EA040031B1 - COMPOUND TARGETING TO IL-23A AND B-LYMPHOCYTE ACTIVATION FACTOR (BAFF) AND ITS USE - Google Patents

Description

Cross-reference to related applications

This application claims priority by filing date under 35 U.S.C. §119 on U.S. Provisional Applications No. 62/196170, filed July 23, 2015; No. 62/201067 filed on August 4, 2015; and No. 62/355302, filed June 27, 2016, all entitled Compound Targeting IL-23A and B-lymphocyte Activating Factor (BAFF) and its uses, the entire contents of each of which are incorporated herein by reference.

State of the art

Inflammation involves the innate and adaptive immune response needed to fight infection. However, when inflammation becomes an uncontrolled autoimmune or autoinflammatory disease, neurodegenerative disease and even cancer can develop. Pro-inflammatory cytokines such as IL1, FAFL (B-lymphocyte activating factor, BAFF), TNF (tumor necrosis factor)a, IL6, IL12, IL17, IL18 and IL23 are known to be involved in inflammation and specific pathways that activate immune cells. . However, it is not clear whether or how inhibition of one or more of these cytokines can affect the treatment of autoimmune or autoimmune diseases.

Interleukin 23 (IL23) is a heterodimeric cytokine consisting of two subunits, p40 and p19. The p19 subunit is also referred to as IL-23A. Although the p19 subunit is unique to IL23, the p40 subunit is also used by the cytokine IL12. IL23 is becoming known as a key regulator of pathogenic T helper 17 (Th-17 (Th17)), γδ T, and innate lymphoid cells (ILCs) that control the production of IL17, IL22 and other cytokines that lead to local inflammation and tissue damage. . IL23 enhances the expression of matrix metalloprotease MMP9, increases angiogenesis, reduces the infiltration of CD8+ T cells, and is involved in the development of malignant tumors. In addition, IL23 in combination with IL6 and TGFe1 stimulates naive CD4+ T-lymphocytes to differentiate into Th-17 (Th17) cells. In turn, Th-17 (Th17) cells produce IL17, a pro-inflammatory cytokine that enhances T-lymphocyte priming and stimulates the production of pro-inflammatory cytokines such as IL1, IL6, TNFa, NOS-2, and also induces the expression of chemokines leading to inflammation and disease pathogenesis. IL23 exerts its effects through a cell surface receptor consisting of the IL12b1 subunit of the IL12 receptor working in conjunction with the unique IL23R subunit. IL23R expression is limited to specific immune cell populations and occurs mainly on T cell subclasses (αβ and γδ TCR+) and NK (natural killer) cells.

In mice, genetic ablation of the IL23p19 gene results in a selective loss of IL23 function, accompanied by significantly impaired T-dependent immune responses, including reduced production of antigen-specific immunoglobulins and impaired delayed-type hypersensitivity responses (Ghilardi N. et al. (2004) J. Immunol. 172(5 ):2827-33). Knockout mice lacking either IL23p40 or IL23p19 or any IL23 receptor subunit (IL23R and IL12b 1) exhibit less severe symptoms of multiple sclerosis, arthritis, and inflammatory bowel disease in animal models. Similar results were obtained using an antibody specific for IL23p19 in EAE (experimental autoimmune encephalomyelitis) and a model of T-cell colitis further confirming the role of IL23 in these disease conditions (Chen Y. et al. (2006) J. Clin. Invet. 116(5): 1317-26; Elson CO. et al. (2007) Gastroenterology 132(7): 2359-70). This highlights the importance of IL23 in chronic inflammation (Langowski et al. (2006) Nature 442 (7101): 461-5; Kikly K, et al. (2006) Curr. Opin. Immunol. 18(6):670-5). In addition, increased production of IL23 is considered to be a major factor in inflammatory arthritis and inflammatory autoimmune diseases (Adamopoulos et al. (2011) J. Immunol. 187: 593-594; and Langris et al. (2005) J. Exp. Med. 201: 233240). The association between IL23, its downstream cytokine IL22, and bone formation has been published in a mouse model system in which IL23 is overexpressed (Sherlock et al. (2012) Nat. Med. 18:1069-76).

B-lymphocyte activating factor (FAFL (BAFF)) is a cytokine that belongs to the tumor necrosis factor (TNF) ligand superfamily and acts as a ligand for the FAVLR (BAFF-R (BR3)) receptors, TACI (transmembrane activator and calcium modulator, and the cyclophilin ligand interactor, TACI) and ASWL (B-lymphocyte maturation antigen, BCMA). Interaction between FAVL (BAFF) and its receptors causes the signals necessary for the formation and maintenance of B cells, which, in turn, synthesize immunoglobulins in response to the penetration of a foreign substance. Appropriate levels of FAFL (BAFF) in a patient help maintain normal levels of immunity, while inappropriate levels can lead to immunodeficiency, and excessive levels can lead to abnormally high levels of antibody production. When a patient shows an autoimmune reaction, they produce antibodies against tissues or organs in their own body. Autoimmune diseases, including lupus erythematosus and rheumatoid arthritis, are the result of excessive levels of BAFF in the body. Thus, it is important to modulate the production of FAFL (BAFF) for the treatment of patients suffering from such diseases.

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FAVL (BAFF) can exist in three forms: membrane-bound (msFAVL (mbBAFF)), trimeric soluble FAVL (rFAVL (sBAFF)) and a multimeric form consisting of 60 FAVL monomers (FALV 60mer (BAFF 60mer)). The relative importance of the various forms of BAFF in normal and disease physiology is not well understood. As noted, FAVL (BAFF) binds to three receptors: FAVLR (BAFFR (BR3)), TACI (TACI) and ASVL (BCMA). Proliferation-inducing ligand (IPL (APRIL)), a related member of the TNF receptor ligand family, has been shown to bind with high affinity to TACI (TACI) and ASVL (BCMA). In contrast to the high affinity IPL (APRIL):ASVL (BCMA) interaction, the FAFF (BAFF):ASVL (BCMA) interaction has a low affinity (1-2 μM) and is not believed to play an important role in vivo (Bossen and Schneider, 2006).

Soluble FAVL (sBAFF) is expressed at high levels in individuals with systemic lupus erythematosus (SLE) and in inflamed target organs such as the kidney. Soluble FAVL (sBAFF) is a critical factor for B cell homeostasis and survival (Kalled et al., 2005; Mackay et al., 2003; Smith and Cancro, 2003; Patke et al., 2004). The formation of autoantibodies FAFL (BAFF)-dependent B lymphocytes leads to the appearance of glomerular IR deposits (IR - immune complex), initially on the glomerular basement membrane (GBM (GBM)), mesangium and interstitial tissue in proximal tubular epithelial cells (PTEC (PTEC)) . Such IR deposits lead to complement fixation and neutrophil activation, leading to local damage to the kidney. Inflammatory mediators (eg, IL6, IL8, MCP-1) produced by damaged renal cells (MK (mesangial cells, MS), PTEC (RTEC), renal fibroblasts, endothelial cells) maintain the inflammatory cycle by increasing immune cell infiltration (eg, B-lymphocytes, T-lymphocytes, dendritic cells, neutrophils and macrophages).

The anti-FAFL (BAFF) monoclonal antibody belimumab (Benlysta®) has a proven ability to reduce the formation of autoantibodies and is of significant benefit to patients with systemic lupus erythematosus (SLE). However, the efficacy of belimumab in patients with SLE is moderate at best, and there is significant room for improvement (Furie et al., 2011). Therefore, there remains a need for new compositions with improved efficacy for the treatment and prevention of autoimmune or inflammatory diseases. In addition, the identification of more effective methods of treatment should provide the introduction of reduced doses, as well as lower costs associated with treatment.

Brief description of the essence of the invention

This document provides compounds specific to FAVL (BAFF) and IL23A, compositions containing such compounds, as well as methods for their use and their preparation.

Aspects of the disclosure of the invention relate to a compound containing a first polypeptide and a second polypeptide, wherein:

(A) said first polypeptide contains:

(i) a first immunoglobulin (VL1) light chain variable domain specific for the first target protein;

(ii) a second immunoglobulin heavy chain variable domain (VH2) specific for a second target protein; and (iii) a hinge region, a heavy chain constant region 2 (CH2), and a heavy chain constant region 3 (CH3); and (B) said second polypeptide contains:

(i) a second immunoglobulin (VL2) light chain variable domain specific for said second target protein;

(ii) a first immunoglobulin (VH1) heavy chain variable domain specific for said first target protein; and:

a) said VL1 and VH1 combine to form an attachment site that binds said first target protein;

b) said VL2 and VH2 combine to form an attachment site that binds said second target protein;

c) said heavy chain constant region 2 (CH2) contains tyrosine at position 252, threonine at position 254 and glutamic acid at position 256, numbered according to the EC index, as in Kabat; And

d) said first target protein is FALV (BAFF) and said second target protein is IL-23A, or said first target protein is IL-23A and said second target protein is FALV (BAFF), and wherein :

(i) said VL1 contains SEQ ID NO: 2, said VH1 contains SEQ ID NO: 1, said VL2 contains SEQ ID NO: 4, and said VH2 contains SEQ ID NO: 3; or (ii) said VL1 contains SEQ ID NO: 4, said VH1 contains SEQ ID NO: 3, said VL2 contains SEQ ID NO: 2, and said VH2 contains SEQ ID NO: 1; or (iii) said VL1 contains SEQ ID NO: 85, said VH1 contains SEQ ID NO: 84, indicated

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VL2 contains SEQ ID NO: 4 and said VH2 contains SEQ ID NO: 4; or (iv) said VL1 contains SEQ ID NO: 4, said VH1 contains SEQ ID NO: 3, indicated

VL2 contains SEQ ID NO: 85 and said VH2 contains SEQ ID NO: 84; or (v) said VL1 contains SEQ ID NO: 87, said VH1 contains SEQ ID NO: 86, indicated

VL2 contains SEQ ID NO: 4 and said VH2 contains SEQ ID NO: 3; or (vi) said VL1 contains SEQ ID NO: 4, said VH1 contains SEQ ID NO: 3, said VL2 contains SEQ ID NO: 87, and said VH2 contains SEQ ID NO: 86; or (vii) said VL1 contains SEQ ID NO: 89, said VH1 contains SEQ ID NO: 88, said VL2 contains SEQ ID NO: 4, and said VH2 contains SEQ ID NO: 3; or (viii) said VL1 has SEQ ID NO: 4, said VH1 has SEQ ID NO: 3, said VL2 has SEQ ID NO: 89, and said VH2 has SEQ ID NO: 88; or (ix) said VL1 contains SEQ ID NO: 91, said VH1 contains SEQ ID NO: 90, said VL2 contains SEQ ID NO: 4, and said VH2 contains SEQ ID NO: 3; or (x) said VL1 contains SEQ ID NO: 4, said VH1 contains SEQ ID NO: 3, said VL2 contains SEQ ID NO: 91, and said VH2 contains SEQ ID NO: 90.

In some embodiments, said first polypeptide further comprises a first linker between said VL1 and said VH2, and said second polypeptide further comprises a second linker between said VL2 and said VH1. In some embodiments, said first linker or said second linker comprises the amino acid sequence GGGSGGGG (SEQ ID NO: 69). In some embodiments, said first linker and said second linker comprise the amino acid sequence GGGSGGGG (SEQ ID NO: 69). In some embodiments, said first polypeptide further comprises a third linker between said VH2 or said VL1 and the hinge region, and said second polypeptide further comprises a fourth linker after said VH1 (at its C-terminus) or said VL2 (at its C-terminus). In some embodiments, said third linker of said first polypeptide contains the amino acid sequence GGCGGGEVAACEKEVAALEKEVAALEKEVAALEK (SEQ ID NO: 82) and said fourth linker of said second polypeptide contains the amino acid sequence GGCGGGKVAACKEKVAALKEKVAALKEKVAALKE (SEQ ID NO: 83). In other embodiments, said third linker of said first polypeptide comprises the amino acid sequence GGCGGGKVAACKEKVAALKEKVAALKEKVAALKE (SEQ ID NO: 83) and said fourth linker of said second polypeptide comprises the amino acid sequence GGCGGGEVAACEKEVAALEKEVAALEKEVAALEK (SEQ ID NO: 82). In some embodiments, said third linker of said first polypeptide comprises the amino acid sequence GGCGGGEVAACEKEVAALEKEVAALEKEVAALEK (SEQ ID NO: 82) or the amino acid sequence GGCGGGKVAACKEKVAALKEKVAALKEKVAALKE (SEQ ID NO: 83). In other embodiments, said fourth linker of said second polypeptide comprises the amino acid sequence GGCGGGEVAACEKEVAALEKEVAALEKEVAALEK (SEQ ID NO: 82) or the amino acid sequence GGCGGGKVAACKEKVAALKEKVAALKEKVAALKE (SEQ ID NO: 83). In some embodiments, said third linker contains the amino acid sequence VEPKSC (SEQ ID NO: 72) or the amino acid sequence FNRGEC (SEQ ID NO: 71). In some embodiments, said fourth linker contains the amino acid sequence FNRGEC (SEQ ID NO: 71) or the amino acid sequence VEPKSC (SEQ ID NO: 72). In some embodiments, said third linker contains the amino acid sequence VEPKSC (SEQ ID NO: 72) and said fourth linker contains the amino acid sequence FNRGEC (SEQ ID NO: 71).

In some embodiments, said first polypeptide further comprises a heavy chain constant region 1 (CH1) domain between said VH2 or said VL1 (depending on configuration) and the hinge region, and said second polypeptide further comprises a light chain constant region (CL) domain at C -end VH1 or VL2 (depending on the configuration), while the specified CL and the specified CH1 linked together through a disulfide bond to form a C1 domain. In some embodiments, said first linker (between said VL1 and said VH2) or said second linker (between said VL2 and said VH1) contains the amino acid sequence GGGSGGGG (SEQ ID NO: 69). In some embodiments, said first linker and said second linker comprise the amino acid sequence GGGSGGGG (SEQ ID NO: 69). In some embodiments, said first polypeptide further comprises a third linker between said VH2 or said VL1 (depending on configuration) and said CH1, and said second polypeptide further comprises a fourth linker between said VH1 or said VL2 (depending on configuration) and said CL . In some embodiments, said third linker or said fourth linker comprises the amino acid sequence LGGGSG (SEQ ID NO: 70). In some embodiments, said third linker and said fourth linker comprise an amino acid

- 3 040031 sequence LGGGSG (SEQ ID NO: 70). In some embodiments, said third linker and/or said fourth linker contains an optional cysteine residue. In some embodiments, said third linker and/or said fourth linker comprise the amino acid sequence GGCGGG (SEQ ID NO: 135) or LGGCGGGS (SEQ ID NO: 136).

In some embodiments, said heavy chain constant region 2 (CH2) contains alanine at positions 234 and alanine at position 235, numbered according to the Kabat EC index for a representative antibody.

In some embodiments, the amino acid sequence of said hinge region, said heavy chain constant region 2 (CH2), or said heavy chain constant region 3 (CH3) is derived from IgG1 or from IgG4. In some embodiments, said hinge region contains the amino acid sequence EPKSCDKTHTCPPCP (SEQ ID NO: 76). The hinge region of SEQ ID NO: 76 is present, for example, in the polypeptide of SEQ ID NO: 5. In other embodiments, the hinge region contains the amino acid sequence LEPKSSDKTHTCPPCP (SEQ ID NO: 130). The hinge region of SEQ ID NO: 130 is present, for example, in the polypeptide of SEQ ID NO: 9. In other embodiments, the hinge region contains the amino acid sequence ESKYGPPCPPCP (SEQ ID NO: 134). The hinge region of SEQ ID NO: 134 is present, for example, in the polypeptide of SEQ ID NO: 13.

In some embodiments, said compound comprises two said first polypeptides and two said second polypeptides, said two first polypeptides being linked together via at least one disulfide bond. In some embodiments, said compound comprises two of said first polypeptides and two of said second polypeptides, wherein said two first polypeptides are linked together via at least one disulfide bond, and wherein each of said first polypeptide is linked to one of said second polypeptides via at least one disulfide bond.

In some embodiments of the invention:

(i) said first polypeptide contains the amino acid sequence of SEQ ID NO: 5, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 6; or (ii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 7, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 8; or (iii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 9, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 10; or (iv) said first polypeptide contains the amino acid sequence of SEQ ID NO: 11 and said second polypeptide contains the amino acid sequence of SEQ ID NO: 12; or (v) said first polypeptide contains the amino acid sequence of SEQ ID NO: 13, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 14; or (vi) said first polypeptide contains the amino acid sequence of SEQ ID NO: 15, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 16; or (vii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 17, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 18; or (viii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 19, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 20; or (ix) said first polypeptide contains the amino acid sequence of SEQ ID NO: 21, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 22; or (x) said first polypeptide contains the amino acid sequence of SEQ ID NO: 23, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 24; or (xi) said first polypeptide contains the amino acid sequence of SEQ ID NO: 25, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 26; or (xii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 27, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 28; or (xiii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 29, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 30; or (xiv) said first polypeptide contains the amino acid sequence of SEQ ID NO: 31, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 32; or (xv) said first polypeptide contains the amino acid sequence of SEQ ID NO: 33, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 34; or (xvi) said first polypeptide contains the amino acid sequence of SEQ ID NO: 35 and said second polypeptide contains the amino acid sequence of SEQ ID NO: 36; or (xvii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 37, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 38; or (xviii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 39, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 40; or (xix) said first polypeptide contains the amino acid sequence of SEQ ID NO: 41,

- 4 040031 and the specified second polypeptide contains the amino acid sequence of SEQ ID NO: 42; or (xx) said first polypeptide contains the amino acid sequence of SEQ ID NO: 43, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 44; or (xxi) said first polypeptide contains the amino acid sequence of SEQ ID NO: 45, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 46; or (xxii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 47, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 48; or (xxiii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 49, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 50; or (xxiv) said first polypeptide contains the amino acid sequence of SEQ ID NO: 51, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 52; or (xxv) said first polypeptide contains the amino acid sequence of SEQ ID NO: 53, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 54; or (xxvi) said first polypeptide contains the amino acid sequence of SEQ ID NO: 55, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 56; or (xxvii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 57, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 58; or (xxviii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 59, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 60; or (xxix) said first polypeptide contains the amino acid sequence of SEQ ID NO: 61, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 62; or (xxx) said first polypeptide contains the amino acid sequence of SEQ ID NO: 63, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 64; or (xxxi) said first polypeptide contains the amino acid sequence of SEQ ID NO: 65, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 66; or (xxxii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 67 and said second polypeptide contains the amino acid sequence of SEQ ID NO: 68.

In some embodiments, in which said compound contains two said first polypeptides and two said second polypeptides, said two first polypeptides are linked together via at least one disulfide bond.

In some embodiments, said compound comprises two of said first polypeptides and two of said second polypeptides, wherein the hinge, CH2 and CH3 of one of the first polypeptides is combined with the hinge, CH2 and CH3 of the other first polypeptide to form a tetravalent molecule. In some embodiments, said compound contains two of said first polypeptides and two of said second polypeptides, wherein each of said first polypeptides contains CH1, hinge, CH2, and CH3, and each of said second polypeptides contains CL, and wherein said hinge, CH2, and CH3 of one of the first polypeptides combines with the hinge, CH2 and CH3 of the other of the first polypeptides, and the CH1 of each of said first polypeptides combines with the CL of one of the said second polypeptides to form a tetravalent molecule (e.g., monomer, monomeric antibody, as described in the Examples section) (e.g. , compounds E and V). In some embodiments, said compound contains two of said first polypeptides and two of said second polypeptides, wherein each of said first polypeptides comprises a third linker, a hinge, CH2 and CH3, and each of said second polypeptides comprises a fourth linker, and wherein said hinge, CH2 and The CH3 of one of the first polypeptides is combined with the hinge, CH2 and CH3 of the other of the first polypeptides, and the third linker of each of the said first polypeptides is combined with the fourth linker of one of the said second polypeptides to form a tetravalent molecule (e.g., monomer, monomeric antibody, as described in section Examples) (eg compounds U and T).

Other aspects of the disclosure relate to a first compound that competes with a second compound for binding to IL-23A and FAVL (BAFF), said first compound comprising a third polypeptide and a fourth polypeptide, wherein:

(A) said third polypeptide contains:

(i) a first immunoglobulin (VL1) light chain variable domain specific for the first target protein;

(ii) a second immunoglobulin heavy chain variable domain (VH2) specific for a second target protein; and (iii) a hinge region, a heavy chain constant region 2 (CH2), and a heavy chain constant region 3 (CH3); and (B) said fourth polypeptide contains:

(i) a second immunoglobulin (VL2) light chain variable domain specific for said second target protein;

(ii) a first immunoglobulin (VH1) heavy chain variable domain specific for said first target protein; and wherein:

- 5 040031 (i) said VL1 and VH1 combine to form an attachment site that binds said first target protein;

(ii) said VL2 and VH2 combine to form an attachment site that binds said second target protein;

(iii) said first target protein is FALV (BAFF) and said second target protein is IL-23A, or said first target protein is IL23A and said second target protein is FALV (BAFF), and wherein said second compound contains a first polypeptide and a second polypeptide, wherein:

(i) said first polypeptide contains the amino acid sequence of SEQ ID NO: 5, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 6; or (ii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 7, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 8; or (iii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 9, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 10; or (iv) said first polypeptide contains the amino acid sequence of SEQ ID NO: 11 and said second polypeptide contains the amino acid sequence of SEQ ID NO: 12; or (v) said first polypeptide contains the amino acid sequence of SEQ ID NO: 13, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 14; or (vi) said first polypeptide contains the amino acid sequence of SEQ ID NO: 15, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 16; or (vii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 17, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 18; or (viii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 19, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 20; or (ix) said first polypeptide contains the amino acid sequence of SEQ ID NO: 21, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 22; or (x) said first polypeptide contains the amino acid sequence of SEQ ID NO: 23, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 24; or (xi) said first polypeptide contains the amino acid sequence of SEQ ID NO: 25, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 26; or (xii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 27, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 28; or (xiii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 29, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 30; or (xiv) said first polypeptide contains the amino acid sequence of SEQ ID NO: 31, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 32; or (xv) said first polypeptide contains the amino acid sequence of SEQ ID NO: 33, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 34; or (xvi) said first polypeptide contains the amino acid sequence of SEQ ID NO: 35, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 36; or (xvii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 37, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 38; or (xviii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 39, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 40; or (xix) said first polypeptide contains the amino acid sequence of SEQ ID NO: 41, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 42; or (xx) said first polypeptide contains the amino acid sequence of SEQ ID NO: 43, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 44; or (xxi) said first polypeptide contains the amino acid sequence of SEQ ID NO: 45, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 46; or (xxii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 47, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 48; or (xxiii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 49, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 50; or (xxiv) said first polypeptide contains the amino acid sequence of SEQ ID NO: 51, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 52; or (xxv) said first polypeptide contains the amino acid sequence of SEQ ID NO: 53, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 54; or (xxvi) said first polypeptide contains the amino acid sequence of SEQ ID NO: 55, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 56; or (xxvii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 57, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 58; or

- 6 040031 (xxviii) said first polypeptide contains the amino acid sequence of SEQ ID NO:

59, and the specified second polypeptide contains the amino acid sequence of SEQ ID NO: 60; or (xxix) said first polypeptide contains the amino acid sequence of SEQ ID NO:

61, and the specified second polypeptide contains the amino acid sequence of SEQ ID NO: 62; or (xxx) said first polypeptide contains the amino acid sequence of SEQ ID NO: 63, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 64; or (xxxi) said first polypeptide contains the amino acid sequence of SEQ ID NO: 65, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 66; or (xxxii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 67 and said second polypeptide contains the amino acid sequence of SEQ ID NO: 68.

Other aspects of the disclosure relate to a compound containing two first polypeptides and two second polypeptides, wherein said two first polypeptides are linked together via at least one disulfide bond, and each said first polypeptide is linked to one said second polypeptide via at least one a disulfide bond, wherein each of said first polypeptides contains:

(i) a first immunoglobulin (VL1) light chain variable domain specific for the first target protein;

(ii) a second immunoglobulin heavy chain variable domain (VH2) specific for a second target protein;

(iii) heavy chain constant region 1 (CH1), hinge region, heavy chain constant region 2 (CH2), and heavy chain constant region 3 (CH3); and wherein each of said second polypeptides contains:

(i) a second immunoglobulin (VL2) light chain variable domain specific for said second target protein;

(ii) a first immunoglobulin (VH1) heavy chain variable domain specific for said first target protein;

(iii) a light chain constant region (CL) domain;

wherein the hinge, CH2 and CH3 of one of the first polypeptides combines with the hinge, CH2 and CH3 of another of the first polypeptides, and the CH1 of each of said first polypeptides combines with the CL of each of the second polypeptides to form a tetravalent molecule; wherein:

a) said VL1 and VH1 combine to form an attachment site that binds said first target protein;

b) said VL2 and VH2 combine to form an attachment site that binds said second target protein;

c) said heavy chain constant region 2 (CH2) contains tyrosine at position 252, threonine at position 254 and glutamic acid at position 256, numbered according to the EC index, as in Kabat; And

d) said first target protein is FALV (BAFF) and said second target protein is IL-23A, or said first target protein is IL-23A and said second target protein is FALV (BAFF), and :

(i) said VL1 contains SEQ ID NO: 2, said VH1 contains SEQ ID NO: 1, said VL2 contains SEQ ID NO: 4, and said VH2 contains SEQ ID NO: 3; or (ii) said VL1 contains SEQ ID NO: 4, said VH1 contains SEQ ID NO: 3, said VL2 contains SEQ ID NO: 2, and said VH2 contains SEQ ID NO: 1; or (iii) said VL1 contains SEQ ID NO: 85, said VH1 contains SEQ ID NO: 84, said VL2 contains SEQ ID NO: 4, and said VH2 contains SEQ ID NO: 4; or (iv) said VL1 has SEQ ID NO: 4, said VH1 has SEQ ID NO: 3, said VL2 has SEQ ID NO: 85, and said VH2 has SEQ ID NO: 84; or (v) said VL1 contains SEQ ID NO: 87, said VH1 contains SEQ ID NO: 86, said VL2 contains SEQ ID NO: 4, and said VH2 contains SEQ ID NO: 3; or (vi) said VL1 contains SEQ ID NO: 4, said VH1 contains SEQ ID NO: 3, said VL2 contains SEQ ID NO: 87, and said VH2 contains SEQ ID NO: 86; or (vii) said VL1 contains SEQ ID NO: 89, said VH1 contains SEQ ID NO: 88, said VL2 contains SEQ ID NO: 4, and said VH2 contains SEQ ID NO: 3; or (viii) said VL1 has SEQ ID NO: 4, said VH1 has SEQ ID NO: 3, said VL2 has SEQ ID NO: 89, and said VH2 has SEQ ID NO: 88; or (ix) said VL1 contains SEQ ID NO: 91, said VH1 contains SEQ ID NO: 90, said VL2 contains SEQ ID NO: 4, and said VH2 contains SEQ ID NO: 3; or (x) said VL1 contains SEQ ID NO: 4, said VH1 contains SEQ ID NO: 3, said VL2 contains SEQ ID NO: 91, and said VH2 contains SEQ ID NO: 90.

