sharks, and, but not limited to, ankylin-based domains, finomers, avimers, anticarin, binding domains based on alternative scaffolds including fibronectin, and binding sites incorporated into the constant region of the antibody (e.g., F-star's Modular Antibody Technology®), preferably scFV, and is any of the antibodies listed above.

[0134] 21. The antibody of item 20, 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 the scFv has the amino acid sequence of SEQ ID NO: 24; or (ii) an amino acid sequence selected from the group consisting of SEQ ID NO: 61 and SEQ ID NO: 62, preferably the scFv has the amino acid sequence of SEQ ID NO: 61.

[0135] 22. The isolated antibody of item 20, wherein the antibody is an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4, and preferably the antibody is IgG1 or IgG4.

[0136] 23. An isolated antibody of any of the above items, which has been humanized.

[0137] 24. One antibody from items 1-23 that is a multispecific molecule.

[0138] 25. Antibodies of item 24, including single-stranded diabody (scDb), tandem scDb (Tandab), linear dimer scDb (LD-scDb), cyclic dimer scDb (CD-scDb), bispecific T cell engager (BiTE; tandem di-scFv), tandem tri-scFv, tribody (Fab-(scFv)2) or vibody (Fab-(scFv)1), Fab, Fab-Fv2, Morrison (IgG CH3-scFv fusion (Morrison L) or IgG) CL-scFv fusion (Morrison H), triabody, scDb-scFv, bispecific Fab2, dimini antibody, tetrabody, scFv-Fc-scFv fusion, scFv-HSA-scFv fusion, didiabody, DVD-Ig, COVD, IgG-scFab, scFab-dsscFv, Fv2-Fc, IgG-scFv fusion, e.g., 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 ( The above antibody is selected from the group consisting of 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 antibody (KiH);Fv, scFv, scDb, tandem-di-scFv, tandem-tri-scFv, Fab-(scFv)2, Fab-(scFv)1, Fab, Fab-Fv2, COVD, MATCH, and duobodies fused to the N-terminus and / or C-terminus of a heterodimeric Fc domain or any other heterodimerizing domain.

[0139] 26. A pharmaceutical composition comprising one antibody from item 1 to 25 and a pharmaceutically acceptable carrier.

[0140] 27. One antibody from items 1 to 25, or a pharmaceutical composition of item 26, for use as a drug.

[0141] 28. One antibody from item 1 to 25, or a pharmaceutical composition of item 26, for use in the treatment of disorders mediated by IL-17A, or disorders that can be treated by inhibiting GRO-α secretion.

[0142] 29. One antibody from items 1 to 25, or a pharmaceutical composition of item 26, for use in the treatment of an inflammatory condition or autoimmune disease.

[0143] 30. Any antibody from items 1-25, or a pharmaceutical composition of item 26, for use in the treatment of any of the following diseases: cancer, arthritis, rheumatoid arthritis, osteoarthritis of the elbow, reactive arthritis, psoriasis, chronic obstructive pulmonary disease, systemic lupus erythematosus (SLE), lupus nephritis, autoimmune inflammatory bowel disease, asthma, multiple sclerosis, cystic fibrosis, bone loss, airway hypersensitivity, demyelinating disorders, skin hypersensitivity, acute transplant rejection, allograft rejection, graft-versus-host disease, systemic sclerosis, urinary tract inflammatory diseases, cardiovascular diseases, vasculitis, periodic fever, glucose metabolism disorders, lung diseases, periodontitis, hepatic interstitial keratitis, allergies, inflammatory pain, spondyloarthropathy, sepsis, septic or endotoxin shock, meningitis, surgical trauma, autoimmune hematological disorders, Alzheimer's disease, sarcoidosis, cirrhosis, hepatitis, glomerulonephritis, or dyslipidemia.

[0144] 31. Use of any one antibody from items 1 to 25 or a pharmaceutical composition from item 26 in the manufacture of a drug for use in the treatment of a disorder mediated by IL-17A or a disorder that can be treated by inhibiting Gro-α secretion.

[0145] 32. Use of any one antibody from items 1 to 25, or a pharmaceutical composition from item 26, in the manufacture of a drug for use in the treatment of an inflammatory condition or autoimmune disease.

[0146] 33. Use of any one antibody from items 1-25 or a pharmaceutical composition of item 26 in the manufacture of a drug for use in the treatment of any of the following diseases: cancer, arthritis, rheumatoid arthritis, osteoarthritis of the elbow, reactive arthritis, psoriasis, chronic obstructive pulmonary disease, systemic lupus erythematosus (SLE), lupus nephritis, autoimmune inflammatory bowel disease, asthma, multiple sclerosis, cystic fibrosis, bone loss, airway hypersensitivity, demyelinating disorders, skin hypersensitivity, acute transplant rejection, allograft rejection, graft-versus-host disease, systemic sclerosis, urinary tract inflammatory diseases, cardiovascular diseases, vasculitis, periodic fever, glucose metabolism disorders, lung diseases, periodontitis, hepatic interstitial keratitis, allergies, inflammatory pain, spondyloarthropathy, sepsis, septic or endotoxin shock, meningitis, surgical trauma, autoimmune hematological disorders, Alzheimer's disease, sarcoidosis, cirrhosis, hepatitis, glomerulonephritis, or dyslipidemia.

[0147] 34. A method for treating an IL-17A-mediated disorder, comprising administering an effective amount of one antibody from item 1 to 25 or a pharmaceutical composition of item 26, such that the condition is alleviated.

[0148] 35. The method of item 34, in which the disorder mediated by IL-17A is an inflammatory condition or an autoimmune disease.

[0149] 36. The method of item 34, wherein the disorder mediated by IL-17A is cancer, arthritis, rheumatoid arthritis, osteoarthritis of the elbow, 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 hyperresponsiveness, demyelinating disorders, skin hypersensitivity, acute transplant rejection, allograft rejection, graft-versus-host disease, systemic sclerosis, urinary tract inflammatory diseases, cardiovascular diseases, vasculitis, periodic fever, glucose metabolism disorders, lung diseases, periodontitis, hepatic interstitial keratitis, allergies, inflammatory pain, spondyloarthropathy, sepsis, septic or endotoxin shock, meningitis, surgical trauma, autoimmune hematological disorders, Alzheimer's disease, sarcoidosis, cirrhosis, hepatitis, glomerulonephritis, or dyslipidemia.

[0150] 37. Nucleic acids encoding antibodies for items 1-25.

[0151] 38. A vector containing nucleic acids from item 37.

[0152] 39. Host cells containing the nucleic acid of item 37 or the vector of item 38.

[0153] 40. A method for producing antibodies according to items 1 to 25, comprising the step of culturing host cells containing nucleic acids or vectors encoding antibodies according to items 1 to 25.

[0154] 41. A kit containing one antibody from items 1-25, or the pharmaceutical composition of item 26.

[0155] This disclosure intends to include all combinations of any one or more of the embodiments described above, as well as any one or more of the embodiments described in the Detailed Description and Examples.

[0156] Other features, purposes, and advantages of the compositions and methods described herein will become apparent from the description and drawings, as well as from the claims. [Brief explanation of the drawing]

[0157] [Figure 1] This study demonstrates the suitability of labeled IL-17A for use in the sorting process. The biological activity of labeled IL-17A in the HT-29 assay was also investigated. Three-fold serial dilutions of labeled and unlabeled IL-17A were tested in parallel for their potential to induce GRO-α secretion in HT-29 cells. The EC50 values ​​for labeled (IL-17A-RPE) and unlabeled (IL-17A) IL-17A were 82.8 ng / ml and 55 ng / ml, respectively. [Figure 2] The titers of 27-07-G02 rabbit IgG (A) and 27-31-CO4 IgG (B) for neutralizing IL-17A in the HT-29 assay are shown. [Figure 3]The titers of 27-07-G02 rabbit IgG (A) and 27-31-CO4 IgG (B) are shown to inhibit the interaction between IL-17A and IL-17RA. [Figure 4] The titers of humanized cFv A1 and A2(A), and PRO571 and PRO592(B) for neutralizing IL-17A in the HT-29 assay are shown. [Figure 5] The titers (ELISA) of anti-IL-17A scFv A1 and A2(A), as well as PRO571 and PRO592(B), for inhibiting the interaction between human IL-17A and IL-17RA are shown. [Figure 6-I] This shows the target specificity of scFv A1 (A), scFv PRO571, and PRO592 (B). The potential for inhibiting the interaction between biotinylated IL-17A and scFv by IL-17B to IL-17F was analyzed by competitive ELISA. Dose-dependent effects of IL-17A and IL-17B to IL-17F were demonstrated. [Figure 6-II] This shows the target specificity of scFv A1 (A), scFv PRO571, and PRO592 (B). The potential for inhibiting the interaction between biotinylated IL-17A and scFv by IL-17B to IL-17F was analyzed by competitive ELISA. Dose-dependent effects of IL-17A and IL-17B to IL-17F were demonstrated. [Figure 7] The thermal evolution curves from DSF measurements of scFv A1 and A2(A), and scFv PRO571 and PRO592(B) are shown. The obtained Tm values ​​are determined by fitting the data to the Boltzmann equation and obtaining the midpoint of the transition. [Figure 8A] This shows the storage stability tests of scFvA1(A), scFv PRO571, and PRO592(B) conducted for 4 weeks at concentrations of >10 mg / mL at three temperatures (37°C, 4°C, and -80°C). Monomer content over time at various storage temperatures is shown on the left, and protein concentration over time at various 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 protein concentration was calculated by UV280 measurement. [Figure 8B] This shows the storage stability tests of scFvA1(A), scFv PRO571, and PRO592(B) conducted for 4 weeks at concentrations of >10 mg / mL at three temperatures (37°C, 4°C, and -80°C). Monomer content over time at various storage temperatures is shown on the left, and protein concentration over time at various 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 protein concentration was calculated by UV280 measurement. [Figure 9] This shows the tracking of monomer content in scFv A1(A), scFv PRO571, and PRO592(B) over five freeze-thaw cycles. [Figure 10] The triple specificity form is shown. A. Rearrangement variants of the three domains of the Fab-(scFv)2 molecule were designed. The ScFv fusion at positions CL and CH1 in the tribody form are considered equivalent, resulting in three variants of this form. B. Three scDb-scFv domain rearrangement variants were designed. Domain specificity is shown below. The Gly-Ser linker connecting the variable domain and interdomain disulfide bonds is shown as a gray key shape. [Figure 11] The general process for lead compound preparation is shown. A. Overlay of SE-HPLC traces of the final A3-A5 (left graph) and A6-A8 (right graph) samples. Peaks with retention times of 7-8 minutes correspond to the apparent molecular weight of the monomers of each molecule. Peaks with retention times greater than 10 minutes are artifacts related to the buffer and salt. B. SDS-PAGE analysis of A3-A8 under non-reducing (left) and reducing (right) buffer conditions. Molecular weight standards were added to the center lane. The bands under non-reducing conditions correspond to predicted molecular weights of approximately 100 kDa for A3-A5 and approximately 75 kDa for A6-A8, respectively. As predicted, in the case of the heterodimer Fab-(scFv)2 antibody form, the (A3-A5) band shifts to approximately 50 kDa under reducing conditions, but the A6-A8 band with a molecular weight of approximately 75 kDa is also observable under reducing conditions. [Figure 12]Figure 12 shows the titer for neutralizing TNFα in the L929 assay. Absorbance, measured using cell counting kit-8, is expressed as a function of the concentrations (in nM) of the triple-specific molecules A3-A8. A13 (parental bispecificity, HSA binder, and TNFα inhibitor) was used as a reference. [Figure 13] This section compares the neutralizing titers of human TNFα and cynomolgus monkey TNFα. Absorbance measured using cell counting kit-8 in the presence of human TNFα or cynomolgus monkey TNF-α is shown as a function of A5 and A7 concentrations. [Figure 14] This study demonstrates the simultaneous inhibition of TNF-α and IL-17A in the presence of HSA in an HT-29 assay. In vitro simultaneous inhibition 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 inhibitor) and A13 (parental bispecificity, HSA binder, and TNFα inhibitor) were used as references. The obtained GRO-α secretion data are expressed as a function of molecular concentration nM (A, C, and E) and ng / ml (B, D, and F). "Without TNFα" represents GRO-α secretion with IL-17A added only, corresponding to the maximum effect of TNFα inhibition. "Without IL-17A" represents GRO-α secretion with TNFα added only, corresponding to the maximum effect of IL-17A inhibition. "Without IL-17a, without TNFα" indicates background GRO-α secretion when IL-17A and TNFα are not added, corresponding to the maximum effect achieved by simultaneously inhibiting TNFα and IL-17A. [Figure 15] This shows the neutralization of IL-17A binding to IL-17RA in a competitive ELISA. The absorbance measured by the competitive ELISA evaluating IL-17A binding to IL-17RA is expressed as a function of increasing concentrations of six trispecific molecules (A3-A8). Secukinumab (an IL-17A inhibitor) was used as a reference. [Figure 16]Shows simultaneous binding to human TNFα, human IL-17A, and HSA by SPR. Six possible order injections of different analytes (human TNFα, human IL-17A, and HSA) were performed on a MAAS-1 SPR instrument, and the resulting sensorgrams are shown. The trispecific molecule was immobilized on the sensor chip (A3 in channel 1B, Ch1B, A4 in channel 2B, Ch2B, A5 in channel 3B, Ch3B, A6 in channel 4B, Ch4B, A7 in channel 5B, Ch5B, A8 in channel 6B, Ch6B), and the antigens were injected sequentially. [Figure 17] Shows the storage stability test performed for 4 weeks at a protein concentration of 10 mg / mL at temperatures of 37 °C, 4 °C, and -80 °C. The changes over time in the monomer content % and monomer loss % were recorded at d0, d2, d7, d14, d21, and d28. [Figure 18] Shows the overlay of SE-HPLC traces of the d0 (black, shaded) and d28 (gray) stability samples of A5 (left), A7 (center), and A8 (right). [Figure 19] Shows the pharmacokinetic profiles of A7 in cynomolgus monkeys after intravenous (n = 3) and subcutaneous (n = 3) administrations and ADA analysis. [Figure 20] Shows the schematic structures of Morrison L constructs A14 and A15. [Figure 21] Shows the average size of the soluble complex determined by dynamic light scattering for scDb-scFv A7. [Figure 22] Shows the average size of the soluble complex determined by dynamic light scattering for A14. [Figure 23] Shows the average size of the soluble complex determined by dynamic light scattering for A15. [Figure 24] Shows the protein concentration recovery of the soluble complex of scDb-scFv A7. <0^000911> [Figure 25] Shows the recovery of the protein concentration of the soluble complex of A14. [Figure 26] Shows the recovery of the protein concentration of the soluble complex of A15.

Mode for Carrying Out the Invention

[0158] This disclosure is based on the discovery of multispecific antibody molecules that specifically bind to IL-17A and TNFα and possess improved affinity, efficacy, and selectivity. Furthermore, the multispecific antibodies of this disclosure have an improved safety profile, as the inventors have demonstrated that the antibodies do not form immune complexes with TNFα and are therefore potentially less immunogenic. Divalent binding of other multispecific agents (e.g., Covagen, Abbvie, etc.) increases the likelihood of immune complex formation, which can lead to immunogenicity or other adverse effects. In contrast, the monovalent bivalent and trispecific constructs of this disclosure are less likely to form such complexes and are therefore less likely to cause anti-drug antibody and immune-related adverse effects. Moreover, the multispecific antibodies of this disclosure have improved biophysical properties, such as developability and high productivity with relatively few impurities, as well as excellent stability.

[0159] This disclosure further provides antibodies that specifically bind to the human IL-17A protein, as well as pharmaceutical compositions, methods for producing such antibodies and pharmaceutical compositions, and methods for using such antibodies and pharmaceutical compositions.

[0160] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art to whom this invention relates.

[0161] The terms “comprising” and “including” are used herein in their unrestricted, non-limiting sense unless otherwise specified. Therefore, with respect to such latter embodiments, the term “comprising” includes the narrower term “consisting of.”

[0162] In the context describing the present invention (particularly in the context of the following claims), the terms “a,” “an,” “the,” and similar references should be interpreted as encompassing both singular and plural forms unless otherwise stated herein or unless the context clearly contradicts this interpretation. For example, the term “cell” includes multiple cells, including mixtures thereof. Where the plural form is used for compounds, salts, etc., it should be interpreted as also meaning a single compound, salt, etc.

[0163] Multispecific antibodies of this disclosure In one embodiment, the disclosure provides an isolated multispecific antibody comprising a first domain that specifically binds to IL-17A and a second domain that specifically binds to TNFα.

[0164] The terms “TNFα,” “TNF-α,” or “tumor necrosis factor” refer specifically to human TNFα. TNFα is found as a soluble protein and as a precursor called transmembrane TNFα, which is expressed as a type II polypeptide on the cell surface. Transmembrane TNFα is processed by a metalloproteinase such as TNFα-converting enzyme (TACE) between residues Ala76 and Val77, releasing the soluble form of TNFα, which consists of 157 amino acid residues. Soluble TNFα is a homotrimer of 17 kDa cleaved monomers. Transmembrane TNFα also exists as a homotrimer of 26 kDa uncleaved monomers. As used herein, the term “TNFα” encompasses both the soluble and transmembrane forms. The term “TNFα” refers specifically to human transmembrane TNFα having UniProt ID number P01375, replicated herein as Sequence ID No. 134. The term "TNFα" specifically refers to soluble transmembrane TNFα having UniProt ID number P01375, which is replicated herein as Sequence ID No. 135.

[0165] Appropriately, the antibodies in this disclosure are isolated antibodies. As used herein, the term “isolated antibody” refers to an antibody that substantially does not contain other antibodies with different antigen specificities (for example, an isolated antibody that specifically binds only to IL-17A and TNFα substantially does not contain 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 cross-react to other antigens, such as IL-17A and TNFα molecules from other species. Furthermore, an isolated antibody may substantially not contain other cellular material and / or chemical substances.

[0166] Appropriately, the antibodies in this disclosure are monoclonal antibodies. As used herein, the terms “monoclonal antibody” or “monoclonal antibody composition” refer to an antibody that is substantially identical to or derived from an amino acid sequence of the same genetic source. Monoclonal antibody compositions exhibit binding specificity and affinity to a particular epitope, or binding specificity and affinity to a particular epitope.

[0167] The antibodies disclosed herein include, but are not limited to, chimeric antibodies and humanized antibodies.

[0168] The term "chimeric antibody" refers to an antibody molecule in which (a) the constant region or part thereof has been modified, substituted, or exchanged, so that the antigen-binding site (variable region) is linked to the constant region of a different or modified class, effector function, and / or species, or to a completely different molecule that gives the chimeric antibody new characteristics, such as an enzyme, toxin, hormone, growth factor, or drug; or (b) the variable region or part thereof has been modified, substituted, or exchanged with a variable region having a different or modified antigen specificity. For example, a mouse antibody can be modified by replacing its constant region with a constant region from human immunoglobulin. By substituting with a human constant region, the chimeric antibody can retain its specificity for recognizing an antigen while reducing its antigenicity in humans compared to the original mouse antibody.

[0169] As used herein, “humanized” antibodies are antibodies that retain the reactivity of non-human antibodies while exhibiting low immunogenicity in humans. This can be achieved, for example, by retaining non-human CDRs and replacing the rest of the antibody with their human counterparts (i.e., the framework portions of the constant and variable regions). Additional framework region modifications can be made within the human framework sequence, as well as within the CDR sequence derived from the germline of another mammalian species. The humanized antibodies of this disclosure may contain amino acid residues not encoded by human sequences (e.g., mutations introduced by random or site-directed mutagenesis in vitro, somatic mutations in vivo, or conservative substitutions to enhance stability or production). See, for example, Morrison et al., Proc. Natl. Acad. Sci. USA, 81:6851-6855, 1984; Morrison and Oi, Adv. Immunol., 44:65-92, 1988; Verhoeyen et al., Science, 239: 1534-1536, 1988; Padlan, Molec. Immun., 28:489-498, 1991; and Padlan, Molec. Immun., 31: 169-217, 1994. Other examples of anthropogenic technology include, but are not limited to, the Xoma technology disclosed in U.S. Patent No. 5,766,886.

[0170] As used herein, the term “recombinant humanized antibody” includes all humanized antibodies in the Disclosure prepared, expressed, created, or isolated by recombinant means, such as antibodies isolated from animals (e.g., rabbits), antibodies expressed using recombinant expression vectors transfected into host cells, antibodies isolated from recombinant combinatorial human antibody libraries, or antibodies prepared, expressed, created, or isolated by any other means, including splicing human immunoglobulin gene sequences with other DNA sequences. Such recombinant human antibodies have variable and constant regions (if any) derived from human germline immunoglobulin sequences. However, such antibodies are susceptible to in vitro mutagenesis (or, if transgenic animals for human Ig sequences are used, in vivo somatic mutagenesis), and therefore the amino acid sequences of the VH (antibody heavy chain variable region) and VL (antibody light chain variable region) of recombinant antibodies are derived from and related to the VH and VL sequences of the human germline, but may not be naturally present in the human antibody germline.

[0171] Appropriately, the antibodies or their binding domains of this disclosure are humanized. Appropriately, the antibodies or their binding domains of this disclosure are humanized and contain rabbit-derived CDRs.

[0172] As used herein, the term "multispecific antibody" refers to an antibody that binds to two or more different epitopes on at least two different targets (e.g., IL-17A and TNFα), or to two or more different epitopes of the same target. The term "multispecific antibody" of the present disclosure has two or more binding domains, e.g., two or three binding domains. The term "multispecific antibody" includes bispecific, trispecific, tetravalent, pentavalent, and hexavalent. As used herein, the term "bispecific antibody" refers to an antibody that binds to two different epitopes on, for example, two different targets (e.g., IL-17A and TNFα), or on the same target. As used herein, the term "trispecific antibody" refers to an antibody that binds to three different epitopes on, for example, three different targets (e.g., IL-17A, TNFα, and HSA), or on the same target.

[0173] The term "multivalent antibody" refers to a single binding molecule having multiple valences, where "valence" is described as the number of antigen-binding portions that bind to epitopes on the same target molecule. "Valence" refers to the presence of a specific number of binding domains specific for an antigen within a molecule. Thus, the terms "monovalent", "divalent", "tetravalent", and "hexavalent" refer to the presence of one, two, four, and six binding domains specific for an antigen within a molecule, respectively. As used herein, the term "monovalent antibody" refers to an antibody having a single antigen-binding portion that binds to a single epitope on a target molecule such as IL-17A or TNFα. As used herein, the term "divalent antibody" refers to an antibody having two antigen-binding portions that each bind to the same epitope.

[0174] The multispecific antibodies of the present disclosure can be monovalent or multivalent, e.g., divalent, trivalent, or tetravalent, preferably monovalent, for binding to IL-17A.

[0175] The multispecific antibodies of this disclosure may be monovalent or polyvalent, for example, divalent, trivalent, or tetravalent, preferably monovalent, for binding to TNFα. Since TNFα forms trimers, they are potentially trivalent and can form three-dimensional immune complexes with antibodies having several domains that specifically bind to TNFα, such as divalent, trivalent, or polyvalent antibodies for binding to TNFα. For illustrative purposes, studies of the size of immune complexes formed between TNF and infliximab (a chimeric TNFα IgG antibody) and etanercept (a TNFR2 dimer fusion protein with IgG1 Fc) at different antigen / antibody ratios have shown that each antibody generates an immune complex with a unique size profile (Kim MS, et al., J Mol Biol. 2007;374:1374-1388). Therefore, potential high immunogenicity is one of the concerns for therapeutic antibodies targeting TNFα. Accordingly, in preferred embodiments, the multispecific antibodies of this disclosure are monovalent for binding to TNFα.

[0176] Preferably, the multispecific antibodies of the Disclosure include a single domain that specifically binds to IL-17A and / or a single domain that specifically binds to TNFα. In a preferred embodiment, the multispecific antibodies of the Disclosure include a single domain that specifically binds to TNFα. Preferably, the multispecific antibodies of the Disclosure include a single domain that specifically binds to IL-17A and a single domain that specifically binds to TNFα. In a particular embodiment, the multispecific antibodies of the 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 particular embodiment, an optional polypeptide linker is present, and the antibody consists of a polypeptide having 4 to 25 amino acid residues.

[0177] Appropriately, the multispecific antibodies of this disclosure can advantageously neutralize the biological activity of human TNFα and human IL-17A. The term “neutralize” as used herein will be understood to refer to a reduction in biological signaling activity, which may be partial or complete. Appropriate assays for determining neutralization are known in the Art, and some such assays are provided in the examples herein.

[0178] In one embodiment, the antibody or its first domain selectively binds to human IL-17A, more so than human IL-17B, IL-17C, IL-17D, IL-17E, and IL-17F, as measured specifically by ELISA. As used herein, the term “selectively binds” means that the antibody, composition, formulation, etc., does not significantly bind to IL-17B / C / D / E / F but binds to IL-17A. Selective binding is high affinity (or low K). D ) and low to medium ICs 50 Characterized by this, which is usually low affinity (or high K) D ) with moderate to high IC 50 This is distinct from nonspecific binding, which has 10 -7 Less than M K D When binding occurs, the binding is considered selective. Appropriately, the antibody or its first domain, as measured by SPR, has a higher affinity or lower K to bind to human IL-17B, IL-17C, IL-17D, IL-17E, IL-17F. D It then binds to human IL-17A. Appropriately, the antibody of this disclosure or its first domain, when measured by ELISA, exhibits IC50 against IL-17B, IL-17C, IL-17D, IL-17E, and IL-17F. 50 This is the IC for IL-17A 50 An IC that is at least 100 times larger than, for example, at least 200 times larger, at least 300 times larger, and at least 400 times larger. 50The antibody or its first domain, as measured in detail by SPR and / or ELISA, binds to human IL-17A but not to human IL-17B, IL-17C, IL-17D, IL-17E, and IL-17F.

[0179] In further embodiments, the multispecific antibodies of the Disclosure further include a third domain having specificity to antigens different from IL-17A and TNFα. Preferably, the multispecific antibodies of the Disclosure are tripspecific antibodies. As used herein, “tripspecific antibody” refers to an antibody molecule having three antigen-binding domains, for example, one binding domain binding to human TNFα, another binding domain binding to human IL-17A, and yet another binding domain binding to an antigen that can extend the half-life of the antibody molecule, such as human serum albumin.

[0180] More specifically, the multispecific antibodies of this disclosure further include a third domain that specifically binds to human serum albumin (HSA).

[0181] To our surprise, we have found that the addition of a third domain that specifically binds to human serum albumin to the multispecific antibody of this disclosure, which includes a first domain that specifically binds to IL-17A and a second domain that specifically binds to TNFα, has the following beneficial effects: (i) extension of the serum half-life of the multispecific antibody of this disclosure comprising at least one domain that specifically binds to human serum albumin; and (ii) The addition of the human serum albumin-binding domain to the multispecific antibody of this disclosure is compatible with the function of other binding domains, such as the neutralizing activity of the Il-17A and TNFα-binding domains.

[0182] The term "HSA" specifically refers to human serum albumin, UniProt ID number P02768. Human serum albumin (HSA) is a 66.4 kDa abundant protein (50% of total protein) in human serum, composed of 585 amino acids (Sugio, Protein Eng, Vol. 12, 1999, 439-446). The multifunctional nature of the HSA protein is attributed to its structure, which allows for the binding and transport of many metabolites, including fatty acids, metal ions, bilirubin, and several drugs (Fanali, Molecular Aspects of Medicine, Vol. 33, 2012, 209-290). Serum HSA concentrations are approximately 3.5–5 g / dL. Albumin-binding antibodies can be used, for example, to extend the in vivo serum half-life of a drug or a protein bound to it.

[0183] Appropriately, in one embodiment, the multispecific antibody of the Disclosure comprises a first domain that specifically binds to IL-17A, a second domain that specifically binds to TNFα, and a third domain that specifically binds to human serum albumin. The multispecific antibody of the Disclosure may be monovalent or polyvalent, for example, divalent, trivalent, or tetravalent, preferably monovalent, in order to bind to human serum albumin. Appropriately, the multispecific antibody of the Disclosure comprises a single domain that specifically binds to IL-17A, a single domain that specifically binds to TNFα, and a single domain that specifically binds to human serum albumin. In a particular embodiment, the multispecific antibody of the Disclosure comprises 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 particular embodiment, an optional polypeptide linker is present and comprises a polypeptide having 4 to 25 amino acid residues.

[0184] Advantageously, the multispecific antibodies of this disclosure comprise a first domain that specifically binds to IL-17A, a second domain that specifically binds to TNFα, and a third domain that optionally specifically binds to human serum albumin, wherein the domains can simultaneously bind to each of these antigens or receptors.

[0185] The domains of the multispecific antibody of this 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 the VNAR structure of sharks, and, but not limited to, ankyrin-based domains, finomers, avimers, antikalin, fibronectin, and binding domains based on alternative scaffolds including binding sites incorporated into the constant region of an antibody (e.g., -star's Modular Antibody Technology®), preferably from the group consisting of Fab, Fv, and scFv, more preferably, the first domain and / or the second domain and / or the third domain being Fv or scFv.

[0186] The multispecific antibodies of this disclosure may be in any suitable form. In one embodiment, the multispecific antibodies of this disclosure may be single-chain diabodies (scDb), tandem scDb (Tandab), linear dimer scDb (LD-scDb), cyclic dimer scDb (CD-scDb), bispecific T cell engagers (BiTE; tandem di-scFv), tandem tri-scFv, tribodies (Fab-(scFv)2) or vibodies (Fab-(scFv)1), Fab, Fab-Fv2, Morrison (IgG CH3-scFv fusion (Morrison L) or IgG) CL-scFv fusion (Morrison H), triabody, scDb-scFv, bispecific Fab2, dimini antibody, tetrabody, scFv-Fc-scFv fusion, scFv-HSA-scFv fusion, didiabody, DVD-Ig, COVD, IgG-scFab, scFab-dsscFv, Fv2-Fc, IgG-scFv fusion, e.g., 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) The antibody is a bispecific antibody based on a linked scFv, Ts2Ab (dsscFv linked to the C-terminus of the heavy chain), a heterodimeric Fc domain, for example, a Knob-into-Hole antibody (KiH); a form selected from the group consisting of Fv, scFv, scDb, tandem-di-scFv, tandem-tri-scFv, Fab-(scFv)2, Fab-(scFv)1, Fab, Fab-Fv2, COVD, MATCH, and a duobody fused to the N-terminus and / or C-terminus of any heterodimeric Fc domain or any other heterodimerizing domain, preferably a tribody or scDb-scFv.

[0187] The term "diabody" refers to an antibody fragment having two antigen-binding sites, where these fragments contain VH linked to VL within the same polypeptide chain (VH-VL). 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 a complementary domain on another chain, creating two antigen-binding sites. Diabodies can be bivalent or bispecific. Diabodies are described in more detail, for example, EP404097, WO93 / 01161, Hudson et al., Nat. Med. 9:129-134 (2003), and Holliger et al., Proc. Natl. Acad. Sci. USA 90: 6444-6448 (1993). Triabodies and tetrabodies are also described in Hudson et al., Nat. Med. 9:129-134 (2003). Bispecificity scDb, more specifically the bispecificity monomer scDb, comprises more specifically two variable heavy chain domains (VH) or fragments thereof and two variable light chain domains (VL) or fragments thereof, which are linked by linkers L1, L2, and L3 to form VHA-L1-VLB-L2-VHB-L3-VLA, VHA-L1-VHB-L2-VLB-L3-VLA, VLA-L1-VLB-L2-VHB-L3-VHA, VLA-L1-V Linkers are linked in the order 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, where the VLA and VHA domains together form the antigen-binding site for the first antigen, and the VLB and VHB together form the antigen-binding site for the second antigen. Linker L1 is a peptide of 2-10 amino acids, more specifically 3-7 amino acids, and most specifically 5 amino acids, and linker L3 is a peptide of 1-10 amino acids, more specifically 2-7 amino acids, and most specifically 5 amino acids. Intermediate linker L2 is a peptide of 10-40 amino acids, more specifically 15-30 amino acids, and most specifically 20-25 amino acids.

[0188] In one embodiment, the multispecific antibodies of this disclosure comprise an immunoglobulin Fc region polypeptide. In this specification, the term “Fc region” 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. A preferred FcR is a native sequence human FcR. Furthermore, preferred FcRs are those that bind to IgG antibodies (gamma receptors) and include the FcγRI, FcyRII, and FcγRIII subclass receptors, including allelic variants and alternatively spliced ​​forms of these receptors, where the FcγRII receptor includes FcγRIIA (“activating receptor”) and FcγRIIB (“inhibitory receptor”), which have similar amino acid sequences, primarily differing in their cytoplasmic domains. The activating receptor FcγRIIA contains an immunoreceptor tyrosine-based activation motif (ITAM) in its cytoplasmic domain. The inhibitory receptor FcγRIIB contains an immunoreceptor tyrosine-based inhibitory motif (ITIM) in its cytoplasmic domain (see M. Daeron, Annu. Rev. Immunol. 5:203-234 (1997)). FCRs are described in the reviews 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). Other FcRs, including those to be identified in the future, are included in the term “FcR” herein. The term “Fc receptor” or “FcR” also includes the neonatal receptor FcRn, which is involved in the transport of maternal IgG to the fetus. See 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); WO2004 / 92219 (Hinton et al)). In vivo binding to FcRn and the serum half-life of human FcRn high affinity binding polypeptides can be assayed, for example, in transgenic mice or transfected human cell lines expressing human FcRn, or in primates administered with polypeptides containing variant Fc regions. WO2004 / 42072 (Presta) describes antibody variants with improved or reduced binding to FcR. For example, see Shields et al, J. Biol. Chem. 9(2): 6591-6604 (2001).

[0189] To increase the number of specificities / functionalities at the same or smaller molecular weight, it is advantageous to use antibodies containing antibody fragments such as Fv, Fab, Fab', and F(ab')2 fragments, and other antibody fragments. These smaller molecules may retain the antigen-binding activity of the whole antibody and may also exhibit improved tissue penetration and pharmacokinetic properties compared to the whole immunoglobulin molecule. While such fragments appear to offer many advantages over the whole immunoglobulin, they lack the Fc domain, which gives a long half-life in vivo, resulting in a faster clearance rate from serum (Medasan et al., 1997, J. Immunol. 158:2211-2217). Because the low molecular weight molecules penetrate target tissues (such as solid tumors) more efficiently, efficacy may be improved at the same or lower doses. Appropriately, the antibodies of this disclosure do not contain the immunoglobulin Fc region polypeptide and, optionally, more specifically, the CH1 and CL regions, if the multispecific antibody contains a third domain that specifically binds to human serum albumin.

[0190] Preferably, the antibodies of this disclosure may be in a tribody form (Fab-(scFv)2). Preferably, the first domain and / or the second domain and / or the third domain are Fab or scFv domains. In particular, the multispecific antibodies of this disclosure have one Fab domain and two scFv domains, more specifically, the scFv domains are fused to the carboxyl terminus of each chain of the Fab domain. The inventors have tested the optimal relative positions of the individual binding domains in the trispecific form from the viewpoint of pharmacodynamic and biophysical properties and have found, surprisingly, that the multispecific antibodies of this disclosure have advantageous properties when, in the tribody form, 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 domain.

[0191] Preferably, the antibodies of this disclosure are in the scDb-scFv form. The term "scDb-scFv" refers to an antibody form in which a single-stranded Fv (scFv) fragment is fused to a single-stranded diabody (scDb) by a flexible Gly-Ser linker. Preferably, the first and / or second and / or third domains are Fv or scFv domains. More specifically, when the multispecific antibody of this disclosure is in the scDb-scFv form, it has one scFv domain fused at the C-terminus to an scDb containing two other domains. When the multispecific antibody of this disclosure is in the scDb-Fv form, it can be represented by the following formula:

[0192] VLA-L1-VHC-L2-VLC-L3-VHA-L4-VLB-L5-VHB, or VLB-L1-VHA-L2-VLA-L3-VHB-L4-VLC-L5-VHC, or VLC-L1-VHB-L2-VLB-L3-VHC-L4-VLA-L5-VHA, or VLA-L1-VHB-L2-VLB-L3-VHA-L4-VLC-L5-VHC, Preferably, VLB-L1-VHA-L2-VLA-L3-VHB-L4-VLC-L5-VHC, or VLA-L1-VHB-L2-VLB-L3-VHA-L4-VLC-L5-VHC, more, VLB-L1-VHA-L2-VLA-L3-VHB-L4-VLC-L5-VHC,

[0193] Here, VLA and VHA are the light chain variable region and heavy chain variable region of the first domain (which specifically binds to IL-17A), respectively; VLB and VHB are the light chain variable region and heavy chain variable region of the second domain (which specifically binds to TNFα), respectively; and VLC and VHC are the light chain variable region and heavy chain variable region of the third domain (which specifically binds to HSA), respectively, where L1, L2, L3, L4, and L5 are polypeptide linkers.

[0194] In the context of this disclosure, the term “polypeptide linker” refers to a linker consisting of a chain of amino acid residues linked by peptide bonds connecting two domains, each attached to one end of the linker. The polypeptide linker must be long enough to link two molecules so that the two molecules conform correctly to each other and thus maintain the desired activity. In certain embodiments, the 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). Furthermore, the amino acid residues selected to be included in the polypeptide linker must exhibit properties that do not significantly interfere with the activity of the polypeptide. Therefore, the linker peptide as a whole should not exhibit a charge that would be inconsistent with the polypeptide activity, interfere with internal folding, or form binding or other interactions with one or more monomer amino acid residues that would significantly hinder the binding of the monomer domain. In certain embodiments, the polypeptide linker is an unstructured polypeptide. Useful linkers include glycine-serine or GS linkers. "Gly-Ser" or "GS" linkers include polymers of glycine and serine in series (e.g., (Gly-Ser)n, (GSGGS)n(GGGGS)n and (GGGS)n, where n is an integer of at least 1), glycine-alanine polymers, alanine-serine polymers, and other flexible linkers such as shaker potassium channel tethers, and various other flexible linkers as will be understood by those skilled in the art. Glycine-serine polymers are preferred because both amino acids in glycine-serine polymers are relatively unstructured and therefore can potentially function as neutral tethers between components. Secondly, because serine is hydrophilic, it can solubilize what may be spherical glycine chains. Thirdly, similar chains have been shown to be effective in binding subunits of recombinant proteins, such as single-chain antibodies.

[0195] Appropriately, L1 and L3 in this disclosure are those described in Sequence ID No. 132. Appropriately, L2, L4 and L5 in this disclosure are those described in Sequence ID No. 23.

[0196] The inventors, from the viewpoint of pharmacodynamic and biophysical properties, tested the optimal relative positions of the individual binding domains in the triple-specificity form and, surprisingly, found that in the scDb-scFv form of the multispecificity antibody of this disclosure, the first and second domains that specifically bind to IL-17A and TNFα, respectively, form scDb, and the third domain that specifically binds to HSA forms scFv.

[0197] The multispecific antibodies of this disclosure have the following advantageous properties: (a) Having the ability to neutralize IL-17A at a titer (relative titer) greater than 2, e.g., greater than 5, greater than 10, greater than 15, greater than 20, greater than 25, greater than 30, greater than 35, greater than 40, greater than 45, preferably greater than 50, compared to the titer of secukinumab determined by measuring Gro-α secretion in the HT-29 assay, where the relative titer is the IC of secukinumab determined by the HT-29 assay. 50 The IC50 value (ng / mL) and the IC50 value of the multispecific antibody determined by the HT-29 assay. 50 This is the ratio to the value (ng / mL); and

[0198] (b) Having the ability to neutralize TNFα at a titer (relative titer) of at least 1, for example, 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, compared to the titer of scDb of SEQ ID NO: 149(A13) as determined by measuring Gro-α secretion in the HT-29 assay, where the relative titer is the IC2 of the scDb of SEQ ID NO: 149(A13) as determined by the HT-29 assay. 50 The nM value and the IC50 of the multispecific antibody determined by the HT-29 assay. 50It is the ratio to the value (nM); and

[0199] (c) Optionally, a titer 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 (relative titer) compared to the titer of secukinumab determined by ELISA assay, wherein the relative titer is the IC of secukinumab determined by ELISA. 50 The IC50 value (ng / mL) and the IC50 of the multispecific antibody determined by ELISA. 50 This is the ratio to the value (ng / mL); and

[0200] (d) Optionally, having the ability to neutralize TNFα at a titer (relative titer) of at least 0.4, for example at least 0.5, preferably at least 1, compared to the titer of scDb of SEQ ID NO: 149(A13) as determined by the L929 assay, where the relative titer is the IC of the scDb of SEQ ID NO: 149 as measured by the L929 assay. 50 The nM value and the IC50 of the multispecific antibody measured by the L929 assay. 50 It is the ratio to the value (nM); and / or

[0201] (e) When measured by surface plasmon resonance, the dissociation constant (K) is less than 5 nM, for example less than 4 nM, less than 3 nM, less than 2 nM, less than 1 nM, preferably less than 0.5 nM. D ) When bound to human IL-17A, and optionally measured by surface plasmon resonance, the K is less than 5 nM, for example less than 4 nM, less than 3 nM, less than 2 nM, less than 1 nM, preferably less than 0.5 nM. D And it binds to IL-17A in cynomolgus monkeys;

[0202] (f) When measured by surface plasmon resonance, the dissociation constant (K) is less than 5 nM, for example 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. D ) binds to human TNFα; and

[0203] (g) When measured by surface plasmon resonance, the dissociation constant (K) is less than 5 nM, for example less than 4 nM, less than 3 nM, preferably less than 2 nM. D ) Binds to human serum albumin, and when optionally measured by surface plasmon resonance, the dissociation constant (K) is less than 5 nM, for example less than 4 nM, less than 3 nM, preferably less than 2 nM. D ) binds to cynomolgus monkey serum albumin.

[0204] 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 antigens at multiple sites via weak non-covalent bonds. The more interactions there are, the stronger the affinity.

[0205] "Binding affinity" generally refers to the combined strength of non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless otherwise specified, "binding affinity," "binding," "binding," or "bound" as used herein refers to the intrinsic binding affinity that reflects the 1:1 interaction between the members of the binding pair (e.g., antibody and antigen). The affinity of molecule X for partner Y is generally expressed by the dissociation constant (K). D ) can be expressed as. Affinity can be measured by common methods known in the art, including those described herein. Low-affinity antibodies generally tend to bind slowly to antigens and dissociate easily, while high-affinity antibodies generally tend to bind quickly to antigens and remain bound for longer. Various methods for measuring binding affinity are known in the art, and any of them can be used for the purposes of this disclosure. Specific descriptive and exemplary embodiments for measuring binding affinity, i.e., binding strength, are described below.

[0206] The term "k" used in this specification assoc ", ka, or "k onThe term "k" is intended to refer to the association rate of a particular antibody-antigen interaction, whereas "k" as used herein refers to the association rate of a particular antibody-antigen interaction. dis "kd", or "k off The term "K" is intended to refer to the dissociation rate of a particular antibody-antigen interaction. In one embodiment, "K" as used herein is used to mean "K D The term "K" is intended to refer to the dissociation constant, which is derived from the ratio of kd to ka (i.e., kd / ka) and expressed as molar concentration (M). D " or "K D The "value" or "KD" or "KD value" is measured in one embodiment by using a surface plasmon resonance assay with a MAAS-1 SPR instrument (SierraSensors), as described in the examples. The antibody binding affinity is measured, for example, by the dissociation constant (K D This can be determined by a lower K. A stronger affinity is better. D This is represented by a lower affinity, and a lower affinity corresponds to a higher K D It is represented by [this].

[0207] Therefore, in a suitable embodiment, the antibodies of this disclosure, when measured by surface plasmon resonance, have dissociation constants (K) of 1 pM to 10 nM, 1 pM to 7 nM, 1 pM to 5 nM, 1 pM to 4 nM, 1 pM to 3 nM, 1 pM to 2.5 nM, 1 pM to 2 nM, 1 pM to 1.5 nM, 1 pM to 1 nM, preferably 1 pM to 0.5 nM. D ) binds to human IL-17A. In a suitable embodiment, the antibody of this disclosure, when measured by surface plasmon resonance, has a dissociation constant (K) of 1 to 500 pM. D ) binds to human IL-17A. In a suitable embodiment, the antibody of this disclosure has a dissociation constant (K) 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. D ) binds to human IL-17A. Appropriately, the antibodies of this disclosure have a dissociation constant (K) less than 1 nM. D It binds to human IL-17A at ). Appropriately, the antibodies of this disclosure have a dissociation constant (K) less than 0.5 nM. D) binds to human IL-17A. In further embodiments, the antibodies of the present disclosure, when measured by surface plasmon resonance (SPR), have a K content of less than 10 nM, for example, 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. D It binds to IL-17A in cynomolgus monkeys.

[0208] Appropriately, the antibodies of this disclosure, when measured by surface plasmon resonance (SPR), have dissociation constants (K) of 1 pM to 10 nM, 1 pM to 7 nM, 1 pM to 5 nM, 1 pM to 4 nM, 1 pM to 3 nM, 1 pM to 2.5 nM, 1 pM to 2 nM, 1 pM to 1.5 nM, 1 pM to 1 nM, preferably 1 pM to 0.5 nM, and more preferably 1 pM to 0.25 nM. D ) binds to human TNFα. In a suitable embodiment, the antibody of this disclosure, when measured by surface plasmon resonance, has a dissociation constant (K) of 1 to 500 pM, preferably 1 to 250 pM. D ) binds to human TNFα. In a suitable embodiment, the antibody of this disclosure, when measured by surface plasmon resonance, has a dissociation constant (K) 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. D It binds to human TNFα at ). Appropriately, the antibodies of this disclosure have a dissociation constant (K) less than 0.5 nM. D It binds to human TNFα at ). Appropriately, the antibodies of this disclosure have a dissociation constant (K) less than 0.25 nM. D It binds to human TNFα.

[0209] In a suitable embodiment, the antibodies of this disclosure, when measured by surface plasmon resonance, have dissociation constants (K) of 1 pM to 10 nM, 1 pM to 7 nM, 1 pM to 5 nM, 1 pM to 4 nM, 1 pM to 3 nM, preferably 1 pM to 2 nM. D ) binds to human serum albumin. In a suitable embodiment, the antibody of this disclosure, when measured by surface plasmon resonance, has a dissociation constant (K) of 1 to 2000 pM. D) binds to human serum albumin. In a suitable embodiment, the antibody of this disclosure has a dissociation constant (K) less than 5 nM, less than 4 nM, less than 3 nM, preferably less than 2 nM, as measured by surface plasmon resonance. D It binds to human serum albumin at ). Appropriately, the antibodies of this disclosure have a dissociation constant (K) of less than 2 nM. D ) binds to human serum albumin. In a further embodiment, the antibody of the present disclosure has a K content of 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). D It then binds to cynomolgus monkey serum albumin.