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In some embodiments relating to the above aspect, each of said first polypeptides further comprises a first linker between said VL1 and said VH2, and each of said second polypeptides further comprises a second linker between said VL2 and said VH1. In some embodiments, said first linker or said second linker comprises the amino acid sequence GGGSGGGG (SEQ ID NO: 69). In some embodiments, said first linker and said second linker comprise the amino acid sequence GGGSGGGG (SEQ ID NO: 69). In some embodiments, each of said first polypeptides further comprises a third linker between said VH2 (or said VL1) and said CH1, and each of said second polypeptides further comprises a fourth linker between said VH1 (or said VL2) and said CL. In some embodiments, said third linker or said fourth linker comprises the amino acid sequence LGGGSG (SEQ ID NO: 70). In some embodiments, said third linker and said fourth linker comprise the amino acid sequence LGGGSG (SEQ ID NO: 70). In some embodiments, said third linker and/or said fourth linker contains an optional cysteine residue. In some embodiments, said third linker and/or said fourth linker comprise the amino acid sequence GGCGGG (SEQ ID NO: 135) or LGGCGGGS (SEQ ID NO: 136). In some embodiments, said heavy chain constant region 2 (CH2) contains alanine at positions 234 and alanine at position 235, numbered according to the EC index, as in Kabat. In some embodiments, the amino acid sequence of said hinge region, said heavy chain constant region 2 (CH2), or said heavy chain constant region 3 (CH3) is derived from IgG1 or from IgG4. In some embodiments, said hinge region comprises the amino acid sequence EPKSCDKTHTCPPCP (SEQ ID NO: 76), the amino acid sequence LEPKSSDKTHTCPPCP (SEQ ID NO: 130), or the amino acid sequence ESKYGPPCPPCP (SEQ ID NO: 134).

In some embodiments of the invention:

(i) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 5, and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 6; or (ii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 7, and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 8; or (iii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 13, and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 14; or (iv) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 15, and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 16; or (v) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 21 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 22; or (vi) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 25, and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 26; or (vii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 29, and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 30; or (viii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 33, and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 34; or (ix) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 37, and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 38; or (x) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 41 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 42; or (xi) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 45 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 46; or (xii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 49, and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 50; or

- 8 040031 (xiii) each of said first polypeptides contains the amino acid sequence

SEQ ID NO: 53, and each of these second polypeptides contains the amino acid sequence of SEQ ID NO: 54; or (xiv) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 55, and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 56; or (xv) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 57, and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 58; or (xvi) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 59, and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 60; or (xvii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 61, and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 62; or (xviii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 63, and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 64; or (xix) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 65, and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 66; or (xx) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 67 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 68.

Other aspects of the disclosure of the invention relate to a compound containing two first polypeptides and two second polypeptides; wherein said two first polypeptides are linked together via at least one disulfide bond, and wherein each said first polypeptide is linked to one said second polypeptide via at least one disulfide bond; and wherein (i) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 5, and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 6; or (ii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 7, and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 8; or (iii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 13, and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 14; or (iv) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 15, and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 16; or (v) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 21 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 22; or (vi) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 25, and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 26; or (vii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 29, and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 30; or (viii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 33, and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 34; or (ix) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 37, and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 38; or (x) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 41 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 42; or (xi) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 45 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 46; or

- 9 040031 (xii) each of said first polypeptides contains the amino acid sequence

SEQ ID NO: 49, and each of these second polypeptides contains the amino acid sequence of SEQ ID NO: 50; or (xiii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 53, and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 54; or (xiv) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 55, and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 56; or (xv) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 57, and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 58; or (xvi) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 59, and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 60; or (xvii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 61, and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 62; or (xviii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 63, and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 64; or (xix) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 65, and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 66; or (xx) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 67 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 68.

Other aspects of the disclosure relate to a compound containing two first polypeptides and two second polypeptides, wherein said two first polypeptides are linked together via at least one disulfide bond, and wherein each said first polypeptide is linked to one said second polypeptide via at least one a disulfide bond, wherein each of said first polypeptides contains:

(iv) a first immunoglobulin (VL1) light chain variable domain specific for the first target protein; (v) a second immunoglobulin heavy chain variable domain (VH2) specific for a second target protein; (vi) hinge region, heavy chain constant region 2 (CH2), and heavy chain constant region 3 (CH3); and wherein each of said second polypeptides contains:

(i) a second immunoglobulin (VL2) light chain variable domain specific for said second target protein; and (ii) a first immunoglobulin heavy chain variable domain (VH1) specific for said first target protein, wherein the hinge, CH2 and CH3 of one of the first polypeptides is combined with the hinge, CH2 and CH3 of another of the first polypeptides, and; wherein

e) said VL1 and VH1 combine to form an attachment site that binds said first target protein;

f) said VL2 and VH2 combine to form an attachment site that binds said second target protein;

g) said heavy chain constant region 2 (CH2) contains tyrosine at position 252, threonine at position 254 and glutamic acid at position 256, numbered according to the EC index, as in Kabat; And

h) said first target protein is FALV (BAFF) and said second target protein is IL-23A, or said first target protein is IL-23A and said second target protein is FAFL (BAFF), and wherein :

(i) said VL1 contains SEQ ID NO: 2, said VH1 contains SEQ ID NO: 1, said VL2 contains SEQ ID NO: 4, and said VH2 contains SEQ ID NO: 3; or (ii) said VL1 contains SEQ ID NO: 4, said VH1 contains SEQ ID NO: 3, said VL2 contains SEQ ID NO: 2, and said VH2 contains SEQ ID NO: 1; or (iii) said VL1 contains SEQ ID NO: 85, said VH1 contains SEQ ID NO: 84, said VL2 contains SEQ ID NO: 4, and said VH2 contains SEQ ID NO: 4; or (iv) said VL1 contains SEQ ID NO: 4, said VH1 contains SEQ ID NO: 3, said VL2 contains SEQ ID NO: 85, and said VH2 contains SEQ ID NO: 84; or

- 10 040031 (v) specified VL1 contains SEQ ID NO: 87, specified VH1 contains SEQ ID NO: 86, specified

VL2 contains SEQ ID NO: 4 and said VH2 contains SEQ ID NO: 3; or (vi) said VL1 contains SEQ ID NO: 4, said VH1 contains SEQ ID NO: 3, indicated

VL2 contains SEQ ID NO: 87 and said VH2 contains SEQ ID NO: 86; or (vii) said VL1 contains SEQ ID NO: 89, said VH1 contains SEQ ID NO: 88, said VL2 contains SEQ ID NO: 4, and said VH2 contains SEQ ID NO: 3; or (viii) said VL1 has SEQ ID NO: 4, said VH1 has SEQ ID NO: 3, said VL2 has SEQ ID NO: 89, and said VH2 has SEQ ID NO: 88; or (ix) said VL1 contains SEQ ID NO: 91, said VH1 contains SEQ ID NO: 90, said VL2 contains SEQ ID NO: 4, and said VH2 contains SEQ ID NO: 3; or (x) said VL1 contains SEQ ID NO: 4, said VH1 contains SEQ ID NO: 3, said VL2 contains SEQ ID NO: 91, and said VH2 contains SEQ ID NO: 90.

In some embodiments relating to the above aspect, each of said first polypeptides further comprises a first linker between said VL1 and said VH2, and each of said second polypeptides further comprises a second linker between said VL2 and said VH1. In some embodiments, said first linker or said second linker comprises the amino acid sequence GGGSGGGG (SEQ ID NO: 69). In some embodiments, said first linker and said second linker comprise the amino acid sequence GGGSGGGG (SEQ ID NO: 69). In some embodiments, each of said first polypeptides further comprises a third linker between said VH2 or said VL1 and said hinge region, and each of said second polypeptides further comprises a fourth linker at the C-terminus of said VH1 or said VL2. In some embodiments, said third linker of said first polypeptide contains the amino acid sequence GGCGGGEVAACEKEVAALEKEVAALEKEVAALEK (SEQ ID NO: 82) and said fourth linker of said second polypeptide contains the amino acid sequence GGCGGGKVAACKEKVAALKEKVAALKEKVAALKE (SEQ ID NO: 83). In other embodiments, said third linker of said first polypeptide comprises the amino acid sequence GGCGGGKVAACKEKVAALKEKVAALKEKVAALKE (SEQ ID NO: 83) and said fourth linker of said second polypeptide comprises the amino acid sequence GGCGGGEVAACEKEVAALEKEVAALEKEVAALEK (SEQ ID NO: 82). In some embodiments, said third linker of said first polypeptide comprises the amino acid sequence GGCGGGEVAACEKEVAALEKEVAALEKEVAALEK (SEQ ID NO: 82) or the amino acid sequence GGCGGGKVAACKEKVAALKEKVAALKEKVAALKE (SEQ ID NO: 83). In other embodiments, said fourth linker of said second polypeptide comprises the amino acid sequence GGCGGGEVAACEKEVAALEKEVAALEKEVAALEK (SEQ ID NO: 82) or the amino acid sequence GGCGGGKVAACKEKVAALKEKVAALKEKVAALKE (SEQ ID NO: 83). In some embodiments, said third linker contains the amino acid sequence VEPKSC (SEQ ID NO: 72) or the amino acid sequence FNRGEC (SEQ ID NO: 71). In some embodiments, said fourth linker contains the amino acid sequence FNRGEC (SEQ ID NO: 71) or the amino acid sequence VEPKSC (SEQ ID NO: 72). In some embodiments, said third linker contains the amino acid sequence VEPKSC (SEQ ID NO: 72) and said fourth linker contains the amino acid sequence FNRGEC (SEQ ID NO: 71). In some embodiments, said heavy chain constant region 2 (CH2) contains alanine at positions 234 and alanine at position 235, numbered according to the EC index, as in Kabat. In some embodiments, the amino acid sequence of said hinge region, said heavy chain constant region 2 (CH2), or said heavy chain constant region 3 (CH3) is derived from IgG1 or from IgG4. In some embodiments, said hinge region comprises the amino acid sequence EPKSCDKTHTCPPCP (SEQ ID NO: 76), the amino acid sequence LEPKSSDKTHTCPPCP (SEQ ID NO: 130), or the amino acid sequence ESKYGPPCPPCP (SEQ ID NO: 134). In some embodiments:

(i) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 9, and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 10; or (ii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 11, and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 12; or (iii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 17, and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 18; or (iv) each of said first polypeptides contains the amino acid sequence

- 11 040031

SEQ ID NO: 19, and each of these second polypeptides contains the amino acid sequence of SEQ ID NO: 20; or (v) each of said first polypeptides contains the amino acid sequence

SEQ ID NO: 23, and each of these second polypeptides contains the amino acid sequence of SEQ ID NO: 24; or (vi) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 27 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 28; or (vii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 31 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 32; or (viii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 35, and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 36; or (ix) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 39, and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 40; or (x) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 43, and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 44; or (xi) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 47 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 48; or (xii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 51 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 52.

However, other aspects of the disclosure of the invention relate to a pharmaceutical composition containing the compound described herein, such as the compound described above.

Other aspects of the invention relate to a method of treating an autoimmune or inflammatory disease, comprising administering to a subject a compound described herein, such as a compound described above, or a pharmaceutical composition containing said compound.

However, other aspects of the disclosure of the invention relate to the compound described herein, such as the compound described above, for use in medicine. In some embodiments, said use relates to the treatment of an autoimmune or inflammatory disease.

Other aspects of the disclosure of the invention relate to a pharmaceutical composition containing the compound described herein, such as the compound described above, for use in medicine. In some embodiments, said use relates to the treatment of an autoimmune or inflammatory disease.

However, other aspects of the disclosure relate to a nucleic acid comprising a nucleotide sequence encoding a polypeptide described herein, such as the polypeptide described above. Other aspects of the disclosure of the invention relate to a vector containing the specified nucleic acid. In some embodiments, the vector contains a promoter operably linked to said nucleic acid. Other aspects of the disclosure of the invention relate to a cell containing the specified nucleic acid or the specified vector.

Other aspects of the invention relate to a method for producing a compound or a polypeptide as described herein, such as a polypeptide described above, comprising obtaining a cell described herein, such as a cell described above, and expressing a nucleic acid, as described herein, in the specified cell. In some embodiments, the method further comprises isolating and purifying said polypeptide or compound.

Details of one or more embodiments of the invention are set forth in the description below. Other features or advantages of the present disclosure will be apparent from the following drawings and the detailed description of several embodiments, as well as from the appended claims.

Brief description of the drawings

The following drawings form part of this specification and are included to further demonstrate certain aspects of the present disclosure, which may be better understood by reference to one or more of these drawings in conjunction with a detailed description of specific embodiments provided herein.

Fig. 1A is a drawing of an exemplary compound specific for FAVL (BAFF) and IL23A. The first polypeptide chain contains the CH3, CH2, VH ₂ (VHn) and YL1 (VLi) domains. The second polypeptide chain contains the VH ₁ (VHI) and VL2 (VLII) domains. VL ₁ and VH are specific for the first target protein (FAVL (BAFF) or IL23A), and VL2 and VH2 are specific for the second target protein (IL23A or FALV (BAFF)). The top panel shows each polypeptide chain separately. The bottom panel shows a tetravalent compound (eg, monomer, monomeric antibody as described in the Examples section) formed by combining the CH2 and CH3 domains of one first polypeptide with the CH2 and CH3 domains of another first polypeptide. The binding domains for the first and second target proteins are formed by combining VH ₁ and VL ₁ , and by combining VH2 and VL2, respectively;

fig. 1B is a drawing of another exemplary compound specific for FAVL (BAFF) and IL23A. The first polypeptide chain contains the CH3, CH2, CH1, VH ₂ (VHII) and VL ₁ (VLI) domains. The second polypeptide chain contains the CL, VH1 (VHI) and VL2 (VLII) domains. VL1 and VH1 are specific for the first target protein (FALV (BAFF) or IL23A), and VL2 and VH2 are specific for the second target protein (IL23A or FAFL (BAFF)). The top panel shows each polypeptide chain separately. The bottom panel shows a tetravalent compound (eg, monomer, monomeric antibody as described in the Examples section) formed by combining the CH2 and CH3 domains of one first polypeptide with the CH2 and CH3 domains of another first polypeptide. The binding domains for the first and second target proteins are formed by combining VH ₁ and VL ₁ , and by combining VH2 and VL2, respectively. The connection is further combined through interactions between the CL and CH1 domains;

fig. 2 is a graph showing serum concentrations (mean ± CO) for Compound A (solid squares), Compound B (open squares), Compound C (solid triangles), and Compound D (open triangles) in male cynomolgus monkeys after a single intravenous dose of 1 u /kg, as described in example 10;

fig. 3 shows the predicted Compound B concentration versus time curve in human serum following a 100 mg subcutaneous dose given once every two weeks as described in Example 11. The dotted line represents the target C _min (30 nM);

fig. 4 is a graphical summary of percentage monomer versus Compound B concentration as described in Example 14;

fig. 5 shows that Compound B maintains functional activity against anti-FAVL in vivo. Mice were dosed equimolarly with either anti-FAFL (BAFF) or Compound B and exposed to human mini-circular FAFL (BAFF) to induce expansion of B cells. On day 14, spleens were harvested and analyzed by flow cytometry for the presence of murine B220+ B lymphocytes as a measure of FAFL functional blockade (BAFF);

fig. 6 shows that Compound B maintains functional activity against anti-IL23 in vivo. Compound B evaluation in IL23-induced cytokinin assay. Mice were dosed equimolarly with either anti-IL23 or Compound B and exposed to human IL23 twice to induce ear inflammation. Twenty-four hours after the final injection, the ears were harvested and analyzed for mouse IL17A and mouse IL22 as a measure of functional blockade of IL23.

Detailed Description of the Invention

This document describes compounds that bind to both FAFL (BAFF) and IL23A (also called IL23p19 or IL23A). To date, there are no compounds approved for use in medicine that target both FAVL (BAFF) and IL23A. There are limited studies with the simultaneous neutralization of two/more key inflammatory mediators using a biotherapeutic approach, and these studies have not shown an improvement in clinical outcomes that have been evaluated in rheumatoid arthritis (RA). In addition, such combinations may increase side effects such as the risk of infection (see, for example, Genovese M.C., Cohen S., Moreland L., Lium D., Robbins S. et al. (2004). Arth. Rheum 50, 1412-9; Genovese M. C., Cohen S., Moreland L., Lium D., Robbins S., et al. (2004) Arth. Rheum 50, 1412-9; and Weinblatt M., Schiff M., Goldman A. Kremer J, Luggen M. et al (2007) Ann Rheum Dis 66, 228-34). In addition, such bispecific compounds are difficult to design due to solubility (eg, aggregation) and stability (eg, poor pharmacokinetics) problems.

Surprisingly, some of the compounds described herein that bind to FAFL (BAFF) and IL-23A have been found to have similar or improved properties compared to individual antibodies that target either IL-23A or FAFL (BAFF ). Some compounds have also been found to have suitable pharmacokinetics and are soluble in suitable dosage ranges. In addition, in some embodiments, there are advantages of single administration over serial administration of a single dose in terms of side effects of individual therapy, and a lower dose. In addition, in some embodiments, the CMC (Connection Map for Compounds (CMC)) properties of the compounds showed that some compounds have high melting points (Tm) and low aggregation. In one aspect, one exemplary compound showed particularly low aggregation at high concentrations.

- 13 040031

The compounds described herein are believed to have one or more advantageous properties, such as reduced side effects, increased ease and safety of administration, increased half-life, increased binding affinity, or increased inhibitory activity, compared to standard antibody molecules, such as IgG or antigen-binding fragment (eg Fab).

Accordingly, aspects of the disclosure relate to compounds specific to both FAFL (BAFF) and IL-23A, as well as methods of using and preparing such compounds. In one aspect, the subject technology relates to compositions with improved efficacy for the treatment and prevention of autoimmune or inflammatory diseases such as systemic lupus erythematosus (SLE (SLE)), systemic lupus erythematosus with kidney damage/lupus nephritis (LN (LN)), associated with ANCA (anti-neutrophil cytoplasmic antibody, ANCA) vasculitis (AAV (AAV)), primary Sjogren's syndrome (PSS (pSS)), chronic graft-versus-host disease (CCVHD (cGVHD)), systemic sclerosis (SSc)), rheumatoid arthritis (RA (RA)), psoriasis (Ps (Ps)), ankylosing spondylitis (AS (AS)) and psoriatic arthritis (PA (RA)). The BAFF/IL23A dual antagonist will inhibit both the autoantibody/immune complex and IL23 cascade-mediated end-organ damage, and may provide better induction and maintenance of a clinical response than the current Standard of Care for the Treatment of SLE (SLE) and LN ( LN). Compared with co-administration of FAFL antibody (BAFF) and IL-23 antibody to simultaneously inhibit both pathways, the dual FALV (BAFF)/IL23A antagonist is more patient-friendly, which may result in improved adherence and reduced pain. The dual antagonist FAVL (BAFF)/IL23A will allow the introduction of reduced doses, as well as reduce the costs associated with treatment. In addition, a BAFF/IL23A antagonist may also be useful in the treatment of a group of diseases associated with misregulated B cells/autoantibody and IL23-mediated tissue damage, including primary Sjögren's syndrome (PSS), chronic graft-versus-host disease (CCVHD), systemic sclerosis (SS); and ANCA-associated vasculitis (AAV).

It is difficult to develop a dual-targeted therapeutic molecule that combines two pharmacological entities and maintains the functional efficacy of each component, while at the same time possessing biophysical properties suitable for large-scale production. The development of dual targeting molecules has been hampered by many in vitro and in vivo stability issues such as poor expression, aggregation, limited shelf life, poor serum stability, and poor in vivo pharmacokinetic properties (Demarest S.J., Glaser S.M. (2008). Curr. Opin Drug Discov Devel 11, 675-87).

Here we disclose a method for designing dual targeting molecules that inhibit both BAFF and IL23 function. The dual-targeting molecules of this technology have useful and unexpected properties such as high melting points (Tm), low aggregation at high concentrations, and predictive properties of human FA that allow biweekly or less frequent subcutaneous administration.

Connections.

Aspects of the disclosure relate to a compound that is specific for both FAVL (BAFF) and IL23A. An exemplary protein sequence for FAFL (BAFF) and an exemplary protein sequence for IL23A are shown below. 1

MDDSTEREQSRLTSCLKKREEMKLKECVSILPRKESPSVRSSKDGKLLAATLLLALLSCC LTVVSFYQVAALQGDLASLRAELQGHHAEKLPAGAGAPKAGLEEAPAVTAGLKIFEPP APGEGNSSQNSRNKRAVQGPEETVTQDCLQLIADSETPTIQKGSYTFVPWLLSFKRGSAL EEKENKILVKETGYFFIYGQVLYTDKTYAMGHLIQRKKVHVFGDELSLVTLFRCIQNMP ETLPNNSCYSAGIAKLEEGDELQLAIPRENAQISLDGDVTFFGALKLL(SEQ ID NO:80) >NP_057668.1 -ИЛ23А [Homo sapiens]

MLGSRAVMLLLLLPWTAQGRAVPGGSSPAWTQCQQLSQKLCTLAWSAHPLVGHMDL REEGDEETTNDVPHIQCGDGCDPQGLRDNSQFCLQRIHQGLIFYEKLLGSDIFTGEPSLLP DSPVGQLHASLLGLSQLLQPEGHHWETQQIPSLSPSQPWQRLLLRFKILRSLQAFVAVAA RVFAHGAATLSP (аминокислоты 1-19 представляют собой предсказанную сигнальную последовательность)(8Е0 ID NO:81)

In some embodiments, the compound comprises a first polypeptide and a second polypeptide. In some embodiments, the first polypeptide contains: (i) a variable domain

- 14 040031 light chain of the first immunoglobulin (VL1), specific for the first target protein; (ii) a second immunoglobulin heavy chain variable domain (VH2) specific for a second target protein; and (iii) a hinge region, a heavy chain constant region 2 (CH2), and a heavy chain constant region 3 (CH3). In some embodiments, the first polypeptide further comprises heavy chain constant region 1 (CH1). In some embodiments, the second polypeptide comprises: (i) a second immunoglobulin light chain (VL2) variable domain specific for a second target protein; (ii) a first immunoglobulin (VH1) heavy chain variable domain specific for the first target protein. In some embodiments, the second polypeptide further comprises a light chain constant region (CL).

It should be understood that the variable domains and constant domains/regions of the first polypeptide may be in any order and the variable domains and constant domains/regions (if any) of the second polypeptide may be in any order. Below are a few illustrative configurations for domains/regions on the first and second polypeptides from the N-terminus to the C-terminus.

Configuration 1 of the first polypeptide: N-VL1-VH2-hinge-CH2-CH3-C.

Configuration 2 of the first polypeptide: N-VH2-VL1-hinge-CH2-CH3-C.

Configuration 3 of the first polypeptide: N-VL1-VH2-CH1-hinge-CH2-CH3-C.

Configuration 4 of the first polypeptide: N-VH2-VL1-CH1-hinge-CH2-CH3-C.

Configuration 1 of the second polypeptide: N-VL2-VH1-C.

Configuration 2 of the second polypeptide: N-VH1-VL2-C.

Configuration 3 of the second polypeptide: N-VL2-VH1-CL-C.

Configuration 4 of the second polypeptide: N-VH1-VL2-CL-C.

Exemplary connection configurations are shown in FIG. 1A and 1B.

In some embodiments, the variable regions of the first polypeptide and the second polypeptide associate with each other to form a binding site for the first target protein and a binding site for the second target protein. In some embodiments, VL1 of the first polypeptide and VH1 of the second polypeptide associate to form an attachment site that binds the first target protein, and VL2 of the second polypeptide and VH2 of the first polypeptide associate to form an attachment site that binds the second target protein. In some embodiments, the first target protein is FAFL (BAFF) and the second target protein is IL23A. In other embodiments, the first target protein is IL23A and the second is FAFL (BAFF) target protein. It should be understood that the terms first and second are not meant to indicate the degree of importance of any of the target proteins.

Exemplary sequence combinations for each of VL1, VH1, VL2, and VH2 are shown in Table 1 below. 1, as well as in table. 2 in example 1.