[0210] Appropriately, the antibodies of this disclosure possess beneficial biophysical properties. (a) Having a melting temperature (Tm) of at least 55°C, preferably at least 58°C, and more preferably at least 60°C in a pH 6.4, 150 mM NaCl phosphate-citrate buffer, as determined by differential scanning fluorescence assay; (b) When the multispecific antibody is at an initial concentration of 10 mg / ml in phosphate-buffered saline (PBS), pH 7.4, it has a loss of monomer content of less than 5%, for example less than 4%, less than 3%, less than 2%, preferably less than 1%, after 5 consecutive freeze-thaw cycles; (c) When the multispecific antibody is at an initial concentration of 10 mg / ml in phosphate-buffered saline (PBS), pH 7.4, it has a loss of monomer content of less than 10%, preferably less than 5%, after being stored at 4°C for at least 2 weeks, more specifically at least 4 weeks; and / or (d) When the multispecific antibody is at an initial concentration of 10 mg / ml in phosphate-buffered saline (PBS), pH 7.4, it has a loss of monomer content of less than 20%, preferably less than 15%, after being stored at 37°C for at least 2 weeks, more specifically at least 4 weeks.

[0211] The melting temperature (Tm) is determined by differential scanning fluorescence (DSF) as previously described (Egan, et al., MAbs, 9(1) (2017), 68-84; Niesen, et al., Nature Protocols, 2(9) (2007) 2212-2221). The midpoint of the thermal evolution transition is determined by differential scanning fluorescence (DSF) using the fluorescent dye SYPRO® Orange (see Wong & Raleigh, Protein Science 25 (2016) 1834-1840). The sample in phosphate-citrate buffer at pH 6.4 is prepared to a final protein concentration of 50 μg / mL and a final concentration of 5×SYPRO® Orange in a total volume of 100 μl. 25 microliters of the prepared sample is added in triple count to a white-walled AB gene PCR plate. The assay is performed using a qPCR instrument as a thermal cycler, and fluorescence emission is detected using a custom dye calibration routine in the software. PCR plates containing the test samples are subjected to a temperature gradient from 25°C to 96°C in 1°C increments, with a 30-second pause after each temperature increase. The total assay time is approximately 2 hours. Tm is calculated using the GraphPad Prism software, employing a mathematical second derivative method to determine the inflection point of the curve. The reported Tm is the average of three measurements.

[0212] The loss of monomer content is determined by SE-HPLC. SE-HPLC is a separation technique based on a solid stationary phase and a liquid mobile phase, as outlined in Chapter 621 of the USP. This method utilizes a hydrophobic stationary phase and an aqueous mobile phase to separate molecules based on size and shape. Molecular separation occurs between the void capacity (V0) and total permeation capacity (VT) of a particular column. Measurements by SE-HPLC are performed using a ChromasterHPLC system (Hitachi High-Technologies Corporation) equipped with automated sample injection and a UV detector set to a detection wavelength of 280 nm. This instrument is controlled by the software EZChrom Elite (Agilent Technologies, version 3.3.2 SP2), which also supports the analysis of the resulting chromatograms. Protein samples are clarified by centrifugation, kept at a temperature of 6°C in an autosampler, and then injected. For the analysis of scFv samples, a Shodex KW403-4F column (Showa Denko K.K., #F6989202) was used with a standardized buffered saline mobile phase (50 mM sodium phosphate, pH 6.5, 300 mM sodium chloride) at a recommended flow rate of 0.35 mL / min. The target sample load per injection was 5 μg. The sample was detected with a 280 nm UV detector, and the data was recorded using an appropriate software suite. The resulting chromatograms were analyzed in the V0–VT range, and large matrix-related peaks with elution times exceeding 10 minutes were excluded.

[0213] Exemplary domains that specifically bind to IL-17A The multispecific antibody of this 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), wherein (a) the VH comprises, in order, three complementarity-determining regions HCDR1, HCDR2, and HCDR3, and (b) the VL comprises, in order, three complementarity-determining regions LCDR1, LCDR2, and LCDR3.

[0214] Suitable domains for use in the multispecific antibodies of this disclosure that specifically bind to IL-17A include, but are not limited to, the following: • A humanized monoclonal antibody or its binding domain, as presented in the following section, “Anti-IL-17A Antibodies of the Disclosure,” the sequence of which is shown in Table 1; • AIN457 (also known as secukinumab, disclosed in U.S. Patent No. 7,807,155 and WO2006 / 013107, which are incorporated herein by reference in their entirety) or its antigen-binding fragment; • LY2439821 (also known as ixekizumab; disclosed in U.S. Patents 7,838,638 and 8,110,191, and WO2007 / 070750, which are incorporated herein by reference in their entirety) or its antigen-binding fragment; • SCH900117 or its antigen-binding fragment (Merck); • RG4943 or its antigen-binding fragment (Roche); • Anti-IL-17A antibody or its antigen-binding fragment is used in WO2006 / 013107, WO2006 / 054059, WO2007 / 070750, WO2007 / 149032, WO2008 / 001063, WO2008 / 021156, WO2010 / 034443, WO2010 / 102251, WO2012 / 018767, WO2014 Disclosed in / 161570, WO2014 / 001368, WO2014 / 122613, WO2015 / 070697, WO2015 / 137843, WO2016 / 048188, WO2016 / 113557, WO2016 / 138842, and WO2017 / 068472, which are incorporated herein by reference in their entirety.

[0215] Preferred domains for use in the multispecific antibodies of this disclosure that specifically bind to IL-17A include, but are not limited to, the humanized monoclonal antibodies or their binding domains presented below, the sequences of which are listed in Table 1.

[0216] Accordingly, in one embodiment, the present disclosure provides a first domain that specifically binds to human IL-17A and a second domain that specifically binds to TNFα, wherein the first domain comprises a set of CDRs (HCDR1, HCDR2, HCDR3, LDCR1, LDCR2, and LDCR3), wherein the set of CDRs is a set of CDRs [where (i) HCDR1' is a sequence selected from any one of SEQ ID NOs: 1, 4, and 7, preferably from SEQ ID NO: 1; HCDR2' is an amino acid sequence selected from any one of SEQ ID NOs: 2, 5, and 8, preferably from SEQ ID NO: 2; HCDR3' is an amino acid sequence selected from any one of SEQ ID NOs: 3, 6, and 9, preferably from SEQ ID NO: 3; LCDR1' is an amino acid sequence selected from any one of SEQ ID NOs: 12, 15, and 18, preferably from SEQ ID NO: 12; LCDR2' is an amino acid sequence selected from any one of SEQ ID NOs: 13, 16, (ii) where HCDR1' is an amino acid sequence selected from any one of 19, preferably from SEQ ID NO: 13; LCDR3' has an amino acid sequence selected from any one of 14, 17, and 20, preferably from SEQ ID NO: 14; or (ii) where HCDR1' is an amino acid sequence selected from any one of 39, 42, and 45, preferably from SEQ ID NO: 39; HCDR2' is an amino acid sequence selected from any one of 40, 43, and 46, preferably from SEQ ID NO: 40; HCDR3' has an amino acid sequence selected from any one of 41, 44, and 47, preferably from SEQ ID NO: 40; LCDR1' is an amino acid sequence selected from any one of 50, 53, and 56, preferably from SEQ ID NO: 50; LCDR2' has an amino acid sequence selected from any one of 51, 54, and 57, preferably from SEQ ID NO: 51;And LCDR3' has 10 or fewer amino acid substitutions from any one of SEQ ID NOs. 52, 55, and 58, preferably selected from SEQ ID NO. 52. [For example, 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.]

[0217] In particular, this 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 one of the amino acid sequences of the VH CDRs listed in Table 1. In particular, the domain comprises (or consists of) one, two, three, or more VH CDRs having one of the amino acid sequences of the VH CDRs listed in Table 1.

[0218] Preferably, the first domain that specifically binds to IL-17A includes a heavy chain variable region (VH), where the VH comprises, in order: (i) three complementarity-determining regions HCDR1, HCDR2, and HCDR3, wherein HCDR1 has an amino acid sequence selected from any one of SEQ ID NOs: 1, 4, and 7; 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. In particular, the present disclosure provides an antibody having binding specificity to human IL-17A and comprising the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs. 1, 2, and 3, respectively; or (ii) three complementarity-determining regions HCDR1, HCDR2, and HCDR3, wherein HCDR1 has an amino acid sequence selected from any one of SEQ ID NOs. 39, 42, and 45; HCDR2 has an amino acid sequence selected from any one of SEQ ID NOs. 40, 43, and 46; and 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 having binding specificity to human IL-17A and comprising the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs. 39, 40, and 41, respectively.

[0219] This 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 one of the amino acid sequences of the VL CDRs listed in Table 1. In particular, the first domain that specifically binds to IL-17A comprises (or consists of) one, two, three, or more VL CDRs having one of the amino acid sequences of the VL CDRs listed in Table 1.

[0220] Preferably, the first domain that specifically binds to IL-17A comprises a light chain variable region (VL), where the VL comprises, in order: (i) three complementarity-determining regions LDCR1, LDCR2, and LDCR3, wherein LDCR1 has an amino acid sequence selected from any one of SEQ ID NOs: 12, 15, and 18; LCDR2 has an amino acid sequence selected from any one of SEQ ID NOs: 13, 16, and 19; and LCDR3 has an amino acid sequence selected from any one of SEQ ID NOs: 14, 17, and 20. In particular, the present disclosure provides an antibody having binding specificity to human IL-17A and comprising the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs. 12, 13, and 14, respectively; or (ii) three complementarity-determining regions LCDR1, LCDR2, and LCDR3, wherein LCDR1 has an amino acid sequence selected from any one of SEQ ID NOs. 50, 53, and 56, LCDR2 has an amino acid sequence selected from any one of SEQ ID NOs. 51, 54, and 57, and LDCR3 has an amino acid sequence selected from any one of SEQ ID NOs. In particular, the present disclosure provides an antibody having binding specificity to human IL-17A and comprising the LDCR1, LDCR2, and LDCR3 sequences of SEQ ID NOs. 50, 51, and 52, respectively.

[0221] Appropriately, this 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). (i) Here (a) The VH comprises, in order, three complementarity-determining regions HCDR1, HCDR2, and HCDR3, wherein HCDR1 has an amino acid sequence selected from any one of SEQ ID NOs: 1, 4, and 7; 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 (b) The VL comprises, in order, three complementarity-determining regions LCDR1, LCDR2, and LCDR3, wherein LCDR1 has an amino acid sequence selected from any one of SEQ ID NOs: 12, 15, and 18; LCDR2 has an amino acid sequence selected from any one of SEQ ID NOs: 13, 16, and 19; and LCDR3 has an amino acid sequence selected from any one of SEQ ID NOs: 14, 17, and 20; or (ii) Here (a) The VH comprises, in order, three complementarity-determining regions HCDR1, HCDR2, and HCDR3, wherein HCDR1 has an amino acid sequence selected from any one of SEQ ID NOs. 39, 42, and 45; HCDR2 has an amino acid sequence selected from any one of SEQ ID NOs. 40, 43, and 46; and HCDR3 has an amino acid sequence selected from any one of SEQ ID NOs. 41, 44, and 47; and (b) The VL comprises, in order, three complementarity-determining regions LCDR1, LCDR2, and LCDR3, wherein LCDR1 has an amino acid sequence selected from any one of SEQ ID NOs. 50, 53, and 56; LCDR2 has an amino acid sequence selected from any one of SEQ ID NOs. 51, 54, and 57; and LCDR3 has an amino acid sequence selected from any one of SEQ ID NOs. 52, 55, and 58.

[0222] 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) the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs. 1, 2, and 3, respectively, and (b) the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs. 12, 13, and 14, respectively, or (ii) (a) the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs. 39, 40, and 41, respectively, and (b) the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs. 50, 51, and 52, respectively.

[0223] Other domains of the Disclosure that specifically bind to IL-17A include mutated amino acids in their CDR region that have at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identity with the CDR region shown in the sequences listed in Table 1. Preferably, other domains of the Disclosure that specifically bind to IL-17A include mutant amino acid sequences in which, compared to the CDR region shown in the sequences listed in Table 1, 1, 2, 3, 4, 5, or 10 or fewer amino acids in the CDR region are mutated by amino acid deletion, insertion, or substitution. The mutation, e.g., substitution, may occur at any residue within the set of CDRs, and may be within CDR1, CDR2, and / or CDR3.

[0224] The term "amino acid" refers to naturally occurring amino acids, synthetic amino acids, and amino acid analogs and mimetic compounds that function similarly to naturally occurring amino acids. Naturally occurring amino acids are those encoded by the genetic code, as well as those that are later modified, such as hydroxyproline, γ-carboxyglutamic acid, and O-phosphoserine. The terms "polypeptide" and "protein" are used interchangeably herein and refer to polymers of amino acid residues. This term applies to amino acid polymers in which one or more amino acid residues are artificial chemical mimics of corresponding natural amino acids, as well as natural and non-natural amino acid polymers. Unless otherwise specified, a particular polypeptide sequence also implicitly includes its conservatively modified variants.

[0225] Techniques necessary to induce mutations, e.g., substitutions, in the CDR, antibody VH or VL domain, and the amino acid sequence of the antibody are generally available in the art. Mutations, e.g., substitutions are used to create variant sequences that may or may not be expected to have a minimal or beneficial effect on activity, and are tested or not tested for their ability to bind to and / or neutralize IL-17A or TNFα or human serum albumin, and / or other desirable properties. Since appropriate mutations, e.g., substitutions in the CDR do not result in loss of function, the antibody of this disclosure or its binding domain containing such a mutated amino acid sequence retains the ability to bind to 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 unmodified domain of this disclosure, as measured by assays, e.g., described herein. Preferably, the domain of this disclosure of an antigen-binding fragment containing such a mutated amino acid sequence may have an improved ability to bind to and / or neutralize IL-17A or TNFα or human serum albumin. Appropriately, the first domain of the present disclosure, which specifically binds to IL-17A and thus contains a mutated amino acid sequence, has the ability to neutralize IL-17A at a titer (relative titer) greater than 2, e.g., greater than 5, greater than 10, greater than 15, greater than 20, greater than 25, greater than 30, greater than 35, greater than 40, greater than 45, preferably greater than 50, compared to the titer of secukinumab determined by measuring Gro-α secretion in the HT-29 assay, where the relative titer is the IC of secukinumab determined by the HT-29 assay. 50 The IC50 value (ng / mL) and the IC50 value of the multispecific antibody determined by the HT-29 assay. 50 This is the ratio to the value (ng / mL).

[0226] Furthermore, appropriate mutations, such as substitutions, within the CDR do not result in a loss of solubility, stability, and high-yield productivity of the multispecific antibodies of this disclosure; therefore, the multispecific antibodies of this disclosure or their binding domains containing such mutated amino acid sequences retain their biophysical properties. For example, they may retain the same high-yield productivity and / or stability as the unmodified multispecific antibodies of this disclosure, as measured, for example, by the assay described herein. Preferably, the multispecific antibodies or their binding domains containing such mutated amino acid sequences may have improved biophysical properties.

[0227] The terms “identical” or “identical” refer to two or more sequences or subsequences that are identical in relation to two or more nucleic acid or polypeptide sequences. “Percent (%) sequence identity” and “homology” for nucleic acid, peptide, polypeptide, or antibody sequences are defined as the percentage of nucleotides or amino acid residues in a candidate sequence that are identical to amino acid residues in a particular peptide or polypeptide sequence, after the sequences have been aligned and gaps introduced, and conservative substitutions have not been considered as part of the sequence identity, in order to achieve the maximum percentage sequence identity, as is necessary. Alignment for determining percentage amino acid sequence identity can be achieved in various ways by those skilled in the art, using publicly available computer software such as BLAST, BLAST-2, or ALIGN software. Those skilled in the art can determine appropriate parameters for measuring alignment, including any algorithm necessary to achieve the maximum alignment over the entire length of the sequences being compared.

[0228] In sequence comparison, typically one sequence acts as a reference sequence and is compared to the test sequence. When using a sequence comparison algorithm, the test sequence and reference sequence are input into the computer, sub-sequence coordinates are specified as needed, and parameters for the sequence algorithm program are specified. Default program parameters can be used, or different parameters can be specified. The sequence comparison algorithm then calculates the percentage sequence identity of the test sequence to the reference sequence based on the program parameters.

[0229] Two examples of algorithms suitable for determining sequence identity percentage and sequence similarity are the BLAST and BLAST2.0 algorithms, described in Altschul et al., Nuc. AcidsRes. 25:3389-3402, 1977 and Altschul et al., J. Mol. Biol. 215:403-410, 1990, respectively. Software for performing BLAST analysis is available through the National Center for Biotechnology Information.

[0230] The percentage of identity between two amino acid sequences can be determined using the algorithm by E. Meyers and W. Miller (Comput. Appl. Biosci., 4: 11-17, 1988), which is incorporated into the ALIGN program (version 2.0), using a PAM120 weighting table, a gap length penalty of 12, and a gap penalty of 4. Furthermore, the percentage of identity between two amino acid sequences can be determined using the algorithm by Needleman and Wunsch (J. Mol, Biol. 48:444-453, 1970), which is incorporated into the GAP program of the GCG software package (available at www.gcg.com), using either a Blossom62 matrix or a PAM250 matrix, and gap weights of 16, 14, 12, 10, 8, 6, or 4, and length weights of 1, 2, 3, 4, 5, or 6.

[0231] Appropriately, the first domain of the multispecific antibody of this disclosure that specifically binds to IL-17A comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein, (i) (a) The VH includes, in order, three complementarity determination regions HCDR1, HCDR2, and HCDR3, 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: 1, 4, and 7; 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: 2, 5, and 8; The HCDR3 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to any one of SEQ ID NOs: 3, 6, and 9, preferably SEQ ID NO: 3; and / or (b) The VL includes, in order, three complementarity determination regions LCDR1, LCDR2, and LCDR3, 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 sequence numbers 12, 15, and 18; 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; The LCDR3 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to any one of SEQ ID NOs: 14, 17, and 20; or

[0232] (ii) (a) The VH includes, in order, three complementarity determination regions HCDR1, HCDR2, and HCDR3, 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; 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, 40, 43, and 46; The HCDR3 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to any one of SEQ ID NOs: 41, 44, and 47; and / or (b) The VL includes, in order, three complementarity determination regions LCDR1, LCDR2, and LCDR3, 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 sequence numbers 50, 53, and 56; 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 sequence numbers 51, 54, and 57; 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 sequence numbers 52, 52, 55, 58, 52

[0233] Appropriately, the first domain that specifically binds to IL-17A includes: (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 NOs. 1, 2, and 3, respectively, and / or LCDR1 having at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identity with the sequences of SEQ ID NOs. 12, 13, and 14, respectively. LCDR2 and LCDR3; 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 sequence numbers 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 sequence numbers 50, 51 and 52, respectively.

[0234] In a further embodiment, the first domain that specifically binds to IL-17A includes a heavy chain variable region VHA and a light chain variable region VLA.

[0235] In the context of this 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. For example, methods for determining sequence homology and grouping sequences by homology, such as using homology search matrices like BLOSUM (Henikoff, S. & Henikoff, JG, Proc. Natl. Acad. Sci. USA 89 (1992) 10915-10919), are well known to those skilled in the art. For VH, Vκ, and Vλ, different subfamilies can be identified, as shown, for example, in Knappik et al., J. Mol. Biol. 296 (2000) 57-86, which groups VH into VH1A, VH1B, VH2-VH6, Vκ into Vκ1-Vκ4, and Vλ into Vλ1-Vλ3. In vivo, antibody Vκ, Vλ, and VH chains are the result of random rearrangement of the V and J segments of the germline κ chain, the V and J segments of the germline λ chain, and the heavy chain V, D, and J segments, respectively. The subfamily to which a particular antibody variable chain belongs is determined by the corresponding V segment, specifically the framework regions FR1-FR3. Accordingly, in this application, any VH sequence characterized by a specific set of framework regions HFR1 to HFR3 can be combined with any HFR4 sequence, for example, an HFR4 sequence taken from one of the heavy chain germline J segments, or an HFR4 sequence taken from a rearranged VH sequence.

[0236] Suitablely, the first domain of the multispecific antibody of the present disclosure that specifically binds to IL-17A comprises a heavy chain variable region VHA, where 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, where 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, where VHA is VH3.

[0237] Suitablely, the first domain of the multispecific antibody of this disclosure that specifically binds to IL-17A comprises a light chain variable region VLA, the VLA comprising Vκ framework FR1, FR2, and FR3, more specifically Vκ1 or Vκ3 FR1-FR3, preferably Vκ1 framework FR1-FR3, and framework FR4, the framework FR4 being selected from Vκ FR4, more specifically Vκ1 FR4, Vκ3 FR4, and Vλ FR4. Suitable Vλ FR4s are those described in SEQ ID NOs: 26-32. In one embodiment, the first domain that specifically binds to IL-17A comprises a Vλ FR4 having at least 60, 70, 80, or 90% identity with an amino acid sequence selected from any of SEQ ID NOs: 26-32, preferably SEQ ID NOs: 26 or 27, more preferably SEQ ID NOs: 27. Preferably, the first domain that specifically binds to IL-17A includes a Vλ FR4 comprising an amino acid sequence selected from any of SEQ ID NOs. 26 to 32, preferably the Vλ FR4 described in SEQ ID NOs. 26 or 27, more preferably the Vλ FR4 described in SEQ ID NOs. 27.

[0238] Accordingly, 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: (i)(a) the HCDR1, HCDR2, and HCDR3 sequences of sequence numbers 1, 2, and 3, respectively, and the LCDR1, LCDR2, and LCDR3 sequences of sequence numbers 12, 13, and 14, respectively; or (i)(b) the HCDR1, HCDR2, and HCDR3 sequences of sequence numbers 39, 40, and 41, respectively, and the LCDR1, LCDR2, and LCDR3 sequences of sequence numbers 50, 51, and 52, respectively; (ii) VH3 or VH4 domain, preferably VH3 domain; and (iii) A VL domain comprising a VL framework including Vκ framework FR1, FR2, and FR3, more specifically Vκ1 or Vκ3 FR1~FR3, preferably Vκ1 FR1~FR3, and framework FR4, wherein framework FR4 is selected from Vκ FR4, more specifically Vκ1 FR4, Vκ3 FR4, and Vλ FR4, more specifically Vλ FR4 comprising an amino acid sequence having at least 60, 70, 80, or 90% identity with an amino acid sequence selected from any one of SEQ ID NOs: 26 to 32, preferably Vλ FR4 described in an amino acid sequence selected from any one of SEQ ID NOs: 26 to 32, more preferably Vλ FR4 described in SEQ ID NO: 27.

[0239] Suitablely, 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 the VH shown in Table 1. Suitablely, 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 (or alternatively, consists of) the VH amino acid sequence shown in Table 1, wherein about 20 or fewer amino acids, preferably 10 or fewer amino acids, in the framework sequence (e.g., a sequence that is not a CDR) are mutated (where the mutations are additions, substitutions, or deletions, in a variety of non-limiting examples). The other domain of the present disclosure that specifically binds to IL-17A is mutated but has amino acids in the VH region that have at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identity with the VH region shown in the sequences shown in Table 1.

[0240] Suitablely, 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 the VL domain described in Table 1. Suitablely, 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 comprises) the VL amino acid sequence described in Table 1, and wherein about 20 or fewer amino acids, preferably about 10 or fewer amino acids, are mutated in the framework sequence (e.g., a sequence that is not a CDR) (where the mutations are additions, substitutions, or deletions, in a variety of non-limiting examples). Other domains of the present disclosure that specifically bind to IL-17A contain mutated amino acids in the VL region that have at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identity with the VL region shown in the sequences described in Table 1.

[0241] In one embodiment, the present disclosure provides a multispecific antibody comprising a first domain that specifically binds to IL-17A and a second domain that specifically binds to TNFα, wherein the first domain comprises a heavy chain variable region comprising an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90%, identical to amino acid sequence SEQ ID NO: 10 or SEQ ID NO: 11, in particular the domain comprises the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs: 1, 2, and 3, respectively. In a further embodiment, the first domain that specifically binds to IL-17A comprises a heavy chain variable region comprising an amino acid sequence that is at least 90% identical to amino acid sequence SEQ ID NO: 11, wherein the heavy chain variable region comprises Q14K, G16E, and G56A (AHo numbering).

[0242] 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 amino acid sequence SEQ ID NO: 48, and in particular, the domain comprises the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NO: 39, 40, and 41, respectively. In a further 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 amino acid sequence SEQ ID NO: 49, and the heavy chain variable region comprises R20T and Q141P (AHo numbering).

[0243] 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 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 (i) amino acid sequence SEQ ID NO: 21 or SEQ ID NO: 22, preferably SEQ ID NO: 21, and in particular the antibody is such Each comprises the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs. 12, 13, and 14; or (ii) a light chain variable region comprising an amino acid sequence that is identical to amino acid sequence SEQ ID NOs. 59 or 60, preferably SEQ ID NOs. 59, by at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90%, and in particular the antibody comprises the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs. 50, 51, and 52, respectively.

[0244] 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 comprising an amino acid sequence that is at least 90% identical to amino acid sequence SEQ ID NO: 22, wherein the light chain variable region comprises A51P (AHo numbering).

[0245] In a further 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 heavy chain variable region comprising an amino acid sequence that is identical to amino acid sequence SEQ ID NO: 10 by at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90%, and amino acid sequence SEQ ID NO: 21 by at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or (ii) a light chain variable region comprising an amino acid sequence that is 99%, preferably at least 90%, identical to amino acid sequence SEQ ID NO: 48, and 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 amino acid sequence SEQ ID NO: 59, 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 amino acid sequence.

[0246] Preferably, the first domain of the multispecific antibody of the present disclosure that specifically binds to IL-17A comprises (i) a heavy chain variable region comprising an amino acid sequence that is identical to amino acid sequence SEQ ID NO: 10 by at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90%; and amino acid sequence SEQ ID NO: 21 by at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably The antibody comprises a light chain variable region containing an amino acid sequence that is at least 90% identical, wherein the antibody comprises the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs. 1, 2, and 3, and / or the LDCR1, LDCR2, and LDCR3 sequences of SEQ ID NOs. 12, 13, and 14, respectively; in particular, wherein the antibody comprises the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs. 1, 2, and 3, and the LDCR1 sequences of SEQ ID NOs. 12, 13, and 14, respectively. (ii) a heavy chain variable region comprising an amino acid sequence that is identical to amino acid sequence SEQ ID NO: 48 by at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90%; and an amino acid sequence that is identical to amino acid sequence SEQ ID NO: 59 by at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90% The antibody comprises a light chain variable region, wherein the antibody comprises the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs. 39, 40, and 41, and / or the LDCR1, LDCR2, and LDCR3 sequences of SEQ ID NOs. 50, 51, and 52, respectively; in particular, the antibody comprises the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs. 39, 40, and 41, and the LDCR1, LDCR2, and LDCR3 sequences of SEQ ID NOs. 50, 51, and 52, respectively.

[0247] In certain embodiments, 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 containing an amino acid sequence selected from the group consisting of SEQ ID NOs: 10 and 11; and / or a VL containing an amino acid sequence selected from the group consisting of SEQ ID NOs: 21 and 22; or (ii) a VH containing an amino acid sequence selected from the group consisting of SEQ ID NOs: 48 and 19; and / or a VL containing an amino acid sequence selected from the group consisting of SEQ ID NOs: 59 and 60.

[0248] In a particular embodiment, the first domain that specifically binds to IL-17A includes (i) the VH sequence of SEQ ID NO: 10 and the VL sequence of SEQ ID NO: 21, or (ii) the VH sequence of SEQ ID NO: 48 and the VL sequence of SEQ ID NO: 59. In yet another particular embodiment, the first domain that specifically binds to IL-17A includes (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.

[0249] In one embodiment, the domain that specifically binds to human IL-17A is the domain listed in Table 1. In one embodiment, the domain that specifically binds to human IL-17A includes 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 (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 the one described in (i) SEQ ID NOs. 24 or 25, preferably SEQ ID NO. 24, or (ii) SEQ ID NOs. 61 or 62, preferably SEQ ID NO. 61.

[0250] Other domains of the present disclosure having binding specificity to human IL-17A include domains in which the amino acids or nucleic acids encoding the amino acids are mutated, but which have at least 60, 70, 80, 90, or 95% identity with the sequences listed in Table 1. In one embodiment, this includes a mutant amino acid sequence in which one, two, three, four, or five or fewer amino acids are mutated in the variable region compared to the variable region shown in the sequences listed in Table 1, but substantially the same activity is retained. As used herein, the term “substantially identical activity” refers to activity represented by substantially identical activity, which is at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 98%, or at least 100%, at least 110%, at least 120%, at least 130%, at least 140%, at least 150%, at least 160%, at least 170%, at least 180%, or at least 190%, for example up to 200%, of the activity determined for the multispecific antibody of this disclosure, e.g., the multispecific antibody of this disclosure, particularly comprising a first domain that specifically binds to human IL-17A as described in Table 1, and / or a second domain that specifically binds to human TNFα as described in Table 1.

[0251] 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, the first domain binds to human IL-17A and retains the desirable functional properties of the domain described in Table 1.

[0252] In one embodiment, the domain of the Disclosure 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 LDCR1, LDCR2, and LDCR3 sequences, wherein one or more of these CDR sequences have a specific amino acid sequence based on the domain described herein or its conserved modifications, and the domain retains the desired functional properties of the antibody of the Disclosure.

[0253] The terms “conservatively modified variant” or “conservative variant” apply to both amino acid sequences and nucleic acid sequences. With respect to a particular nucleic acid sequence, a conservatively modified variant refers to a nucleic acid that codes for the same or essentially the same amino acid sequence, or does not code for the amino acid sequence to an essentially identical sequence. Due to the degeneracy of the genetic code, a large number of functionally identical nucleic acids code for any given protein. For example, the codons GCA, GCC, GCG, and GCU all code for the amino acid alanine. Therefore, at all positions where alanine is specified by the codon, the codon can be changed to any of the corresponding codons listed without altering the coded polypeptide. Such nucleic acid variants are “silent variants,” which are a type of conservatively modified variant. All nucleic acid sequences herein that code for polypeptides also describe all possible silent variants of the nucleic acid. Those skilled in the art will recognize that each codon of a nucleic acid (except AUG, usually the sole codon for methionine, and TGG, usually the sole codon for tryptophan) can be modified to produce functionally identical molecules. Thus, each silent variant of a nucleic acid that codes for a polypeptide is implied in each listed sequence.

[0254] In the case of polypeptide sequences, a “conservatively modified variant” or “conservative variant” includes individual substitutions, deletions, or additions to a polypeptide sequence that result in the substitution of an amino acid by a chemically similar amino acid. Tables of conservative substitutions that provide functionally similar amino acids are well known in the art. Such conservatively modified variants (i.e., having one or more “conservative modifiers”) are added to, and not excluded from, the polymorphic variants, interspecific homologs, and alleles of this disclosure. The following eight groups include amino acids that are conservedly 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, for example, Creighton, Proteins (1984)). In one embodiment, the term “conservative sequence modification” is used to refer to amino acid modifications that do not significantly affect or alter the antibody binding properties, including the amino acid sequence.

[0255] Accordingly, this 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 comprises):

[0256] (i) A heavy chain variable region (VH) comprising, in order, three complementarity-determining regions HCDR1, HCDR2, and HCDR3, wherein HCDR1 is amino acid sequence SEQ ID NO: 1 or a conserved variant thereof; HCDR2 is amino acid sequence SEQ ID NO: 2 or a conserved variant thereof; HCDR3 is an amino acid sequence selected from any one of SEQ ID NO: 3 or a conserved variant thereof; and A light chain variable region (VL) comprising, in order, three complementarity-determining regions LCDR1, LCDR2, and LCDR3, wherein LCDR1 is amino acid sequence number 12 or a conserved variant thereof; LCDR2 is amino acid sequence number 13 or a conserved variant thereof; LCDR3 has amino acid sequence number 14 or a conserved variant thereof; or (ii) A heavy chain variable region (VH) comprising, in order, three complementarity-determining regions HCDR1, HCDR2, and HCDR3, wherein HCDR1 is amino acid sequence SEQ ID NO: 39 or a conserved variant thereof; HCDR2 is amino acid sequence SEQ ID NO: 40 or a conserved variant thereof; HCDR3 is an amino acid sequence selected from any one of SEQ ID NO: 41 or a conserved variant thereof; and A light chain variable region (VL) comprising, in order, three complementarity-determining regions LCDR1, LCDR2, and LCDR3, wherein LCDR1 is amino acid sequence number 50 or a conserved variant thereof; LCDR2 is amino acid sequence number 51 or a conserved variant thereof; and LCDR3 has amino acid sequence number 52 or a conserved variant thereof. Here, the antibody specifically binds to human IL-17A and / or neutralizes IL-17A.

[0257] The anti-IL-17A antibody disclosed herein This disclosure is based on the discovery of antibody molecules that specifically bind to human IL-17A and possess improved affinity, efficacy, and selectivity. Furthermore, the antibodies of this disclosure have improved biophysical properties, such as improved solubility, developability, relatively low impurity and high productivity (>98%, specifically >99% monomer when detected by SE-HPLC), and stability.

[0258] In one embodiment, the present disclosure provides isolated antibodies having binding specificity to human IL-17A and comprising a set of CDRs (HCDR1, HCDR2, HCDR3, LDCR1, LDCR2, and LDCR3), wherein the set of CDRs is a set of CDRs [where, (i) HCDR1' is an amino acid sequence selected from any one of SEQ ID NOs: 1, 4, and 7, preferably from SEQ ID NO: 1; HCDR2' is an amino acid sequence selected from any one of SEQ ID NOs: 2, 5, and 8, preferably from SEQ ID NO: 2; HCDR3' is an amino acid sequence selected from any one of SEQ ID NOs: 3, 6, and 9, preferably from SEQ ID NO: 3; LCDR1' is an amino acid sequence selected from any one of SEQ ID NOs: 12, 15, and 18, preferably from SEQ ID NO: 12; LCDR2' is an amino acid sequence selected from any one of SEQ ID NOs: 13, 16, and 19, preferably from SEQ ID NO: 13; LCDR3' has an amino acid sequence selected from any one of SEQ ID NOs: 14, 17, and 20, preferably from SEQ ID NO: 14; or (ii) HCDR1' is an amino acid sequence selected from any one of SEQ ID NOs: 39, 42, and 45, preferably from SEQ ID NO: 39; HCDR2' is an amino acid sequence selected from any one of SEQ ID NOs: 40, 43, and 46, preferably from SEQ ID NO: 40; HCDR3' is an amino acid sequence selected from any one of SEQ ID NOs: 41, 44, and 47, preferably from SEQ ID NO: 40; LCDR1' is an amino acid sequence selected from any one of SEQ ID NOs: 50, 53, and 56, preferably from SEQ ID NO: 50; LCDR2' is an amino acid sequence selected from SEQ ID NOs: 51, 54 The amino acid sequence is one of , and 57, preferably selected from SEQ ID NO: 51; and LCDR3' has 10 or fewer amino acid substitutions, for example, 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.

[0259] The term "IL-17A" or "IL17A" specifically refers to human IL-17A having UniProt ID number Q16552, which is reproduced herein as Sequence ID No. 33. The term "cynomolgus IL-17A" or "cynomolgus monkey IL-17A" refers to cynomolgus monkey (Macaca fascicularis) IL-17A having UniProt ID number G1QUS7.

[0260] The term "IL-17B" specifically refers to a human IL-17B having UniProt ID number Q9UHF5, reproduced herein as Sequence ID No. 34. The term "IL-17C" specifically refers to a human IL-17C having UniProt ID number Q9P0M4, reproduced herein as Sequence ID No. 35. The term "IL-17D" specifically refers to a human IL-17D having UniProt ID number Q8TAD2, reproduced herein as Sequence ID No. 36. The term "IL-17E" specifically refers to a human IL-17E having UniProt ID number Q9H293, reproduced herein as Sequence ID No. 37. The term "IL-17F" specifically refers to a human IL-17F having UniProt ID number Q96PD4, reproduced herein as Sequence ID No. 38.

[0261] The term "epitope" refers to a local region of an antigen to which an antibody can specifically bind. An epitope can be, for example, an adjacent amino acid of a polypeptide, or it can be formed by combining, for example, two or more non-adjacent regions of more than one polypeptide.

[0262] As used herein, terms such as “antibody” include whole antibodies; any antigen-binding fragment (i.e., “antigen-binding portion”) of a whole antibody or its single chain; and molecules containing antibody CDRs, VH regions, or VL regions (including, but not limited to, multispecific antibodies). Naturally occurring “whole antibodies” are glycoproteins containing 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 called CL. The VH and VL regions can be further subdivided into hypervariable regions called complementarity-determining regions (CDRs), and interspersed with more conserved regions called framework regions (FRs). Each VH and VL consists of three CDRs and four FRs, arranged in the order FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4 from the amino terminus to the carboxyl terminus. The variable regions of the heavy and light chains contain binding domains that interact with the antigen. The constant region of the 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.

[0263] As used herein, the term “isotype” refers to an antibody class provided by a heavy chain constant region gene (e.g., IgM, IgE, IgD, IgA, IgY, and IgG such as IgG1 or IgG4). Isotypes also include modified versions of one of these classes, which are modified to alter Fc function, for example, to enhance or reduce effector function or binding to the Fc receptor. Preferably, the antibodies of this disclosure are IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4. More preferably, the antibodies of this disclosure are IgG1 or IgG4.

[0264] As used herein, terms such as “antigen-binding fragment” and “antigen-binding moiety” 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 included in the term “antigen-binding moiety” of an antibody include: Fab fragments, which are monovalent fragments consisting of VL, VH, CL, and CH1 domains; F(ab)2 fragments, which are bivalent fragments containing two Fab fragments linked by disulfide crosslinking at a hinge region; Fd fragments, which consist of a VH domain and a CH1 domain; Fv fragments, which consist of the VL and VH domains of a single arm of the antibody; and single-domain antibody (dAb) fragments, which consist of a VH domain (Ward et al., 1989 Nature). 341:544-546); including isolated complementarity-determining regions (CDRs), dsFv, scAb, STAB, single-domain antibodies (sdAb or dAb), single-domain heavy-chain antibodies, and single-domain light-chain antibodies, VHH, VNAR, single-domain antibodies based on shark VNAR structures, and, but not limited to, ankyrin-based domains, finomers, avimers, anticarin, fibronectin-based binding domains, and binding sites incorporated into the constant region of antibodies (e.g., F-star's Modular Antibody Technology®).

[0265] The term “complementarity-determining region” (“CDR”) refers to an amino acid sequence with boundaries determined using one of several well-known schemes, e.g., 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., Dev. Comp. Immunol., 27, 55-77 (2003) (“IMGT” numbering scheme), and Honegger & Pluckthun, J. Mol. Biol. 309 (2001) This includes the numbering scheme described in 657-670 ("AHo" numbering). For example, in the classical form, in Kabat, the CDR amino acid residues of the heavy chain variable domain (VH) are numbered 31-35 (HCDR1), 50-65 (HCDR2), and 95-102 (HCDR3), and the CDR amino acid residues of the light chain variable domain (VL) are numbered 24-34 (LCDR1), 50-56 (LCDR2), and 89-97 (LCDR3). In Chothia, the CDR amino acids of VH are numbered 26-32 (HCDR1), 52-56 (HCDR2), and 95-102 (HCDR3), and the amino acid residues of VL are numbered 24-34 (LCDR1), 50-56 (LCDR2), and 89-97 (LCDR3).Combining the CRR definitions of both Kabat and Chothia, the CDR consists of amino acid residues 26-35 (HCDR1), 50-65 (HCDR2), and 95-102 (HCDR3) in human VH, and 24-34 (LCDR1), 50-56 (LCDR2), and 89-97 (LCDR3) in human VL. In IMGT, the CDR amino acid residues of VH are numbered approximately 26-35 (HCDR1), 51-57 (HCDR2), and 93-102 (HCDR3), and the CDR amino acid residues of VL are numbered approximately 27-32 (LCDR1), 50-52 (LCDR2), and 89-97 (LCDR3) (numbering by "Kabat"). In IMGT, the CDR of an antibody can be determined using the program IMGT / DomainGap Align.

[0266] In the context of this invention, unless otherwise specified, the numbering system proposed by Honegger & Pluuckthun ("AHo") is used (Honegger & Pluuckthun, J. Mol. Biol. 309 (2001) 657-670). Furthermore, the following residues are defined as CDRs according to the AHo numbering scheme: LCDR1 (also called CDR-L1): L24~L42; LCDR2 (also called CDR-L2): L58~L72; LCDR3 (also called CDR-L3): L107~L138; HCDR1 (also called CDR-H1): H27~H42; HCDR2 (also called CDR-H2): H57~H76; HCDR3 (also called CDR-H3): H108~H138. To clarify, the numbering system by Honegger & Pluuckthun takes into account the length diversity found in antibodies naturally present in both the various VH and VL subfamilies, particularly CDR, providing gaps in the sequence. Therefore, in a given antibody variable domain, not all positions from 1 to 149 are typically occupied by amino acid residues.

[0267] Preferably, the "antigen-binding region" comprises at least amino acid residues 4-138 of the variable light chain (VL) and amino acid residues 5-138 of the variable heavy chain (VH) (in either case, numbered by Honegger & Pluuchthun), more preferably amino acid residues 3-144 of VL and 4-144 of VH, and particularly preferably the complete VL and VH chains (amino acid positions 1-149 of VL and 1-149 of VH). The antigen-binding region can also be incorporated into single-domain antibodies, maxi-bodies, mini-bodies, intra-bodies, dia-bodies, tria-bodies, tetra-bodies, v-NARs, and bis-scFvs (see, for example, Holliger and Hudson, 2005, Nature Biotechnology, 23, 9, 1 Hel l 36). The antigen-binding portion of an antibody can be transplanted onto a polypeptide-based scaffold, such as fibronectin type III (Fn3) (see U.S. Patent No. 6,703,199, which describes a fibronectin polypeptide monobody). The antigen-binding portion can be incorporated into a single-chain molecule containing a pair of tandem Fv segments (VH-CH1-VH-CH1) that, together with a complementary light chain polypeptide, form a pair of antigen-binding domains (Zapata et al., 1995 Protein Eng. 8 (10): 1057-1062; and U.S. Patent No. 5,641,870).

[0268] As used herein, terms such as “domain,” “domain that specifically binds to X,” “binding domain,” “antigen-binding fragment,” and “antigen-binding moiety” of an antibody refer to one or more fragments of the whole intact antibody that retain the ability to specifically bind to a given antigen (e.g., IL-17A, TNFα, HSA). The antigen-binding function of the antibody is performed by fragments of the intact antibody. In some embodiments, the binding domain of the multispecific antibody of this disclosure is a Fab fragment, which is a monovalent fragment consisting of VL, VH, CL, and CH1 domains; an F(ab)2 fragment, which is a bivalent fragment containing two Fab fragments linked by disulfide crosslinking at a hinge region; an Fd fragment consisting of a VH domain and a CH1 domain; an Fv fragment consisting of the VL and VH domains of a single arm of the antibody; and a single-domain antibody (dAb) fragment consisting of a VH domain (Ward et al., 1989 Nature). 341:544-546); including isolated complementarity-determining regions (CDRs), dsFv, scAb, STAB, single-domain antibodies (sdAb or dAb), single-domain heavy-chain antibodies, and single-domain light-chain antibodies, VHH, VNAR, single-domain antibodies based on shark VNAR structures, and, but not limited to, ankyrin-based domains, finomers, avimers, anticarin, fibronectin-based alternative scaffold-based binding domains, and binding sites incorporated into the constant region of antibodies (e.g., F-star's Modular Antibody Technology®). Preferably, the binding domain of the Disclosure is an Fv fragment (Fv). Preferably, the binding domain of the Disclosure is a single-stranded Fv fragment (scFv). Preferably, the binding domain of the Disclosure is a Fab fragment.

[0269] Preferably, the “domain” or “domain that specifically binds to X” or “binding domain,” its “antigen-binding fragment,” or “antigen-binding moiety” comprises at least amino acid residues 4-138 of the variable light chain (VL) and amino acid residues 5-138 of the variable heavy chain (VH) (in either case, numbered by Honegger & Pluckthun), more preferably amino acid residues 3-144 of the VL and amino acid residues 4-144 of the VH, and particularly preferably the complete VL and VH chains (amino acid positions 1-149 of the VL and amino acid positions 1-149 of the VH). The antigen-binding moiety can also be incorporated into single-domain antibodies, maxibodies, minibodies, intrabodies, diabodies, triabodies, tetrabodies, v-NARs, and bis-scFvs (see, for example, Holliger and Hudson, 2005, Nature Biotechnology, 23, 9, 1 Hel l 36). The antigen-binding portion of an antibody can be transplanted onto a polypeptide-based scaffold, such as fibronectin type III (Fn3) (see U.S. Patent No. 6,703,199, which describes fibronectin polypeptide monobodies). The antigen-binding portion can be incorporated into a single-chain molecule containing a pair of tandem Fv segments (VH-CH1-VH-CH1) that, together with a complementary light chain polypeptide, form a pair of antigen-binding domains (Zapata et al., 1995 Protein Eng. 8 (10): 1057-1062; and U.S. Pat. No. 5,641,870; and U.S. Patent No. 5,641,870).

[0270] As used herein, the terms “binding specificity” or “specifically binding” refer to the ability of an individual antibody or antibody domain to react with one antigenic determinant rather than with different antigenic determinants. As used herein, the terms “specifically binding to” or “specific to” refer to a measurable and reproducible interaction, such as the binding of an antibody or antibody domain to a target, which determines the presence of the target in the presence of a heterogeneous molecular 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 binds to this target with higher affinity, binding strength, more readily, and / or for a longer duration than it would to bind to other targets. In its most common form (and unless a defined reference is cited), “specific binding” refers to the ability of an antibody or antibody domain to distinguish a target of interest from an unrelated molecule, as determined, for example, by specificity assay methods known in the art. Such methods include, but are not limited to, Western blotting, ELISA, RIA, ECL, IRMA, SPR (surface plasmon resonance) assays, and peptide scans. For example, a standard ELISA assay can be performed. Scoring can be performed by standard colorimetric assays (e.g., secondary antibodies using horseradish peroxide, and tetramethylbenzidine using hydrogen peroxide). Reactions in specific wells are scored by optical density at, for example, 450 nm. A typical background (=negative reaction) is approximately 0.1 OD. A typical positive reaction may be approximately 1 OD. This means that the ratio of positive to negative scores can be more than 10-fold. In further examples, an SPR assay can be performed, where a difference of at least 10-fold, preferably at least 100-fold, between the background and the signal indicates specific binding. Typically, the determination of binding specificity is performed using a set of about 3-5 unrelated molecules, such as milk powder or transferrin, rather than a single reference molecule. The specific antibodies or antibody domains of this disclosure have binding specificity to human IL-17A or human TNFα.

[0271] The multispecific antibodies of this disclosure comprise a first domain that specifically binds to IL-17A and a second domain that specifically binds to TNFα, and thus have binding specificity to IL-17A and TNFα, and in particular binding specificity to human IL-17A and human TNFα. In one embodiment, the antibody of this disclosure has binding specificity to human IL-17A and cynomolgus monkey (Macaca fascicularis) (also known as Cynomolgus monkey or "Cynomolgus") IL-17A. In one embodiment, the antibody of this disclosure has binding specificity to human TNFα and cynomolgus monkey (Macaca fascicularis) (also known as Cynomolgus monkey or "Cynomolgus") TNFα.

[0272] In another embodiment, the present disclosure relates to an antibody or antibody domain having binding specificity to human IL-17A and to cynomolgus monkey (Macaca fascicularis) (also known as Cynomolgus monkey or "Cynomolgus") IL-17A.