Table 1. Exemplary sequence combinations for VL1, VH1, VL2 and VH2

Combination number Sequence VL1 VH1 sequence Sequence VL2 VH2 sequence 1 SEQ ID NO:2 SEQ ID NO: 1 SEQID NOG SEQID NOG 2 SEQ ID NO:4 SEQID NOG SEQID NOG SEQ ID NO: 1 3 SEQ ID NO:85 SEQ ID NO :84 SEQID NOG SEQ ID NO: 3 4 SEQ ID NO:4 SEQID NOG SEQIDNO:85 SEQ ID NO 84 5 SEQ ID NO :87 SEQ ID NO:86 SEQID NOG SEQID NOG 6 SEQ ID NO:4 SEQID NOG SEQ ID NO:87 SEQ ID NO 86 7 SEQID NO:89 SEQ ID NO:88 SEQID NOG SEQID NOG 8 SEQ ID NO:4 SEQID NOG SEQ ID NO:89 SEQ ID NO: 88 9 SEQIDNO:91 SEQID NO:90 SEQID NOG SEQID NOG 10 SEQ ID NO:4 SEQID NOG SEQ ID NOG 1 SEQID NO:90

In some embodiments, the compound comprises a VL1 sequence containing the CDR1, CDR2 and CDR3 of the first light chain and a VH1 sequence containing the CDR1, CDR2 and CDR3 of the first heavy chain, a VL2 sequence containing the CDR1, CDR2 and CDR3 of the second light chain, and a VH2 sequence, containing CDR1, CDR2 and CDR3 of the second heavy chain. In some embodiments, the CDRs are the CDRs of one or more of the VL1, VH1, VL2, and VH2 sequences shown in Table 1. 1. Exemplary light chain and heavy chain CDR sequences for the VL1, VH1, VL2, and VH2 sequences shown in Table 1. 1 shown below

SEQ ID NO: 1 CDR: GGTFNNNAIN (SEQ ID NO:92) (CDR1), GIIPMFGTAKYSQNFQG (SEQ ID NO:93) (CDR2), SRDLLLFPHHALSP (SEQ ID NO:94) (CDR3)

SEQ ID NO: 2 CDR: QGDSLRSYYAS (SEQ ID NO:95) (CDR1), GKNNRPS (SEQ ID NO:96) (CDR2), SSRDSSGNHWV (SEQ ID NO:97) (CDR3)

SEQ ID NO: 3 CDR: GYTFTDQTIH (SEQ ID NO:98) (CDR1), YIYPRDDSPKYNENFKG (SEQ ID NO:99) (CDR2), PDRSGYAWFIY (SEQ ID NO: 100) (CDR3)

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SEQ ID NO: 4 CDR: KASRDVAIAVA (SEQ ID NO: 101) (CDR1), WASTRHT (SEQ ID

NO: 102) (CDR2), HQYSSYPFT (SEQ ID NO: 103) (CDR3)

SEQ ID NO: 84 CDR: DHIFSIHWMQ (SEQ ID NO: 104) (CDR1), EIFPGSGTTDYNEKFKG (SEQ ID NO: 105) (CDR2), GAFDY (SEQ ID NO: 106) (CDR3)

SEQ ID NO: 85 CDR: RASQDIGNRLS (SEQ ID NO: 107) (CDR1 ATSSLDS (SEQ ID

NO: 108) (CDR2), LQYASSPFT (SEQ ID NO: 109) (CDR3)

SEQ ID NO: 86 CDR: DHIFSIHWMQ (SEQ ID NO: 110) (CDR1), EIFPGSGTTDYNEKFKG (SEQ ID NO: 111) (CDR2), GAFDY (SEQ ID NO: 112) (CDR3)

SEQ ID NO: 87 CDR: RASQDIGNRLN (SEQ ID NO: 112) (CDR1 ATSSLDS (SEQ ID

NO: 113) (CDR2), LQYASSPFT (SEQ ID NO: 114) (CDR3)

SEQ ID NO: 88 CDR: GYSFSTFFIH (SEQ ID NO: 115) (CDR1), RIDPNSGATKYNEKFES (SEQ ID NO: 116) (CDR2), GEDLLIRTDALDY (SEQ ID NO: 117) (CDR3)

SEQ ID NO: 89 CDR: KASQNAGIDVA (SEQ ID NO: 118) (CDR1), SKSNRYT (SEQ ID NO:

119) (CDR2), LQYRSYPRT (SEQ ID NO: 120) (CDR3)

SEQ ID NO: 90 CDR: GYSFSTFFIH (SEQ ID NO: 121) (CDR1), GRIDPNSGATKYNEKFES (SEQ ID NO: 122) (CDR2), GEDLLIRTDALDY (SEQ ID NO: 123) (CDR3)

SEQ ID NO: 91 CDR: KASQNAGIDVA (SEQ ID NO: 124) (CDR1), SKSNRYT (SEQ ID

NO: 125) (CDR2), LQYRSYPRT (SEQ ID NO: 126) (CDR3)

In some embodiments, the compound contains a VH1, VL1, VH2, and/or VL2 that contains a sequence that is at least 80% (e.g., 85, 90, 95, 96, 97, 98, or 99%) identical (residue to residue over the entire length of the sequence) of the sequence described in table. 1. The percent identity of two amino acid sequences is determined using the Karlin and Altschul Proc algorithm. Natl. Acad. sci. USA 87:2264-68, 1990, as amended in Karlin and Altschul Proc. Natl. Acad. sci. USA 90:5873-77, 1993. Such an algorithm is included in the NBLAST and XBLAST programs (version 2.0) Altschul et al. J. Mol. Biol. 215:403-10, 1990. BLAST protein searches can be performed using the XBLAST program, score 50, word length 3 to obtain amino acid sequences homologous to protein molecules of interest. Gapped BLAST can be used where there are gaps in the alignment of two sequences, as described in Altschul et al., Nucleic Acids Res. 25(17):3389-3402, 1997. When using the BLAST and Gapped BLAST programs, the default settings of the respective programs (eg, XBLAST and NBLAST) may be applied.

In some embodiments, the compound contains VH1, VL1, VH2, and/or VL2, which contains a sequence containing one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) mutations in the sequence described in table. 1. Such mutations may be conservative amino acid substitutions. As used herein, the term conservative amino acid substitution refers to an amino acid substitution that does not change the relative charge or size characteristics of the protein in which the amino acid substitution occurs. Conservative amino acid substitutions include, for example, substitutions made between amino acids among the following groups: a) M, I, L, V; b) F, Y, W; c) K, R, H; d) A, G; e) S, T; e) Q, N; and g) E, D.

The hinge, CH2, and CH3 amino acid sequences of a compound (and optionally CH1 and CL if the compound contains such regions) can be obtained from any suitable source, such as an antibody constant region, such as IgG1, IgG2, IgG3, or IgG4. The amino acid sequences of the constant regions of the light and heavy chains of an antibody are well known in the art, for example such sequences are provided in the IMGT database (www.imgt.org) or www.vbase2.org/vbstat.php., both incorporated herein by links. In addition, in some expression systems, the C-terminal lysine residue of the CH3 domain may be deleted after translation. Accordingly, the C-terminal lysine residue of the CH3 domain is an optional amino acid residue. It is in the present invention that molecules are provided that do not have a C-terminal lysine residue of the CH3 domain. Also contemplated by the present invention are such constructs containing a C-terminal lysine residue of the CH3 domain. In some embodiments, the CH2 and CH3 amino acid sequences are derived from IgG1 (eg, SEQ ID NO: 75) or IgG4 (eg, SEQ ID NO: 73). In some embodiments, CL comprises the amino acid sequence kappa CL or lambda CL. In some embodiments, the implementation of the hinge region contains the amino acid sequence EPKSCDKTHTCPPCP (SEQ ID NO: 76). The hinge region of SEQ ID NO: 76 is present, for example, in the polypeptide of SEQ ID NO: 5. In other embodiments, the hinge region contains the amino acid sequence LEPKSSDKTHTCPPCP (SEQ ID NO: 130). The hinge region of SEQ ID NO: 130 is present, for example, in the polypeptide of SEQ ID

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NO: 9. The hinge region of SEQ ID NO: 130 is also present at the start of the Fc domain of SEQ ID NO: 129. In still other embodiments, the hinge region contains the amino acid sequence ESKYGPPCPPCP (SEQ ID NO: 134). The hinge region of SEQ ID NO: 134 is present, for example, in the polypeptide of SEQ ID NO: 13. The hinge region of SEQ ID NO: 134 is also present at the beginning of the domain

Fc SEQ ID NO: 127.

In some embodiments, CH2 and/or CH3 compounds (and optionally CH1 and CL if the compound contains such regions) may contain one or more amino acid substitutions other than wild-type CH2 or CH3, for example, one or more amino acid substitutions in CH2 or CH3 IgG1 wild type, or one or more amino acid substitutions in CH2 or CH3 IgG4 wild type (SEQ ID NO: 39 is suggested as an example of IgG1 wild type). Such substitutions are known in the art (see, for example, US 7704497, US 7083784, US 6821505, US 8323962, US 6737056 and US 7416727).

In some embodiments, CH2 contains an amino acid substitution at 234, 235, 252, 254, and/or 256, numbered according to the Kabat EC index for a conventional antibody (Kabat et al. Sequences of Proteins of Immunological Interest, U.S. Department of Health and Human Services, 1991, which is hereby incorporated by reference in its entirety). It should be understood that all amino acid positions described herein are numbered according to the EC index as in Kabat for conventional antibodies. In some embodiments, CH2 contains an amino acid substitution at position 252, 254, and/or 256. In some embodiments, the amino acid at position 252 is tyrosine, phenylalanine, serine, tryptophan, or threonine. In some embodiments, the amino acid at position 254 is threonine. In some embodiments, the amino acid at position 254 is serine, arginine, glutamine, glutamic acid, or aspartic acid. In some embodiments, CH2 contains tyrosine at position 252, threonine at position 254, and glutamic acid at position 256 (referred to herein as the YTE mutant). In some embodiments, CH2 contains an amino acid substitution at position 234 and/or 235. In some embodiments, CH2 contains alanine at position 234 and alanine at position 235, also referred to herein as a KO mutant. In some embodiments, CH2 contains tyrosine at position 252, threonine at position 254, glutamic acid at position 256, alanine at position 234, and alanine at position 235, also referred to herein as the KO-YTE mutant.

In some embodiments, one or more linkers can be used to link domains/regions together on the first and/or second polypeptide. For example, the first polypeptide may contain a linker between VL1 and VH2. The first polypeptide further comprises a linker between VL1 or VH2 (depending on the configuration, as discussed above) and a hinge (eg, -VL1-linker-hinge or -VH2-linker-hinge). If the first polypeptide contains CH1, for example, the first polypeptide may contain a linker between VL1 or VH2 (depending on the configuration, as discussed above) and CH1 (for example, -VL1-linker-CH1 or -VH2-linker-CH1). In another example, the second polypeptide may contain a linker between VL2 and VH1. The second polypeptide may further comprise a linker after the VL2 or VH1 (depending on the configuration as discussed above, for example, -VL2 linker or -VH1 linker) at the C-terminus of the polypeptide chain. If the second polypeptide further comprises CL, the second polypeptide may further comprise a linker between VL2 or VH1 (depending on the configuration, as discussed above) and CL (such as in -VL2-linker-CL or -VH1-linker-CL). It should be understood that any number of linkers can be used to connect any domain or region to any other domain or region on the first polypeptide, and/or any number of linkers can be used to connect any domain or region to any other domain or region on the second polypeptide .

Any suitable linker known in the art is contemplated herein. In some embodiments, the implementation of the linker is a peptide linker. In some embodiments, the implementation of the peptide linker contains at least two amino acids, for example, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acids. In some embodiments, the peptide linker has no more than 50, 40, 30, 25, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4 , 3, or 2 amino acids in length. In some embodiments, the peptide linker is between 2 and 50, 2 and 40, 2 and 30, 2 and 20, 2 and 10, 2 and 9, 2 and 8, 2 and 7, or 2 and 6 amino acids in length. In some embodiments, the peptide linker comprises the amino acid sequence GGGSGGGG (SEQ ID NO: 69), LGGGSG (SEQ ID NO: 70), FNRGEC (SEQ ID NO: 71), VEPKSC (SEQ ID NO: 72), GGCGGGEVAACEKEVAALEKEVAALEKEVAALEK (SEQ ID NO: : 82), GGCGGGKVAACKEKVAALKEKVAALKEKVAALKE (SEQ ID NO: 83), or combinations thereof. In some embodiments, the implementation of the peptide linker may contain multiple copies of the linker sequence, for example, multiple copies of the sequence GGGSGGGG (SEQ ID NO: 69), LGGGSG (SEQ ID NO: 70), FNRGEC (SEQ ID NO: 71), VEPKSC (SEQ ID NO: 71), : 72), or combinations thereof.

In some embodiments, the first and second polypeptides have the following configurations.

Configuration 1 of the first polypeptide: N-VL1-VH2-hinge-CH2-CH3-C.

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Configuration 2 of the first polypeptide: N-VH2-VL1-hinge-CH2-CH3-C.

Configuration 1 of the second polypeptide: N-VL2-VH1-C.

Configuration 2 of the second polypeptide: N-VH1-VL2-C, wherein the first linker between VL1 and VH2 of the first polypeptide or the second linker between VL2 and VH1 of the second polypeptide contains the amino acid sequence GGGSGGGG (SEQ ID NO: 69). In some embodiments, said first linker and said second linker comprise the amino acid sequence GGGSGGGG (SEQ ID NO: 69). In some embodiments, the first polypeptide further comprises a third linker between VH2 or VL2 and said hinge region, and the second polypeptide further comprises a fourth linker after said VH1 or VL2 (at its C-terminus). In some embodiments, said third linker of said first polypeptide contains the amino acid sequence GGCGGGEVAACEKEVAALEKEVAALEKEVAALEK (SEQ ID NO: 82) and said fourth linker of said second polypeptide contains the amino acid sequence GGCGGGKVAACKEKVAALKEKVAALKEKVAALKE (SEQ ID NO: 83). In other embodiments, said third linker of said first polypeptide comprises the amino acid sequence GGCGGGKVAACKEKVAALKEKVAALKEKVAALKE (SEQ ID NO: 83) and said fourth linker of said second polypeptide comprises the amino acid sequence GGCGGGEVAACEKEVAALEKEVAALEKEVAALEK (SEQ ID NO: 82). In some embodiments, said third linker of said first polypeptide comprises the amino acid sequence GGCGGGEVAACEKEVAALEKEVAALEKEVAALEK (SEQ ID NO: 82) or the amino acid sequence GGCGGGKVAACKEKVAALKEKVAALKEKVAALKE (SEQ ID NO: 83). In other embodiments, said fourth linker of said second polypeptide comprises the amino acid sequence GGCGGGEVAACEKEVAALEKEVAALEKEVAALEK (SEQ ID NO: 82) or the amino acid sequence GGCGGGKVAACKEKVAALKEKVAALKEKVAALKE (SEQ ID NO: 83). In some embodiments, said third linker contains the amino acid sequence VEPKSC (SEQ ID NO: 72) or the amino acid sequence FNRGEC (SEQ ID NO: 71). In some embodiments, said fourth linker contains the amino acid sequence FNRGEC (SEQ ID NO: 71) or the amino acid sequence VEPKSC (SEQ ID NO: 72). In some embodiments, said third linker contains the amino acid sequence VEPKSC (SEQ ID NO: 72) and said fourth linker contains the amino acid sequence FNRGEC (SEQ ID NO: 71). The amino acid sequence of FNRGEC (SEQ ID NO: 71) is the last six amino acid residues of the CL domain (SEQ ID NO: 77) and the amino acid of VEPKSC (SEQ ID NO: 72) includes the last amino acid of CH1 and the first five amino acid residues of the hinge region of SEQ ID NO: 76 (as in SEQ ID NO: 5).

In some embodiments, the first and second polypeptides have the following configurations.

Configuration 3 of the first polypeptide: N-VL1-VH2-CH1-hinge-CH2-CH3-C.

Configuration 4 of the first polypeptide: N-VH2-VL1-CH1-hinge-CH2-CH3-C.

Configuration 3 of the second polypeptide: N-VL2-VH1-CL-C.

Second polypeptide configuration 4: N-VH1-VL2-CL-C, wherein the first linker between said VL1 and said VH2 of the first polypeptide, or said second linker between said VL2 and said VH1 of the second polypeptide, contains the amino acid sequence GGGSGGGG (SEQ ID NO: 69) . In some embodiments, said first linker and said second linker comprise the amino acid sequence GGGSGGGG (SEQ ID NO: 69). In some embodiments, said first polypeptide further comprises a third linker between said VH2 or VL1 and said CH1, and said second polypeptide further comprises a fourth linker between said VH1 or said VL2 and said CL. In some embodiments, said third linker or said fourth linker comprises the amino acid sequence LGGGSG (SEQ ID NO: 70). In some embodiments, said third linker and said fourth linker comprise the amino acid sequence LGGGSG (SEQ ID NO: 70). In some embodiments, said third linker and/or said fourth linker contains an optional cysteine residue. In some embodiments, said third linker and/or said fourth linker comprise the amino acid sequence GGCGGG (SEQ ID NO: 135) or LGGCGGGS (SEQ ID NO: 136).

In some embodiments, said heavy chain constant region 2 (CH2) contains alanine at positions 234 and alanine at position 235, numbered according to the Kabat EC index for a representative antibody.

In some embodiments, the connection contains two first polypeptides and two second polypeptides. In some embodiments, the hinge, CH2, and CH3 of one of the first polypeptides combine with the hinge, CH2, and CH3 of another of the first polypeptides to form a tetravalent molecule (e.g., the first two polypeptides dimerize through bonds between their respective

- 18 040031 hinge, CH2 and CH3 domains to form a tetravalent molecule containing two binding sites specific to the first target protein and two binding sites specific to the second target protein), monomer or monomeric antibody, as described in the Examples section. If the first polypeptide additionally contains a CH1 domain and the other polypeptide additionally contains a CL domain, the CH1 and CL domains can also participate in the formation of a tetravalent molecule (for example, the first two polypeptides dimerize through links between their respective hinge, CH2 and CH3 domains, and the CH1 of each specified of the first polypeptides are combined with the CL of one of the specified second polypeptides to form a tetravalent molecule containing two binding sites to the first target protein, and two binding sites to the second target protein), monomer, monomeric antibody, as described in the Examples section. In some embodiments, the first two polypeptides are linked together via at least one disulfide bond. In some embodiments, said compound comprises two of said first polypeptides and two of said second polypeptides, wherein each of said first polypeptides comprises a third linker, a hinge, CH2 and CH3, and each of said second polypeptides comprises a fourth linker, and wherein said hinge, CH2 and The CH3 of one of the first polypeptides is combined with the hinge, CH2 and CH3 of the other of the first polypeptides, and the third linker of each of the said first polypeptides is combined with the fourth linker of one of the said second polypeptides to form a tetravalent molecule (e.g., monomer, monomeric antibody, as described in section Examples) (eg compounds U and T).

In some embodiments, the disclosure relates to a compound comprising two first polypeptides and two second polypeptides, wherein said two first polypeptides are linked together via at least one disulfide bond, and wherein each of said first polypeptide is linked to one of said second polypeptides via at least at least one disulfide bond, wherein each of said first polypeptides contains:

(i) a first immunoglobulin (VL1) light chain variable domain specific for the first target protein;

(ii) a second immunoglobulin heavy chain variable domain (VH2) specific for a second target protein;

(iii) heavy chain constant region 1 (CH1), hinge region, heavy chain constant region 2 (CH2), and heavy chain constant region 3 (CH3); and wherein each of said second polypeptides contains:

(i) a second immunoglobulin (VL2) light chain variable domain specific for said second target protein;

(ii) a first immunoglobulin (VH1) heavy chain variable domain specific for said first target protein;

(iii) a light chain constant region (CL) domain, wherein the hinge, CH2 and CH3 of one of the first polypeptides combines with the hinge, CH2 and CH3 of another of the first polypeptides, and the CH1 of each of said first polypeptides combines with the CL of each of the second polypeptides to form a tetravalent molecule; wherein

a) said VL1 and VH1 combine to form an attachment site that binds said first target protein;

b) said VL2 and VH2 combine to form an attachment site that binds said second target protein;

c) said heavy chain constant region 2 (CH2) contains tyrosine at position 252, threonine at position 254 and glutamic acid at position 256, numbered according to the EC index, as in Kabat; And

d) said first target protein is FALV (BAFF) and said second target protein is IL-23A, or said first target protein is IL-23A and said second target protein is FALV (BAFF), and wherein :

(i) said VL1 contains SEQ ID NO: 2, said VH1 contains SEQ ID NO: 1, said VL2 contains SEQ ID NO: 4, and said VH2 contains SEQ ID NO: 3; or (ii) said VL1 contains SEQ ID NO: 4, said VH1 contains SEQ ID NO: 3, said VL2 contains SEQ ID NO: 2, and said VH2 contains SEQ ID NO: 1; or (iii) said VL1 contains SEQ ID NO: 85, said VH1 contains SEQ ID NO: 84, said VL2 contains SEQ ID NO: 4, and said VH2 contains SEQ ID NO: 4; or (iv) said VL1 has SEQ ID NO: 4, said VH1 has SEQ ID NO: 3, said VL2 has SEQ ID NO: 85, and said VH2 has SEQ ID NO: 84; or (v) said VL1 contains SEQ ID NO: 87, said VH1 contains SEQ ID NO: 86, said VL2 contains SEQ ID NO: 4, and said VH2 contains SEQ ID NO: 3; or (vi) said VL1 contains SEQ ID NO: 4, said VH1 contains SEQ ID NO: 3, said VL2 contains SEQ ID NO: 87, and said VH2 contains SEQ ID NO: 86; or

- 19 040031 (vii) specified VL1 contains SEQ ID NO: 89, specified VH1 contains SEQ ID NO: 88, specified

VL2 contains SEQ ID NO: 4 and said VH2 contains SEQ ID NO: 3; or (viii) said VL1 contains SEQ ID NO: 4, said VH1 contains SEQ ID NO: 3, indicated

VL2 contains SEQ ID NO: 89 and said VH2 contains SEQ ID NO: 88; or (ix) said VL1 contains SEQ ID NO: 91, said VH1 contains SEQ ID NO: 90, said VL2 contains SEQ ID NO: 4, and said VH2 contains SEQ ID NO: 3; or (x) said VL1 contains SEQ ID NO: 4, said VH1 contains SEQ ID NO: 3, said VL2 contains SEQ ID NO: 91, and said VH2 contains SEQ ID NO: 90.

In some embodiments relating to the above aspect, each of said first polypeptides further comprises a first linker between said VL1 and said VH2, and each of said second polypeptides further comprises a second linker between said VL2 and said VH1. In some embodiments, said first linker or said second linker comprises the amino acid sequence GGGSGGGG (SEQ ID NO: 69). In some embodiments, said first linker and said second linker comprise the amino acid sequence GGGSGGGG (SEQ ID NO: 69). In some embodiments, each of said first polypeptides further comprises a third linker between said VH2 or said VL1 and said CH1, and each of said second polypeptides further comprises a fourth linker between said VH1 or said VL2 and said CL. In some embodiments, said third linker or said fourth linker comprises the amino acid sequence LGGGSG (SEQ ID NO: 70). In some embodiments, said third linker and said fourth linker comprise the amino acid sequence LGGGSG (SEQ ID NO: 70). In some embodiments, said third linker and/or said fourth linker contains an optional cysteine residue. In some embodiments, said third linker and/or said fourth linker comprise the amino acid sequence GGCGGG (SEQ ID NO: 135) or LGGCGGGS (SEQ ID NO: 136). In some embodiments, said heavy chain constant region 2 (CH2) contains alanine at positions 234 and alanine at position 235, numbered according to the EC index, as in Kabat. In some embodiments, the amino acid sequence of said hinge region, said heavy chain constant region 2 (CH2), or said heavy chain constant region 3 (CH3) is derived from IgG1 or from IgG4. In some embodiments, said hinge region comprises the amino acid sequence EPKSCDKTHTCPPCP (SEQ ID NO: 76), the amino acid sequence LEPKSSDKTHTCPPCP (SEQ ID NO: 130), or the amino acid sequence ESKYGPPCPPCP (SEQ ID NO: 134). In some embodiments, (i) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 5 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 6; or (ii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 7 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 8; or (iii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 13 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 14; or (iv) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 15 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 16; or (v) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 21 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 22; or (vi) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 25 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 26; or (vii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 29 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 30; or (viii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 33 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 34; or (ix) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 37 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 38; or (x) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 41 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 42; or

- 20 040031 (xi) each of said first polypeptides contains the amino acid sequence

SEQ ID NO: 45 and each of these second polypeptides contains the amino acid sequence of SEQ ID NO: 46; or (xii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 49 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 50; or (xiii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 53 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 54; or (xiv) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 55 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 56; or (xv) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 57 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 58; or (xvi) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 59 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 60; or (xvii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 61 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 62; or (xviii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 63 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 64; or (xix) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 65 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 66; or (xx) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 67 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 68.

Other aspects of the disclosure of the invention relate to a compound containing two first polypeptides and two second polypeptides; wherein said two first polypeptides are linked together via at least one disulfide bond, and wherein each said first polypeptide is linked to one said second polypeptide via at least one disulfide bond; and wherein (i) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 5, and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 6; or (ii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 7 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 8; or (iii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 13 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 14; or (iv) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 15 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 16; or (v) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 21 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 22; or (vi) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 25 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 26; or (vii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 29 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 30; or (viii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 33 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 34; or (ix) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 37 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 38; or (x) each of said first polypeptides contains the amino acid sequence

- 21 040031

SEQ ID NO: 41 and each of these second polypeptides contains the amino acid sequence of SEQ ID NO: 42; or (xi) each of said first polypeptides contains the amino acid sequence

SEQ ID NO: 45 and each of these second polypeptides contains the amino acid sequence of SEQ ID NO: 46; or (xii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 49 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 50; or (xiii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 53 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 54; or (xiv) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 55 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 56; or (xv) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 57 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 58; or (xvi) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 59 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 60; or (xvii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 61 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 62; or (xviii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 63 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 64; or (xix) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 65 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 66; or (xx) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 67 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 68.

Also contemplated in this document are other compounds that compete for binding to a compound as described herein, such as a test compound that competes with a compound as described herein for binding to FAFL (BAFF) and IL23A. In some embodiments, a test compound may have at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity (amino acid to amino acid over the entire sequence) with a compound as described herein. Competitive binding can be determined using any assay known in the art, such as equilibrium binding, ELISA, surface plasmon resonance, or spectroscopy.

In some embodiments, a compound described herein specifically binds to both FAVL (BAFF) and IL23A. A compound that specifically binds to an antigen or epitope is a term well known in the art, and methods for determining such specific binding are also well known in the art. A molecule is said to exhibit specific binding if it reacts or binds more frequently, faster, for longer duration, and/or with greater affinity to a particular target antigen than to alternative targets. A compound binds specifically to a target antigen or epitope if it binds with greater affinity, avidity, more rapidly, and/or longer duration than it binds to other substances. For example, a compound that specifically (or selectively) binds to an antigen (e.g., BAFF or IL23A) or an antigenic epitope on that antigen is one that binds to that target antigen with greater affinity, avidity, easier and/or longer than it binds to other antigens or other epitopes on the same antigen. Reading this definition, it should also be understood that, for example, a compound that specifically binds to a first target antigen may or may not specifically or selectively bind to a second target antigen. Thus, specific binding or selective binding does not necessarily require (although it may include) exclusive binding. In general, as a rule, but not necessarily, reference to binding means selective binding. In some examples, a compound that specifically binds to a target antigen, or an epitope of that antigen, may not bind to other antigens or other epitopes on the same antigen.

In some embodiments, a compound as described herein has an appropriate affinity for FAVL (BAFF) or IL23A, or antigenic epitopes thereof. How is it applied in

- 22 040031 herein, the binding affinity refers to the conditional binding constant or CA. KA is the opposite of the dissociation constant (Kd). Compounds as described herein may have a binding affinity (Kd) of at least 10 ^-5 , 10 ^-6 , 10 ^-7 , 10 ^-8 , 10 ^-9 , 10 ^-10 , 10 ^-11 , 10 ^-12 M or less for one or both target antigens or antigenic epitopes. Increased binding affinity corresponds to reduced Kd. In some embodiments, the compounds described herein have a binding affinity (Kd) of at least 10 ^-11 M or less for one or both target antigens or antigenic epitopes. The high binding affinity of a compound for the first antigen and the second antigen for the third antigen can be indicated by a higher CA (or lower Kd value) for binding the first and second antigen than the CA (or Kd value) for binding the third antigen. In such cases, the compound has specificity for a first antigen and a second antigen (e.g., a first protein in a first conformation or the like, and a second protein in a first conformation or the like) with respect to a third antigen (e.g., the same first or second protein in second conformation or similar, or a third protein). The difference in binding affinity (eg, specificity or other factors) may be at least 1.5, 2, 3, 4, 5, 10, 15, 20, 37.5, 50, 70, 80, 91, 100, 500, 1000, 10000 or 10 ⁵ times.

Binding affinity (or binding specificity) can be determined by various methods, including equilibrium binding, ELISA, surface plasmon resonance, or spectroscopy (eg, using fluorescence analysis). Typical conditions for assessing binding affinity are HBS-P buffer (10 mM HEPES pH 7.4, 150 mM NaCl, 0.005% (v/v) Surfactant P20). These methods can be used to measure the concentration of the bound binding protein as a function of the concentration of the target protein. The concentration of bound binding protein, ([Bound]) depends on the concentration of free target protein ([Free]) and the concentration of binding sites for the binding protein on the target, where (N) is the number of binding sites per target molecule, is described by the following equation:

[Bound] = [N][Free]/(K ₄ + [Free])

It is not always necessary to accurately determine K _a , however, since sometimes it is sufficient to obtain a quantitative estimate of affinity, for example, calculated using a method such as ELISA or FCS (fluorescence excited cell sorting, FACS) analysis, which is proportional to K _a , and therefore , can be used for comparisons such as: determining if the affinity is higher, for example 2 times higher, to obtain a qualitative estimate of affinity, or to predict affinity, for example, by activity in a functional assay, for example, in assays in vitro or in vivo.

In some embodiments, the implementation of the connection contains the first and second polypeptide, as defined in table. 2A. In some embodiments, the compound contains:

(i) said first polypeptide contains the amino acid sequence of SEQ ID NO: 5, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 6; or (ii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 7, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 8; or (iii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 9, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 10; or (iv) said first polypeptide contains the amino acid sequence of SEQ ID NO: 11 and said second polypeptide contains the amino acid sequence of SEQ ID NO: 12; or (v) said first polypeptide contains the amino acid sequence of SEQ ID NO: 13, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 14; or (vi) said first polypeptide contains the amino acid sequence of SEQ ID NO: 15, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 16; or (vii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 17, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 18; or (viii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 19, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 20; or (ix) said first polypeptide contains the amino acid sequence of SEQ ID NO: 21, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 22; or (x) said first polypeptide contains the amino acid sequence of SEQ ID NO: 23, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 24; or (xi) said first polypeptide contains the amino acid sequence of SEQ ID NO: 25, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 26; or (xii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 27, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 28; or (xiii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 29, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 30; or (xiv) said first polypeptide contains the amino acid sequence of SEQ ID NO: 31,

- 23 040031 and the specified second polypeptide contains the amino acid sequence of SEQ ID NO: 32; or (xv) said first polypeptide contains the amino acid sequence of SEQ ID NO: 33, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 34; or (xvi) said first polypeptide contains the amino acid sequence of SEQ ID NO: 35, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 36; or (xvii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 37, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 38; or (xviii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 39, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 40; or (xix) said first polypeptide contains the amino acid sequence of SEQ ID NO: 41, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 42; or (xx) said first polypeptide contains the amino acid sequence of SEQ ID NO: 43, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 44; or (xxi) said first polypeptide contains the amino acid sequence of SEQ ID NO: 45, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 46; or (xxii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 47, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 48; or (xxiii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 49, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 50; or (xxiv) said first polypeptide contains the amino acid sequence of SEQ ID NO: 51, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 52; or (xxv) said first polypeptide contains the amino acid sequence of SEQ ID NO: 53, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 54; or (xxvi) said first polypeptide contains the amino acid sequence of SEQ ID NO: 55, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 56; or (xxvii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 57, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 58; or (xxviii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 59, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 60; or (xxix) said first polypeptide contains the amino acid sequence of SEQ ID NO: 61, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 62; or (xxx) said first polypeptide contains the amino acid sequence of SEQ ID NO: 63, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 64; or (xxxi) said first polypeptide contains the amino acid sequence of SEQ ID NO: 65, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 66; or (xxxii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 67 and said second polypeptide contains the amino acid sequence of SEQ ID NO: 68.

Methods for obtaining compounds, nucleic acids, vectors and cells.

Aspects of the disclosure also include nucleic acids encoding compounds described herein or polypeptides as described herein (eg, the first or second polypeptides as described herein), which may be encoded together or separately. Polynucleotides encoding the compounds described herein or polypeptides described herein can be obtained and the nucleotide sequences of the polynucleotides determined by any method known in the art.

In some embodiments, the implementation of the nucleic acid is contained in a vector, such as an expression vector. In some embodiments, the vector contains a promoter that is operably linked to the nucleic acid.

Various promoters can be used to express the compounds described herein or the polypeptides described herein, which include, but are not limited to: cytomegalovirus (CMV) early intermediate promoter, viral LTR, such as Rous sarcoma virus LTR, HIV-LTR, HTLV-1 LTR, simian virus 40 early promoter (SV40), E. coli UV5 lac promoter and herpes simplex virus tk promoter.

Regulated promoters may also be used. Such regulated promoters include those that use the E. coli lac repressor as a transcriptional modulator to regulate transcription from mammalian cell promoters that carry a lac operator [Brown M. et al., Cell, 49: 603-612 (1987)], and those that use the tetracycline repressor (tetR) [Gossen M., and Bujard H., Proc. Natl. Acad. sci. USA 89: 5547-5551 (1992); Yao F. et al., Human Gene Therapy, 9: 1939-1950 (1998); Shockelt P. et al., Proc. Natl. Acad. sci. USA, 92: 6522-6526 (1995)]. Other systems include FK506 dimer, VP 16 or p65 using astradiol, RU486, murislerone diphenol, or rapamycin. Inducible systems are available from Invitrogen, Clontech and Ariad.

Regulated promoters that contain the repressor with an operon can be used. In one embodiment, the Escherichia coli lacquer repressor may function as

- 24 040031 transcription modulator for regulating transcription through mammalian cell promoters that carry the lac operator [M. Brown et al., Cell, 49: 603-612 (1987)]; Gossen & Bujart (1992) [M. Gossen et al., Natl. Acad. sci. USA, 89: 5547-5551 (1992)] combined a tetracycline repressor (tetR) with a transcriptional activator (VP 16) to create a mammalian cell transcription activator tetR fusion protein, tTa (tetR-VP 16), with a tetO-carrying minimal promoter. derived from the major intermediate-early promoter of human cytomegalovirus (hCMV (hCMV)) in order to create a tetR-tet operator system to control gene expression in mammalian cells. In one embodiment, an inducible tetracycline switch is used (Yao et al., Human Gene Therapy; Gossen et al., Natl. Acad. Sci. USA, 89: 5547-5551 (1992); Shockett et al., Proc. Natl Acad. Sci. USA, 92: 6522-6526 (1995)).

Additionally, the vector may contain, for example, some or all of the following: a selectable marker gene, such as a neomycin gene, for selection of stable or transient transfectants in mammalian cells; enhancer/promoter sequences from the human CMV immediate early gene for high levels of transcription; transcription termination and RNA processing signals with SV40 for mRNA stability; origins of replication SV40 and ColE1 for proper episomal replication; internal ribosome binding sites (IRSC), polylinker; and T7 and SP6 RNA promoters for in vitro transcription of sense and antisense RNA. Suitable vectors and methods for producing vectors containing transgenes are well known and available in the art.

An expression vector containing a nucleic acid can be transferred into a host cell using conventional methods (e.g., electroporation, liposome transfection, and calcium phosphate precipitation), and the transfected cells are then cultured using conventional methods for preparing compounds, which are described herein. In some embodiments, expression of the compounds described herein is regulated by constitutive, inducible, or tissue-specific promoters.

The host cells used to express the compounds described herein or the polypeptides described herein can be either bacterial or Escherichia coli cells, or preferably eukaryotic cells. In particular, mammalian cells such as Chinese hamster ovary (CHO) cells, in combination with a vector such as the core promoter element of the intermediate early gene from human cytomegalovirus, is an efficient expression system for immunoglobulins (Foecking et al. (1986) Powerful And Versatile Cockett et al (1990) High Level Expression Of Tissue Inhibitor Of Metalloproteinases In Chinese Hamster Ovary Cells Using Glutamine Synthetase Gene Amplification, Biotechnology 8:662-667).

A variety of host-expressing vector systems can be used to express the compounds described herein or the polypeptides described herein. Such host-expression systems are vehicles by which the coding sequences for the compounds described herein or the polypeptides described herein can be produced and subsequently purified, but are also cells that can, when transformed or transfected with using the appropriate coding nucleotide sequences to express the compounds described herein in situ. These include, but are not limited to: microorganisms such as bacteria (eg E. coli and B. subtilis) by transformation with recombinant bacteriophage DNA, plasmid DNA or cosmid DNA expression vectors containing the coding sequences for the compounds described herein; yeast (eg, Saccharomyces pichia) transformed with recombinant yeast expression vectors containing sequences encoding the compounds described herein; insect cell systems infected with recombinant viral expression vectors (eg, baclovirus) containing sequences encoding the compounds described herein; plant cell systems infected with recombinant viral expression vectors (eg, cauliflower mosaic virus (CaMV) and tobacco mosaic virus (TMV)) or transformed with recombinant plasmid expression vectors (eg, Te plasmids) containing sequences encoding molecules of the compounds described herein document; or mammalian cell systems (e.g., COS, CHO, BHK, 293, 293T, 3T3 cells, lymphocytic cells (see US Pat. No. 5,807,715), Per C.6 cells (human retinal cells developed by Crucell) carrying recombinant expression constructs containing promoters derived from the genome of mammalian cells (eg, the metallothionein promoter) or mammalian viruses (eg, adenovirus late promoter; vaccinia virus 7.5 K promoter).

In bacterial systems, a number of expression vectors can be selected to great advantage depending on the application intended for the compound being expressed. For example, when a large amount of such a protein is to be produced, vectors driving the expression of high levels of easily purified fusion protein products may be desirable to prepare pharmaceutical compositions of the compounds described herein. Such

- 25 040031 vectors include, but are not limited to: E. coli expression vector pUR278 (Ruther et al. (1983) Easy Identification Of cDNA Clones, EMBO J. 2: 1791-1794), in which the coding sequence can be ligated separately into a framed vector with a lac Z coding region such that a fusion protein is produced; pIN vectors (Inouye et al. (1985) Up-Promoter Mutations In The Ipp Gene Of Escherichia Coli, Nucleic Acids Res. 13: 3101-3110; Van Heeke et al. (1989) Expression Of Human Asparagine Synthetase In Escherichia Coli, J Biol Chem 24: 5503-5509) and the like. pGEX vectors can also be used to express polypeptides as glutathione S-transferase (GST) fusion proteins. In general, such fusion proteins are soluble and can be readily purified from lysed cells by adsorption and binding to matrix glutathione garose beads, followed by elution in the presence of free glutathione. The pGEX vectors are designed to include thrombin or Xa protease cleavage sites so that the cloned target gene product can be freed from the GST portion.

In insect systems, the Autographa califomica nuclear polyhedrosis virus (AcNPV) is used as a vector for the expression of foreign genes. The virus replicates in the cells of Spodoptera frugiperda. The coding sequence can be cloned separately into non-essential regions (eg the polyhedrin gene) of the virus and placed under the control of an AcNPV promoter (eg the polyhedrin promoter).

A number of virus-based expression systems can be used in mammalian host cells. Where an adenovirus is used as an expression vector, the coding sequence of interest may be ligated to a transcription/translation control complex, such as a late promoter and a tripartite leader sequence. This chimeric gene can be inserted into the adenovirus genome by in vitro or in vivo recombination. Insertion into a non-essential region of the viral genome (eg, the E1 or E3 region) will result in a recombinant virus that is viable and capable of expressing the immunoglobulin molecule in infected hosts (eg, see Logan et al. (1984) Adenovirus Tripartite Leader Sequence Enhances Translation Of mRNAs Late After Infection, Proc. Natl. Acad. Sci. USA 81:3655-3659). Specific initiation signals may also be required for efficient translation of the inserted sequences encoding the antibody(s). These signals include the ATG start codon and adjacent sequences. In addition, the initiation codon must be in phase with the reading frame of the desired coding sequence in order to ensure translation of the entire insert. These exogenous translation control signals and initiation codons can be of various origins, both natural and synthetic. Expression efficiency can be increased by including appropriate transcription elements, enhancer elements, transcription terminators, etc. (See Bitter et al. (1987) Expression And Secretion Vectors For Yeast, Methods in Enzymol. 153: 516-544).

In addition, a host cell strain can be chosen that modulates the expression of the inserted sequences, or modifies and processes the gene product in the specific manner desired. Such modifications (eg, glycosylation) and processing (eg, cleavage) of protein products may be important for the function of the protein. For example, in some embodiments, the compounds as described herein may be expressed as a single gene product (e.g., as a single polypeptide chain, i.e., as a polyprotein precursor) that requires proteolytic cleavage by native or recombinant cellular mechanisms in order to to form individual polypeptides of the compounds described herein. Thus, the disclosure of the invention encompasses the design of a nucleotide sequence for encoding a precursor polyprotein molecule containing polypeptides of the compounds described herein, containing coding sequences capable of directing post-translational cleavage of said polyprotein precursor. Post-translational cleavage of the polyprotein precursor results in the formation of the polypeptides of the compounds described herein. Post-translational cleavage of a precursor molecule containing the polypeptides of the compounds described herein may occur in vivo (i.e., within the host cell by native or recombinant cellular systems/mechanisms, e.g., furin cleavage at the appropriate site) or may occur in vitro ( for example, incubation of said polypeptide chain in a composition containing proteases or peptidases of known activity and/or in a composition containing conditions or reagents known to promote the desired proteolytic activity). Purification and modification of recombinant proteins are well known in the art, and thus the design of a polyprotein precursor may include a number of embodiments of the invention that can be readily recognized by the skilled artisan. Any known proteases or peptidases that are known in the art can be used to modify the precursor molecule described, for example, thrombin or factor Xa (Nagai et al. (1985) Oxygen Binding Properties Of Human Mutant Hemoglobins Synthesized In Escherichia Coli, Proc. Nat Acad. Sci. USA 82: 7252-7255 and reviewed in Jenny et al. (2003) A Critical Review Of The Methods For Cleavage Of Fusion Proteins With Thrombin And Factor Xa, Protein Expr. Purif 31: 1-11 , each of which is incorporated herein in its entirety by reference

- 26 040031 ki)), enterokinase (Collins-Racie et al. (1995) Production Of Recombinant Bovine Enterokinase Catalytic Subunit In Escherichia Coli Using The Novel Secretory Fusion Partner DsbA, Biotechnology 13: 982-987 is hereby incorporated into this document in its entirety by reference)), furin, and AcTEV (Parks et al. (1994) Release Of Proteins And Peptides From Fusion Proteins Using A Recombinant Plant Virus Proteinase, Anal. Biochem. 216: 413-417 are hereby incorporated herein in their entirety by references)) and FMD protease C3.

Various host cells have characteristic and specific mechanisms for post-translational processing and modification of proteins and gene products. Appropriate cell lines or host systems can be selected to ensure correct modification and processing of the foreign expressed protein. To this end, eukaryotic host cells that possess the cellular machinery for proper processing of the primary transcript, glycosylation, and phosphorylation of the gene product can be used. Such mammalian host cells include, but are not limited to: CHO, VERY, BHK, HeLa, COS, MDCK, 293, 293T, 3T3, WI38, BT483, Hs578T, HTB2, BT20 and T47D, CRL7030 and Hs578Bst.

Stable expression is desirable for long term, high yield production of recombinant proteins. For example, cell lines can be developed that stably express the compounds described herein. Instead of using expression vectors containing viral origins of replication, host cells can be transformed with DNA, which is controlled by appropriate expression control elements (eg, promoter, enhancer sequences, transcription terminators, polyadenylation sites, etc.) and a selection marker . After the introduction of foreign DNA, engineered cells can be allowed to grow for 1-2 days in enriched medium and then switched to selective medium. The selection marker in the recombinant plasmid confers resistance to the selection agent and allows cells to stably integrate the plasmid into their chromosomes and grow into foci, which in turn can be cloned and expanded into cell lines. This method can be advantageously used to develop cell lines expressing the compounds described herein. Such engineered cell lines may be particularly useful for screening and evaluating compounds that interact directly or indirectly with the compounds described herein.

A number of selection systems can be applied, including but not limited to: herpes simplex virus thymidine kinase gene (Wigler et al. (1977) Transfer Of Purified Herpes Virus Thymidine Kinase Gene To Cultured Mouse Cells, Cell 11: 223-232), gene hypoxanthine-guanine phosphoribosyltransferase (Szybalska et al. (1992) Use Of The HPRT Gene And The HAT Selection Technique In DNA-Mediated Transformation Of Mammalian Cells First Steps Toward Developing Hybridoma Techniques And Gene Therapy, Bioessays 14: 495-500), and the gene adenine phosphoribosyltransferases (Fowy et al. (1980) Isolation Of Transforming DNA: Cloning The Hamster aprt Gene, Cell 22: 817-823) can be used in tk, hgprt or aprt cells, respectively. In addition, antimetabolite resistance can be used as a basis for selection for the following genes: dhfr conferring methotrexate resistance (Wigler et al. (1980) Transformation Of Mammalian Cells With An Amplifiable Dominant-Acting Gene, Proc. Natl. Acad. Sci. USA 77: 3567-3570 O'Hare et al (1981) Transformation Of Mouse Fibroblasts To Methotrexate Resistance By A Recombinant Plasmid Expressing A Prokaryotic Dihydrofolate Reductase, Proc Natl Acad Sci USA 78: 1527-1531); gpt conferring resistance to mycophenolic acid (Mulligan et al. (1981) Selection For Animal Cells That Express The Escherichia coli Gene Coding For Xanthine-Guanine Phosphoribosyltransferase, Proc. Natl. Acad. Sci. USA 78: 2072-2076); neo conferring resistance to aminoglycoside G-418 (Tolstoshev (1993) Gene Therapy, Concepts, Current Trials And Future Directions, Ann. Rev. Pharmacol. Toxicol. 32: 573-596; Mulligan (1993), The Basic Science Of Gene Therapy , Science 260: 926932; and Morgan et al. (1993), Human Gene Therapy, Ann. Rev. Biochem. 62: 191-217) and hygro conferring resistance to hygromycin (Santerre et al. (1984) Expression Of Prokaryotic Genes For Hygromycin B And G418 Resistance As Dominant-Selection Markers In Mouse L Cells, Gene 30: 147-156). Methods well known in the art of recombinant DNA technology, and which can be applied, are described in Ausubel et al. (Eds.), 1993, Current Protocols in Molecular Biology, John Wiley & Sons, NY; Kriegler, 1990, Gene Transfer and Expression, A Laboratory Manual, Stockton Press, NY; and in Chapters 12 and 13 Dracopoli et al. (Eds), 1994, Current Protocols in Human Genetics, John Wiley & Sons, NY; Colberre-Garapin et al. (1981) A New Dominant Hybrid Selective Marker for Higher Eukaryotic Cells, J. Mol. Biol. 150:1-14.

Expression levels of the compounds described herein or the polypeptides described herein can be increased using an amplification vector (see Bebbington and Hentschel, The use of vectors based on gene amplification for the expression of cloned genes in mammalian cells in DNA cloning, Vol.3 (Academic Press, New York, 1987) When a marker in a vector system expressing a compound described herein can be amplified, increasing the level of inhibitor present in the host cell culture will increase the copy number of the marker. Because the region of amplification is associated with the nucleotide sequence of a compound described herein or a polypeptide described herein,

- 27 040031 production of the polypeptide will also increase (Crouse et al. (1983) Expression And Amplification Of Engineered Mouse Dihydrofolate Reductase Minigenes, Mol. Cell. Biol. 3: 257-266).

The host cell can be co-transfected with two expression vectors, a first vector encoding a first compound polypeptide as described herein and a second vector encoding a second compound polypeptide as described herein. The two vectors may contain identical selection markers allowing the same expression of both polypeptides. Alternatively, a single vector encoding both polypeptides may be used. The coding sequences for the polypeptides of the compounds described herein may comprise cDNA or genomic DNA.

Once a compound described herein or a polypeptide described herein has been recombinantly expressed, it can be purified by any method known in the art for the purification of polypeptides, polyproteins or antibodies (e.g. similar to antibody purification schemes based on antigen selectivity), e.g. chromatography (e.g. ion exchange, affinity, in particular affinity for a specific antigen (if necessary after selection with protein A, where the compound contains an Fc domain (or its part)) and size exclusion chromatography), centrifugation, differential solubility, or any other standard method for purifying polypeptides or antibodies.

Other aspects of the invention are related to a cell containing the nucleic acid described in this document, or the vector described in this document. The cell may be a prokaryotic or eukaryotic cell. In some embodiments, the cell is a mammalian cell. Exemplary cell types are described herein.

Yet other aspects of the invention relate to a method for producing a compound described herein or a polypeptide described herein (e.g., a first polypeptide or a second polypeptide), a method comprising obtaining cells described herein, and expressing a nucleic acid described in this document, in the specified cell. In some embodiments, the method further comprises isolating and purifying a compound described herein or a polypeptide described herein.

Methods of treatment and compositions for use in medicine.

Other aspects of the disclosure of the invention relate to methods of treatment and compositions for use in medicine. Non-limiting examples of compounds for use in such methods and compositions are those containing:

(i) said first polypeptide contains the amino acid sequence of SEQ ID NO: 5, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 6; or (ii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 7, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 8; or (iii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 9, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 10; or (iv) said first polypeptide contains the amino acid sequence of SEQ ID NO: 11 and said second polypeptide contains the amino acid sequence of SEQ ID NO: 12; or (v) said first polypeptide contains the amino acid sequence of SEQ ID NO: 13, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 14; or (vi) said first polypeptide contains the amino acid sequence of SEQ ID NO: 15, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 16; or (vii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 17, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 18; or (viii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 19, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 20; or (ix) said first polypeptide contains the amino acid sequence of SEQ ID NO: 21, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 22; or (x) said first polypeptide contains the amino acid sequence of SEQ ID NO: 23, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 24; or (xi) said first polypeptide contains the amino acid sequence of SEQ ID NO: 25, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 26; or (xii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 27, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 28; or (xiii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 29, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 30; or (xiv) said first polypeptide contains the amino acid sequence of SEQ ID NO: 31, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 32; or (xv) said first polypeptide contains the amino acid sequence of SEQ ID NO: 33, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 34; or (xvi) said first polypeptide contains the amino acid sequence of SEQ ID NO: 35,

- 28 040031 and the specified second polypeptide contains the amino acid sequence of SEQ ID NO: 36; or (xvii) said first polypeptide contains the amino acid sequence of SEQ ID NO:

37, and the specified second polypeptide contains the amino acid sequence of SEQ ID NO: 38; or (xviii) said first polypeptide contains the amino acid sequence of SEQ ID NO:

39, and the specified second polypeptide contains the amino acid sequence of SEQ ID NO: 40; or (xix) said first polypeptide contains the amino acid sequence of SEQ ID NO: 41, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 42; or (xx) said first polypeptide contains the amino acid sequence of SEQ ID NO: 43, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 44; or (xxi) said first polypeptide contains the amino acid sequence of SEQ ID NO: 45, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 46; or (xxii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 47, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 48; or (xxiii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 49, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 50; or (xxiv) said first polypeptide contains the amino acid sequence of SEQ ID NO: 51, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 52; or (xxv) said first polypeptide contains the amino acid sequence of SEQ ID NO: 53, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 54; or (xxvi) said first polypeptide contains the amino acid sequence of SEQ ID NO: 55, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 56; or (xxvii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 57, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 58; or (xxviii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 59, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 60; or (xxix) said first polypeptide contains the amino acid sequence of SEQ ID NO: 61, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 62; or (xxx) said first polypeptide contains the amino acid sequence of SEQ ID NO: 63, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 64; or (xxxi) said first polypeptide contains the amino acid sequence of SEQ ID NO: 65, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 66; or (xxxii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 67 and said second polypeptide contains the amino acid sequence of SEQ ID NO: 68.

In some embodiments, the method of treatment or use is a method of treating an autoimmune or inflammatory disease, or use in such a method. In some embodiments, the method includes administering a compound described herein, or a pharmaceutical composition containing said compound, to a subject, such as a subject who has or is at risk of acquiring an autoimmune or inflammatory disease.

The subject being treated with the methods described herein may be a mammal, more preferably a human. Mammals include, but are not limited to, farm animals, sport animals, pets, primates, horses, dogs, cats, mice, and rats. The human subject in need of treatment may be a human subject who has, or is at risk of acquiring, or is suspected of having a disease. A subject having diseases can be detected by a routine medical examination, such as a physical examination, laboratory testing, organ function testing, CT scan, or ultrasound. A subject who is suspected of having any such disease may exhibit one or more of the symptoms of the disease. Signs and symptoms of diseases, such as autoimmune and inflammatory diseases, are well known to those skilled in the art. A subject at risk of acquiring a disease may be a subject having one or more risk factors for the disease.

Non-limiting examples of autoimmune diseases include lupus nephritis (LN)) (systemic lupus erythematosus (SLE (SLE)) with kidney damage), systemic lupus erythematosus (SLE (SLE)), primary Sjögren's syndrome (PSS (pSS)), Sjögren, graft-versus-host disease (GVHD) (e.g., chronic graft-versus-host disease (cGVHD))), systemic sclerosis (SSc), antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV )), rheumatoid arthritis, psoriasis, type 1 diabetes, systemic lupus erythematosus, transplant rejection, autoimmune thyroid disease (Hashimoto's disease), sarcoidosis, scleroderma, granulomatous vasculitis, Crohn's disease, ulcerative colitis, Sjögren's disease, ankylosing spondylitis, psoriatic arthritis , polymyositis dermatomyositis, periarteritis nodosa, immunologically mediated blistering skin disease, Behçet's syndrome, multiple sclerosis, systemic scleroderma, b Goodpasture's disease, or immunologically mediated glomerulonephritis.