[0273] More precisely, the anti-IL-17A antibody of this disclosure is an isolated antibody.

[0274] Appropriately, the anti-IL-17A antibody of this disclosure is a monoclonal antibody.

[0275] The anti-IL-17A antibodies of this disclosure include, but are not limited to, chimeric antibodies and humanized antibodies.

[0276] Appropriately, the anti-IL-17A antibody of this disclosure is humanized. Appropriately, the anti-IL-17A antibody of this disclosure is humanized and contains rabbit-derived CDR.

[0277] The antibodies of this disclosure include, but are not limited to, isolated humanized monoclonal antibodies described herein, including those in the examples. Examples of such anti-human IL-17A antibodies are those whose sequences are listed in Table 1. Additional details regarding the production and characterization of the antibodies described herein are provided in the examples.

[0278] The isolated antibody of this disclosure having binding specificity to human IL-17A comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein (a) the VH comprises, in order, three complementarity-determining regions HCDR1, HCDR2, and HCDR3, and (b) the VL comprises, in order, three complementarity-determining regions LDCR1, LDCR2, and LDCR3.

[0279] This disclosure provides an antibody that specifically binds to the IL-17A protein, wherein the antibody comprises a VH CDR having one of the amino acid sequences of the VH CDRs listed in Table 1. In particular, this disclosure provides an antibody that specifically binds to the IL-17A protein, wherein the antibody comprises one, two, three or more VH CDRs having one of the amino acid sequences of the VH CDRs listed in Table 1.

[0280] This disclosure provides an antibody having binding specificity to human IL-17A, comprising a heavy chain variable region (VH), wherein the VH comprises, in order: (i) three complementarity-determining regions HCDR1, HCDR2, and HCDR3, wherein HCDR1 has an amino acid sequence selected from any one of SEQ ID NOs: 1, 4, and 7; 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. In particular, the present disclosure provides an antibody having binding specificity to human IL-17A and comprising the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs. 1, 2, and 3, respectively; or (ii) three complementarity-determining regions HCDR1, HCDR2, and HCDR3, wherein HCDR1 has an amino acid sequence selected from any one of SEQ ID NOs. 39, 42, and 45; HCDR2 has an amino acid sequence selected from any one of SEQ ID NOs. 40, 43, and 46; and 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 having binding specificity to human IL-17A and comprising the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs. 39, 40, and 41, respectively.

[0281] This disclosure also provides an antibody that specifically binds to the IL-17A protein, wherein the antibody comprises a VL CDR having one of the amino acid sequences of the VL CDRs listed in Table 1. In particular, this disclosure provides an antibody that specifically binds to the IL-17A protein, wherein the antibody comprises one, two, three or more VL CDRs having one of the amino acid sequences of the VL CDRs listed in Table 1.

[0282] This disclosure provides an antibody having binding specificity to human IL-17A, comprising a light chain variable region (VL), wherein the VL comprises, in order: (i) three complementarity-determining regions LDCR1, LDCR2, and LDCR3, wherein LDCR1 has an amino acid sequence selected from any one of SEQ ID NOs: 12, 15, and 18; LDCR2 has an amino acid sequence selected from any one of SEQ ID NOs: 13, 16, and 19; and LDCR3 has 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 the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs. 12, 13, and 14, respectively; or (ii) three complementarity-determining regions LCDR1, LCDR2, and LCDR3, wherein LCDR1 has an amino acid sequence selected from any one of SEQ ID NOs. 50, 53, and 56, LCDR2 has an amino acid sequence selected from any one of SEQ ID NOs. 51, 54, and 57, and LDCR3 has an amino acid sequence selected from any one of SEQ ID NOs. In particular, the present disclosure provides an antibody having binding specificity to human IL-17A and comprising the LDCR1, LDCR2, and LDCR3 sequences of SEQ ID NOs. 50, 51, and 52, respectively.

[0283] Appropriately, this disclosure provides an antibody having binding specificity to human IL-17A, comprising a heavy chain variable region (VH) and a light chain variable region (VL).

[0284] (i) Here (a) The VH comprises, in order, three complementarity-determining regions HCDR1, HCDR2, and HCDR3, wherein HCDR1 has an amino acid sequence selected from any one of SEQ ID NOs: 1, 4, and 7; 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 (b) The VL comprises, in order, three complementarity-determining regions LCDR1, LCDR2, and LCDR3, wherein LCDR1 has an amino acid sequence selected from any one of SEQ ID NOs: 12, 15, and 18; LCDR2 has an amino acid sequence selected from any one of SEQ ID NOs: 13, 16, and 19; and LCDR3 has an amino acid sequence selected from any one of SEQ ID NOs: 14, 17, and 20; or

[0285] (ii) Here (a) The VH comprises, in order, three complementarity-determining regions HCDR1, HCDR2, and HCDR3, wherein HCDR1 has an amino acid sequence selected from any one of SEQ ID NOs. 39, 42, and 45; HCDR2 has an amino acid sequence selected from any one of SEQ ID NOs. 40, 43, and 46; and HCDR3 has an amino acid sequence selected from any one of SEQ ID NOs. 41, 44, and 47; and (b) The VL comprises, in order, three complementarity-determining regions LCDR1, LCDR2, and LCDR3, wherein LCDR1 has an amino acid sequence selected from any one of SEQ ID NOs. 50, 53, and 56; LCDR2 has an amino acid sequence selected from any one of SEQ ID NOs. 51, 54, and 57; and LCDR3 has an amino acid sequence selected from any one of SEQ ID NOs. 52, 55, and 58.

[0286] In particular, the present disclosure provides antibodies having binding specificity to human IL-17A and comprising (i) (a) the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs. 1, 2, and 3, respectively, and (b) the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs. 12, 13, and 14, respectively; or (ii) (a) the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs. 39, 40, and 41, respectively, and (b) the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs. 50, 51, and 52, respectively.

[0287] Other antibodies in this disclosure include mutated amino acids in their CDR region that have at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identity with the CDR region shown in the sequences listed in Table 1. Preferably, other antibodies in this disclosure include mutated amino acid sequences in which, compared to the CDR region shown in the sequences listed in Table 1, 1, 2, 3, 4, 5, or 10 or fewer amino acids are mutated by amino acid deletion, insertion, or substitution in the CDR region. Mutations, such as substitutions, may occur at any residue within the set of CDRs, and may be within CDR1, CDR2, and / or CDR3.

[0288] Suitablely, an antibody of the present disclosure comprising a mutant amino acid sequence can inhibit the activity of 1 ng of human IL-17A by 50% at a concentration of 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, and even more preferably 0.2 ng / ml or less, the inhibitory activity of which is determined by measuring human IL-17A-induced GRO-α secretion in an HT-29 assay in the presence of 50 pg / ml TNFα.

[0289] Appropriately, an isolated antibody of this disclosure having binding specificity to human IL-17A comprises a heavy chain variable region (VH) and a light chain variable region (VL), where,

[0290] (i) (a) The VH includes, in order, three complementarity determination regions HCDR1, HCDR2, and HCDR3, 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: 1, 4, and 7; 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: 2, 5, and 8; The HCDR3 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to any one of SEQ ID NOs: 3, 6, and 9, preferably SEQ ID NO: 3; and / or (b) The VL includes, in order, three complementarity determination regions LCDR1, LCDR2, and LCDR3, 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 sequence numbers 12, 15, and 18; 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; The LCDR3 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to any one of SEQ ID NOs: 14, 17, and 20; or

[0291] (ii) (a) The VH includes, in order, three complementarity determination regions HCDR1, HCDR2, and HCDR3, 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; 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, 40, 43, and 46; The HCDR3 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to any one of SEQ ID NOs: 41, 44, and 47; and / or (b) The VL includes, in order, three complementarity determination regions LCDR1, LCDR2, and LCDR3, 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 sequence numbers 50, 53, and 56; 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 sequence numbers 51, 54, and 57; 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 sequence numbers 52, 52, 55, 58, 52

[0292] Appropriately, isolated antibodies of the present disclosure having binding specificity to human IL-17A include: (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 NOs. 1, 2, and 3, respectively; LCDR1, LCDR2, and LCDR3; 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 sequence numbers 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 sequence numbers 50, 51, and 52, respectively.

[0293] 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.

[0294] Suitablely, this 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 this disclosure comprises a VH4 domain. In a preferred embodiment, the isolated antibody of this disclosure comprises a VH3 domain.

[0295] Suitablely, the present disclosure provides isolated antibodies that specifically bind to human IL-17A, wherein the antibody comprises Vκ framework FR1, FR2, and FR3, more particularly Vκ1 or Vκ3 FR1-FR3, preferably Vκ1 framework FR1-FR3, and framework FR4, wherein framework FR4 is selected from Vκ FR4, more particularly Vκ1 FR4, Vκ3 FR4, and Vλ FR4. Suitable Vλ FR4 are those described in SEQ ID NOs: 26-32. In one embodiment, the present disclosure provides isolated antibodies that specifically bind to human IL-17A, wherein the antibody comprises Vλ FR4 having at least 60, 70, 80, or 90% identity with an amino acid sequence selected from any of SEQ ID NOs: 26-32, preferably SEQ ID NOs: 26 or 27, more preferably SEQ ID NOs: 27. Appropriately, this disclosure provides an isolated antibody that specifically binds to human IL-17A, wherein the antibody comprises a Vλ FR4 having an amino acid sequence selected from any of SEQ ID NOs. 26 to 32, preferably the Vλ FR4 described in SEQ ID NOs. 26 or 27, more preferably the Vλ FR4 described in SEQ ID NOs. 27.

[0296] Therefore, in one embodiment, this disclosure provides an antibody comprising:

[0297] (i)(a) the HCDR1, HCDR2, and HCDR3 sequences of sequence numbers 1, 2, and 3, respectively, and the LCDR1, LCDR2, and LCDR3 sequences of sequence numbers 12, 13, and 14, respectively; or (i)(b) the HCDR1, HCDR2, and HCDR3 sequences of sequence numbers 39, 40, and 41, respectively, and the LCDR1, LCDR2, and LCDR3 sequences of sequence numbers 50, 51, and 52, respectively; (ii) VH3 or VH4 domain, preferably VH3 domain; and (iii) A VL domain comprising a VL framework comprising Vκ framework FR1, FR2, and FR3, more particularly Vκ1 or Vκ3 FR1 to FR3, preferably Vκ1 FR1 to FR3, and framework FR4, wherein framework FR4 is selected from Vκ FR4, more particularly Vκ1 FR4, Vκ3 FR4, and Vλ FR4, more particularly Vλ FR4 comprising an amino acid sequence having at least 60, 70, 80, or 90% identity with an amino acid sequence selected from any of SEQ ID NOs. 26 to 32, preferably Vλ FR4 described in an amino acid sequence selected from any of SEQ ID NOs. 26 to 32, more preferably Vλ FR4 described in SEQ ID NO. 27.

[0298] Appropriately, this disclosure provides isolated antibodies that specifically bind to human IL-17A, wherein the antibodies include the VH domains listed in Table 1.

[0299] Appropriately, the Disclosure also provides an isolated antibody that specifically binds to human IL-17A, wherein the antibody comprises the VH amino acid sequence listed in Table 1, wherein approximately 10 or fewer amino acids in the framework sequence (e.g., a sequence other than the CDR sequence) are mutated (mutations are additions, substitutions, or deletions, to name a variety of non-limiting examples).

[0300] Appropriately, the Disclosure also provides isolated antibodies that specifically bind to human IL-17A, wherein the antibodies comprise the VH amino acid sequence described in Table 1, wherein approximately 20 or fewer amino acids in the framework sequence (e.g., a sequence other than the CDR sequence) are mutated (mutations are additions, substitutions, or deletions, to name a variety of non-limiting examples).

[0301] Other antibodies in this disclosure are mutated but contain the VH region shown in the sequences listed in Table 1 and amino acids that have at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identity within the VH region.

[0302] Appropriately, this disclosure provides isolated antibodies that specifically bind to human IL-17A, wherein the antibodies include the VL domains listed in Table 1.

[0303] Appropriately, the Disclosure also provides an isolated antibody that specifically binds to human IL-17A, wherein the antibody comprises the VL amino acid sequence listed in Table 1, wherein approximately 10 or fewer amino acids in the framework sequence (e.g., a sequence that is not a CDR sequence) are mutated (mutations are additions, substitutions, or deletions, to name a variety of non-limiting examples).

[0304] Appropriately, the Disclosure also provides isolated antibodies that specifically bind to human IL-17A, wherein the antibodies comprise the VL amino acid sequence listed in Table 1, wherein approximately 20 or fewer amino acids in the framework sequence (e.g., a sequence that is not a CDR sequence) are mutated (mutations are additions, substitutions, or deletions, to name a variety of non-limiting examples).

[0305] Other antibodies in this disclosure include mutated but VL regions shown in the sequences listed in Table 1, and amino acids in the VL region that have at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identity.

[0306] In one embodiment, the disclosure provides an isolated antibody that specifically binds to human IL-17A, wherein the antibody comprises a heavy chain variable region containing 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 amino acid sequence SEQ ID NO: 10 or SEQ ID NO: 11, and in particular the antibody comprises the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs: 1, 2, and 3, respectively. In a further embodiment, the disclosure provides an isolated antibody that specifically binds to human IL-17A, wherein the antibody comprises a heavy chain variable region containing an amino acid sequence that is at least 90% identical to amino acid sequence SEQ ID NO: 11, wherein the heavy chain variable region comprises Q14K, G16E, and G56A (AHo numbering).

[0307] 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 containing 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 amino acid sequence SEQ ID NO: 48, and in particular the antibody comprises the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs: 39, 40, and 41, respectively. In a further embodiment, the disclosure provides an isolated antibody that specifically binds to human IL-17A, wherein the antibody comprises a heavy chain variable region containing an amino acid sequence that is at least 90% identical to amino acid sequence SEQ ID NO: 49, wherein the heavy chain variable region comprises R20T and Q141P (AHo numbering).

[0308] In another embodiment, the Disclosure provides an isolated antibody that specifically binds to human IL-17A, wherein the antibody comprises a light chain variable region containing 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 (i) the amino acid sequence SEQ ID NO: 21 or SEQ ID NO: 22, preferably SEQ ID NO: 21, and in particular the antibody comprises SEQ ID NOs: 12, 13, and 14, respectively. (ii) comprising the LCDR1, LCDR2, and LCDR3 sequences; or (ii) comprising a light chain variable region comprising an amino acid sequence that is identical to 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 sequence number 59 or sequence number 60, preferably sequence number 59, and in particular the antibody comprising the LCDR1, LCDR2, and LCDR3 sequences of sequence numbers 50, 51, and 52, respectively.

[0309] In a further embodiment, the present disclosure provides an isolated antibody that specifically binds to human IL-17A, wherein the antibody comprises a light chain variable region having an amino acid sequence that is at least 90% identical to amino acid sequence SEQ ID NO: 22, wherein the light chain variable region comprises A51P (AHo numbering).

[0310] In a further embodiment, the present disclosure provides an antibody that specifically binds to human IL-17A, wherein (i) 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 amino acid sequence SEQ ID NO: 10, and 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 amino acid sequence SEQ ID NO: 21. (ii) The antibody comprises a light chain variable region containing a certain amino acid sequence; or (ii) the antibody comprises a heavy chain variable region containing an amino acid sequence that is identical to amino acid sequence SEQ ID NO: 48 by at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90%, and a light chain variable region containing an amino acid sequence that is identical to amino acid sequence SEQ ID NO: 59 by at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90%.

[0311] Therefore, this disclosure provides an isolated antibody that specifically binds to human IL-17A, and here, (i) The antibody comprises a heavy chain variable region containing an amino acid sequence that is identical to amino acid sequence SEQ ID NO: 10 by at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90%, and a light chain variable region containing an amino acid sequence that is identical to amino acid sequence SEQ ID NO: 21 by at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90%. The region includes, where the antibody comprises the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs. 1, 2, and 3, and / or the LDCR1, LDCR2, and LDCR3 sequences of SEQ ID NOs. 12, 13, and 14, respectively; in particular, the antibody comprises the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs. 1, 2, and 3, respectively, and the LDCR1, LDCR2, and LDCR3 sequences of SEQ ID NOs. 12, 13, and 14, respectively; or, (ii) The antibody comprises a heavy chain variable region containing an amino acid sequence that is identical to amino acid sequence SEQ ID NO: 48 by at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90%, and a light chain variable region containing an amino acid sequence that is identical to amino acid sequence SEQ ID NO: 59 by at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90%. The region includes, where the antibody comprises the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs. 39, 40, and 41, and / or the LDCR1, LDCR2, and LDCR3 sequences of SEQ ID NOs. 50, 51, and 52, respectively; in particular, the antibody comprises the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs. 39, 40, and 41, respectively, and the LDCR1, LDCR2, and LDCR3 sequences of SEQ ID NOs. 50, 51, and 52, respectively.

[0312] In a further embodiment, the present disclosure provides an isolated antibody that specifically binds to human IL-17A, wherein the antibody comprises a heavy chain variable region containing 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 amino acid sequence SEQ ID NO: 49, and a light chain variable region containing 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 amino acid sequence SEQ ID NO: 60, wherein the heavy chain variable region particularly includes R20T and Q141P (AHo numbering).

[0313] Accordingly, this disclosure provides an isolated antibody that specifically binds to human IL-17A, wherein the antibody comprises a heavy chain variable region containing 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 amino acid sequence SEQ ID NO: 49, and a light chain variable region containing 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 amino acid sequence SEQ ID NO: 60. In particular, the heavy chain variable region includes R20T and Q141P (AHo numbering), where the antibody includes the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs. 39, 40, and 41, and / or the LDCR1, LDCR2, and LDCR3 sequences of SEQ ID NOs. 50, 51, and 52, respectively. In particular, where the antibody includes the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs. 39, 40, and 41, respectively, and the LDCR1, LDCR2, and LDCR3 sequences of SEQ ID NOs. 50, 51, and 52, respectively.

[0314] In certain embodiments, the Disclosure provides isolated antibodies that specifically bind to human IL-17A and comprise (i) a VH containing an amino acid sequence selected from the group consisting of SEQ ID NOs: 10 and 11, and / or its VL containing an amino acid sequence selected from the group consisting of SEQ ID NOs: 21 and 22; or (ii) a VH containing an amino acid sequence selected from the group consisting of SEQ ID NOs: 48 and 49, and / or its VL containing an amino acid sequence selected from the group consisting of SEQ ID NOs: 59 and 60. In certain embodiments, the antibody of the Disclosure comprises (i) the VH sequence of SEQ ID NOs: 10 and the VL sequence of SEQ ID NOs: 21; or (ii) the VH sequence of SEQ ID NOs: 48 and the VL sequence of SEQ ID NOs: 59. In yet another particular embodiment, the antibody of the Disclosure comprises (i) the VH sequence of SEQ ID NOs: 11 and the VL sequence of SEQ ID NOs: 22; or (ii) the VH sequence of SEQ ID NOs: 49 and the VL sequence of SEQ ID NOs: 60.

[0315] In one embodiment, the antibody that specifically binds to human IL-17A is the antibody listed in Table 1. In one embodiment, the antibody that specifically binds to human IL-17A includes an amino acid sequence that is identical to at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90%, of an amino acid sequence selected from the group consisting of (i) SEQ ID NOs. 24 and 25, preferably SEQ ID NO. 24, or (ii) SEQ ID NOs. 61 or 62, preferably SEQ ID NO. 61.

[0316] Other antibodies in the present disclosure having binding specificity to human IL-17A include those in which amino acids or nucleic acids encoding amino acids are mutated but have at least 60, 70, 80, 90, or 95% identity with the sequences listed in Table 1. In one embodiment, this includes mutant amino acid sequences in which one, two, three, four, or five or fewer amino acids are mutated in the variable region compared to the variable region shown in the sequences listed in Table 1, but which retain substantially the same activity. As used herein, the term “substantially identical activity” refers to an activity that is substantially identical to the activity determined for the parent antibody, e.g., the antibody of this disclosure, in particular the antibody of this disclosure listed in Table 1, and is at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 98%, or at least 100%, at least 110%, at least 120%, at least 130%, at least 140%, at least 150%, at least 160%, at least 170%, at least 180%, or at least 190%, e.g., up to 200%, of that activity.

[0317] In yet another embodiment, the present disclosure provides an antibody comprising an amino acid sequence homologous to the sequence described in Table 1, wherein the antibody binds to human IL-17A and retains the desired functional properties of the antibody described in Table 1.

[0318] In one embodiment, the antibody of the 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 specific amino acid sequence based on the antibody described herein or a conserved modification thereof, and the antibody retains the desired functional properties of the antibody of the Disclosure.

[0319] Therefore, this disclosure provides a monoclonal antibody comprising: (i) A heavy chain variable region (VH) comprising, in order, three complementarity-determining regions HCDR1, HCDR2, and HCDR3, wherein HCDR1 is amino acid sequence SEQ ID NO: 1 or a conserved variant thereof, HCDR2 is amino acid sequence SEQ ID NO: 2 or a conserved variant thereof, and HCDR3 is an amino acid sequence selected from any one of SEQ ID NO: 3 or a conserved variant thereof; and A light chain variable region (VL) comprising, in order, three complementarity-determining regions LCDR1, LCDR2, and LCDR3, wherein LCDR1 is amino acid sequence number 12 or a conserved variant thereof, LCDR2 is amino acid sequence number 13 or a conserved variant thereof, and LCDR3 is amino acid sequence number 14 or a conserved variant thereof; or (ii) A heavy chain variable region (VH) comprising, in order, three complementarity-determining regions HCDR1, HCDR2, and HCDR3, wherein HCDR1 is amino acid sequence SEQ ID NO: 39 or a conserved variant thereof, HCDR2 is amino acid sequence SEQ ID NO: 40 or a conserved variant thereof, and HCDR3 is an amino acid sequence selected from any one of SEQ ID NO: 41 or a conserved variant thereof; and A light chain variable region (VL) comprising, in order, three complementarity-determining regions LCDR1, LCDR2, and LCDR3, wherein LCDR1 is amino acid sequence number 50 or a conserved variant thereof, LCDR2 is amino acid sequence number 51 or a conserved variant thereof, and LCDR3 is amino acid sequence number 52 or a conserved variant thereof; Here, the antibody specifically binds to human IL-17A and / or neutralizes IL-17A.

[0320] In one embodiment, the antibody of the present disclosure is optimized for expression in mammalian cells and has a heavy chain variable region and a light chain variable region, wherein one or more of these sequences have a specific amino acid sequence based on the antibody described herein, and wherein the antibody retains the desired functional properties of the antibody of the present disclosure. Accordingly, the present disclosure provides a monoclonal antibody optimized for expression in mammalian cells and 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 (i) either SEQ ID NOs: 10 and 11 and their conserved modifiers, or (ii) either SEQ ID NOs: 48 and 49 and their conserved modifiers; the light chain variable region comprises an amino acid sequence selected from (i) either SEQ ID NOs: 21 and 22 and their conserved modifiers, or (ii) either SEQ ID NOs: 59 and 60 and their conserved modifiers; wherein the antibody specifically binds to and / or neutralizes human IL-17A.

[0321] In one embodiment, the antibody of the Disclosure is optimized for expression in mammalian cells and has a full-length heavy chain sequence and a full-length light chain sequence, one or more of which have a specific amino acid sequence based on the antibody or a conserved modification thereof described herein, wherein the antibody retains the desired functional properties of the antibody of the Disclosure.