- 29 040031

Non-limiting examples of inflammatory diseases include rheumatoid arthritis, systemic lupus erythematosus, alopecia areata, ancolizing spondylitis, antiphospholipid syndrome, autoimmune Addison's disease, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune inner ear disease, autoimmune lymphoproliferative syndrome (ALPS), autoimmune thrombocytopenic purpura ), Behcet's disease, bullous pemphigoid, cardiomyopathy, abdominal sprue dermatitis, chronic fatigue syndrome, immunodeficiency syndrome (CFIDS), chronic inflammatory demyelinating polyneuropathy, cicatricial pemphigus, cold lectin disease, Thibierge-Weissenbach syndrome, Crohn's disease, Dego's disease, dermatomyositis, juvenile dermatomyositis , discoid lupus, significant mixed cryoglobulinemia, fibromyalgo-fibromyositis, Graves' disease, Guillain-Barré syndrome, Hashimoto's thyroiditis, idiopathic pulmonary fibrosis, Werlhof's disease (ITP), IgA nephropathy, insulin-dependent diabetes mellitus diabetes (type I), insulin-dependent diabetes (type I), juvenile arthritis, Meniere's disease, mixed connective tissue disease, multiple sclerosis, myasthenia gravis, pemphigus vulgaris, pernicious anemia, periarteritis nodosa, polychondritis, pluriglandular syndromes, polymyalgia rheumatica, polymyositis, and dermatomyositis, primary agammaglobulinemia, primary biliary cirrhosis, psoriasis, Raynaud's phenomenon, Reiter's syndrome, rheumatic fever, sarcoidosis, scleroderma, Sjögren's syndrome, muscle stiffness syndrome, Takayasu's arteritis, temporal arteritis/giant cell arteritis, ulcerative colitis, uveitis, vasculitis, vitiligo , and Wegener's granulomatosis. In some embodiments, the autoimmune or inflammatory disease is Crohn's disease, ankylosing spondylitis, or psoriatic arthritis.

In order to practice the method described herein, an effective amount of a compound or pharmaceutical composition as described herein may be administered to a subject (eg, human) that requires treatment. Various delivery systems are known and can be used to administer compounds of this technology. Routes of administration include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, and oral routes. The compounds of this technology can be administered, for example, by infusion, bolus or injection, and they can be administered together with other biologically active agents such as anti-inflammatory agents. The introduction can be systemic or local. In the desired embodiments of the invention, the introduction is performed by subcutaneous injection. The drug for such injections can be prepared in, for example, pre-filled syringes that can be administered once every two weeks.

An effective amount, as used herein, refers to the amount of each compound that is necessary to provide a therapeutic effect to a subject, either alone or in combination with one or more other compounds. Effective amounts may vary, as determined by those skilled in the art, depending on the particular condition being treated, the severity of the condition, the individual parameters of the subject, including age, physical condition, size, sex, weight, duration of treatment, nature of concomitant therapy (if there is one), the specific route of administration, and similar factors that are within the knowledge and experience of the clinician. These factors are well known to those skilled in the art and can be detected with no more than routine experimentation. It is desirable, in general, as a rule, that the maximum dose of the individual components or their combinations be used, that is, the highest safe dose from a medical point of view. Those skilled in the art will appreciate that a subject may insist on a lower dose or tolerable dose for medical reasons, psychological reasons, or any other reason.

Empirical factors such as half-life will generally guide dosage determination. For example, compounds compatible with the human immune system, such as compounds containing regions from humanized antibodies or fully human antibodies, can be used to prolong the half-life of the compound and prevent the compound from being attacked by the host's immune system. The frequency of administration can be determined and adjusted during treatment, usually, but not necessarily, based on treatment and/or inhibition and/or improvement and/or slowing of the disease. Alternatively, continuous sustained release formulations of the compound may be useful. Various dosage forms and devices are known in the art to achieve sustained release.

In some embodiments, the dosage is daily, every two days, every three days, every four days, every five days, or every six days. In some embodiments, the dosing frequency is weekly, every 2 weeks, every 4 weeks, every 5 weeks, every 6 weeks, every 7 weeks, every 8 weeks, every 9 weeks, or 10 weeks, or once a month, every 2 months or every 3 months or longer. The progress of this therapy can be easily monitored using conventional methods and assays. The dosage regimen (including the compounds used) may change over time.

In some embodiments, a normal weight adult subject may be administered doses that range from about 0.01 to 1000 mg/kg. In some embodiments, the implementation of the dose is from 1 to 200 mg. The specific dosing regimen, i.e., dose, time, and repetition, will depend on the individual subject and that subject's medical history, as well as the properties of the compound (e.g., compound half-life and other factors that are well known to those skilled in the art).

In the context of this invention, the appropriate dose of a compound as described herein will depend on the particular compound (or composition thereof) involved, formulation and route of administration, type and severity of disease, whether the compound is being administered prophylactically or therapeutically, prior therapy, the subject's clinical history and responses to the antagonist, and at the discretion of the attending physician. Typically, the physician will administer the compound until the prescribed dose is reached to achieve the desired result. Administration of one or more compounds may be continuous or intermittent, depending on, for example, the physiological state of the recipient, whether the purpose of administration is therapeutic or prophylactic, and other factors known to those skilled in the art. The administration of the compound may be substantially continuous over a preselected period of time, or may be divided into a number of time-separated doses, eg before, during or after the onset of the disease.

As used herein, the term treatment refers to the use or administration of a compound, or a composition containing a compound, to a subject having a disease, symptoms of a disease, or a predisposition to a disease, for the purpose of curing, healing, alleviating, alleviating, modifying, correcting, improving, or affecting a disease, a symptom of a disease, or a predisposition to a disease.

Relief of a disease includes slowing down the development or progression of a disease or reducing the severity of a disease. Relief of the disease does not necessarily require curative results. As used herein, to slow the progression of a disease means to delay, interfere with, retard, stabilize, and/or delay the progression of a disease. This delay may be of varying duration depending on the medical history and/or individuals being treated. A method of slowing or alleviating the development of a disease, or delaying the onset of a disease, is a method that reduces the likelihood of developing one or more symptoms of a disease in a given specific period of time and / or reduces the severity of symptoms over a certain period of time, compared with the situation when the method is not applied . Such comparisons are usually based on clinical studies using a sufficient number of subjects to obtain a statistically significant result.

The development or progression of a disease means the initial manifestations and/or subsequent progression of the disease. Disease progression can be detected and assessed using standard clinical techniques that are well known in the art. However, development also refers to progression, which may not be noticeable. For the purposes of this disclosure, development or progression refers to the biological course of symptoms. Development includes emergence, recurrence, and onset. As used herein, the onset or onset of a disease includes initial onset and/or relapse.

In some embodiments, a compound described herein is administered to a subject in need of treatment in an amount sufficient to inhibit the activity of one or both BAFF or IL23A by at least 20% (e.g., 30-40, 50, 60, 70, 80, 90% or more) in vivo or in vitro. Methods for determining the inhibitory ability of a compound are known in the art. Exemplary FAVL (BAFF) or IL23A inhibition assays are provided in the examples.

Conventional methods known to those skilled in the art may be used to administer a compound or pharmaceutical composition to a subject, depending on the type of disease to be treated or the location of the disease. The composition may also be administered by other conventional routes, such as orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally, or via an implanted reservoir. The term parenteral as used herein includes subcutaneous, intradermal, intravenous, intramuscular, intraarticular, intraarterial, intrasynovial, spinal, lesion, and intracranial injection or infusion methods. In addition, it can be administered to the subject by administering a slow absorbent, as is used with 1-, 3-, or 6-month slow absorbers or biodegradable materials and methods.

pharmaceutical compositions.

Still other aspects of the disclosure of the invention are related to a pharmaceutical composition containing the compound described in this document. A composition containing a compound of the technology in question (e.g., a compound that is specific for both FAVL (BAFF) and IL23A) may be administered to a subject who has or is at risk of acquiring an autoimmune or

- 31 040031 inflammatory disease. The subject technology further relates to the use of a compound of the subject technology in the manufacture of a medicament for the treatment of an autoimmune or inflammatory disease. The compounds may be administered either alone or in combination with other compositions for the prevention or treatment of an autoimmune or inflammatory disease. Non-limiting examples of compounds of this technology for use in such pharmaceutical compositions are those that include:

(i) said first polypeptide contains the amino acid sequence of SEQ ID NO: 5, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 6; or (ii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 7, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 8; or (iii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 9, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 10; or (iv) said first polypeptide contains the amino acid sequence of SEQ ID NO: 11 and said second polypeptide contains the amino acid sequence of SEQ ID NO: 12; or (v) said first polypeptide contains the amino acid sequence of SEQ ID NO: 13, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 14; or (vi) said first polypeptide contains the amino acid sequence of SEQ ID NO: 15, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 16; or (vii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 17, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 18; or (viii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 19, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 20; or (ix) said first polypeptide contains the amino acid sequence of SEQ ID NO: 21, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 22; or (x) said first polypeptide contains the amino acid sequence of SEQ ID NO: 23, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 24; or (xi) said first polypeptide contains the amino acid sequence of SEQ ID NO: 25, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 26; or (xii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 27, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 28; or (xiii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 29, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 30; or (xiv) said first polypeptide contains the amino acid sequence of SEQ ID NO: 31, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 32; or (xv) said first polypeptide contains the amino acid sequence of SEQ ID NO: 33, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 34; or (xvi) said first polypeptide contains the amino acid sequence of SEQ ID NO: 35, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 36; or (xvii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 37, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 38; or (xviii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 39, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 40; or (xix) said first polypeptide contains the amino acid sequence of SEQ ID NO: 41, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 42; or (xx) said first polypeptide contains the amino acid sequence of SEQ ID NO: 43, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 44; or (xxi) said first polypeptide contains the amino acid sequence of SEQ ID NO: 45, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 46; or (xxii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 47, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 48; or (xxiii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 49, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 50; or (xxiv) said first polypeptide contains the amino acid sequence of SEQ ID NO: 51, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 52; or (xxv) said first polypeptide contains the amino acid sequence of SEQ ID NO: 53, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 54; or (xxvi) said first polypeptide contains the amino acid sequence of SEQ ID NO: 55, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 56; or (xxvii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 57, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 58; or (xxviii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 59, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 60; or

- 32 040031 (xxix) said first polypeptide contains the amino acid sequence of SEQ ID NO:

61, and the specified second polypeptide contains the amino acid sequence of SEQ ID NO: 62; or (xxx) said first polypeptide contains the amino acid sequence of SEQ ID NO:

63, and the specified second polypeptide contains the amino acid sequence of SEQ ID NO: 64; or (xxxi) said first polypeptide contains the amino acid sequence of SEQ ID NO: 65, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 66; or (xxxii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 67 and said second polypeptide contains the amino acid sequence of SEQ ID NO: 68.

As used herein, the term pharmaceutical composition refers to the formulation of a compound described herein in combination with a pharmaceutically acceptable carrier. The pharmaceutical composition may further contain additional agents (eg, for specific delivery, half-life extension, or other therapeutic compounds).

As used herein, the term pharmaceutically acceptable carrier means a pharmaceutically acceptable substance, composition, or vehicle such as a liquid or solid excipient, solvent, excipient, excipients (e.g., lubricant, talc, calcium or zinc stearate, or stearic acid), or a solvent encapsulating material that is involved in transferring or transporting the compound from one site of the body (eg, sites of administration) to another site (eg, organ, tissue, or body part). A pharmaceutically acceptable carrier is acceptable in the sense of being compatible with the other components of the drug and does not harm the tissues of the subject (eg, is physiologically compatible, sterile, has a physiological pH, etc.). Some examples of substances that may serve as pharmaceutically acceptable carriers include: 1) sugars such as lactose, glucose and sucrose; 2) starches such as corn starch and potato starch; 3) cellulose and its derivatives such as sodium carboxymethylcellulose, methylcellulose, ethylcellulose, microcrystalline cellulose and cellulose acetate; 4) powdered tragacanth; 5) malt; 6) gelatin; 7) lubricants such as magnesium stearate, sodium lauryl sulfate and talc; 8) excipients such as cocoa butter and suppository waxes; 9) oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; 10) glycols such as propylene glycol; 11) polyols such as glycerol, sorbitol, mannitol and polyethylene glycol (PEG); 12) esters such as ethyl oleate and ethyl laurate; 13) agar; 14) buffer substances such as magnesium hydroxide and aluminum hydroxide; 15) alginic acid; 16) pyrogen-free water; 17) isotonic saline; 18) Ringer's solution; 19) ethyl alcohol; 20) buffered pH solutions; 21) polyesters, polycarbonates and/or polyanhydrides; 22) bulk-creating substances such as polypeptides and amino acids; 23) a serum component such as serum albumin, HDL and LDL; 22) C ₂ -C ₁₂ alcohols such as ethanol; and 23) other non-toxic compatible substances used in pharmaceutical preparations. Other ingredients may include: wetting agents, coloring agents, release agents, coating agents, sweeteners, flavors, flavors, preservatives and antioxidants. Terms such as excipient, carrier, pharmaceutically acceptable carrier, or the like are used interchangeably herein.

In some embodiments, the compound of the technology in question is administered in the composition by injection, via a catheter, via a suppository, or via an implant, wherein the implant may be a porous, non-porous, or jelly-like substance, including a membrane such as a rubber membrane or fiber. As a rule, during the introduction of the composition, materials are used on which the compound of the studied technology is not absorbed.

In other embodiments, the compounds of this technology are delivered in a controlled release system. In one embodiment, a pump may be used (see, for example, Langer, 1990, Science 249: 1527-1533; Sefton, 1989, CRC Crit. Ref. Biomed. Eng. 14: 201; Buchwald et al., 1980, Surgery 88: 507; Saudek et al., 1989, N. Engl. J. Med. 321: 574). In another embodiment of the invention, polymeric materials can be used. (See, for example, Medical Applications of Controlled Release (Langer and Wise eds., CRC Press, Boca Raton, Fla., 1974); Controlled Drug Bioavailability, Drug Product Design and Performance (Smolen and Ball eds., Wiley, New York , 1984); Ranger and Peppas, 1983, Macromol. Sci. Rev. Macromol. Chem. 23:61. See also Levy et al., 1985, Science 228: 190; During et al., 1989, Ann. Neurol. 25 : 351; Howard et al., 1989, J. Neurosurg. 71: 105). Other controlled release systems are discussed, for example, in Langer, supra.

Compounds of the technology under study may be administered as pharmaceutical compositions containing a therapeutically effective amount of a binding agent and one or more pharmaceutically compatible ingredients.

In exemplary embodiments of the invention, the pharmaceutical composition is formulated according to routine procedures as a pharmaceutical composition adapted for intravenous or subcutaneous administration to a subject, such as a human. Typically, compositions for administration by injection are solutions in sterile isotonic aqueous buffer. IN

- 33 040031 if necessary, the pharmaceutical composition may also contain a substance that increases solubility, and a local anesthetic, such as lidocaine, to relieve pain at the injection site. In general, as a rule, the components are supplied either separately or in a mixture in a dosage form, for example, in the form of a dry lyophilized powder or anhydrous concentrate in a hermetically sealed container, such as an ampoule or sachet, indicating the amount of active substance. When the pharmaceutical composition is to be administered by infusion, it may be prepared in the form of an infusion bottle containing sterile pharmaceutical grade water or saline. When the pharmaceutical composition is administered by injection, an ampoule of sterile water for injection or saline may be provided so that the components can be mixed prior to administration.

The pharmaceutical composition for systemic administration may be a liquid, such as sterile saline, lactose Ringer's solution, or Hank's solution. In addition, the pharmaceutical composition may be in solid form, and redissolved or resuspended immediately prior to use. Freeze-dried forms are also contemplated.

The pharmaceutical composition may be contained within lipid particles or vesicles, such as liposomes or microcrystals, which are also suitable for parenteral administration. The particles may have any suitable structure, such as single or multilayer, as long as the compositions are contained therein. Compounds can be entrapped in stabilized plasmid-lipid particles (PLPs) that contain the fusion lipid dioleyoylphosphatidilethanolamine (DOPE), low levels (5-10 ml%) of cationic lipid, and that are stabilized with a polyethylene glycol (PEG) coating (Zhang Y.P. et al., Gene Ther. 1999, 6: 1438-47). Positively charged lipids such as K-[1-(2,3-dioleoyloxy)propyl]-K,K,K-trimethylammonium methyl sulfate, or DOTAP are particularly desirable for such particles and vesicles. The preparation of such lipid particles is well known. See, for example, US Pat. Nos. 4,880,635; 4906477; 4911928; 4917951; 4920016 and 4921757.

Pharmaceutical compositions of the present invention may be administered or packaged, for example, as a single dose. The term unit dose, when applied to a pharmaceutical composition of this disclosure, refers to physically discrete units suitable as a unit dosage for a subject, each unit containing a predetermined amount of the active substance, calculated to produce the desired therapeutic effect, in combination with the necessary diluent, for example, a carrier or carrier.

In some embodiments, a compound described herein may be combined with a therapeutic moiety, such as an anti-inflammatory agent. Methods for linking such therapeutic moieties to polypeptides, including, for example, Fc domains, are well known; see, for example, Amon et al., Monoclonal Antibodies For Immunotargeting Of Drugs In Cancer Therapy, in Monoclonal Antibodies And Cancer Therapy, Reisfeld et al. (Eds.), 1985, pp. 243-56, Alan R. Liss, Inc.); Hellstrom et al., Antibodies For Drug Delivery, in Controlled Drug Delivery (2nd Ed.), Robinson et al. (Eds.), 1987, pp. 623-53, Marcel Dekker, Inc.); Thorpe, Antibody Carriers Of Cytotoxic Agents In Cancer Therapy: A Review, in Monoclonal Antibodies '84: Biological And Clinical Applications, Pinchera et al. (Eds.), 1985, pp. 475-506) Analysis, Results, And Future Prospective Of The Therapeutic Use Of Radiolabeled Antibody In Cancer Therapy, in Monoclonal Antibodies For Cancer Detection And Therapy, Baldwin et al. (Eds.), 1985, pp. 303-16, Academic Press; and Thorpe et al. (1982) The Preparation And Cytotoxic Properties Of Antibody-Toxin Conjugates, Immunol. Rev., 62: 119-158.

Additionally, the pharmaceutical composition may be provided as a pharmaceutical kit, which contains: a) a container containing the compound of the present invention in lyophilized form; and b) a second container containing a pharmaceutically acceptable solvent (eg sterile water) for injection. A pharmaceutically acceptable solvent may be used to reconstitute or dilute the lyophilized compound of the technology in question. Optionally, such container(s) may be accompanied by a message in the form specified by a government agency regulating the manufacture, use, or sale of pharmaceutical or biological products, for which the message reflects approval by the body of manufacture, use, or sale for human administration.

In another aspect of the invention, an article of manufacture is included that contains materials useful in the treatment of the diseases described above. In some embodiments, an article of manufacture comprises a container and a label. Suitable containers include, for example, bottles, vials, syringes and test tubes. Containers can be made from various materials such as glass or plastic. In some embodiments, the container contains a composition that is effective in treating the diseases described herein and may have a sterile access port. For example, the container may be an intravenous solution bag or a vial having a stopper that is pierced by a hypodermic needle. The active substance in the composition is a compound according to this technology. In some embodiments, the implementation of the label on the container, or that is attached to the container, indicates that the composition is used to treat the disease of choice. The article of manufacture may further comprise a second container containing a pharmaceutically acceptable buffer such as phosphate buffered saline, Ringer's solution, or glucose solution. It may further contain other materials commercially desired by the user, including other buffers, diluents, filters, needles, syringes, and inserts with instructions for use.

Without going into details, it is believed that a person skilled in the art, based on the above description of the invention, uses this disclosure of the invention in full. As a consequence, the following specific embodiments are to be considered as illustrative only and not as limiting the rest of the disclosure in any form. All publications cited in this document are incorporated by reference for the purposes or subject matter of the invention mentioned in this document.

Examples

Example 1 Construction of Exemplary Compounds Targeting IL23A and FAVL (BAFF).

Compounds of this technology were prepared by recombinant methods known in the art (see, for example, PCT publications WO 2006/113665, WO 2008/157379 and WO 2010/080538, the contents of which are incorporated herein by reference). In table. 2A shows exemplary compounds that bind to both IL23A and FAVL (BAFF) used in the examples below. Briefly, plasmids encoding the first and second polypeptides for each compound were transfected together into CHO-S cells using the FreeStyle MAX Reagent (CHO). The cells were cultured for 13-14 days and the compounds that were produced by the cells were purified using Protein-A chromatography. Compounds were further purified by size exclusion chromatography.

- 35 040031

Table 2A. Exemplary IL23A and FAVL Binding Compounds (BAFF)

Connection ID vL big chain vH of the big chain vL small circuit vH small circuit 3rd and/or 4th linker types Constant domain isotype SEQ ID NO: (1st/2nd polypeptides) Compound E BAFF (1) VL (SEQ ID NO:2) IL23A(1) VH (SEQ ID NO:3) IL23A (1) VL (SEQ ID NO:4) BAFF (1) VH (SEQ ID NO:1) LGGGSG (SEQ ID NO:70) IgGlKO- YTE (SEQ ID NO:128) 5/6 Compound V IL23A (1) VL (SEQ ID NO:4) FAVL (BAFF)(1) VH (SEQ IDNO:1) BAFF (1) VL (SEQ ID NO:2) IL23A (1) VH (SEQ ID NO:3) LGGGSG (SEQ ID NO:70) IgGlKO- YTE (SEQ ID NO: 128) 7/8 Connection U BAFF (1) VL (SEQ ID NO:2) IL23A(1) VH (SEQ ID NO:3) IL23A (1) VL (SEQ ID NO:4) BAFF (1) VH (SEQ ID NO:1) E(C-l) (SEQ ID NO:82), K-helix (C-l) (SEQ ID NO:83) FcIgGlKOYTE (SEQ ID NO:129) 9/10 Connection T IL23A (1)VL (SEQ ID NO:4) FAVL (BAFF)(1) VH (SEQ IDNO:1) BAFF (1) VL (SEQ ID NO:2) IL23A (1)VH (SEQ ID NO:3) E(C-l) (SEQ ID NO:82), K-helix( C-l) (SEQ ID NO:83) FcIgGlKOYTE (SEQ ID NO:129) 11/12 Compound X BAFF (1) VL (SEQ ID NO:2) IL23A(1) VH (SEQ ID NO:3) IL23A (1) VL (SEQ ID NO:4) BAFF (1) VH (SEQ ID NO:1) LGGGSG (SEQ ID NO: 70) IgG4-Pro- YTE (SEQ ID NO: 74) 13/14 Connection F IL23A (1) VL (SEQ ID NO:4) FAVL (BAFF)(1) VH (SEQ IDNO:1) BAFF (1) VL (SEQ ID NO:2) IL23A (1) VH (SEQ ID NO:3) LGGGSG (SEQ ID NO: 70) IgG4-Pro- YTE (SEQ ID NO: 74) 15/16 Connection W BAFF (1) VL (SEQ ID NO:2) IL23A(1) VH (SEQ ID NO:3) IL23A (1) VL (SEQ ID NO:4) BAFF (1) VH (SEQ ID NO:1) E(C-l) (SEQ ID NO:82), K-helix( C-l) (SEQ ID NO:83) Fc-IgG4Pro-YTE (SEQ ID NO: 127) 17/18 Connection S IL23A(1) VL (SEQ ID NO:4) FAVL (BAFF)(1) VH (SEQ IDNO:1) BAFF (1) VL (SEQ ID NO:2) IL23A (1) VH (SEQ ID NO:3) E(C-l) (SEQ ID NO:82), K-helix( C-l) (SEQ ID NO:83) Fc-IgG4Pro-YTE (SEQ ID NO:127) 19/20 Connection G IL23A(1) VL (SEQ ID NO:4) FAVL (BAFF) (4) VH FAVL (BAFF) (4)VL IL23A (1) VH (SEQ ID LGGGSG (SEQ ID NO: 70) IgGlKO- YTE (SEQ ID) 21/22

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(SEQ ID NO:84) (SEQ ID NO: 85) NO:3) NO:128) AA connection IL23A(1) VL (SEQ ID NO:4) BAFF (4) VH (SEQ ID NO:84) BAFF (4) VL (SEQ ID NO: 85) IL23A (1) VH (SEQ ID NO:3) E(C-l) (SEQ ID NO:82), K-helix( C-l) (SEQ ID NO:83) FcIgGlKOYTE (SEQ ID NO:129) 23/24 Compound I IL23A(1) VL (SEQ ID NO:4) BAFF (4) VH (SEQ ID NO:84) BAFF (4) VL (SEQ ID NO: 85) IL23A (1) VH (SEQ ID NO:3) LGGGSG (SEQ ID NO: 70) IgG4-Pro- YTE (SEQ ID NO: 74) 25/26 AB connection IL23A(1) VL (SEQ ID NO:4) BAFF (4) VH (SEQ ID NO:84) BAFF (4) VL (SEQ ID NO: 85) IL23A (1) VH (SEQ ID NO:3) E(C-l) (SEQ ID NO:82), K-helix (C-l) (SEQ ID NO:83) FcIgGlKOYTE (SEQ ID NO:129) 27/28 Connection K IL23A(1) VL (SEQ ID NO:4) BAFF (5) VH (SEQ ID NO:86) BAFF (5) VL (SEQ ID NO:87) IL23A (1) VH (SEQ ID NO:3) LGGGSG (SEQ ID NO: 70) IgGlKO- YTE (SEQ ID NO:128) 29/30 AC connection IL23A(1) VL (SEQ ID NO:4) BAFF (5) VH (SEQ ID NO:86) BAFF (5) VL (SEQ ID NO:87) IL23A (1) VH (SEQ ID NO:3) E(C-l) (SEQ ID NO:82), K-helix (C-l) (SEQ ID NO:83) FcIgGlKOYTE (SEQ ID NO:129) 31/32 Connection M IL23A(1) VL (SEQ ID NO:4) BAFF (5) VH (SEQ ID NO:86) BAFF (5) VL (SEQ ID NO:87) IL23A (1) VH (SEQ ID NO:3) LGGGSG (SEQ ID NO: 70) IgG4-Pro- YTE (SEQ ID NO: 74) 33/34 AD connection IL23A(1) VL (SEQ ID NO:4) BAFF (5) VH (SEQ ID NO:86) BAFF (5) VL (SEQ ID NO:87) IL23A (1) VH (SEQ ID NO:3) E(C-l) (SEQ ID NO:82), K-helix (C-l) (SEQ ID NO:83) Fc-IgG4Pro-YTE (SEQ ID NO:127) 35/36 Connection C IL23A(1) VL (SEQ ID NO:4) FAVL (BAFF) (6) VH (SEQID NO:88) FAVL (BAFF) (6) VL (SEQ ID NO:89) IL23A (1) VH (SEQ ID NO:3) LGGGSG (SEQ ID NO: 70) IgGlKO- YTE (SEQ ID NO:128) 37/38 AE connection IL23A(1) VL (SEQ ID NO:4) BAFF (6) VH (SEQ ID NO:88) BAFF (6) VL (SEQ ID NO:89) IL23A (1) VH (SEQ ID NO:3) E(C-l) (SEQ ID NO:82), K-helix( C-l) (SEQ ID NO:83) FcIgGlKOYTE (SEQ ID NO:129) 39/40