[0322] As used herein, the term “optimized” means that a nucleotide sequence has been modified to encode an amino acid sequence using preferred codons in a producing cell or organism, generally a eukaryotic cell, such as a Pichia cell, a Chinese hamster ovary cell (CHO), or a human cell. An optimized nucleotide sequence is engineered to retain, to the greatest extent possible, the amino acid sequence originally encoded by an initiation nucleotide sequence, also known as the “parent” sequence. The optimized sequences herein are engineered to have preferred codons in mammalian cells. However, optimized expression of these sequences in other eukaryotic or prokaryotic cells is also assumed herein. The amino acid sequence encoded by an optimized nucleotide sequence is also referred to as optimized.

[0323] Another type of variable region modification involves mutating 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 antibody of interest, known as "affinity maturation." Mutations can be introduced by site-directed mutagenesis or PCR-mediated mutagenesis, and the effect on antibody binding, or other functional properties of interest, can be evaluated by in vitro or in vivo assays as provided herein in the examples. Conservative modifications (as described above) can be introduced. Mutations may be amino acid substitutions, additions, or deletions. More typically, one, two, three, four, or five or fewer residues within the CDR region are altered.

[0324] Affinity-matured antibodies have one or more modifications in one or more variable domains, which results in improved affinity of the antibody to an antigen compared to parental antibodies without these modifications. In one embodiment, affinity-matured antibodies may even have nanomolar or picomolar affinity for the target antigen. Affinity-matured antibodies are produced by procedures known in the art. For example, Marks et al, Bio / Technology 10:779-783 (1992) describe 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 Nat. 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). Accordingly, this disclosure provides the affinity-matured antibody.

[0325] The antibodies of this disclosure can further be prepared using antibodies having one or more VH and / or VL sequences as shown herein as starting materials for engineering modified antibodies, which may have altered properties from the starting antibodies. Antibodies can be engineering by modifying one or more residues within one or both variable regions (i.e., VH and / or VL), for example, within one or more CDR regions, and / or within one or more framework regions. Additionally or alternatively, antibodies can be engineering by modifying residues within constant regions to change, for example, the effector function of the antibody.

[0326] One type of variable region engineering that can be implemented is CDR implantation. Antibodies interact with target antigens primarily through amino acid residues located in six heavy and light chain complementarity-determining regions (CDRs). For this reason, the amino acid sequences within CDRs are more diverse among individual antibodies than the sequences outside of CDRs. Since CDR sequences are involved in most antibody-antigen interactions, recombinant antibodies that mimic the properties of specific native antibodies can be expressed by constructing expression vectors containing the CDR sequences of specific native antibodies transplanted onto the framework sequences of different antibodies with different properties (see, for example, Riechmann, L. et al., 1998 Nature 332:323-327; Jones, P. et al., 1986 Nature 321:522-525; Queen, C. et al., 1989 Proc. Natl. Acad. Sci. USA 86: 10029-10033; Winter's U.S. Patent No. 5,225,539, and Queen et al.'s U.S. Patents No. 5,530,101, 5,585,089, 5,693,762, and 6,180,370).

[0327] Such framework sequences can be obtained from publicly available DNA databases or published references containing germline antibody gene sequences or reconstituted antibody sequences. For example, germline DNA sequences of human heavy and light chain variable region genes are found in the "VBase" human germline sequence database (available on the internet at www.mrc-cpe.cam.ac.uk / vbase), as well as in Kabat, EA, et al., 1991 Sequences of Proteins of Immunological Interest, Fifth Edition, US Department of Health and Human Services, NIH Publication No. 91-3242; Tomlinson, IM, et al., 1992 J. fol. Biol. 227:776-798; and Cox, JPL et al., 1994 Eur. J Immunol. 24:827-836, the contents of which are expressly incorporated herein by reference. For example, germline DNA sequences of human heavy and light chain variable region genes, and reconstituted antibody sequences can be found in the "IMGT" database (available on the internet at www.imgt.org; see Lefranc, MP et al., 1999 Nucleic Acids Res. 27:209-212; the contents of each of these are expressly incorporated herein by reference).

[0328] Examples of framework sequences for use with the antibodies of this disclosure are structurally similar to framework sequences used by selected antibodies of this disclosure, such as consensus sequences and / or framework sequences used by monoclonal antibodies of this disclosure. The VH CDR1, 2, and 3 sequences, and the VL CDR1, 2, and 3 sequences, can be transplanted into framework regions having sequences identical to those found in the germline immunoglobulin genes from which the framework sequences originate, or the CDR sequences can be transplanted into framework regions containing one or more mutations compared to the germline sequences. For example, in certain cases, it has been found beneficial to mutate residues within the framework region to maintain or enhance the antigen-binding ability of the antibody (see, e.g., Queen et al., U.S. Patents 5,530,101, 5,585,089, 5,693,762, and 6,180,370).

[0329] A wide variety of antibody / immunoglobulin frameworks or scaffolds can be used, as long as the resulting polypeptide contains at least one binding region that specifically binds to IL-17A. Such frameworks or scaffolds include the five major idiotypes of human immunoglobulins, their antigen-binding fragments, and preferably immunoglobulins of other animal species having humanized embodiments.

[0330] In one embodiment, the Disclosure relates to a method for generating non-immunoglobulin-based antibodies using a non-immunoglobulin scaffold to which the CDR of the Disclosure can be implanted. Known or future non-immunoglobulin frameworks and scaffolds can be used, insofar as they contain a binding region specific to the target IL-17A protein. Known non-immunoglobulin frameworks or scaffolds include, but are not limited to, fibronectin (Compound Therapeutics, Inc., Waltham, Mass.), ankyrin (Molecular Partners AG, Zurich, Switzerland), domain antibodies (Domantis, Ltd., Cambridge, Mass., and Ablynx nv, Zwijnaarde, Belgium), lipocalin (Pieris Proteolab AG, Freising, Germany), small modular immunopharmaceuticals (Trubion Pharmaceuticals Inc., Seattle, Wash.), maxibody (Avidia, Inc., Mountain View, Calif), protein A (Affibody AG, Sweden), and affilin (gammacrystallin or ubiquitin) (Scil Proteins GmbH, Halle, Germany).

[0331] The antibodies described herein possess valuable properties that are expected to be beneficial for human patients requiring human IL-17A targeted therapy. The antibodies described herein are characterized by one or more of the following properties (determined in Examples 1 and 2):

[0332] The antibody specifically binds to human IL-17A, and then: (a) Having binding specificity to cynomolgus monkey IL-17A, (b) When measured by ELISA, it is found that it selectively binds to human IL-17A more than human IL-17B, IL-17C, IL-17D, IL-17E, and IL-17F. (c) Inhibit or block the binding between IL-17A and its receptor (IL-17RA), (d) Reduce or neutralize IL-17A activity, (e) When evaluated in vitro using the HT-29 assay, it is possible to inhibit GRO-α secretion.

[0333] (f) Having the ability to inhibit the interaction between IL-17A and IL-17RA at a relative titer greater than 5, more specifically greater than 10, greater than 15, and more specifically greater than 20, compared to the titer of secukinumab determined by ELISA assay, where the relative titer is the IC of secukinumab determined by ELISA. 50 The IC of the antibody in scFv format, determined by the value (ng / mL) and ELISA. 50 This is the ratio to the value (ng / mL).

[0334] (g) Having the ability to neutralize IL-17A at a titer (relative titer) greater than 50, more specifically greater than 100, and more specifically greater than 150, compared to the titer (relative titer) of secukinumab determined by measuring GRO-α secretion by the HT-29 assay, where the relative titer is the IC of secukinumab as measured by the HT-29 assay. 50 The IC50 of the antibody of the present invention in scFv format, measured by the HT-29 assay, and the IC50 value (ng / mL). 50 This is the ratio to the value (ng / mL).

[0335] (h) The activity of 1 ng of human IL-17A can be inhibited by 50% at concentrations of ≤1 ng / mL, more specifically ≤0.5 ng / mL, and more specifically ≤0.2 ng / mL, and the inhibitory activity is determined by measuring the GRO-α secretion induced by human IL-17A in the presence of 50 pg / ml TNFα using an HT-29 assay.

[0336] (i) When measured by surface plasmon resonance, preferably in a direct setting, the dissociation constant (K) is less than 5 nM, more specifically less than 1 nM, less than 0.5 nM, less than 0.2 nM, more specifically less than 100 pM, and more specifically less than 50 pM. D ) and binds to human IL-17A,

[0337] (j) When measured by surface plasmon resonance, specifically when measured by surface plasmon resonance with capture settings, K is less than 10 nM, e.g., less than 7 nM, less than 5 nM, less than 2 nM, less than 1 nM, specifically less than 0.5 nM D And it binds to IL-17A in cynomolgus monkeys.

[0338] In the case of (k)scFv format, the melting temperature (Tm) determined by differential scanning fluorescence assay is at least 60°C, more specifically at least 62°C, more specifically at least 65°C, and even more specifically at least 70°C, in particular here, the antibody is in pH 6.4, 150 mM NaCl phosphate-citrate buffer.

[0339] (l) If in scFv format, after 5 consecutive freeze-thaw cycles, if the initial concentration of the antibody of the present invention is 10 mg / ml, there is a loss of monomer content of less than 5%, more specifically less than 3%, and more specifically less than 1%, in particular here, the antibody is in phosphate-buffered saline (PBS) at pH 7.4, and / or

[0340] If in (m)scFv format, after storage at 4°C for at least 2 weeks, more specifically at least 4 weeks, if the initial concentration of the antibody of this disclosure is 10 mg / ml, the loss of monomer content is 5% or less, more specifically less than 4%, less than 3%, less than 2%, more specifically less than 1%, in particular herein, the antibody is in phosphate-buffered saline (PBS), pH 7.4; and / or (n) If the initial concentration of the antibody of this disclosure is 10 mg / ml, there will be a loss of less than 5% monomer content after storage at 37°C for at least 2 weeks, and more specifically, at least 4 weeks.

[0341] In one embodiment, the antibodies of this disclosure selectively bind to human IL-17A rather than human IL-17B, IL-17C, IL-17D, IL-17E, and IL-17F, as measured by ELISA. As used herein, the term “selectively binding” means that the antibody, composition, formulation, etc., does not significantly bind to IL-17B / C / D / E / F but binds to IL-17A. Selective binding typically results in a moderate to high IC50. 50 To distinguish it from nonspecific binding with low affinity, high affinity (or low K) D ) and low to medium ICs 50 Characterized by: Typically, the antibody is 10 -7 Less than M K D When binding occurs, the binding is considered selective. Appropriately, the antibodies of this disclosure, when measured by SPR, have a higher affinity, i.e., a lower K, for binding to human IL-17B, IL-17C, IL-17D, IL-17E, and IL-17F. D It then binds to human IL-17A. Appropriately, the antibodies of this disclosure, when measured by ELISA, show IC25 for IL-17B, IL-17C, IL-17D, IL-17E, and IL-17F, and IC25 for IL-17A. 50 ICs that are at least 100 times larger, and more specifically, at least 200 times, at least 300 times, and at least 400 times larger. 50 It has a value.

[0342] The antibodies of this disclosure specifically bind to IL-17A, where binding to IL-17A (a) inhibits or blocks the binding of IL-17A to its receptor (IL-17RA), and (b) reduces or neutralizes IL-17A activity.

[0343] As used herein, the term “neutralizing antibody” describes an antibody that can neutralize the biological signaling activity of IL-17A by, for example, blocking the binding of IL-17A to one or more of its receptors, and more specifically, by blocking the binding of IL-17A to IL-17RA. The antibodies of this disclosure are IL-17A neutralizing antibodies. As used herein, the term “neutralizing” will be understood to refer to a reduction in biological signaling activity, whether partial or complete. Neutralization of IL-17A can be determined by various assays, examples of which are described elsewhere herein.

[0344] Accordingly, the antibodies of this disclosure can inhibit GRO-α secretion when evaluated in vitro in an HT-29 assay (as described in Examples 1 and 2). In one embodiment, the antibodies of this disclosure have the ability to neutralize IL-17A at a titer greater than 50, preferably greater than 100, and more preferably greater than 150 (relative titer) compared to the titer of secukinumab determined by measuring GRO-α secretion in an HT-29 assay, where the relative titer is the IC50 of secukinumab determined by the HT-29 assay. 50 The IC50 value (ng / mL) and the IC50 value of the antibody of this disclosure in scFv format determined by the HT-29 assay. 50 This is a ratio to the value (ng / mL). In a further embodiment, the antibody of this disclosure can inhibit 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, and more preferably 0.2 ng / mL or less, the inhibitory activity being determined by measuring the GRO-α secretion induced by human IL-17A in an HT-29 assay in the presence of 50 pg / ml TNFα.

[0345] In one embodiment, the antibody of the present disclosure has the ability to block the interaction between IL-17A and IL-17RA with a titer (relative titer) greater than 5, preferably greater than 10, more preferably greater than 15, and more specifically greater than 20, compared to the titer of secukinumab determined in an ELISA assay, where the relative titer is the ratio of the IC 50 value (ng / mL) of secukinumab measured by ELISA to the IC 50 value (ng / mL) of the antibody of the present disclosure in scFv form measured by ELISA.

[0346] As used herein, the term "affinity" refers to the strength of the interaction between an antibody and an antigen at a single antigenic site. Within each antigenic site, the variable regions of the "arms" of the antibody interact with the antigen at multiple sites via weak non-covalent bonds, and the greater the number of interactions, the stronger the affinity.

[0347] "Binding affinity" generally refers to the total strength of non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless otherwise specified, as used herein, "binding affinity", "binds to", "binds with", or "is bound to" refers to the intrinsic binding affinity that reflects a 1:1 interaction between members of a binding pair (e.g., an antibody fragment and an antigen). The affinity of molecule X for partner Y can generally be represented by the dissociation constant (K D ). Affinity can be measured by common methods known in the art, including those described herein. Generally, low-affinity antibodies tend to bind antigens slowly and dissociate easily, while high-affinity antibodies generally bind antigens quickly and maintain the binding for a long time. Various methods for measuring binding affinity are known in the art, and any of them can be used for the purposes of the present invention. Specific exemplary embodiments for measuring binding affinity, i.e., binding strength, are described below.

[0348] As used herein, "K assoc ", "Ka", or "K onThe term "___" is intended to refer to the association rate of a particular antibody-antigen interaction, while the terms "K dis ", "Kd" or "K off " are intended to refer to the dissociation rate of a particular antibody-antigen interaction. In one embodiment, the term "K D " as used herein is intended to refer to the dissociation constant, which is obtained from the ratio of Kd to Ka (i.e., Kd / Ka) and is expressed as molar concentration (M). The "K D " or "K D value" or "KD" or "KD value" according to the present disclosure is, in one embodiment, measured using a T200 instrument (Biacore, GE Healthcare). To measure the affinity of a humanized scFv for human IL-17A, biotinylated human IL-17A is captured using Biacore's Biotin-CAPture kit. After each analyte injection cycle, the CAP sensor chip is regenerated and a new antigen is captured. The scFv is injected as an analyte using a dose-response multi-cycle reaction rate assay at analyte concentrations in the range of 0.35 - 90 nM diluted in running buffer. The resulting sensorgram is fitted using a 1:1 binding model. Alternatively or additionally, the affinity of the humanized scFv can be measured and analyzed using alternative SPR assay settings: IL-17A is immobilized on a CM5 sensor chip (GE Healthcare) by amine coupling, and serial dilutions of the scFv from 0.35 - 90 nM are reinjected onto the immobilized IL-17A.

[0349] Appropriately, the affinity of the antibody of this disclosure for IL-17A may be higher than the affinity of IL-17A for IL-17RA. It is understood that a higher affinity of the antibody of this disclosure compared to the affinity of IL-17A for IL-17RA may be particularly useful for dissociating or neutralizing pre-formed IL-17RA / IL-17A complexes. In one embodiment, the antibody of this disclosure neutralizes the IL-17RA / IL-17A interaction. In another embodiment, the antibody of this disclosure inhibits or blocks the binding of IL-17A to its receptor (IL-17RA). In one embodiment, the antibody of this disclosure neutralizes IL-17A activity.

[0350] Appropriately, the affinity of the antibodies of this disclosure for IL-17A may be comparable to, or higher than, preferably higher than, the affinity of secukinumab for IL-17A. In one embodiment, the antibodies of this disclosure neutralize IL-17A activity at a titer equivalent to or higher than, preferably higher than, secukinumab. In a further embodiment, the antibodies of this disclosure neutralize IL-17RA / IL-17A interaction at a titer equivalent to or higher than, preferably higher than, secukinumab.

[0351] Antibody binding affinity is, for example, determined by the dissociation constant (K). D This can be determined by a lower K. A stronger affinity is better. D This is represented by a lower affinity, and a lower affinity corresponds to a higher K D It is represented by [this].

[0352] Therefore, in a suitable embodiment, the antibodies of this disclosure, preferably measured by surface plasmon, and more preferably by surface plasmon resonance in a direct setting, yield K levels of 1-10000 pM, 1-7000 pM, 1-5000 pM, 1-2500 pM, 1-2000 pM, 1-1000 pM, 1-750 pM, 1-500 pM, 1-400 pM, 1-300 pM, 1-200 pM, 1-100 pM, and 1-50 pM. DIt may have the following characteristics. In a suitable embodiment, the antibody of this disclosure, when measured by surface plasmon resonance in a direct setting, has a K content of 1-200 pM, more specifically 1-100 pM. D It has the following characteristics. In certain embodiments, the antibodies of this disclosure, when measured by surface plasmon resonance in a direct setting, have a K of 1 to 50 pM. D It has the following characteristics. In a suitable embodiment, the antibody of the present disclosure, when measured preferably by surface plasmon, and more preferably by surface plasmon resonance in a direct setting, has a K content of less than 5 nM, less than 4 nM, less than 3 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. D It may have the following characteristics. Appropriately, the antibodies of this disclosure have a K content of less than 1 nM, more specifically less than 100 pM. D It has. Appropriately, the antibodies of this disclosure have a K content of less than 0.5 nM, more specifically less than 50 pM. D It has the following characteristics. More precisely, the antibodies of this disclosure, when measured by surface plasmon resonance in a direct setting, have a K content of less than 0.2 nM, more specifically less than 50 pM. D It has.

[0353] In a further embodiment, when measured by surface plasmon resonance (SPR) in a capture setting, the antibodies present disclosure have a K content of less than 10 nM, e.g., less than 7 nM, less than 5 nM, less than 2 nM, less than 1 nM, and more specifically less than 0.5 nM in cynomolgus monkey IL-17A. D They are joined together.

[0354] Appropriately, the antibodies of this disclosure, when measured by surface plasmon resonance (SPR), preferably in a direct setting, yield at least 10 3 M -1 s -1 At least 10 4 M -1 s -1 In summary, at least 5 × 10 4 M -1 s -1 At least 10 5 M -1 s-1 In summary, at least 5 × 10 5 M -1 s -1 At least 10 6 M -1 s -1 The above K on It binds to human IL-17A at a rapid rate. Preferably, the antibody of this disclosure, when measured by SPR, preferably by direct setting by surface plasmon resonance, has a rate of at least 10 5 M -1 s -1 In detail, at least 5 × 10 5 M -1 s -1 In more detail, see at least 10 6 M -1 s -1 The above K on It has speed.

[0355] Appropriately, when the antibodies of this disclosure are measured by surface plasmon resonance (SPR), preferably by direct measurement by surface plasmon resonance, they react with human IL-17A at a concentration of 5 × 10⁻¹⁴. -3 s -1 Below, 3 x 10 -3 s -1 The following 10 -3 s -1 Below, 5 x 10 -4 s -1 Below, 3 x 10 -4 s -1 The following 10 -4 s -1 Below, 5 x 10 -5 s -1 The following K off It binds at a rate. Preferably, the antibody of this disclosure, when measured by SPR, preferably by direct setting by surface plasmon resonance, is 10 -4 s -1 For more details, see below: 5×10 -5 s -1 The following K off It has speed.

[0356] Appropriately, the antibodies of this disclosure possess beneficial biophysical properties.

[0357] Appropriately, the antibodies of this disclosure, when expressed in scFv (single-chain variable fragment) antibody form, have a melting temperature (Tm) of at least 60°C, preferably at least 62°C, more preferably at least 65°C, and even more preferably at least 70°C, as determined by differential scanning fluorescence (DSF) assay as already described (Egan, et al., MAbs, 9(1) (2017), 68-84; Niesen, et al., Nature Protocols, 2(9) (2007) 2212-2221). The midpoint of the thermal expansion transition of the scFv construct is determined by differential scanning fluorescence assay using the fluorescent dye SYPRO® Orange (see Wong & Raleigh, Protein Science 25 (2016) 1834-1840). The sample is prepared in pH 6.4 phosphate-citrate buffer to a final protein concentration of 50 μg / mL, with a total volume of 100 μl containing the final concentration of 5×SYPRO® Orange. 25 μl of the prepared sample is added in triple count to a white-walled AB gene PCR plate. The assay is performed using a qPCR instrument used as a thermal cycler, and fluorescence emission is detected using a custom dye calibration routine in the software. The PCR plate containing the test sample is subjected to a temperature gradient from 25°C to 96°C in 1°C increments, with a 30-second pause after each temperature increase. The total assay time is approximately 2 hours. Tm is calculated using the GraphPad Prism software with a mathematical second derivative method to determine the inflection point of the curve. The reported Tm is the average of three measurements.

[0358] The antibodies of this disclosure, particularly when expressed in scFv (single-chain variable fragment) antibody form, and especially when the initial concentration of the antibodies of this disclosure is 10 mg / ml, are characterized in that the loss of monomer content after five consecutive freeze-thaw cycles is less than 5%, more specifically less than 3%, and more specifically less than 1%.

[0359] After storage for at least two weeks, and more specifically at 4°C for at least four weeks, the antibodies of this disclosure, particularly when expressed in scFv (single-chain variable fragment) antibody form, are characterized by a loss of monomer content of 5% or less, more specifically less than 4%, less than 3%, less than 2%, and preferably less than 1%, when the initial concentration of the antibodies of this disclosure 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 outlined in Chapter 621 of the USP. This method utilizes a hydrophobic stationary phase and an aqueous mobile phase to separate molecules based on size and shape. Molecular separation occurs between the void volume (V0) and total permeation volume (VT) of a particular column. Measurements by SE-HPLC are performed using a Chromaster HPLC system (Hitachi High-Technologies Corporation) equipped with automated sample injection and a UV detector set to a detection wavelength of 280 nm. This instrument is controlled by the software EZChrom Elite (Agilent Technologies, version 3.3.2 SP2), which also supports the analysis of the resulting chromatograms. Protein samples are clarified by centrifugation and kept at a temperature of 4–6°C in the autosampler before injection. For the analysis of scFv samples, a Shodex KW403-4F column (Showa Denko Inc., #F6989202) is used with a standardized buffered saline mobile phase (50 mM sodium phosphate pH 6.5, 300 mM sodium chloride) at a recommended flow rate of 0.35 mL / min. The target sample load per injection was 5 μg. Samples are detected with a 280 nm UV detector, and data are recorded with the appropriate software suite. The resulting chromatograms are analyzed in the range from V0 to VT, so matrix-related peaks with elution times greater than 10 minutes are excluded.

[0360] Appropriately, the isolated antibodies of this disclosure are selected from the group consisting of monoclonal antibodies, chimeric antibodies, Fab, Fv, scFv, dsFv, scAb, STAB, single-domain antibodies (sdAb or dAb), single-domain heavy-chain antibodies, and single-domain light-chain antibodies, VHH, VNAR, single-domain antibodies based on the VNAR structure of sharks, and, but not limited to, ankyrin-based domains, finomers, avimers, anticarin, binding domains based on alternative scaffolds including fibronectin, and binding sites incorporated into the constant region of the antibody (e.g., F-star's Modular Antibody Technology®), preferably scFv.

[0361] Suitablely, the antibody of this disclosure is Fv. Suitablely, the antibody of this disclosure is an scFv antibody fragment. A "single-chain Fv" or "scFv" or "sFv" antibody fragment comprises the VH and VL domains of the antibody, and these domains are present on a single polypeptide chain. Generally, the Fv polypeptide further comprises a polypeptide linker between the VH and VL domains, thereby enabling the sFv to form a desired structure for target binding. A "single-chain Fv" or "scFv" antibody fragment comprises the VH and VL domains of the antibody, and these domains are present on a single polypeptide chain. Generally, scFv polypeptides further contain a polypeptide linker between the VH and VL domains, which allows the scFv to form a structure desirable for antigen binding (see, for example, Pluckthun, The pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds., (Springer-Verlag, New York, 1994), pp. 269-315). In certain embodiments, the functional fragment is in scFv form including the linker according to SEQ ID NO: 23. In one embodiment, the antibody that specifically binds to human IL-17A is the antibody listed in Table 1. In one embodiment, an 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 NOs. 24 and SEQ ID NOs. 25; or (ii) the group consisting of SEQ ID NOs. 61 and SEQ ID NOs. 62. In a further embodiment, the antibody of the present disclosure is a single-stranded variable fragment (scFv) as shown in (i) SEQ ID NOs. 24 or SEQ ID NOs. 25, or (ii) SEQ ID NOs. 61 or SEQ ID NOs. 62.

[0362] Appropriately, the antibody of the Disclosure is an IgG antibody. In one embodiment, the antibody of the Disclosure is an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4, preferably IgG1.

[0363] Exemplary domains that specifically bind to TNFα The multispecific antibody of this disclosure comprises a second domain that specifically binds to TNFα, the domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein (a) the VH comprises, in order, three complementarity-determining regions HCDR1, HCDR2, and HCDR3, and (b) the VL comprises, in order, three complementarity-determining regions LDCR1, LDCR2, and LDCR3.

[0364] Suitable domains for specific binding to TNFα for use in the multispecific antibodies of this disclosure include, but are not limited to, the following: • The sequence is that of a humanized monoclonal antibody or its binding domain listed in Table 1 (described in WO2017 / 158101, the entirety of which is incorporated herein by reference); • Infliximab (Remicade®; U.S. Patents 6,277,969, 6,284,471, 6,790,444, and 6,835,823, all incorporated herein by reference); • Adalimumab / D2E7 (Humira®; described in U.S. Patent No. 6,090,382, which is incorporated herein by reference in its entirety); • Certolizumab, PEGylated Fab fragment (Cimzia®; described in U.S. Patents 7,012,135 and 7,186,820, all of which are incorporated herein by reference); • Golimumab (Simponi® registered trademark; published U.S. application No. 2009 / 214528, which is incorporated herein by reference in its entirety).

[0365] Preferred domains for use with the multispecific antibodies of this disclosure that specifically bind to TNFα include, but are not limited to, humanized monoclonal antibodies or their binding domains whose sequences are listed in Table 1 (as described in WO2017 / 158101, the entire text of which is incorporated herein by reference).

[0366] Accordingly, 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, LDCR1, LDCR2, and LDCR3), wherein the set of CDRs is a set of CDRs [where HCDR1' is an amino acid sequence selected from any one of SEQ ID NOs. 25, 28, and 31, preferably from SEQ ID NO. 25; HCDR2' is an amino acid sequence selected from any one of SEQ ID NOs. 26, 29, and 32, preferably from SEQ ID NO. 26; HCDR3' is an amino acid sequence selected from any one of SEQ ID NOs. 27, 30, and 33, preferably from SEQ ID NO. 27] The amino acid sequence is an amino acid sequence; LCDR1' is an amino acid sequence selected from any one of SEQ ID NOs. 38, 41, and 44, preferably from SEQ ID NO. 38; LCDR2' is an amino acid sequence selected from any one of SEQ ID NOs. 39, 42, and 45, preferably from SEQ ID NO. 39; and LCDR3' has an amino acid sequence selected from any one of SEQ ID NOs. 40, 43, and 46, preferably from SEQ ID NO. 40, with 10 or fewer amino acid substitutions, for example, 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.

[0367] In particular, this 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 one of the amino acid sequences of the VH CDRs listed in Table 1. In particular, the second domain that specifically binds to TNFα comprises (or consists of) one, two, three, or more VH CDRs having one of the amino acid sequences of the VH CDRs listed in Table 1.

[0368] Appropriately, the present disclosure provides a second domain that specifically binds to TNFα, wherein the second domain comprises a heavy chain variable region (VH), the VH comprising, in order, three complementarity-determining regions HCDR1, HCDR2, and HCDR3, wherein HCDR1 has an amino acid sequence selected from any one of SEQ ID NOs. 25, 28, and 31; HCDR2 has an amino acid sequence selected from any one of SEQ ID NOs. 26, 29, and 32; and HCDR3 has 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.

[0369] This 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 one of the amino acid sequences of the VL CDRs listed in Table 1. In particular, the second domain that specifically binds to TNFα comprises (or consists of) one, two, three, or more VL CDRs having one of the amino acid sequences of the VL CDRs listed in Table 1.

[0370] More specifically, the second domain that specifically binds to TNFα includes a light chain variable region (VL), where the VL includes, in order, three complementarity-determining regions LDCR1, LDCR2, and LDCR3, where LDCR1 has an amino acid sequence selected from any one of SEQ ID NOs. 38, 41, and 44; LCDR2 has an amino acid sequence selected from any one of SEQ ID NOs. 39, 42, and 45; and LCDR3 has an amino acid sequence selected from any one of SEQ ID NOs. 40, 43, and 46. In particular, the second domain includes the LDCR1, LDCR2, and LDCR3 sequences of SEQ ID NOs. 38, 39, and 40, respectively.

[0371] Appropriately, this 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:

[0372] (a) The VH comprises, in order, three complementarity-determining regions HCDR1, HCDR2, and HCDR3, wherein HCDR1 has an amino acid sequence selected from any one of SEQ ID NOs. 25, 28, and 31; HCDR2 has an amino acid sequence selected from any one of SEQ ID NOs. 26, 29, and 32; and HCDR3 has an amino acid sequence selected from any one of SEQ ID NOs. 27, 30, and 33; and (b) The VL comprises, in order, three complementarity-determining regions LCDR1, LCDR2, and LCDR3, wherein LCDR1 has an amino acid sequence selected from any one of SEQ ID NOs. 38, 41, and 44; LCDR2 has an amino acid sequence selected from any one of SEQ ID NOs. 39, 42, and 45; and LCDR3 has an amino acid sequence selected from any one of SEQ ID NOs. 40, 43, and 46.

[0373] 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) the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs. 25, 26, and 27, respectively, and (b) the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs. 38, 39, and 40, respectively.

[0374] Other domains of the Disclosure that specifically bind to TNFα include mutated amino acids in their CDR region that have at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identity with the CDR region shown in the sequences listed in Table 1. Preferably, other domains of the Disclosure that specifically bind to TNFα include mutant amino acid sequences in which, compared to the CDR region shown in the sequences listed in Table 1, 1, 2, 3, 4, 5, or 10 or fewer amino acids are mutated in the CDR region by amino acid deletion, insertion, or substitution. The mutation, e.g., substitution, may occur at any residue within the set of CDRs and may be within CDR1, CDR2, and / or CDR3.

[0375] Appropriately, the second domain of the multispecific antibody of this disclosure that specifically binds to TNFα comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein, (a) The VH comprises, in order, three complementarity-determining regions HCDR1, HCDR2, and HCDR3, wherein HCDR1 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to any one of SEQ ID NOs. 25, 28, and 31; HCDR2 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to any one of SEQ ID NOs. 26, 29, and 32; HCDR3 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to any one of SEQ ID NOs. 27, 30, and 33; and / or, (b) The VL comprises, in order, three complementarity-determining regions LCDR1, LCDR2, and LCDR3, wherein LCDR1 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to any one of SEQ ID NOs: 38, 41, and 44; LCDR2 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to any one of SEQ ID NOs: 39, 42, and 45; and LCDR3 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to any one of SEQ ID NOs: 40, 43, and 46.

[0376] Preferably, the second domain that specifically binds to TNFα includes 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 NOs. 25, 26, and 27, 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 NOs. 38, 39, and 40, respectively.

[0377] In a further embodiment, the second domain that specifically binds to TNFα includes a heavy chain variable region VHB and a light chain variable region VLB.

[0378] Suitablely, 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, where VHB is VH1A, VH1B, VH3, or VH4. In one embodiment, the second domain of the present disclosure that specifically binds to TNFα comprises a heavy chain variable region VHB, where VHB is VH4. In a preferred embodiment, the second domain of the present disclosure that specifically binds to TNFα comprises a heavy chain variable region VHB, where VHB is VH3.

[0379] Suitablely, 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, the VLB comprising Vκ framework FR1, FR2, and FR3, more specifically Vκ1 or Vκ3 FR1-FR3, preferably Vκ1 framework FR1-FR3, and framework FR4, wherein framework FR4 is selected from Vκ FR4, more specifically Vκ1 FR4, Vκ3 FR4, and Vλ FR4. Suitable Vλ FR4s are those described in SEQ ID NOs: 97-103. In one embodiment, the second domain that specifically binds to TNFα comprises a Vλ FR4 comprising an amino acid sequence having at least 60, 70, 80, or 90% identity with an amino acid sequence selected from any of SEQ ID NOs: 97-103, preferably SEQ ID NOs: 97 or 98, more preferably SEQ ID NOs: 97. Preferably, the second domain that specifically binds to TNFα includes a Vλ FR4 comprising an amino acid sequence selected from any of SEQ ID NOs: 97 to 103, preferably the Vλ FR4 described in SEQ ID NOs: 97 or 98, more preferably the Vλ FR4 described in SEQ ID NOs: 98.

[0380] Accordingly, 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) the HCDR1, HCDR2, and HCDR3 sequences of sequence numbers 25, 26, and 27, respectively, and the LCDR1, LCDR2, and LCDR3 sequences of sequence numbers 38, 39, and 40, respectively; (b) VHB, where VHB is VH3 or VH4, preferably VH3; and (c) A VLB comprising a VL framework comprising Vκ framework FR1, FR2, and FR3, more specifically Vκ1 or Vκ3 FR1 to FR3, preferably Vκ1 FR1 to FR3, and framework FR4, wherein framework FR4 is selected from Vκ FR4, more specifically Vκ1 FR4, Vκ3 FR4, and Vλ FR4, more specifically Vλ FR4 comprising an amino acid sequence having at least 60, 70, 80, or 90% identity with an amino acid sequence selected from any of SEQ ID NOs: 97 to 103, preferably Vλ FR4 described in any of SEQ ID NOs: 97 to 103, preferably Vλ FR4 described in SEQ ID NOs: 97 or 98, more preferably Vλ FR4 described in SEQ ID NOs: 98.

[0381] Suitablely, 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 the VH sequence described in Table 1. Suitablely, the present disclosure also provides a multispecific antibody comprising a first domain that specifically binds to IL-17A and a second domain that specifically binds to TNFα, wherein the second domain comprises (or comprises) the VH amino acid sequence described in Table 1, wherein about 20 or fewer amino acids, preferably about 10 or fewer amino acids, in the framework sequence (e.g., a sequence that is not a CDR) are mutated (wherein mutations are additions, substitutions, or deletions, in a variety of non-limiting examples). Other domains of the present disclosure that specifically bind to TNFα are mutated but contain amino acids that have at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identity with the VH region shown in the sequences described in Table 1.

[0382] Suitablely, 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 the VL shown in Table 1. Suitablely, the present disclosure also provides a multispecific antibody comprising a first domain that specifically binds to IL-17A and a second domain that specifically binds to TNFα, wherein the second domain comprises (or consists of) the VL amino acid sequence shown in Table 1, wherein about 20 or fewer amino acids, preferably about 10 or fewer amino acids, in the framework sequence (e.g., a sequence that is not a CDR) are mutated (where the mutation is, in various non-limiting examples, addition, substitution, or deletion). Other domains of the present disclosure that specifically bind to TNFα are mutated but contain amino acids that have at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identity with the VL region shown in the sequences shown in Table 1.

[0383] 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 heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 34, 35, 36, and 37, preferably SEQ ID NO: 34, and 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, in particular, the domain comprises 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 includes a heavy chain variable region containing an amino acid sequence that is at least 90% identical to amino acid sequence SEQ ID NO: 34, wherein the heavy chain variable region includes G56A, R82L, S85A, K86Q (AHo numbering), and in particular, the domain contains the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs: 25, 26, and 27, respectively. Preferably, the second domain that specifically binds to TNFα may include a heavy chain variable region containing an amino acid sequence that is at least 90% identical to amino acid sequence SEQ ID NO: 36, wherein the heavy chain variable region includes A24K, G56A, R82L (AHo numbering), and in particular, the domain contains the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs: 25, 26, and 27, respectively. Appropriately, the second domain that specifically binds to TNFα may include a heavy chain variable region comprising an amino acid sequence that is at least 90% identical to amino acid sequence SEQ ID NO: 37, wherein the heavy chain variable region comprises G56A, R82L, S85A, and K86Q (AHo numbering), and in particular, the domain comprises the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs: 25, 26, and 27, respectively.

[0384] 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 light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 47, 48, 49, 50, and 51, preferably SEQ ID NO: 47, and an amino acid sequence that is identical to at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90%, and in particular, the domain comprises the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs: 38, 39, and 40, respectively.

[0385] In a preferred embodiment, the second domain that specifically binds to TNFα includes a light chain variable region comprising an amino acid sequence that is at least 90% identical to amino acid sequence SEQ ID NO: 47, wherein the light chain variable region comprises T22N, A51R, F89Y (AHo numbering), and in particular, the domain comprises the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs: 38, 39, and 40, respectively. Preferably, the second domain that specifically binds to TNFα may include a light chain variable region comprising an amino acid sequence that is at least 90% identical to amino acid sequence SEQ ID NO: 49, wherein the light chain variable region comprises T22N, D88E, S95G, E99A (AHo numbering), and in particular, the domain comprises the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs: 38, 39, and 40, respectively. Preferably, the second domain that specifically binds to TNFα may include a light chain variable region containing an amino acid sequence that is at least 90% identical to amino acid sequence SEQ ID NO: 50, wherein the light chain variable region includes T22N, A51R, F89Y, S95G, G141T, L145V (AHo numbering), and in particular, the domain contains the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs: 38, 39, and 40, respectively. Preferably, the second domain that specifically binds to TNFα may include a light chain variable region containing an amino acid sequence that is at least 90% identical to amino acid sequence SEQ ID NO: 51, wherein the light chain variable region includes T22N, K50Q, A51R, F89Y, G141T, L145V (AHo numbering), and in particular, the domain contains the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs: 38, 39, and 40, respectively.

[0386] In a further 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 comprising an amino acid sequence sequence of at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90%, identical to amino acid sequence SEQ ID NO: 34, and a light chain variable region comprising an amino acid sequence of at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90%, identical to amino acid sequence SEQ ID NO: 47.

[0387] Preferably, the second domain that specifically binds to TNFα comprises a heavy chain variable region containing 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 amino acid sequence SEQ ID NO: 34, and a light chain variable region containing 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 amino acid sequence SEQ ID NO: 47. The domain includes a chain variable region, where the domain comprises the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs. 25, 26, and 27, and / or the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs. 38, 39, and 40, respectively, preferably the domain comprises 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.

[0388] Preferably, the second domain that specifically binds to TNFα comprises a heavy chain variable region containing 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 amino acid sequence SEQ ID NO: 37, and 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 amino acid sequence SEQ ID NO: 50 or 51. The domain includes a light chain variable region, where the domain comprises the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs. 25, 26, and 27, and / or the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs. 38, 39, and 40, respectively, preferably the domain comprises 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.

[0389] In specific embodiments, 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 containing an amino acid sequence selected from the group consisting of SEQ ID NOs: 34, 35, 36, and 37, and / or a VL containing an amino acid sequence selected from the group consisting of SEQ ID NOs: 47, 48, 49, 50, and 51. In a particular embodiment, the second domain that specifically binds to TNFα comprises the VH sequence of SEQ ID NO: 34 and the VL sequence of SEQ ID NO: 47. In yet another particular embodiment, the second domain that specifically binds to TNFα comprises the VH sequence of SEQ ID NO: 37 and the VL sequence of SEQ ID NO: 50 or SEQ ID NO: 51.

[0390] In one embodiment, the domain that specifically binds to human TNFα is the domain listed in Table 1. Other domains of the Disclosure having binding specificity to human TNFα include domains in which amino acids or nucleic acids encoding amino acids are mutated but which have at least 60, 70, 80, 90, or 95% identity with the sequences listed in Table 1. In one embodiment, this includes mutant amino acid sequences in which one, two, three, four, or five or fewer amino acids are mutated in the variable region compared to the variable region shown in the sequences listed in Table 1, but which retain substantially the same activity.

[0391] 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, the domain binds to human TNFα, and retains the desirable functional properties of the domain described in Table 1.

[0392] In one embodiment, the domain of the Disclosure that specifically binds to TNFα has a heavy chain variable region comprising HCDR1, HCDR2, and HCDR3 sequences and a light chain variable region comprising LDCR1, LDCR2, and LDCR3 sequences, wherein one or more of these CDR sequences have a specific amino acid sequence based on the domain described herein or its conserved modifications, and the domain retains the desired functional properties of the antibody of the Disclosure.

[0393] Accordingly, this 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 comprises): The heavy chain variable region (VH) comprises three complementarity-determining regions HCDR1, HCDR2, and HCDR3, respectively, wherein HCDR1 is amino acid sequence SEQ ID NO: 25 or a conserved variant thereof, HCDR2 is amino acid sequence SEQ ID NO: 26 or a conserved variant thereof, and HCDR3 is an amino acid sequence selected from any one of SEQ ID NO: 27 or a conserved variant thereof; and The light chain variable region (VL) comprises three complementarity-determining regions, LCDR1, LCDR2, and LCDR3, respectively, wherein LCDR1 is amino acid sequence number 38 or a conserved variant thereof, LCDR2 is amino acid sequence number 39 or a conserved variant thereof, and LCDR3 is amino acid sequence number 40 or a conserved variant thereof; Here, the domain specifically binds to human TNFα and / or neutralizes TNFα.

[0394] Exemplary domains that specifically bind to human serum albumin (HSA). The multispecific antibody of this disclosure comprises a third domain that specifically binds to human serum albumin, the domain comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein (a) the VH comprises, in order, three complementarity-determining regions HCDR1, HCDR2, and HCDR3, and (b) the VL comprises, in order, three complementarity-determining regions LDCR1, LDCR2, and LDCR3.

[0395] Appropriately, the multispecific antibodies of this disclosure may include a third binding domain having a third specificity distinct from the specificities of the first and second domains. Appropriately, the multispecific antibodies of this disclosure may include a third domain that specifically binds to human serum albumin (HSA). Thus, in one embodiment, the multispecific antibody of this disclosure includes 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.

[0396] Suitable domains for use with the multispecific antibodies of this disclosure that specifically bind to human serum albumin include, but are not limited to, humanized monoclonal antibodies or their binding domains whose sequences are listed in Table 1.

[0397] In one embodiment, the domains that specifically bind to human serum albumin include a set of CDRs (HCDR1, HCDR2, HCDR3, LDCR1, LDCR2, and LDCR3), where the set of CDRs is a set of CDRs [where, (a) HCDR1' is the one described in SEQ ID NO: 52, HCDR2' is the one described in SEQ ID NO: 53, HCDR3' is the one described in SEQ ID NO: 54, LCDR1' is the one described in SEQ ID NO: 62, LCDR2' is the one described in SEQ ID NO: 63, and LCDR3' is the one described in SEQ ID NO: 64; or (b) Having 10 or fewer amino acid substitutions from (b)HCDR1' is as described in SEQ ID NO: 73, HCDR2' is as described in SEQ ID NO: 74, HCDR3' is as described in SEQ ID NO: 75, LCDR1' is as described in SEQ ID NO: 83, LCDR2' is as described in SEQ ID NO: 84, and LCDR3' is as described in SEQ ID NO: 85, for example, 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.

[0398] In particular, the domain that specifically binds to human serum albumin comprises a VH CDR having one of the amino acid sequences of the VH CDRs listed in Table 1. 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 one of the amino acid sequences of the VH CDRs listed in Table 1.

[0399] More appropriately, the domain that specifically binds to human serum albumin includes a heavy chain variable region (VH), where the VH includes, in order, three complementarity-determining regions HCDR1, HCDR2, and HCDR3, where (a) The HCDR1 is an amino acid sequence selected from any one of SEQ ID NOs. 52, 55, and 58; the HCDR2 is an amino acid sequence selected from any one of SEQ ID NOs. 53, 56, and 59; and the HCDR3 is an amino acid sequence selected from any one of SEQ ID NOs. 54, 57, and 60; or (b) The HCDR1 is an amino acid sequence selected from any one of SEQ ID NOs. 73, 76, and 79; the HCDR2 is an amino acid sequence selected from any one of SEQ ID NOs. 74, 77, and 80; and the HCDR3 is an amino acid sequence selected from any one of SEQ ID NOs. 75, 78, and 81.

[0400] In a preferred embodiment, the domains that specifically bind to human serum albumin include the HCDR1, HCDR2, and HCDR3 sequences of sequence numbers 52, 53, and 54, respectively. In another embodiment, the domains that specifically bind to human serum albumin include the HCDR1, HCDR2, and HCDR3 sequences of sequence numbers 73, 74, and 75, respectively.

[0401] Appropriately, the domain that specifically binds to human serum albumin comprises a VL CDR having one of the amino acid sequences of the VL 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 VL CDRs having one of the amino acid sequences of the VL CDRs listed in Table 1.

[0402] More appropriately, the domain that specifically binds to human serum albumin includes a light chain variable region (VL), where the VL includes, in order, three complementarity-determining regions LDCR1, LDCR2, and LDCR3, where (a) LCDR1 is an amino acid sequence selected from any one of SEQ ID NOs. 62, 65, and 68; LCDR2 is an amino acid sequence selected from any one of SEQ ID NOs. 63, 66, and 69; and LCDR3 is an amino acid sequence selected from any one of SEQ ID NOs. 64, 67, and 70; or (b) LCDR1 is an amino acid sequence selected from any one of SEQ ID NOs. 83, 86, and 89; LCDR2 is an amino acid sequence selected from any one of SEQ ID NOs. 84, 87, and 90; and LCDR3 is an amino acid sequence selected from any one of SEQ ID NOs. 85, 88, and 91.

[0403] In a preferred embodiment, the domains that specifically bind to human serum albumin include the LDCR1, LDCR2, and LDCR3 sequences of SEQ ID NOs. 62, 63, and 64, respectively. In another embodiment, the domains that specifically bind to human serum albumin include the LDCR1, LDCR2, and LDCR3 sequences of SEQ ID NOs. 83, 84, and 85, respectively.

[0404] Appropriately, this 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), (a) The VH comprises, in order, three complementarity-determining regions HCDR1, HCDR2, and HCDR3, wherein HCDR1 has an amino acid sequence selected from any one of SEQ ID NOs. 52, 55, and 58; HCDR2 has a sequence selected from any one of SEQ ID NOs. 53, 56, and 59; and HCDR3 has an amino acid sequence selected from any one of SEQ ID NOs. 54, 57, and 60; and the VL comprises, in order, three complementarity-determining regions LDCR1, LDCR2, and LDCR3, wherein LDCR1 has an amino acid sequence selected from any one of SEQ ID NOs. 62, 65, and 68; LDCR2 has an amino acid sequence selected from any one of SEQ ID NOs. 63, 66, and 69; and LDCR3 has an amino acid sequence selected from any one of SEQ ID NOs. 64, 67, and 70; or (b) The VH comprises, in order, three complementarity-determining regions HCDR1, HCDR2, and HCDR3, wherein HCDR1 has an amino acid sequence selected from any one of SEQ ID NOs. 73, 76, and 79; HCDR2 has an amino acid sequence selected from any one of SEQ ID NOs. 74, 77, and 80; and HCDR3 has an amino acid sequence selected from any one of SEQ ID NOs. 75, 78, and 81; and the VL comprises, in order, three complementarity-determining regions LDCR1, LDCR2, and LDCR3, wherein LDCR1 has an amino acid sequence selected from any one of SEQ ID NOs. 83, 86, and 89; LDCR2 has an amino acid sequence selected from any one of SEQ ID NOs. 84, 87, and 90; and LDCR3 has an amino acid sequence selected from any one of SEQ ID NOs. 85, 88, and 91.