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Compound D IL23A(1) VL (SEQ ID NO:4) BAFF (6) VH (SEQ ID NO:88) BAFF (6) VL (SEQ ID NO:89) IL23A (1) VH (SEQ ID NOG) LGGGSG (SEQ ID NO: 70) IgG4-Pro- YTE (SEQ ID NO: 74) 41/42 AF connection IL23A(1) VL (SEQ ID NO:4) BAFF (6) VH (SEQ ID NO:88) BAFF (6) VL (SEQ ID NO:89) IL23A (1) VH (SEQ ID NOG) E(C-l) (SEQ ID NO:82), K-helix( C-l) (SEQ ID NO:83) Fc-IgG4Pro-YTE (SEQ ID NO:127) 43/44 Compound A IL23A(1) VL (SEQ ID NO:4) FAVL (BAFF) (7) VH (SEQ ID NO:90) FAVL (BAFF) (7) VL (SEQ ID NO:91) IL23A (1) VH (SEQ ID NOG) LGGGSG (SEQ ID NO: 70) IgGlKO- YTE (SEQ ID NO:128) 45/46 AG connection IL23A(1) VL (SEQ ID NO:4) FAVL (BAFF) (7) VH (SEQ ID NO:90) FAVL (BAFF) (7) VL (SEQ ID NO:91) IL23A (1) VH (SEQ ID NOG) E(C-l) (SEQ ID NO:82), K-helix( C-l) (SEQ ID NO:83) FcIgGlKOYTE (SEQ ID NO:129) 47/48 Connection B IL23A(1) VL (SEQ ID NO:4) FAVL (BAFF) (7) VH (SEQ ID NO:90) FAVL (BAFF) (7) VL (SEQ ID NO:91) IL23A (1) VH (SEQ ID NOG) LGGGSG (SEQ ID NO: 70) IgG4-Pro- YTE (SEQ ID NO:74) 49/50 AN connection IL23A(1) VL (SEQ ID NO:4) FAVL (BAFF) (7) VH (SEQ ID NO:90) FAVL (BAFF) (7) VL (SEQ ID NO:91) IL23A (1) VH (SEQ ID NOG) E(C-l) (SEQ ID NO:82), K-helix( C-l) (SEQ ID NO:83) Fc-IgG4Pro-YTE (SEQ ID NO:127) 51/52 Connection H BAFF (4) VL (SEQ ID NO:85 IL23A(1) VH (SEQ ID NO:3) IL23A (1) VL (SEQ ID NOG) BAFF (4) VH (SEQ ID NO:84) LGGGSG (SEQ ID NO: 70) IgGlKO- YTE (SEQ ID) NO:128) 53/54 Compound J BAFF (4) VL (SEQ ID NO: 85) IL23A(1) VH (SEQ ID NO:3) IL23A (1) VL (SEQ ID NOG) BAFF (4) VH (SEQ ID NO:84) LGGGSG (SEQ ID NO: 70) IgG4-Pro- YTE (SEQ ID NO:74) 55/56 Connection L BAFF (5) VL (SEQ ID NO:87) IL23A(1) VH (SEQ ID NOG) IL23A (1) VL (SEQ ID NOG) BAFF (5) VH (SEQ ID NO:86) LGGGSG (SEQ ID NO: 70) IgGlKO- YTE (SEQ ID NO:128) 57/58 Compound N BAFF (5) VL (SEQ ID NO:87) IL23A(1) VH (SEQ ID NOG) IL23A (1) VL (SEQ ID NOG) BAFF (5) VH (SEQ ID NO:86) LGGGSG (SEQ ID NO: 70) IgG4-Pro- YTE (SEQ ID NO:74) 59/60 Connection O FAVL (BAFF) (6) VL IL23A(1) VH (SEQ ID NOG) IL23A (1) VL (SEQ ID FAVL (BAFF) (6) VH LGGGSG (SEQ ID NO: 70) IgGlKO- YTE (SEQ ID) 61/62

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(SEQ ID NO:89) NO:4) (SEQ ID NO:88) NO:128) Connection P BAFF (6) VL (SEQ ID NO:89) IL23A(1) VH (SEQ ID NO:3) IL23A (1) VL (SEQ ID NO:4) FAVL (BAFF) (6) VH (SEQ ID NO:88) LGGGSG (SEQ ID NO: 70) IgG4-Pro- YTE (SEQ ID NO:74) 63/64 Connection Q FAVL (BAFF) (7) VL (SEQ ID NO:91) IL23A(1) VH (SEQ ID NO:3) IL23A (1)VL (SEQ ID NO:4) BAFF (7) VH (SEQ ID NO:90) LGGGSG (SEQ ID NO: 70) IgGlKO- YTE (SEQ ID NO:128) 65/66 Compound R FAVL (BAFF) (7) VL (SEQ ID NO:91) IL23A(1) VH (SEQ ID NO:3) IL23A (1) VL (SEQ ID NO:4) FAVL (BAFF) (7) VH (SEQ ID NO:90) LGGGSG (SEQ ID NO: 70) IgG4-Pro- YTE (SEQ ID NO:74) 67/68

VL - light chain variable domain, VH - heavy chain variable domain, each chain contains a linker GGGSGGG (SEQ ID NO: 69) between VL and VH.

Control antibodies were also used for comparison purposes. Monoclonal antibodies targeting either FAVL (BAFF) or IL23 served as controls.

Table 2B. Comte] ash antibodies/antagonist Control compounds Subsequence Control antibody 1 (monoclonal antibody to FAVL (BAFF)) (FAVL (BAFF) (D) FAVL (BAFF)(1) VH (SEQ ID NO: 1) FAVL (BAFF)(1) VL (SEQ ID NO:2) Control fusion protein antagonist 2 (selective antagonist FAVL (BAFF) without antibody) (FAFL (BAFF) (2)) BAFF (2) (SEQ ID NO: 131) Control antibody 3 (monoclonal antibody to FAVL (BAFF)) (FAVL (BAFF) (3)) FAVL (BAFF)(3) VH (SEQ ID NO: 132) FAVL (BAFF)(3) VL (SEQ ID) NO:133) Control antibody 4 (monoclonal antibody to IL23A) (IL-23A (1)) IL23A(1) VH (SEQ ID NO:3) IL23A(1) VL (SEQ ID NO:4)

Example 2 Thermal Stability of Compounds.

Ways.

The thermal unfolding and aggregation of 2 mg/ml solutions of the compounds in phosphate buffer from 20 to 110° C. at a scan rate of 60° C./h was monitored using an automatic capillary differential scanning calorimeter (DSC) (MicroCai, LLC, Boston). Two scans were performed with the appropriate buffer in order to adjust the instrument's thermal history and obtain a baseline for each sample, with the average of these scans subtracted from the next protein thermogram to obtain the true heat capacity.

The normalized scan results were subsequently analyzed using Origin 7.0. Baselines before the start of the transition were subtracted for each resulting heat capacity thermogram in order to obtain the resulting excess heat capacity (Cp, ex) as a function of temperature. The specified values of the transition temperatures (Tm) represent the positions of the peak maxima determined by visual inspection of the experimental thermograms.

Results.

The results are shown in table. 3. The data show that exemplary compounds targeting IL23A and FAFL (BAFF) have a first peak transition temperature range (T _mi ) ranging from 51.59 to 71.25°C. The results are unexpected since the exemplary compounds all have the same general structure and contain the same VH and VL gene sequences targeting IL-23A. Compounds with higher transition temperatures are more stable and are expected to have a longer shelf life.

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Table 3. Thermal Unfolding Transition Temperatures for Joints

Connection ID _Tmi (°C) T _t2 (°C) Compound A 69.35 83.63 Connection B 65.87 — Connection C 67.89 82.36 Compound D 64.93 — Compound E 51.59 68.8 Connection F 52.75 64.48 Connection G 70.88 83.21 Connection H 71.25 83.62 Compound I 67.83 — Compound J 68.26 — Connection K 68.33 83.23 Connection L 68.3 82.58 Connection M 64.98 — Compound N 65.52 — Connection O 67.79 83.85 Connection P 65.09 — Connection Q 69.19 81.9 Compound R 65.97 — Compound X 52.36 64.53

Example 3 Surface Plasmon Resonance (SPR) Affinity of Exemplary Compounds.

Test compounds were analyzed by R&D to determine affinity for FAVL (BAFF) and IL23A.

Materials and methods.

R&D experiments were performed on a ProteOn XPR36 instrument (Bio Rad). The GLM chip was pretreated with sequential injections of 60 s of 0.5% SDS (sodium dodecyl sulfate, SDS), 50 mM NaOH solution, and 100 mM HCl at a flow rate of 30 μl/min in both vertical and horizontal directions. The pretreated GLM chip was activated by injecting a mixture of EDC (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide) (76.7mg/ml) and Sulfo-NHS (N-hydroxysulfosuccinimide) (21.7mg/ml ) with a ratio of 1:1 in 6 horizontal channels. IgG goat-anti-human (GAHA) Fc γ (Invitrogen) at 30 μg/ml in 10 mM, pH 5.0 sodium acetate buffer was immobilized to 8,000 resonant units on an activated GLM chip in 6 horizontal channels. The chip was finally deactivated with 1M ethanolamine-HCl in 6 horizontal channels. The prepared GAHA chip was returned to the vertical position to capture test compounds in 5 vertical channels, and the last channel was used as a reference column.

The capture chip was again placed in the horizontal plane for binding. Bound human IL-23 (Boehringer Ingelheim Pharmaceuticals, Inc) at five concentrations, 10.0, 5.00, 2.50, 1.25, and 0.625 nM, was injected horizontally over the surface of the test compounds for 10 min at a flow rate of 40 µl/min in the following run buffer (Bio Rad): phosphate-buffered saline (pH 7.4), 0.005% Tween 20. Dissociation was allowed for 2 h. .85% phosphoric acid (Bio Rad) at a flow rate of 100 µl/min in the horizontal and vertical directions after 10 min of association and 2 h of dissociation. The reconstituted GAHA was ready for another binding cycle. Binding of compounds to cynomolgus monkey IL23, human FAVL (BAFF), or cynomolgus monkey FAVL (BAFF) was performed in a similar manner, but the titration concentrations for binding to human FAVL (BAFF) or cynomolgus FAVL (BAFF) were 6.25, 3.12, 1.56, 0.78 and 0.39 nM.

Results.

The results in the table. 4 show that both test compounds were able to bind FAVL (BAFF) and IL23 with a dissociation constant (Kd) in the picomolar range.

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Table 4. Affinity of compounds that bind to FAVL (BAFF) and IL23

Connection ID K _d for human FAVL (BAFF) (pM) K _d for FAVL (BAFF) cynomolgus macaque (pM) K _d for human IL23 (pM) K _d for cynomolgus monkey IL23 (pM) Compound A less than 15 less than 15 less than 15 less than 15 Connection B less than 15 less than 15 less than 15 less than 15 Connection C less than 15 less than 15 less than 15 less than 15 Compound D less than 15 less than 15 less than 15 less than 15 Compound E less than 15 less than 15 less than 15 less than 15 Connection F less than 15 less than 15 less than 15 less than 15

Example 4 Inhibition of human or cynomolgus macaque trimer-induced NFkB activation in FAVLR (BAFFRVCHO with luciferase reporter) cells.

Materials/methods.

Briefly, human NFkB CHO FAVLR (BAFFR) cells with luciferase reporter were collected, washed, counted and resuspended to a concentration of 1.6x106 cells per ml in assay medium (AM) (v/v) 1% penicillin/streptomycin in X-VIV015 , chemically defined serum-free medium (Lonza). Human or cynomolgus macaque recombinant BAFF trimer (Boynringer Ingelheim Pharmaceuticals, Inc.) was prepared at a single concentration (52 pM) in AM and pre-incubated with AM alone, or with successive titrations of test compound for 30 min at 37°C , 5% CO ₂ in a humidified incubator. After pre-incubation with BAFF plus test compound, 50 µl of the mixture(s) was added to 50 µl of cells and the test plate was further incubated at 37°C (as described) for 24 hours. or recombinant trimer FAVL (BAFF) diluted in AM (stimulated controls). After a 24 hour incubation, the cell suspension was treated with 100 µl STEADY-Glo reagent (Promega) following the manufacturer's instructions and analyzed for luciferase expression. The resulting Relative Luminescence Units (RLUs) were plotted against Log 10 nanomolar test compound concentration, with _IC50 and IC90 values calculated using a 4-parameter logistic model supported by the Excel Xlfit add-in (ID Business Solutions Limited). The IC ₅₀ and IC ₉₀ values of the test compound were calculated as described above, and the geometric averages were calculated over several experiments and are shown in Table 1. 5 and 6.

Results.

The test compounds dose-dependently inhibited NFkB activation induced by the human or cynomolgus monkey trimer (BAFF) in FAVLR (BAFFR)-CHO cells with a luciferase reporter. The results illustrated in table. 5 and 6 show that the geometric mean IC ₅₀ and IC ₉₀ values for the test compounds were comparable to or higher than for the control FAVL antagonists (BAFF).

Table 5 Geometric mean IC ₅₀ and IC ₉₀ values for inhibition of the human FAFL trimer (BAFF) in FAVLR reporter cells (BAFFR)-CHO

Connection ID 1C ₅ o (pM) 1Seo (pM) Compound A 43 180 Connection B 39 256 Connection C 35 110 Compound D 34 172 Connection F 72 293 Connection O 27 119 Connection P 19 88 Connection Q 26 108 Compound R thirty 139 Compound X 106 361 Control antibody 1 (BAFF (1)) 232 743 Control fusion protein antagonist 2 (FAVL (BAFF) (2)) 173 914 Control antibody 3 (BAFF (3)) 70 417

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Table 6. Geometric mean IC50 and IC90 values for trimer inhibition

FAVL (BAFF) of the cynomolgus macaque in FAVLR (BAFFR)-CHO reporter cells

Connection ID IC50 Geometric mean pM IC90 Geometric mean pM Compound A 132 191 Connection B 132 295 Compound D 118 248 Control Antibody 1 (BAFF) (B) 288 635 Control fusion protein antagonist 2 (BAFF (2)) 602 1089 Control antibody 3 (BAFF (3)) 257 588

Example 5 Inhibition of NFkB activation induced by human FAFL trimer (BAFF) in TACI-CHO cells with a luciferase reporter.

Materials/methods.

Briefly, human NFkB CHO TACI (TACI) cells with luciferase reporter were collected, washed, counted and resuspended to a concentration of 1.6x106 cells per ml in assay medium (AM) (v/v) 1% penicillin/streptomycin in X-VIV015 , chemically defined serum-free medium (Lonza). Recombinant human FAFF trimer (BAFF) (Boynringer Ingelheim Pharmaceuticals, Inc.) was prepared at a single concentration (222 pM) in AM and pre-incubated with AM alone, or with successive titrations of test compound for 30 minutes at 37°C, 5% CO2 in an incubator with humidification. After pre-incubation with BAFF plus test compound, 50 µl of the mixture(s) was added to 50 µl of cells and the test plate was further incubated at 37°C (as described) for 24 hours. Control samples received either AM (unstimulated controls), or human recombinant trimer FAVL (BAFF) diluted in AM (stimulated controls). After a 24 hour incubation, the cell suspension was treated with 100 μl STEADY-Glo reagent (Promega) following the manufacturer's instructions and analyzed for luciferase expression. The resulting Relative Luminescence Units (RLUs) were plotted against Log 10 nanomolar test compound concentration, with IC ₅₀ and IC ₉₀ values calculated using a 4-parameter logistic model supported by the Excel Xlfit add-in (ID Business Solutions Limited). The IC ₅₀ and IC ₉₀ values of the test compound were calculated as described above, and the geometric averages were calculated over several experiments and are shown in Table 1. 7.

Results.

The test compounds dose-dependently inhibited NFkB activation induced by the human FAFL trimer (BAFF) in luciferase reporter TACI-CHO cells. The results illustrated in table. 7 show that the geometric mean IC ₅₀ and IC ₉₀ values for the test compound were comparable to or higher than those for the control FALV antagonists (BAFF).

Table 7. Geometric mean IC50 and IC90 values for human BAFF trimer inhibition in TACI-CHO reporter cells

Connection ID IC50 Geometric mean pM IC90 Geometric mean pM Compound A 159 417 Connection B 159 510 Compound D 163 436 Control antibody 1 (BAFF (1)) 273 576 Control fusion protein antagonist 2 (FAVL (BAFF) (2)) 829 1945 Control antibody 3 (BAFF (3)) 241 618

Example 6 Inhibition of NFkB activation induced by 60-mer FAVL (BAFF) in FAVLR (BAFFR)-CHO cells with a luciferase reporter.

Materials/methods.

Briefly, human NFkB CHO FAVLR (BAFFR) cells with luciferase reporter were collected, washed, counted and resuspended to a concentration of 1.6x106 cells per ml in assay medium (AC (AM)) (v/v) 1% penicillin/streptomycin in X-VIV015, chemically defined serum-free medium (Lonza). Recombinant human BAFF 60-mer (Boynringer Ingelheim Phar

- 42 040031 maceuticals, Inc.) was prepared at a single concentration (4.2 pM) in AC (AM) and pre-incubated with AC (AM) alone, or with successive titrations of test compound for 30 min at 37°C, 5 % CO2 in a humidified incubator. After pre-incubation with BAFF plus test compound, 50 µl of mixture(s) was added to 50 µl of cells and the test plate was further incubated at 37°C (as described) for 24 hours. controls), or recombinant human 60-mer FAVL (BAFF) diluted in AC (AM) (stimulated controls). After a 24 hour incubation, the cell suspension was treated with 100 μl STEADY-Glo reagent (Promega) following the manufacturer's instructions and analyzed for luciferase expression. The resulting Relative Luminescence Units (RLUs) were plotted against Log 10 nanomolar test compound concentration, with IC ₅₀ and IC ₉₀ values calculated using a 4-parameter logistic model supported by the Excel Xlfit add-in (ID Business Solutions Limited). The IC ₅₀ and IC ₉₀ values of the test compound were calculated as described above, and the geometric averages were calculated over several experiments and are shown in Table 1. 8.

Results.

The test compounds dose-dependently inhibited NFkB activation induced by human 60-mer FAVL (BAFF) in FAVLR (BAFFR)-CHO cells with a luciferase reporter. The results illustrated in table. 8 show that the geometric mean IC ₅₀ and IC ₉₀ values for test compounds A, B, C, D, O, P, Q and R were higher than for control antagonists FAVL (BAFF).

Table 8. Geometric Mean IC ₅₀ and IC ₉₀ Values for Inhibition of Human 60-mer FAVL (BAFF) in FAVLR Reporter Cells (BAFFR)-CHO

Connection ID IC50 (pM) IC90 (pM) Compound A 6 19 Connection B 6 17 Connection C 6 14 Compound D 4 12 Connection F 1300 7388 Connection O 5 6 Connection P 2 9 Connection Q 5 13 Compound R 5 16 Compound X 1555 6839 Control antibody 1 (BAFF (1)) 24075 180053 Control fusion protein antagonist 2 (FAVL (BAFF) (2)) 43 107 Control antibody 3 (BAFF (3)) 15 38

Example 7 Neutralization of NFkB activation induced by membrane bound FAVL (BAFF) in FAVLR (BAFFR)-CHO cells with a luciferase reporter.

Materials/methods.

CHO-K1 cells expressing human FAVL (BAFF) were counted and resuspended to a concentration of 2x106 cells per ml in standard growth medium. To stop membrane-bound FAFL (BAFF) degradation, cells were treated with 0.125% paraformaldehyde (Electron Microscopy) and incubated at room temperature for one hour. Fixed human cells FAVL (BAFF) CHO-K1 were then washed and resuspended to 2x106 cells per ml in standard growth medium and incubated overnight at 37°C, 5% CO ₂ . Then, fixed human BAFF CHO-K1 cells were collected and resuspended to a concentration of 3.2x106 cells per ml in XVIV015, a chemically defined serum-free assay medium (Lonza) (AC (AM)) containing 1% penicillin/streptomycin (vol. /about.).

Human NFkB CHO FAVLR (BAFFR) cells with luciferase reporter were collected, washed and resuspended to a concentration of 1.6x106 cells per ml in assay medium. Fixed human FAVL (BAFF) CHO-K1 cells, prepared at 3.2x106 cells per ml in AC (AM), and pre-incubated with serial titrations of test compounds for 30 min, were then added to 50 µl of CHO FAVLR (BAFFR) NFkB cells human luciferase reporter cells and additionally incubated at 37°C for 24 h. Control reporter cells received either AC (AM) alone (unstimulated controls) or fixed human FALV (BAFF) CHO-K1 cells diluted in AC (AM) ( stimulated controls). After 24

- 43 040031 hour incubation, samples were treated with 100 µl of STEADY-Glo reagent (Promega) and analyzed for luciferase expression. Relative luminescence units (RLUs) were plotted against Log 10 nanomolar concentrations of test compounds, with IC ₅₀ and IC ₉₀ values calculated using a 4-parameter logistic model supported by the Excel Xlfit add-in (ID Business Solutions Limited). The IC ₅₀ and IC ₉₀ values of the test compounds were calculated as described above, and geometric averages were calculated across several experiments and are shown in Table 1. 9.

Results.

The test compounds dose-dependently inhibited NFkB activation induced by membrane bound human FAVL (BAFF) in FAVLR (BAFFR)-CHO cells with a luciferase reporter. The results illustrated in table. 9 show that the IC ₅₀ and IC ₉₀ values for the test compounds were higher than for all three control FAVL antagonists (BAFF).

Table 9. Geometric mean IC ₅₀ and IC ₉₀ values for inhibition of membrane bound FAVL (BAFF) in FAVLR reporter cells (BAFFR)-CHO

Connection ID 1C ₅ o (pM) IC90 (pM) Compound A 118 1015 Connection B 125 1139 Compound D 104 938 Control antibody 1 (BAFF (1)) 553 10116 Control fusion protein antagonist 2 (FAVL (BAFF) (2)) 581 3678 Control antibody 3 (BAFF (3)) 250 2916

Example 8 Inhibition of human IL-23 activity in B-lymphocyte lymphoblasts (DB cells) with STAT3-luciferase reporter.

Materials/methods.

B lymphocyte lymphoblasts (DB cells, ATCC catalog number CRL-2289) were stably transduced with the lentiviral STAT-3/luciferase reporter gene (Qiagen). The transduced cells were kept under selection using puromycin (Life Technologies). The complete culture medium was RPMI-1640 medium (Life Technologies) supplemented with 10% FBS (Hyclone) and 2 μg/ml puromycin (Life Technologies). The assay medium was RPMI-1640 (Life Technologies) supplemented with 10% FBS (Hyclone).

Engineered DB-STAT3 cells were seeded at 20,000 cells/well, in a 96-well flat-bottomed white plate, in 80 μl/well of assay medium. Test compounds (10x) were prepared in round bottom polypropylene 96-well plates in assay medium and diluted accordingly to achieve dose ranges of 1 μg/mL to 10 pg/mL. 10 µl of diluted test molecules or assay medium (for control wells) was added to each well in triplicate. 10 μl of 10x human IL-23 was added to each well (to a final concentration of 75 ng/ml per plate). Alternatively, 10 µl of medium was added to the control wells. The dose of IL-23 selected for use in the assay is the EC ₆₀ human IL-23 stimulation dose for engineered DB cells as determined in previous studies. The plates were incubated overnight at 37°C in 5% CO ₂ . The ONE-Glo™ (Promega) luciferase assay reagent was prepared and 100 µl was added to each well and mixed. Luminescence was measured on an Envision plate reader and then plotted (y-axis) against antibody concentration (x-axis). Compound IC ₅₀ values were determined by applying the data to a 4-parameter sigmoidal dose-dependent function using GraphPad Prism 6 software. IC ₉₀ values were determined by computing data using Find ECanything using GraphPad Prism 6 software. Geometric means were calculated over several experiments , and are shown in Table. 10.

Results.

The results showed that the test compounds were able to inhibit the activity of human IL-23 in DB cells with STAT3-luciferase reporter.

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Table 10. Geometric means of IC ₅₀ and IC ₉₀ for inhibition of __________ human IL-23 in B cell lymphoblasts __________

Connection ID ICso (pM) IC90 (pM) Compound A 145 1229 Connection B 161 1101 Connection C 189 2330 Compound D 128 3087 Control antibody 4 (IL-23 A (1)) 79 634

Example 9 Inhibition of human and cynomolgus monkey IL-23 activity in mouse splenocytes. Materials and methods.

Mononuclear cells from mouse spleens (female C57BL/6 mice less than 13 weeks old; Jackson Laboratories) were isolated, washed, counted and resuspended to 4 x 10 ⁶ cells/ml in standard T lymphocyte medium (TLC). 100 µl of the mIL-2/splenocyte suspension was added to 96 well microtiter plates. Recombinant human IL-23 (Boehringer Ingelheim Pharmaceuticals, Inc.) or recombinant cynomolgus monkey IL-23 (Boehringer Ingelheim Pharmaceuticals, Inc.) were diluted in TCM (tissue culture medium) and preincubated for 2 h at 37 °C only in TCS, or with test sample titrations. After pre-incubation of the test sample plus IL-23, 100 μl of the mixture was added to the cells and the test plates were incubated at 37°C with 5% CO ₂ -humidified air for 48 hours. Control samples received either TCS (non-stimulated controls) or recombinant IL23 diluted in TCS (stimulated controls). After incubation, mouse IL-17 levels were determined from the supernatant using the Quantikine® Mouse IL-17 Immunoassay according to the manufacturer's instructions (R&D Systems). Interpolated mIL17 pg/ml values were determined for each sample and converted to percent of control (PCR). SOC was plotted against test sample concentration and IC ₉₀ values were calculated using a 4 Parameter Logistic Model that was available through Excel XLfit (Activity Base software, ID Business Solutions, Ltd.). Test compounds were analyzed for IC ₅₀ and IC ₉₀ as described above, and geometric averages were calculated over multiple runs for each test sample, and are shown in Table 1. eleven.

Results.

The results show that the test compounds were able to suppress the release of IL-17 from mouse splenocytes, which was induced by human or cynomolgus monkey IL23.

Table 11 Geometric mean IC50 and IC90 values for human IL-23 inhibition in B cell lymphoblasts

Connection ID human IL23 IL23 cynomolgus monkey IC50 (pM) IC90 (pM) IC50 (pM) IC90 (pM) Compound A 146 1647 155 1309 Connection B 202 1360 200 1480 Connection C 133 761 110 880 Compound D 105 1700 97 1554 Control antibody 4 (IL-23A (1)) 84 366 64 331

Example 10 Pharmacokinetics of compounds in the cynomolgus macaque.

Materials and methods.

Single intravenous (IV(IV)) dose PK studies for test compounds A, B, C and D were performed in male cynomolgus monkeys (n=2 for each molecule). Doses were administered as a slow bolus IV (IV) injection of 1 mg/kg. Whole blood samples were collected pre-dose, and 0.25, 2, and 6 hours post-dose on the day of dosing, and 1, 2, 3, 4, 7, 10, 14, 21, and 28 days post-dose. Serum concentration of dosed molecules was measured using an MSD-based ligand binding assay.

Standard curve and quality control (QC) samples were prepared in 100% pooled cynomolgus monkey serum. Each standard curve consisted of seven non-zero points starting at 512 ng/mL, which were then serially diluted 3-fold. A blank sample (matrix without test sample) was also included. Four QC samples were prepared in the low, mid and high ranges, starting at 256 ng/mL, which were then serially diluted four times to 8 ng/mL, then a twofold dilution was used to prepare the lowest QC of 4 ng/mL . Samples for the standard curve and QC (QC) were included in duplicate at each analytical run. The lower and upper limits of quantification were defined as the lowest and highest QC points having reproducible back calculated concentrations.

- 45 040031 not exceeding 30 percent (%) of the nominal concentration. The acceptance criterion for standard curve points was 30 percent (%) of the nominal concentration.

To measure the concentration of active drug in serum samples, a master mix was prepared combining biotinylated recombinant human FAVL (BAFF) and 0.5 µg/mL sulfo-labeled goat anti-human IgG secondary antibody in 0.5 µg/mL binding buffer (5 % BSA in 1 x PBS (phosphate-buffered saline) with 0.05% Tween 20). The master mix was added to a 96-well non-binding and light-blocking plate at a volume of 50 μl/well. Twenty-five µl of standards and QC (stocks diluted 1:20 in binding buffer) were added to the wells, in duplicate, per non-binding plate containing the master mix. Unknown serum samples were diluted 1:20 in binding buffer and 1:400 in binding buffer containing 5% serum. 25 µl of the diluted samples were added to the wells of the non-binding plate containing the master mix. Non-binding plates were incubated at room temperature on a plate shaker (500 rpm, 1.5 h). In parallel, the Streptavidin GOLD MSD plate was blocked using 150 µl blocking buffer (5% BSA with 1X PBS with 0.05% Tween 20) and incubated at room temperature on a plate shaker (500 rpm, 1.5 h). After incubation, the MSD plates were washed three times with 300 μl/well wash buffer (0.05% Tween 20 in 1X PBS). 50 μl of sample from non-binding plates was added to MSD plates and incubated at room temperature (1.5 h, 600 rpm). After incubation, plates were washed three times with wash buffer and 150 µl of 2x Read Buffer T was added to each well and read immediately on an MSD Sector Imager 2400 instrument. Standard curves were compared to a four parameter logistic equation using MSD Discovery Workbench software. Pharmacokinetic parameters were calculated using non-compartmental analysis in Phoenix WinNonlin 6.3 (Chertara, MD, USA).

Results.

Mean (SD - standard deviation) serum concentration versus time for the test compounds are shown in FIG. 2. Average (SD) pharmacokinetic parameters for the tested compounds are given in table. 12. Serum samples showing a sharp drop in drug concentration over time that were confirmed to be positive for anti-drug antibodies were excluded from pharmacokinetic parameter calculations.

Table 12. Mean (SD) pharmacokinetic parameters of test compounds in male cynomolgus monkeys after 1 mg/kg intravenous dose

Connection ID Peak 0-last (area under the plasma concentration-time curve, AUC olast) (mcg * d / ml) PPK o- infinity (AUC o-inf) (µg*d/ml) CL (clearance) (ml/day/kg) Vss (apparent volume of distribution at steady state) (ml/kg) T (period half-life) (days) Compound A 52.5 (6.6) 54.2 (7.1) 18.6 (2.4) 61.6 (4.3) 3.1 (0.3) Connection B 102.7 (1.8) 127.0 (3.6) 7.9 (0.2) 64.4 (6.6) 6.2 (0.7) Connection C 26.6 (1.4) 27.0 (1.3) 37.1 (1.8) 66.9 (13.2) 2.3 (0.4) Compound D 33.8(1.3) 34.5 (1.8) 29.0 (1.5) 55.7 (5.5) 2.8 (0.8)

Example 11 Human PK Prediction and Human Dose of an Exemplary Compound.

Dedrick's elemental scaling was used to scale mean monkey serum compound B concentrations to those of humans using an allometric score of 1.0 for volume of distribution and 0.85 for clearance. The predicted intravenous human serum concentration versus time curve was compared to a linear two-compartment model. The SC serum concentration versus time curve for humans was predicted by combining the parameters from the 2-compartment IV model with the average SC uptake rate and bioavailability observed for marketed therapeutic mAnts (monoclonal antibodies). It is assumed that the clearance and half-life are 0.34 l / d and 9.9 d in healthy people, respectively. In FIG. 3 shows the predicted serum concentration versus time profile of Compound B in human serum after a dose of 100 mg subcutaneously administered once every two weeks.

The predicted effective dose in humans is 1 mg/kg given subcutaneously every other week. This drug regimen is expected to maintain a Cmin > 30 nM (6 µg/mL) with 2 weeks or less frequent subcutaneous administration. The predicted effective dose can be derived from the PD biomarkers of concentration responses observed for belimumab (Benlysta®), tabalumab, and blisibimod in patients with SLE and RA. In these studies, the maximum inhibition of biomarkers associated with FAVL (BAFF) was compared with a constant Cmin of 30-40 nM. The concentration required to neutralize IL23 is much lower than that required to neutralize FAVL (BAFF) and therefore does not affect the total required Cmin for the dual antagonist.

Example 12 Purification of connections.

Ways.

Compounds were purified using Mab Select SuRe as an affinity purification step.

The elution was performed using sodium acetate buffer, pH 3.5. After purification with Mab Select SuRE, samples were neutralized and loaded onto Poros 50 HS resin and eluted using a sodium chloride gradient in sodium citrate buffer. Peak washout for monomers was at 20 mM NaCitrate and 120 mM NaCl pH 6.0. After ion exchange chromatography, the sample consisted consistently of greater than 95% monomer.

Rapid Sedimentation (SS) studies using Analytical Ultra Centrifugation (AUC) were used to obtain information on the purity of the sample and its state of aggregation.

Samples were centrifuged under optimal conditions in an XL-I (Beckman Coulter, Fullerton, CA) at 20° C. using an An60Ti four-hole rotor at 40,000 rpm.

The precipitation process was monitored by ultraviolet absorption at 280 nm using the appropriate dilution buffer as a reference buffer. Changes in the concentration distribution in the ultracentrifuge cell were recorded over time using the XL-I operating software and analyzed using the c(S) continuous distribution model in the SEDFIT software (version 14.1) to obtain the distribution of the sedimentation factor. The percentage of monomer was calculated from the integrated peak area.

Results.

The results of purification of compounds are given in table. 13. The data show that the compounds are of high purity and uniformity, which indicate good stability.

__________________________________________________________________ Table 13

Parameter name Connections A, B, C, D Monomer Percentage (Sedimentation Rate) 99% monomer 1% aggregate

Example 13 Mass Spectrophotometric Profile of Compounds.

Ways.

native samples.

This procedure provides data on the intact mass of a compound or protein. 0.15 µl of sample was injected into an Agilent PoroShell 300SB-C3, 5 µm column (30x1.0 mm). The column temperature was 80° C. and the flow rate was 150 μl/min. The compound or protein was eluted from the column with a gradient of 10%B at 0 min to 85%B at 6 min. Mobile phase A was water/acetonitrile/formic acid/ammonium acetate (99/1/0.1/2 mM) and mobile phase B was n-propanol/acetonitrile/water/formic acid (70/20/ 10/0.1). The runoff was sent to an Agilent 6224 TOF mass spectrometer, which (the runoff) was scanned from mass 600 to mass 3200. The raw data was recovered using the MassHunter program. Sample with disulfide bonds removed.

This procedure provides data on protein mass, or light chain, and heavy chain mass. 5 µl of the sample was added to 5 µl of a 20:1 mixture of 8M guanidine HCL:TCEP (tris 2-carboxyethylphosphine) and incubated for 15 min at room temperature. 0.15 µl of this sample was injected as above with the following differences: the compound or protein was eluted from the column with a gradient from 5%B at 0 min to 85%B at 6 min, the column temperature was 60°C, and the mass range was 6002000.

deglycosylated sample.

This procedure provides data on the deglycosylated mass of the protein, or light chain and heavy chain. 7.5 μl of sample was added to 3.2 μl of a 20:1 mixture of 400 mM ammonium bicarbonate:peptide-N-glycosidase F, and incubated for 3 hours at 37°C. Then, 10 μl of a 20:1 mixture of 8M guanidine HCL:TCEP was added to the sample and incubated for 15 min at room temperature. This sample was administered as above for the disulfide removed sample.

Peptide mapping by mass spectrometry.

Samples were diluted in a denaturing buffer consisting of 6 M guanidine HCl, 250 mM Tris-HCl pH 7.5 and 10 mM DTT (dithiothreitol) and then incubated at 37°C for 30 min. Samples were then alkylated with iodoacetamide and incubated in the dark at room temperature for 30 min. Reactions were purified and buffer changed to 100 mM Tris-HCl, pH 7.5 using Sephadex G-25 Superfine cartridges. The samples were then digested with trypsin during a 4 hour incubation at 37°C.

- 47 040031

The cleavage reaction of the mixtures was then stopped by adding TFA (trifluoroacetic acid).

The resulting tryptic product was applied to a Phenomenex Jupiter C18 reverse phase chromatography column using a Dionex Ultimate 3000 HPLC autosampler. A gradient solvent system was used, consisting of solvent A: 0.1% formic acid/99% water/1% acetonitrile and solvent B: 0.1% formic acid/5% water/95% acetonitrile. The percentage of solvent B was increased from 0 to 38% within 140 minutes. Chromatographic separation occurred at room temperature with a flow rate of 100 μl/min. Samples were stored in the autosampler at 4°C. After chromatographic separation, the sample was injected into a Thermo Scientific Orbitrap Fusion mass spectrometer operating in positive electrospray ionization mode. The method used included CID activation types with a resolution of 30,000, a minimum signal of 10,000, an isolation width of 1.0, and a normalized collision energy of 35.0 V. The RF level of the S-lens was set to 20%. The acquisition type is a profile for full scan MS (mass spectrometry) and centroid for CID MC/MC data. Data is collected in the mass range 250-2000 Da at a acquisition rate of 1 spectrum per second.

Collected raw LC-MS and LC-MS/MS fragmentation data were analyzed using Proteome Discover 1.4 (Thermo Scientific) against this sequence. Peptides identified containing the N-X-S/T consensus (X is not P) were then analyzed by manual extraction of EIC (extracted ion chromatogram) for glycosylated peptides. The intensity of MC of glycosylated peptides throughout the EIC was used to estimate their percentage of the total content of glycoforms.

Results.

The results are shown in table. 14. Data indicating the predicted amino acid sequence and structure were displayed and reconstructed without unexpected heterogeneity. The glycosylation pattern is typical of conventional antibodies expressed in CHO cells and does not show any atypical structures.

Table 14

Parameter Connection B Mass spectrometry: Molecular weight profile of the intact molecule Intact/Made in sequence Mass spectrometry: Glycosylation profile Similar to CHO-expressed IgG

Example 14 Monomer content at high concentration.

The purpose of describing this process is to evaluate the intrinsic properties of Compound B by evaluating aggregation at elevated concentration. The standard buffer 20 mM NaCitrate, 120 mM NaCl pH 6 is used without composition evaluation to understand the tendency of molecules to aggregate.

Ways.

Compound B was gradually concentrated to the highest possible concentration without observing precipitation using an Amicon Ultra 50,000 Da molecular weight cut-off centrifuge filter (Millipore, Billerike). Then, concentrated protein solutions were analyzed in a sedimentation rate (SS) experiment to obtain information on sample purity and aggregate states. Chromatography was performed using an Agilent 1200 series HPLC system. The system was run at 1.0 ml/min for 23 minutes. Approximately 30 µg of material was injected into a Tosoh Biosciences TSKgel G3000SWXL column (5 µm 250a 7.8 x 30 cm) and the results were read at 280 nm. The run buffer used is 50 mM NaPhosphate, 0.2 M L-arginine, pH 6.8.

Results.

A summary of the SEC (size exclusion chromatography) analytical data for Compound B during the concentration process is shown in Table 1. 15 and in FIG. 4. Data showing the aggregation and purity of the sample with increasing concentration is presented, which indicates that Compound B does not have a tendency to aggregate with increased concentration.

Table 15. Summary of SEC Analytical Data for Compound B during Concentration

Compound B concentration (mg/ml) 10 20 40 EC analytical data 99.6% monomer, 0.4% aggregate 99.5% monomer, 0.5% aggregate 99.6% monomer, 0.4% aggregate 75 99.5% monomer, 0.5% aggregate 100 99.5% monomer, 0.5% aggregate 121 99.5% monomer, 0.5% aggregate

M - monomer, A - aggregate.

Example 15 Valency of Compounds.

- 48 040031

Ways.

Analytical membrane-limited electrophoresis was used to measure the valency of compounds that had previously been dialyzed in 10 mM acetate, 50 mM KCl pH 5.0 buffer overnight. In the experimental protocol, 20 µl of a sample at a concentration of 1 mg/ml was loaded into a 2x2x4 mm ³ quartz cuvette, both ends of which were closed with semi-permeable regenerated cellulose membranes of the BioTech brand with a molecular weight cut-off (MMO) of 10. These membranes retain macromolecules, at the same time time pass water and solvent components. Then, an electric current of 1 mA was passed through the cuvette, establishing an electric field along its length, in which the charged macromolecule moved in the electric field with a continuous flow of fresh buffer. The real-time moving concentration limit was determined using a linear photodiode array (LPDA) that provides intensity readings along the cuvette. The concentration cutoff speed was used to calculate the electrophoretic mobility and hence the effective valency. Then, additional information such as Stokes radius (derived from sedimentation rate in AUC), counter radius (0.122 nm for chloride ion), buffer conductivity (6.35 mS), and ionic strength (0.05 M) were used to determine the major valency of the macromolecule. .

Results.

The valence data (see Table 16) indicates the colloidal stability of the compounds in solution, i.e. protein-protein networking in solution. Compounds with a valence greater than 15 have a strong repulsive network and a high potential for being prepared at high concentrations.

Table 16. Valence data for compounds

Connection ID Valence at pH 5.0 Compound A 24.4±0.2 Connection B 21.5±0.3 Compound D 21.8±0.2

Example 16 Stability of Compounds in Whole Blood

Ways.

Whole blood interference analysis was performed on the Octet RED96 in order to detect the effects of non-specific binding or off-target binding of compounds in the presence of whole blood (CK). Compound solutions in whole blood and 1x kinetic working buffer (1xkb) were incubated at 37°C for 48 h. Kinetic measurements for incubated compound samples were performed on an Octet RED96 equipped with a streptavidin (SA) biosensor probe (ForteBio, Menlo Park, California) at 27°C. The rate signal/binding ratio in buffer and whole blood has been reported. A ratio less than 2 was considered as indicating no interference.

Results.

The results are shown in table. 17. No interference with whole blood was observed for the tested compounds.

Table 17. Whole blood binding results for compounds

Options Binding Signal Ratio in Human CK/Kinetic Buffer with FAVL (BAFF) Signal binding ratio in CK/kinetic buffer with human IL23 Compound A 1.2 1.1 Connection B 1.2 1.5 Connection C 1.1 1.3 Compound D 1.3 1.7 Compound E 1.0 1.1 Connection F 1.0 1.2

Example 17 In silico immunogenicity prediction.

Ways.

The immunogenicity of protein drugs was predicted in silico using the EpiMatrix computational tool, which was developed by EpiVax, Inc. (Providence, Rhode Island).

The EpiMatrix includes predictions of a T helper epitope as well as a regulatory T cell epitope, of which the former induces an immune response while the latter is inhibitory.

Briefly, the protein sequence was first broken down into 9 amino acid long overlapping protein frames that have been proven to be the core binding proteins of the HLA class II genes. The binding potential of 9 amino acid long peptides to each of the eight common HLA class II alleles is estimated based on experimental data or computational prediction. Scoring is given to display the binding potential of 9 amino acid long peptides with

- 49 040031 each HLA allele, and normalization is performed in order to make it possible to compare the binding of any 9-amino acid polypeptide to any of the many HLA alleles, and predict immunogenicity on a global scale. Finally, the program generates an overall immunogenicity score, the tReg Adjusted Epx Score, which determines the likelihood that compounds will elicit an immune response in vivo.

Results.

The results are shown in table. 18. The overall immunogenicity scores for the test compounds are low and suggest that these compounds are not likely to elicit a strong immune response in vivo.

Table 18. EpiVax Scores

Parameter EpiVax (Circuit 1, Circuit 2) Compound A -46.13, -28.39 Connection B -45.77, -28.39 Connection C -41.40, -28.42 Compound D -41.01, -28.42 Compound E -39.81, -41.43 Connection F -42.15, -36.94 Connection G -43.75, -29.30 Connection H -35.19, -43.44 Compound I -43.38, -29.30 Compound J -34.78, -43.44 Connection K -43.75, -33.61 Connection L -37.80, -43.44 Connection M -43.38, -33.61 Compound N -37.40, -43.44 Connection O -34.66, -39.56 Connection P -34.24, -39.56 Connection Q -34.64, -47.37 Compound R -34.22, -47.37 Connection S -40.96, -40.09 Connection T -43.91, -40.09 U connection -41.31, -44.98 Compound V -42.53, -36.94 Connection W -38.33, -44.98 Compound X -39.42, -41.43

Example 18 Human BAFF Inhibition in Primary Human B Cell Proliferation Assay.

Materials and methods.

The normal healthy blood samples (n equals 3 donors) used in this study were purchased from the Biological Specialty Corporation, Philadelphia, PA. The complete culture medium was IMDM (Life Technologies) supplemented with 10% PBS (Life Technologies) plus penic/strept. (Life Technologies). B cell isolation buffer consisted of sterile PBS (Dulbecco's phosphate buffered saline) without Mg ⁺⁺ and Ca ⁺⁺ (Life Technologies), plus 2% PBS, plus 2 mM EDTA (Life Technologies).

Isolation of human B-lymphocytes from whole blood of a healthy person.

400 ml of heparinized whole blood was transferred into a 1000 ml sterile polystyrene bottle and an equal volume of PBS was added. 35 ml of PBS diluted blood was placed on top of a 15 mm Ficoll-Paque™ Plus gradient (GE Healthcare) preloaded in a 50 ml polystyrene round bottom tube. The tubes were centrifuged at 2000 rpm at room temperature for 20 min without interruption. Then, two thirds of the upper supernatant was aspirated and the middle gray layer containing peripheral blood mononuclear cells (PBMCs) was transferred into a 50 ml conical tube. PBS (without Ca ⁺⁺ and Mg ⁺⁺ ) plus 2% FBS was added to a volume of 50 ml. The tubes were centrifuged at 1200 rpm for 10 min. The resulting cell pellet was resuspended in PBS (without Ca ⁺⁺ and Mg ⁺⁺ ) + 2% PBS (wash buffer) and the centrifugation step was repeated (1200 rpm for 10 min). The cell pellet was resuspended in wash buffer to 50 ml. At this point, cell viability and cell concentration were measured.

- 50 040031

The tube was then centrifuged at 1200 rpm for 10 min and the supernatant was removed. The cell pellet was resuspended in complete culture medium and the cell concentration was adjusted to 5 x 10 ⁶ cells/ml. The suspended cells were placed in a 75 cm ² tissue culture flask and incubated in a 5% CO2 incubator (at 37° C.) overnight. B-lymphocytes were then isolated using the Dynabeads® Untouched™ Human B Cells Kit (Life Technologies) according to the manufacturer's protocol. The cell concentration was adjusted to be suitable for the subsequent procedure after measuring the cell concentration and viability.

Assessing B-lymphocyte proliferation using the ³ H-thymidine incorporation assay.

B lymphocytes were plated in a Falcon tissue culture plate, 96 round-bottomed wells (1 x 10 ⁵ cells/100 μl of culture medium per well). Test articles were then prepared in complete culture medium at 4 consecutive concentrations ranging from 0.028 to 20 nM, and B-lymphocyte stimulating substances (antibodies against human IgM and BAFF) at 4 concentrations (8 µg/mL and 20 ng/mL, respectively) . 50 µl of pre-diluted test articles (0.007 to 5 nM, final concentration) was added to the appropriate wells. Then, 50 μl of pre-diluted goat anti-human IgM (2 μg/ml, final concentration) and hFAVL (human FAVL, hBAFF) (5 ng/ml or 98 pM, final concentration) were added to the appropriate wells (see Fig. 6). The lymphocytes were cultured in a humidified incubator, 5% CO ₂ , 37° C. for 72 hours. 18 hours before the end of the incubation, 20 μl of 1 μCi ³ H-thymidine (Pelkin Elmer) was added to each well. A cell harvester was used to transfer the DNA with ³ H-thymidine/well incorporated into a microfiber glass filter plate, which had 30 µl of scintillation enhancer added to the wells after air drying the plate for at least 4 hours. ) of each well was measured using a MicroBeta Topcount and a PIM value (y-axis) versus test article concentration (x-axis) was plotted.

Statistical analysis.

IC ₅₀ and IC ₉₀ values were determined by plotting the data with a 4-parameter sigmoid dose-response function using GraphPad Prism 6 software. Geometric means were calculated from three experiments and are shown in Table 1. 19.

Results.

The results showed that compound B was approximately 2-fold more potent than control antibody 3 when comparing IC ₅₀ and IC ₉₀ values.

Table 19. Geometric mean IC ₅₀ and IC ₉₀ values for inhibition of human FAVL (BAFF) in the Primary B Lymphocyte Proliferation Assay

Connection ID 1C ₅ o (pM) IC90 (pM) Connection B 33.7 160.2 Control antibody 3 (BAFF (3)) 87.3 370.8

Example 19 Suppression of B220+ B Lymphocytes Due to FAVL Overexpression (BAFF) in B10.RIII Mice.

Materials and methods.

Briefly, on the first day, female B10.RIII mice (6-8 weeks old, Jackson Laboratory) were randomly divided into 10 groups, 10 animals/group, and given an intra-abdominal injection of 100 μl, or citrate buffer (20 mM NaCitrate, 115 mM sodium chloride, pH 6.0), or test compounds at an equivalent molar dose of 1.3, 0.4 and 0.13 mg/kg versus 1, 3 and 0.1 mg/kg, respectively. Previously untreated, untreated mice were additional controls. Immediately after day 1 of treatment, mice were injected with a single 3 mg dose (1.5 mg/mL) of human mini-circle DNA FAFF (BAFF) (System Biosciences) by hydrodynamic injection, as opposed to an empty vector (VV) for the control group. . The intraperitoneal treatment with either citrate buffer or test compounds was repeated every 72 hours on days 3, 6, 9 and 12.

On day 14, mice were anesthetized with isoflurane (Butler Schein) and sacrificed via cervical dislocation. The spleens were removed and the cell suspension was analyzed by flow cytometry for the presence of B220+ B lymphocytes. Mean levels for each treated group were determined and statistical significance compared to controls was calculated using one-way analysis of variance followed by Dunnett's multiple comparison test. The results are illustrated in FIG. 5.

Results.

The results showed that treatment for 14 days with Compound B was able to significantly suppress B220+ B lymphocyte proliferation induced by human BAFF human minicircle DNA.

Example 20 Inhibition of human IL23-induced release of IL17A and IL22 in C57/B16 mice.

Materials and methods.

Briefly, female C57BL/6 mice (7-10 weeks old, Charles River) were randomly separated

- 51 040031 per 8 groups, 8 animals/group, and they were given an intra-abdominal injection of 100 μl, or citrate buffer (20 mM NaCitrate, 115 mM sodium chloride, pH 6.0), or test compounds at an equivalent molar dose of 1.3, 0.4 and 0.13 mg/kg versus 1, 3 and 0.1 mg/kg, respectively.

One hour after test compound dosing, mice were anesthetized with isoflurane (Butler Schein) and given 20 μl intradermal injection, or 0.1% BSA (Sigma) control, or 15 μg/ml (0.3 μg) rhIL23 (recombinant human IL23) (own production), diluted in saline (Invitrogen), in both ears. Intradermal injections of antigenic stimuli were repeated daily for 2 consecutive days. 24 hours after the second challenge, mice were sacrificed by cervical dislocation and each ear was removed. Ear tissues were homogenized in 1 ml of homogenization buffer (HBSS (Gibco) 0.4% Triton X-100 (Sigma) 1X SigmaFast Protease Inhibitor (Sigma)) using a Biomedicals Fast-Prep 24 MP homogenizer. Homogenized samples were centrifuged at 4° C. for 10 min and the supernatant was collected. Supernatants were analyzed for mouse IL17A and IL22 using Quantikine® Mouse IL-17 and mouse IL-22 immunoassays according to manufacturer's instructions (R&D Systems). Interpolated cytokine pg/ml values were determined for each sample. Mean pg/mL levels for each treatment group were determined and statistical significance compared to controls was calculated using one-way analysis of variance followed by Dunnett's multiple comparison test.

Results.

The results in FIG. 6 show that treatment with a single intraperitoneal dose of compound B is able to significantly inhibit the release of mouse IL17 and IL22 in the skin induced by consecutive intradermal injections of recombinant human IL23 for two days.