[0405] In a preferred embodiment, the disclosure provides a multispecific antibody comprising a third domain that specifically binds to HSA, wherein the third domain comprises (a) the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs. 52, 53, and 54, respectively, and (b) the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs. 62, 63, and 64, respectively. In another embodiment, the disclosure provides a multispecific antibody comprising a third domain that specifically binds to HSA, wherein the third domain comprises (a) the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs. 73, 74, and 75, respectively, and (b) the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs. 83, 84, and 85, respectively.

[0406] Other domains of the Disclosure that specifically bind to HSA include mutated amino acids in their CDR region that have at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identity with the CDR region shown in the sequences listed in Table 1. Preferably, other domains of the Disclosure that specifically bind to HSA include mutant amino acid sequences in which, compared to the CDR region shown in the sequences listed in Table 1, 1, 2, 3, 4, 5, or 10 or fewer amino acids are mutated by amino acid deletion, insertion, or substitution in the CDR region. Mutations, such as substitutions, may occur at any residue within the set of CDRs, and may be within CDR1, CDR2, and / or CDR3.

[0407] Appropriately, the third domain of the multispecific antibody of this disclosure that specifically binds to HSA comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein, (a) The VH comprises, in order, three complementarity-determining regions HCDR1, HCDR2, and HCDR3, wherein HCDR1 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to any one of SEQ ID NOs. 52, 55, and 58; HCDR2 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to any one of SEQ ID NOs. 53, 56, and 59; and / or (b) The VL comprises, in order, three complementarity-determining regions LCDR1, LCDR2, and LCDR3, wherein LCDR1 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to any one of SEQ ID NOs. 62, 65, and 68; LCDR2 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to any one of SEQ ID NOs. 63, 66, and 69; and LCDR3 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to any one of SEQ ID NOs. 64, 67, and 70.

[0408] Preferably, the multispecific antibody comprises a third domain that specifically binds to HSA, wherein the third domain comprises 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 NOs. 52, 53, and 54, and / or comprises 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 NOs. 62, 63, and 64, respectively.

[0409] Appropriately, the third domain of the multispecific antibody of this disclosure that specifically binds to HSA comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein, (a) The VH comprises, in order, three complementarity-determining regions HCDR1, HCDR2, and HCDR3, wherein HCDR1 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to any one of SEQ ID NOs. 73, 76, and 79; HCDR2 has an amino acid sequence that is at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to any one of SEQ ID NOs. 74, 77, and 80; and / or (b) The VL comprises, in order, three complementarity-determining regions LCDR1, LCDR2, and LCDR3, wherein 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 sequence numbers 83, 86, and 89; 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 sequence numbers 84, 87, and 90; and 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 sequence numbers 85, 88, and 91.

[0410] Preferably, the multispecific antibody comprises a third domain that specifically binds to HSA, the third domain comprising 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 NOs. 73, 74, and 75, and / or comprising 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 NOs. 83, 84, and 85, respectively.

[0411] 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).

[0412] Suitablely, 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, where the VHC is VH1A, VH1B, VH3, or VH4. In one embodiment, the third domain of the present disclosure that specifically binds to HSA comprises a heavy chain variable region VHC, where the VHC is VH4. In a preferred embodiment, the third domain of the present disclosure that specifically binds to HSA comprises a heavy chain variable region VHC, where the VHC is VH3.

[0413] Suitablely, the present disclosure provides a multispecific antibody comprising a third domain that specifically binds to HSA, wherein the third domain comprises a light chain variable region VLC, the VLC comprising Vκ framework FR1, FR2, and FR3, more specifically Vκ1 or Vκ3 FR1-FR3, preferably Vκ1 framework FR1-FR3, and framework FR4, wherein FR4 is selected from Vλ FR4, more specifically Vκ1 FR4, Vκ3 FR4, and Vλ FR4. Suitable Vλ FR4s are those described in SEQ ID NOs: 97-103. In one embodiment, the third domain comprises a Vλ FR4 having at least 60, 70, 80, or 90% identity with an amino acid sequence selected from any of SEQ ID NOs: 97-103, preferably SEQ ID NOs: 97 or 98, more preferably SEQ ID NOs: 97. Preferably, the third domain includes a Vλ FR4 comprising an amino acid sequence selected from any of SEQ ID NOs: 97 to 103, preferably the Vλ FR4 described in SEQ ID NOs: 97 or 98, more preferably the Vλ FR4 described in SEQ ID NOs: 98.

[0414] Accordingly, 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) the HCDR1, HCDR2, and HCDR3 sequences of sequence numbers 52, 53, and 54, respectively, and the LCDR1, LCDR2, and LCDR3 sequences of sequence numbers 62, 63, and 64, respectively; (b) VHC, wherein the VHC is VH3 or VH4, preferably VH3; and (c) A VLC comprising a VL framework comprising Vκ framework FR1, FR2, and FR3, more particularly Vκ1 or Vκ3 FR1~FR3, preferably Vκ1 FR1~FR3, and framework FR4, wherein framework FR4 is selected from Vκ FR4, more particularly Vκ1 FR4, Vκ3 FR4, and Vλ FR4, more particularly Vλ FR4 comprising an amino acid sequence having at least 60, 70, 80, or 90% identity with an amino acid sequence selected from any of SEQ ID NOs: 97 to 103, preferably Vλ FR4 described in any one of SEQ ID NOs: 97 to 103, preferably Vλ FR4 described in SEQ ID NOs: 97 or 98, more preferably Vλ FR4 described in SEQ ID NOs: 98.

[0415] In a further embodiment, this disclosure provides a multispecific antibody comprising a third domain that specifically binds to HSA, wherein the third domain comprises: (i) the HCDR1, HCDR2, and HCDR3 sequences of sequence numbers 73, 74, and 75, respectively, and the LCDR1, LCDR2, and LCDR3 sequences of sequence numbers 83, 84, and 85, respectively; (ii) VHC, wherein the VHC is VH3 or VH4, preferably VH3; and (iii) A VLC comprising a VL framework comprising Vκ frameworks FR1, FR2, and FR3, more specifically Vκ1 or Vκ3 FR1 to FR3, preferably Vκ1 FR1 to FR3, and framework FR4, wherein framework FR4 is selected from Vκ FR4, more specifically Vκ1 FR4, Vκ3 FR4, and Vλ FR4, more specifically Vλ FR4 comprising an amino acid sequence having at least 60, 70, 80, or 90% identity with an amino acid sequence selected from any of SEQ ID NOs. 97 to 103, preferably Vλ FR4 described in an amino acid sequence selected from any one of SEQ ID NOs. 97 to 103, preferably Vλ FR4 of SEQ ID NOs. 97 or 98, more preferably Vλ FR4 described in SEQ ID NOs. 98

[0416] Suitablely, the present disclosure provides a multispecific antibody comprising a third domain that specifically binds to HSA, wherein the third domain comprises the VH sequence described in Table 1. Suitablely, the present disclosure also provides a multispecific antibody comprising a third domain that specifically binds to HSA, wherein the third domain comprises (or comprises) the VH amino acid sequence described in Table 1, wherein about 20 or fewer amino acids, preferably about 10 or fewer amino acids, are mutated in the framework sequence (e.g., a sequence other than the CDR sequence) (mutations are additions, substitutions, or deletions, in a variety of non-limiting examples). Other domains of the present disclosure that specifically bind to HSA are mutated but contain amino acids that have at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identity with the VH region shown in the sequences described in Table 1.

[0417] Suitablely, the present disclosure provides a multispecific antibody comprising a third domain that specifically binds to HSA, wherein the third domain comprises the VL domain described in Table 1. Suitablely, the present disclosure also provides a multispecific antibody comprising a third domain that specifically binds to HSA, wherein the third domain comprises (or comprises) the VL amino acid sequence described in Table 1, wherein about 20 or fewer amino acids, preferably about 10 or fewer amino acids, are mutated in the framework sequence (e.g., a sequence other than the CDR sequence) (mutations are additions, substitutions, or deletions, in a variety of non-limiting examples). Other domains of the present disclosure that specifically bind to HSA are mutated but contain amino acids that have at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identity with the VL region shown in the sequences described in Table 1.

[0418] 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 amino acid sequence SEQ ID NO: 52, 53, and 54, respectively.

[0419] 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 amino acid sequence SEQ ID NOs. 62, 63, and 64, respectively.

[0420] In a further embodiment, the present disclosure provides a multispecific antibody comprising a third domain that specifically binds to HSA, wherein the third domain comprises a heavy chain variable region comprising an amino acid sequence sequence identical to amino acid sequence SEQ ID NO: 61 by at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90%; and a light chain variable region comprising an amino acid sequence sequence identical to amino acid sequence SEQ ID NO: 71 by at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90%.

[0421] In a preferred embodiment, the present disclosure provides a multispecific antibody comprising a third domain that specifically binds to HSA, wherein the third domain comprises a heavy chain variable region comprising an amino acid sequence that is identical to amino acid sequence SEQ ID NO: 61 by at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90%; and amino acid sequence SEQ ID NO: 71 by at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least The light chain variable region comprises an amino acid sequence that is at least 90% identical; where the domain comprises the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs. 52, 53, and 54, and / or the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs. 62, 63, and 64, respectively, preferably the domain comprises the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs. 52, 53, and 54, and the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs. 62, 63, and 64, respectively.

[0422] In certain embodiments, the present disclosure provides a multispecific antibody comprising a third domain that specifically binds to HSA, wherein the third domain comprises a VH containing amino acid sequence SEQ ID NO: 61 and / or a VL containing amino acid sequence SEQ ID NO: 71. In preferred embodiments, the third domain that specifically binds to HSA comprises the VH sequence of SEQ ID NO: 61 and the VL sequence of SEQ ID NO: 71.

[0423] In yet another 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 identical to amino acid sequence SEQ ID NO: 82 and at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90%, and in particular, the domain comprises the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs: 73, 74, and 75, respectively.

[0424] 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 identical to amino acid sequence SEQ ID NO: 92 and at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90%, wherein the domain comprises LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs: 83, 84, and 85, respectively.

[0425] In a further embodiment, the present disclosure provides a multispecific antibody comprising a third domain that specifically binds to HSA, wherein the third domain comprises a heavy chain variable region comprising an amino acid sequence identical to amino acid sequence SEQ ID NO: 82 by at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90%, and a light chain variable region comprising an amino acid sequence identical to amino acid sequence SEQ ID NO: 92 by at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90%.

[0426] In a preferred embodiment, the present disclosure provides a multispecific antibody comprising a third domain that specifically binds to HSA, wherein the third domain comprises a heavy chain variable region comprising an amino acid sequence identical to amino acid sequence SEQ ID NO: 82 by at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90%, and an amino acid sequence identical to amino acid sequence SEQ ID NO: 92 by at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least The domain also includes a light chain variable region containing an amino acid sequence that is 90% identical to the domain, and the domain includes the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs. 73, 74, and 75, and / or the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs. 83, 84, and 85, respectively, preferably the domain includes the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs. 73, 74, and 75, and the LDCR1, LDCR2, and LDCR3 sequences of SEQ ID NOs. 83, 84, and 85, respectively.

[0427] In certain embodiments, the present disclosure provides a multispecific antibody comprising a third domain that specifically binds to HSA, wherein the third domain comprises a VH containing amino acid sequence SEQ ID NO: 82 and / or a VL containing amino acid sequence SEQ ID NO: 92. In preferred embodiments, the third domain that specifically binds to HSA comprises the VH sequence of SEQ ID NO: 82 and the VL sequence of SEQ ID NO: 92.

[0428] In one embodiment, the domain that specifically binds to human serum albumin is the domain listed in Table 1. In one embodiment, the domain that specifically binds to HSA includes an amino acid sequence selected from the group consisting of SEQ ID NOs: 72 and 93, preferably SEQ ID NO: 72, and an amino acid sequence that is identical to at least 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, preferably at least 90%. In one embodiment, the domain that specifically binds to HSA is the one described in SEQ ID NO: 72 or SEQ ID NO: 93, preferably SEQ ID NO: 72.

[0429] Other domains of the present disclosure having binding specificity to HSA include domains in which the amino acids or nucleic acids encoding amino acids are mutated but which have at least 60, 70, 80, 90, or 95% identity with the sequences listed in Table 1. In one embodiment, this includes a mutant amino acid sequence in which one, two, three, four, or five or fewer amino acids are mutated in the variable region compared to the variable region shown in the sequences listed in Table 1, but which retain substantially the same activity.

[0430] In yet another embodiment, the present disclosure provides a multispecific antibody comprising a third domain that specifically binds, wherein the third domain comprises an amino acid sequence homologous to the sequences listed in Table 1, and the domain binds to human serum albumin and retains the desirable functional properties of those domains listed in Table 1.

[0431] In one embodiment, the domain of the Disclosure that specifically binds to HSA has a heavy chain variable region comprising HCDR1, HCDR2, and HCDR3 sequences and a light chain variable region comprising LCDR1, LDCR2, and LDCR3 sequences, wherein one or more of these CDR sequences have a specific amino acid sequence based on the domain described herein or its conserved modifications, and the domain retains the desired functional properties of the antibody of the Disclosure.

[0432] Accordingly, this disclosure provides a multispecific antibody comprising a third domain that specifically binds to HSA, wherein the third domain comprises (or comprises): A heavy chain variable region (VH) comprising, in order, three complementarity-determining regions HCDR1, HCDR2, and HCDR3, wherein HCDR1 is amino acid sequence SEQ ID NO: 52 or a conserved variant thereof, HCDR2 is amino acid sequence SEQ ID NO: 53 or a conserved variant thereof, and HCDR3 is an amino acid sequence selected from any one of SEQ ID NO: 54 or its conserved variant; and A light chain variable region (VL) comprising three complementarity-determining regions, LCDR1, LCDR2, and LCDR3, wherein LCDR1 is amino acid sequence number 62 or a conserved variant thereof, LCDR2 is amino acid sequence number 63 or a conserved variant thereof, and LCDR3 is amino acid sequence number 64 or a conserved variant thereof; Here, the domain specifically binds to human serum albumin.

[0433] This disclosure also provides a multispecific antibody comprising a third ...

Claims

1. An antibody having binding specificity to human IL-17A, wherein the antibody is as follows: (i) the HCDR1, HCDR2, and HCDR3 sequences of sequence numbers 1, 2, and 3, respectively, and LCDR1, LCDR2, and LCDR3 sequences of sequence numbers 12, 13, and 14, respectively. This includes, and here, VH includes an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 10; and VL contains an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 21; or (ii) The HCDR1, HCDR2, and HCDR3 sequences of sequence numbers 39, 40, and 41, respectively, and LCDR1, LCDR2, and LCDR3 sequences of sequence numbers 50, 51, and 52, respectively. It includes, and here VH includes an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 48, and The antibody wherein VL contains an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:

59.

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

3. (i) The VH sequence of sequence number 10 and the VL sequence of sequence number 21; or (ii) VH sequence of sequence number 48 and VL sequence of sequence number 59 The antibody according to claim 2, comprising:

4. (i) The VH sequence of sequence number 11 and the VL sequence of sequence number 22; or (ii) VH sequence of sequence number 49 and VL sequence of sequence number 60 The antibody according to claim 2, comprising:

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

6. The antibody according to any one of claims 1 to 5, wherein, when measured by ELISA, the antibody selectively binds to human IL-17A rather than human IL-17B, IL-17C, IL-17D, IL-17E, and IL-17F.

7. The coupling to IL-17A (a) Suppress or inhibit the binding between IL-17A and its receptor (IL-17RA), and (b) Reduces or neutralizes IL-17A activity, The antibody according to any one of claims 1 to 6.

8. The antibody according to claim 7, wherein the antibody can inhibit GRO-α secretion when evaluated in vitro in an HT-29 assay.

9. The aforementioned antibody (a) When determined by the ELISA assay, it has the ability to inhibit the interaction between IL-17A and IL-17RA with a potency greater than 5 (relative potency) compared to the potency of secukinumab, and here The aforementioned relative potency is measured by IC50 in ng / mL units of secukinumab as measured by ELISA. ５０ The IC50 value and the IC50 value of the antibody of the present invention in scFv format measured by ELISA in ng / mL units. ５０ It is the ratio to the value; and / or (b) The ability to neutralize IL-17A with a potency greater than 50 (relative potency) relative to the potency of secukinumab, as determined by measuring GRO-α secretion in the HT-29 assay, and here, The aforementioned relative potency is measured in the IC50 of secukinumab in ng / mL units in the HT-29 assay. ５０ The value and the IC50 in ng / mL units of the antibody of the present invention in scFv format, as measured in the HT-29 assay. ５０ It is the ratio to the value; and / or (c) The antibody according to any one of claims 1 to 8, which can inhibit the activity of 1 ng of human IL-17A by 50% at a concentration of 1 ng / mL or less, and the inhibitory activity is determined by measuring the GRO-α secretion induced by human IL-17A in an HT-29 assay in the presence of 50 pg / mL of TNFa.

10. The aforementioned antibody is as follows (a) When measured directly by surface plasmon resonance, human IL-17A has a dissociation constant of less than 5 nM (K D ) are joined together, and (b) When measured by surface plasmon resonance with the capture setting, cynomolgus monkey IL-17A has a K of less than 10 nM. D An antibody according to any one of claims 1 to 9, which is bound by [a specific mechanism].

11. When the aforementioned antibody is measured directly by surface plasmon resonance, it has a dissociation constant (K) of less than 0.5 nM for human IL-17A. D The antibody according to claim 10, which is bound by ).

12. The aforementioned antibody (a) If the antibody is in scFv format, and is in pH 6.4, 150 mM NaCl phosphate-citrate buffer, the melting temperature (Tm) determined by differential scanning fluorescence assay is at least 60°C; (b) When the antibody of the present invention is in scFv format, and the initial concentration of the antibody is 10 mg / ml, and the antibody is in phosphate-buffered saline (PBS), pH 7.4, the loss of monomer content is less than 5% after five consecutive freeze-thaw cycles; (c) In the case of scFv format, if the antibody of the present invention has an initial concentration of 10 mg / ml, and the antibody is in phosphate-buffered saline (PBS), pH 7.4, it will have a loss of 5% or less of monomer content after being stored at 4°C for at least two weeks; (d) The antibody according to any one of claims 1 to 11, wherein, when the antibody of the present invention is at an initial concentration of 10 mg / ml, the loss of monomer content is less than 5% after storage at 37°C for at least two weeks.

13. The antibody according to any one of claims 1 to 12, wherein the antibody is selected from the group consisting of a monoclonal antibody, Fab, Fv, scFv, dsFv, and scAb.

14. The antibody according to claim 13, wherein the antibody is scFv.

15. The antibody according to claim 14, wherein the scFv has (i) an amino acid sequence selected from the group consisting of SEQ ID NO: 24 and SEQ ID NO: 25; or (ii) an amino acid sequence selected from the group consisting of SEQ ID NO: 61 and SEQ ID NO:

62.

16. The antibody according to claim 15, wherein the scFv has an amino acid sequence selected from the group consisting of SEQ ID NO: 24 and SEQ ID NO:

61.

17. The antibody according to claim 13, wherein the antibody is an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4.

18. The antibody according to claim 17, wherein the antibody is IgG1 or IgG4.

19. An antibody according to any one of claims 1 to 18, which is a multispecific molecule.

20. The antibody is one of the following: single-chain diabody (scDb), tandem scDb, linear dimer scDb, cyclic dimer scDb, tandem di-scFv, tandem tri-scFv, Fab-(scFv) ２ or Fab-(scFv) １ , Fab, Fab-Fv ２ , IgG CH3-scFv fusion (Morrison L) or IgG CL-scFv fusion (Morrison H), scDb-scFv, bispecific Fab ２ , scFv-Fc-scFv fusion, scFv-HSA-scFv fusion, IgG-scFab, scFab-dsscFv, Fv ２ -Fc, IgG-scFv fusion with scFv linked to the C-terminus of the light chain, IgG-scFv fusion with scFv linked to the N-terminus of the light chain, IgG-scFv fusion with scFv linked to the N-terminus of the heavy chain, IgG-scFv fusion with scFv linked to the C-terminus of the heavy chain, IgG-scFv fusion with scFv linked to the N-termini of both the heavy and light chains, IgG-scFv fusion with dsscFv linked to the C-terminus of the heavy chain, bispecific antibody based on a heterodimeric Fc domain, Knob-into-Hole antibody (KiH); A fragment fused to the N and / or C-terminus of either strand of the heterodimer Fc domain, wherein Fv, scFv, scDb, tandem-di-scFv, tandem-tri-scFv, Fab-(scFv) ２ , Fab-(scFv) １ , Fab, and Fab-Fv ２ Fragments, matches, and duobodies selected from the list. The antibody according to claim 19, which is a format selected from the group consisting of the following.

21. A pharmaceutical composition comprising an antibody according to any one of claims 1 to 20 and a pharmaceutically acceptable carrier.

22. An antibody according to any one of claims 1 to 20, or a pharmaceutical composition according to claim 21, for use as a pharmaceutical agent.

23. An antibody according to any one of claims 1 to 20, or a pharmaceutical composition according to claim 21, for use in the treatment of an inflammatory condition or an autoimmune disease.

24. An antibody according to any one of claims 1 to 20, or a pharmaceutical composition according to claim 21, 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, cystic fibrosis, bone loss, airway hypersensitivity, demyelinating disorders, skin hypersensitivity, acute transplant rejection, allograft rejection, graft-versus-host disease, systemic sclerosis, urinary tract inflammatory diseases, cardiovascular diseases, vasculitis, periodic fever, glucose metabolism disorders, lung diseases, periodontitis, hepatic interstitial keratitis, allergies, inflammatory pain, spondyloarthropathy, sepsis, septic or endotoxin shock, meningitis, surgical trauma, autoimmune hematological disorders, Alzheimer's disease, sarcoidosis, cirrhosis, hepatitis, glomerulonephritis, or dyslipidemia.

25. Use of an antibody according to any one of claims 1 to 20, or a pharmaceutical composition according to claim 21, in the manufacture of a drug for use in the treatment of an inflammatory condition or autoimmune disease.

26. Use of an antibody according to any one of claims 1 to 20, or a pharmaceutical composition according to claim 21, in the manufacture of a drug 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, cystic fibrosis, bone loss, airway hypersensitivity, demyelinating disorders, skin hypersensitivity, acute transplant rejection, allograft rejection, graft-versus-host disease, systemic sclerosis, urinary tract inflammatory diseases, cardiovascular diseases, vasculitis, periodic fever, glucose metabolism disorders, lung diseases, periodontitis, hepatic interstitial keratitis, allergies, inflammatory pain, spondyloarthropathy, sepsis, septic or endotoxin shock, meningitis, surgical trauma, autoimmune hematological disorders, Alzheimer's disease, sarcoidosis, cirrhosis, hepatitis, glomerulonephritis, or dyslipidemia.

27. A nucleic acid encoding an antibody according to any one of claims 1 to 20.

28. A vector comprising the nucleic acid described in claim 27.

29. A host cell comprising the nucleic acid described in claim 27, or the vector described in claim 28.

30. A method for producing an antibody according to any one of claims 1 to 20, wherein the method comprises the step of culturing a host cell containing a nucleic acid or vector encoding the antibody according to any one of claims 1 to 20.

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