Sequences

SEQID NO Subsequence 1 QVQLQQSGAEVKKPGSSVRVSCKASGGTFNNNAINWVRQAPGQGLEWMGGIIP MFGTAKYSQNFQGRVAITADESTGTASMELSSLRSEDTAVYYCARSRDLLLFPHH ALSPWGRGTMVTVSS 2 SSELTQDPAVSVALGQTVRVTCQGDSLRSYYASWYQQKPGQAPVLVIYGKNNRP SGIPDRFSGSSSGNTASLTITGAQAEDEADYYCSSRDSSGNHWVFGGGTELTVL 3 QVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPR DDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIY WGQGTLVTVSS 4 DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRH TGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIK 5 SSELTQDPAVSVALGQTVRVTCQGDSLRSYYASWYQQKPGQAPVLVIYGKNNRP SGIPDRFSGSSSGNTASLTITGAQAEDEADYYCSSRDSSGNHWVFGGGTELTVLG GGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLE WIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRS GYAWFIYWGQGTLVTVSSLGGGSGASTKGPSVFPLAPSSKSTSGGTAALGCLVK DYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN HKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEV TCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTC LVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN VFSCSVMHEALHNHYTQKSLSLSPG 6 DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRH TGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSG GGGQVQLQQSGAEVKKPGSSVRVSCKASGGTFNNNAINWVRQAPGQGLEWMG GIIPMFGTAKYSQNFQGRVAITADESTGTASMELSSLRSEDTAVYYCARSRDLLLF PHHALSPWGRGTMVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASWCLLNN FYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVY ACEVTHQGLSSPVTKSFNRGEC 7 DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRH TGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSG GGGQVQLQQSGAEVKKPGSSVRVSCKASGGTFNNNAINWVRQAPGQGLEWMG

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GIIPMFGTAKYSQNFQGRVAITADESTGTASMELSSLRSEDTAVYYCARSRDLLLF PHHALSPWGRGTMVTVSSLGGGSGASTKGPSVFPLAPSSKSTSGGTAALGCLVK DYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVN HKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEV TCVWDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTVLHQD WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTC LVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN VFSCSVMHEALHNHYTQKSLSLSPG 8 SSELTQDPAVSVALGQTVRVTCQGDSLRSYYASWYQQKPGQAPVLVIYGKNNRP SGIPDRFSGSSSGNTASLTITGAQAEDEADYYCSSRDSSGNHWVFGGGTELTVLG GGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLE WIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRS GYAWFIYWGQGTLVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASWCLLNN FYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVY ACEVTHQGLSSPVTKSFNRGEC 9 SSELTQDPAVSVALGQTVRVTCQGDSLRSYYASWYQQKPGQAPVLVIYGKNNRP SGIPDRFSGSSSGNTASLTITGAQAEDEADYYCSSRDSSGNHWVFGGGTELTVLG GGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLE WIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRS GYAWFIYWGQGTLVTVSSGGCGGGEVAACEKEVAALEKEVAALEKEVAALEKL EPI<SSDI<THTCPPCPAPEAAGGPSVFLFPPI<PI<DTLYITREPEVTCVVVDVSHEDP EVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV SNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAV EWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALH NHYTQKSLSLSPG 10 DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRH TGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSG GGGQVQLQQSGAEVKKPGSSVRVSCKASGGTFNNNAINWVRQAPGQGLEWMG GIIPMFGTAKYSQNFQGRVAITADESTGTASMELSSLRSEDTAVYYCARSRDLLLF PHHALSPWGRGTMVTVSSGGCGGGKVAACKEKVAALKEKVAALKEKVAALKE eleven DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRH TGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSG GGGQVQLQQSGAEVKKPGSSVRVSCKASGGTFNNNAINWVRQAPGQGLEWMG GIIPMFGTAKYSQNFQGRVAITADESTGTASMELSSLRSEDTAVYYCARSRDLLLF PHHALSPWGRGTMVTVSSGGCGGGEVAACEKEVAALEKEVAALEKEVAALEKL EPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVWDVSHEDP EVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV SNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAV EWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALH NHYTQKSLSLSPG 12 SSELTQDPAVSVALGQTVRVTCQGDSLRSYYASWYQQKPGQAPVLVIYGKNNRP SGIPDRFSGSSSGNTASLTITGAQAEDEADYYCSSRDSSGNHWVFGGGTELTVLG GGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLE WIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRS GYAWFIYWGQGTLVTVSSGGCGGGKVAACKEKVAALKEKVAALKEKVAALKE 13 SSELTQDPAVSVALGQTVRVTCQGDSLRSYYASWYQQKPGQAPVLVIYGKNNRP SGIPDRFSGSSSGNTASLTITGAQAEDEADYYCSSRDSSGNHWVFGGGTELTVLG GGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLE WIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRS GYAWFIYWGQGTLVTVSSLGGGSGASTKGPSVFPLAPCSRSTSESTAALGCLVKD YFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVD HKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLYITREPEVTCV WDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWL NGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFS CSVMHEALHNHYTQKSLSLSLG 14 DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRH TGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSG GGGQVQLQQSGAEVKKPGSSVRVSCKASGGTFNNNAINWVRQAPGQGLEWMG GIIPMFGTAKYSQNFQGRVAITADESTGTASMELSSLRSEDTAVYYCARSRDLLLF PHHALSPWGRGTMVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASWCLLNN FYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVY

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ACEVTHQGLSSPVTKSFNRGEC 15 DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRH TGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSG GGGQVQLQQSGAEVKKPGSSVRVSCKASGGTFNNNAINWVRQAPGQGLEWMG GIIPMFGTAKYSQNFQGRVAITADESTGTASMELSSLRSEDTAVYYCARSRDLLLF PHHALSPWGRGTMVTVSSLGGGSGASTKGPSVFPLAPCSRSTSESTAALGCLVKD YFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVD HKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLYITREPEVTCV WDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWL NGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFS CSVMHEALHNHYTQKSLSLSLG 16 SSELTQDPAVSVALGQTVRVTCQGDSLRSYYASWYQQKPGQAPVLVIYGKNNRP SGIPDRFSGSSSGNTASLTITGAQAEDEADYYCSSRDSSGNHWVFGGGTELTVLG GGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLE WIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRS GYAWFIYWGQGTLVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASWCLLNN FYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVY ACEVTHQGLSSPVTKSFNRGEC 17 SSELTQDPAVSVALGQTVRVTCQGDSLRSYYASWYQQKPGQAPVLVIYGKNNRP SGIPDRFSGSSSGNTASLTITGAQAEDEADYYCSSRDSSGNHWVFGGGTELTVLG GGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLE WIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRS GYAWFIYWGQGTLVTVSSGGCGGGEVAACEKEVAALEKEVAALEKEVAALEKE SKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLYITREPEVTCVWDVSQEDPEVQFN WYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGL PSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESN GQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQ KSLSLSLG 18 DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRH TGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSG GGGQVQLQQSGAEVKKPGSSVRVSCKASGGTFNNNAINWVRQAPGQGLEWMG GIIPMFGTAKYSQNFQGRVAITADESTGTASMELSSLRSEDTAVYYCARSRDLLLF PHHALSPWGRGTMVTVSSGGCGGGKVAACKEKVAALKEKVAALKEKVAALKE 19 DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRH TGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSG GGGQVQLQQSGAEVKKPGSSVRVSCKASGGTFNNNAINWVRQAPGQGLEWMG GIIPMFGTAKYSQNFQGRVAITADESTGTASMELSSLRSEDTAVYYCARSRDLLLF PHHALSPWGRGTMVTVSSGGCGGGEVAACEKEVAALEKEVAALEKEVAALEKE SKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLYITREPEVTCVWDVSQEDPEVQFN WYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGL PSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESN GQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQ KSLSLSLG 20 SSELTQDPAVSVALGQTVRVTCQGDSLRSYYASWYQQKPGQAPVLVIYGKNNRP SGIPDRFSGSSSGNTASLTITGAQAEDEADYYCSSRDSSGNHWVFGGGTELTVLG GGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLE WIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRS GYAWFIYWGQGTLVTVSSGGCGGGKVAACKEKVAALKEKVAALKEKVAALKE 21 DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRH TGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSG GGGQVQLVQSGAEVKKPGSSVKVSCKAPDHIFSIHWMQWVRQAPGQGLEWMG EIFPGSGTTDYNEKFKGKVTITVDKSTSTAYMELSSLRSEDTAVYYCASGAFDYW GQGTTVTVSSLGGGSGASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTV SWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKV DKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVWDVS HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPS DIAVE WESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMH EALHNHYTQKSLSLSPG 22 DIQMTQSPSSLSASVGDRVTITCRASQDIGNRLSWLQQEPGKAPKRLIYATSSLDS GVPSRFSGSRSGTEFTLTISSLQPEDFVTYYCLQYASSPFTFGQGTKLEIKGGGSGG

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GGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYI YPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAW FIYWGQGTLVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASWCLLNNFYPRE AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLSSPVTKSFNRGEC 23 DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRH TGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSG GGGQVQLVQSGAEVKKPGSSVKVSCKAPDHIFSIHWMQWVRQAPGQGLEWMG EIFPGSGTTDYNEKFKGKVTITVDKSTSTAYMELSSLRSEDTAVYYCASGAFDYW GQGTTVTVSSGGCGGGEVAACEKEVAALEKEVAALEKEVAALEKLEPKSSDKTH TCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYV DGVEVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPI EKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQP ENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSL SLSPG 24 DIQMTQSPSSLSASVGDRVTITCRASQDIGNRLSWLQQEPGKAPKRLIYATSSLDS GVPSRFSGSRSGTEFTLTISSLQPEDFVTYYCLQYASSPFTFGQGTKLEIKGGGSGG GGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYI YPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAW FIYWGQGTLVTVSSGGCGGGKVAACKEKVAALKEKVAALKEKVAALKE 25 DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRH TGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSG GGGQVQLVQSGAEVKKPGSSVKVSCKAPDHIFSIHWMQWVRQAPGQGLEWMG EIFPGSGTTDYNEKFKGKVTITVDKSTSTAYMELSSLRSEDTAVYYCASGAFDYW GQGTTVTVSSLGGGSGASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVD KRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLYITREPEVTCVWDVSQEDPE VQFNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVS NKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVE WESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHN HYTQKSLSLSLG 26 DIQMTQSPSSLSASVGDRVTITCRASQDIGNRLSWLQQEPGKAPKRLIYATSSLDS GVPSRFSGSRSGTEFTLTISSLQPEDFVTYYCLQYASSPFTFGQGTKLEIKGGGSGG GGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYI YPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAW FIYWGQGTLVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASWCLLNNFYPRE AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLSSPVTKSFNRGEC 27 DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRH TGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSG GGGQVQLVQSGAEVKKPGSSVKVSCKAPDHIFSIHWMQWVRQAPGQGLEWMG EIFPGSGTTDYNEKFKGKVTITVDKSTSTAYMELSSLRSEDTAVYYCASGAFDYW GQGTTVTVSSGGCGGGEVAACEKEVAALEKEVAALEKEVAALEKESKYGPPCPP CPAPEFLGGPSVFLFPPKPKDTLYITREPEVTCVWDVSQEDPEVQFNWYVDGVE VHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISK AKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 28 DIQMTQSPSSLSASVGDRVTITCRASQDIGNRLSWLQQEPGKAPKRLIYATSSLDS GVPSRFSGSRSGTEFTLTISSLQPEDFVTYYCLQYASSPFTFGQGTKLEIKGGGSGG GGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYI YPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAW FIYWGQGTLVTVSSGGCGGGKVAACKEKVAALKEKVAALKEKVAALKE 29 DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRH TGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSG GGGQVQLVQSGAEVKKPGSSVKVSCKAPDHIFSIHWMQWVRQAPGQGLEWMG EIFPGSGTTDYNEKFKGKVTITVDKSTSTAYMELSSLRSEDTAVYYCASGAFDYW GQGTTVTVSSLGGGSGASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTV SWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKV DKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVWDVS HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPS DIAVE WESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMH

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EALHNHYTQKSLSLSPG thirty DIQMTQSPSSLSASVGDRVTITCRASQDIGNRLNWYQQKPGKAPKRLIYATSSLDS GVPSRFSGSRSGTEFTLTISSLQPEDFVTYYCLQYASSPFTFGQGTKLEIKGGGSGG GGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYI YPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAW FIYWGQGTLVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASWCLLNNFYPRE AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLSSPVTKSFNRGEC 31 DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRH TGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSG GGGQVQLVQSGAEVKKPGSSVKVSCKAPDHIFSIHWMQWVRQAPGQGLEWMG EIFPGSGTTDYNEKFKGKVTITVDKSTSTAYMELSSLRSEDTAVYYCASGAFDYW GQGTTVTVSSGGCGGGEVAACEKEVAALEKEVAALEKEVAALEKLEPKSSDKTH TCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVWDVSHEDPEVKFNWYV DGVEVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPI EKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQP ENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSL SLSPG 32 DIQMTQSPSSLSASVGDRVTITCRASQDIGNRLNWYQQKPGKAPKRLIYATSSLDS GVPSRFSGSRSGTEFTLTISSLQPEDFVTYYCLQYASSPFTFGQGTKLEIKGGGSGG GGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYI YPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAW FIYWGQGTLVTVSSGGCGGGKVAACKEKVAALKEKVAALKEKVAALKE 33 DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRH TGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSG GGGQVQLVQSGAEVKKPGSSVKVSCKAPDHIFSIHWMQWVRQAPGQGLEWMG EIFPGSGTTDYNEKFKGKVTITVDKSTSTAYMELSSLRSEDTAVYYCASGAFDYW GQGTTVTVSSLGGGSGASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTKTYTCNVDHKPSNTKVD KRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLYITREPEVTCVWDVSQEDPE VQFNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVS NKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVE WESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHN HYTQKSLSLSLG 34 DIQMTQSPSSLSASVGDRVTITCRASQDIGNRLNWYQQKPGKAPKRLIYATSSLDS GVPSRFSGSRSGTEFTLTISSLQPEDFVTYYCLQYASSPFTFGQGTKLEIKGGGSGG GGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYI YPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAW FIYWGQGTLVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASWCLLNNFYPRE AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLSSPVTKSFNRGEC 35 DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRH TGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSG GGGQVQLVQSGAEVKKPGSSVKVSCKAPDHIFSIHWMQWVRQAPGQGLEWMG EIFPGSGTTDYNEKFKGKVTITVDKSTSTAYMELSSLRSEDTAVYYCASGAFDYW GQGTTVTVSSGGCGGGEVAACEKEVAALEKEVAALEKEVAALEKESKYGPPCPP CPAPEFLGGPSVFLFPPKPKDTLYITREPEVTCVWDVSQEDPEVQFNWYVDGVE VHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISK AKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 36 DIQMTQSPSSLSASVGDRVTITCRASQDIGNRLNWYQQKPGKAPKRLIYATSSLDS GVPSRFSGSRSGTEFTLTISSLQPEDFVTYYCLQYASSPFTFGQGTKLEIKGGGSGG GGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYI YPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAW FIYWGQGTLVTVSSGGCGGGKVAACKEKVAALKEKVAALKEKVAALKE 37 DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRH TGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSG GGGQVQLVQSGAEVKKPGASVKVSCKASGYSFSTFFIHWVRQRPGQGLEWIGRI DPNSGATKYNEKFESKVTLTRDTSISTAYMELSRLRSDDTAVYYCARGEDLLIRT DALDYWGQGTSVTVSSLGGGSGASTKGPSVFPLAPSSKSTSGGTAALGCLVKDY FPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHK PSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTC

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FSCSVMHEALHNHYTQKSLSLSPG 38 DIQMTQSPSSLSASVGDRVSITCKASQNAGIDVAWFQQKPGKAPKLLIYSKSNRY TGVPSRFSGSGSGTDFTLTISSLQPEDFADYYCLQYRSYPRTFGGGTKLEIKGGGS GGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIG YIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGY AWFIYWGQGTLVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASWCLLNNFY PREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYAC EVTHQGLSSPVTKSFNRGEC 39 DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRH TGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSG GGGQVQLVQSGAEVKKPGASVKVSCKASGYSFSTFFIHWVRQRPGQGLEWIGRI DPNSGATKYNEKFESKVTLTRDTSISTAYMELSRLRSDDTAVYYCARGEDLLIRT DALDYWGQGTSVTVSSGGCGGGEVAACEKEVAALEKEVAALEKEVAALEKLEP KSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVWDVSHEDPEV KFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSN KALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEW ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPG 40 DIQMTQSPSSLSASVGDRVSITCKASQNAGIDVAWFQQKPGKAPKLLIYSKSNRY TGVPSRFSGSGSGTDFTLTISSLQPEDFADYYCLQYRSYPRTFGGGTKLEIKGGGS GGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIG YIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGY AWFIYWGQGTLVTVSSGGCGGGKVAACKEKVAALKEKVAALKEKVAALKE 41 DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRH TGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSG GGGQVQLVQSGAEVKKPGASVKVSCKASGYSFSTFFIHWVRQRPGQGLEWIGRI DPNSGATKYNEKFESKVTLTRDTSISTAYMELSRLRSDDTAVYYCARGEDLLIRT DALDYWGQGTSVTVSSLGGGSGASTKGPSVFPLAPCSRSTSESTAALGCLVKDYF PEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTKTYTCNVDHK PSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLYITREPEVTCVVV DVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNG KEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCS VMHEALHNHYTQKSLSLSLG 42 DIQMTQSPSSLSASVGDRVSITCKASQNAGIDVAWFQQKPGKAPKLLIYSKSNRY TGVPSRFSGSGSGTDFTLTISSLQPEDFADYYCLQYRSYPRTFGGGTKLEIKGGGS GGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIG YIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGY AWFIYWGQGTLVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFY PREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYAC EVTHQGLSSPVTKSFNRGEC 43 DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRH TGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSG GGGQVQLVQSGAEVKKPGASVKVSCKASGYSFSTFFIHWVRQRPGQGLEWIGRI DPNSGATKYNEKFESKVTLTRDTSISTAYMELSRLRSDDTAVYYCARGEDLLIRT DALDYWGQGTSVTVSSGGCGGGEVAACEKEVAALEKEVAALEKEVAALEKESK YGPPCPPCPAPEFLGGPSVFLFPPKPKDTLYITREPEVTCVWDVSQEDPEVQFNW YVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGLPS SIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQ PENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKS LSLSLG 44 DIQMTQSPSSLSASVGDRVSITCKASQNAGIDVAWFQQKPGKAPKLLIYSKSNRY TGVPSRFSGSGSGTDFTLTISSLQPEDFADYYCLQYRSYPRTFGGGTKLEIKGGGS GGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIG YIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGY AWFIYWGQGTLVTVSSGGCGGGKVAACKEKVAALKEKVAALKEKVAALKE 45 DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRH TGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSG GGGQVQLVQSGAEVKKPGASVKVSCKASGYSFSTFFIHWVRQAPGQGLEWIGRI

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DPNSGATKYNEKFESRVTMTRDTSISTAYMELSRLRSDDTAVYYCARGEDLLIRT DALDYWGQGTSVTVSSLGGGSGASTKGPSVFPLAPSSKSTSGGTAALGCLVKDY FPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHK PSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTC VWDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCL VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNV FSCSVMHEALHNHYTQKSLSLSPG 46 DIQMTQSPSSLSASVGDRVSITCKASQNAGIDVAWFQQKPGKAPKLLIYSKSNRY TGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQYRSYPRTFGQGTKLEIKGGGS GGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIG YIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGY AWFIYWGQGTLVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASWCLLNNFY PREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYAC EVTHQGLSSPVTKSFNRGEC 47 DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRH TGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSG GGGQVQLVQSGAEVKKPGASVKVSCKASGYSFSTFFIHWVRQAPGQGLEWIGRI DPNSGATKYNEKFESRVTMTRDTSISTAYMELSRLRSDDTAVYYCARGEDLLIRT DALDYWGQGTSVTVSSGGCGGGEVAACEKEVAALEKEVAALEKEVAALEKLEP KSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVWDVSHEDPEV KFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSN KALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEW ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPG 48 DIQMTQSPSSLSASVGDRVSITCKASQNAGIDVAWFQQKPGKAPKLLIYSKSNRY TGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQYRSYPRTFGQGTKLEIKGGGS GGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIG YIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGY AWFIYWGQGTLVTVSSGGCGGGKVAACKEKVAALKEKVAALKEKVAALKE 49 DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRH TGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSG GGGQVQLVQSGAEVKKPGASVKVSCKASGYSFSTFFIHWVRQAPGQGLEWIGRI DPNSGATKYNEKFESRVTMTRDTSISTAYMELSRLRSDDTAVYYCARGEDLLIRT DALDYWGQGTSVTVSSLGGGSGASTKGPSVFPLAPCSRSTSESTAALGCLVKDYF PEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTKTYTCNVDHK PSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLYITREPEVTCWV DVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNG KEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCS VMHEALHNHYTQKSLSLSLG 50 DIQMTQSPSSLSASVGDRVSITCKASQNAGIDVAWFQQKPGKAPKLLIYSKSNRY TGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQYRSYPRTFGQGTKLEIKGGGS GGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIG YIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGY AWFIYWGQGTLVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASWCLLNNFY PREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYAC EVTHQGLSSPVTKSFNRGEC 51 DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRH TGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSG GGGQVQLVQSGAEVKKPGASVKVSCKASGYSFSTFFIHWVRQAPGQGLEWIGRI DPNSGATKYNEKFESRVTMTRDTSISTAYMELSRLRSDDTAVYYCARGEDLLIRT DALDYWGQGTSVTVSSGGCGGGEVAACEKEVAALEKEVAALEKEVAALEKESK YGPPCPPCPAPEFLGGPSVFLFPPKPKDTLYITREPEVTCVWDVSQEDPEVQFNW YVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGLPS SIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQ PENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKS LSLSLG 52 DIQMTQSPSSLSASVGDRVSITCKASQNAGIDVAWFQQKPGKAPKLLIYSKSNRY TGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQYRSYPRTFGQGTKLEIKGGGS GGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIG YIYPRDDSPKYNENFKSTADRSGVITADKSTEDY

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AWFIYWGQGTLVTVSSGGCGGGKVAACKEKVAALKEKVAALKEKVAALKE 53 DIQMTQSPSSLSASVGDRVTITCRASQDIGNRLSWLQQEPGKAPKRLIYATSSLDS GVPSRFSGSRSGTEFTLTISSLQPEDFVTYYCLQYASSPFTFGQGTKLEIKGGGSGG GGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYI YPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAW FIYWGQGTLVTVSSLGGGSGASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEP VTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSN TKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVW DVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNG KEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCS VMHEALHNHYTQKSLSLSPG 54 DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRH TGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSG GGGQVQLVQSGAEVKKPGSSVKVSCKAPDHIFSIHWMQWVRQAPGQGLEWMG EIFPGSGTTDYNEKFKGKVTITVDKSTSTAYMELSSLRSEDTAVYYCASGAFDYW GQGTTVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASWCLLNNFYPREAKV QWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC 55 DIQMTQSPSSLSASVGDRVTITCRASQDIGNRLSWLQQEPGKAPKRLIYATSSLDS GVPSRFSGSRSGTEFTLTISSLQPEDFVTYYCLQYASSPFTFGQGTKLEIKGGGSGG GGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYI YPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAW FIYWGQGTLVTVSSLGGGSGASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEP VTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSN TKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLYITREPEVTCVWDVS QEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEY KCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPS DIAVE WESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMH EALHNHYTQKSLSLSLG 56 DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRH TGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSG GGGQVQLVQSGAEVKKPGSSVKVSCKAPDHIFSIHWMQWVRQAPGQGLEWMG EIFPGSGTTDYNEKFKGKVTITVDKSTSTAYMELSSLRSEDTAVYYCASGAFDYW GQGTTVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASWCLLNNFYPREAKV QWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC 57 DIQMTQSPSSLSASVGDRVTITCRASQDIGNRLNWYQQKPGKAPKRLIYATSSLDS GVPSRFSGSRSGTEFTLTISSLQPEDFVTYYCLQYASSPFTFGQGTKLEIKGGGSGG GGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYI YPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAW FIYWGQGTLVTVSSLGGGSGASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEP VTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSN TKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVW DVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNG KEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCS VMHEALHNHYTQKSLSLSPG 58 DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRH TGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSG GGGQVQLVQSGAEVKKPGSSVKVSCKAPDHIFSIHWMQWVRQAPGQGLEWMG EIFPGSGTTDYNEKFKGKVTITVDKSTSTAYMELSSLRSEDTAVYYCASGAFDYW GQGTTVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASWCLLNNFYPREAKV QWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC 59 DIQMTQSPSSLSASVGDRVTITCRASQDIGNRLNWYQQKPGKAPKRLIYATSSLDS GVPSRFSGSRSGTEFTLTISSLQPEDFVTYYCLQYASSPFTFGQGTKLEIKGGGSGG GGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYI YPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAW FIYWGQGTLVTVSSLGGGSGASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEP VTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTKTYTCNVDHKPSN TKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLYITREPEVTCVWDVS

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QEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEY KCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPS DIAVE WESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMH EALHNHYTQKSLSLSLG 60 DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRH TGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSG GGGQVQLVQSGAEVKKPGSSVKVSCKAPDHIFSIHWMQWVRQAPGQGLEWMG EIFPGSGTTDYNEKFKGKVTITVDKSTSTAYMELSSLRSEDTAVYYCASGAFDYW GQGTTVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASWCLLNNFYPREAKV QWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC 61 DIQMTQSPSSLSASVGDRVSITCKASQNAGIDVAWFQQKPGKAPKLLIYSKSNRY TGVPSRFSGSGSGTDFTLTISSLQPEDFADYYCLQYRSYPRTFGGGTKLEIKGGGS GGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIG YIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGY AWFIYWGQGTLVTVSSLGGGSGASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYF PEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKP SNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCV WDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTVLHQDWL NGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPG 62 DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRH TGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSG GGGQVQLVQSGAEVKKPGASVKVSCKASGYSFSTFFIHWVRQRPGQGLEWIGRI DPNSGATKYNEKFESKVTLTRDTSISTAYMELSRLRSDDTAVYYCARGEDLLIRT DALDYWGQGTSVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASWCLLNNFY PREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYAC EVTHQGLSSPVTKSFNRGEC 63 DIQMTQSPSSLSASVGDRVSITCKASQNAGIDVAWFQQKPGKAPKLLIYSKSNRY TGVPSRFSGSGSGTDFTLTISSLQPEDFADYYCLQYRSYPRTFGGGTKLEIKGGGS GGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIG YIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGY AWFIYWGQGTLVTVSSLGGGSGASTKGPSVFPLAPCSRSTSESTAALGCLVKDYF PEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTKTYTCNVDHK PSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLYITREPEVTCWV DVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNG KEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCS VMHEALHNHYTQKSLSLSLG 64 DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRH TGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSG GGGQVQLVQSGAEVKKPGASVKVSCKASGYSFSTFFIHWVRQRPGQGLEWIGRI DPNSGATKYNEKFESKVTLTRDTSISTAYMELSRLRSDDTAVYYCARGEDLLIRT DALDYWGQGTSVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASWCLLNNFY PREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYAC EVTHQGLSSPVTKSFNRGEC 65 DIQMTQSPSSLSASVGDRVSITCKASQNAGIDVAWFQQKPGKAPKLLIYSKSNRY TGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQYRSYPRTFGQGTKLEIKGGGS GGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIG YIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGY AWFTYWGQGTLVTVSSLGGGSGASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYF PEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKP SNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCV WDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTVLHQDWL NGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPG 66 DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRH TGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSG GGGQVQLVQSGAEVKKPGASVKVSCKASGYSFTFFIHWVRQAPGQGLEWIGRI DPNSGATKYNEKFESRVTMTRDTSTAYMERLSY

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DALDYWGQGTSVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASWCLLNNFY PREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYAC EVTHQGLSSPVTKSFNRGEC 67 DIQMTQSPSSLSASVGDRVSITCKASQNAGIDVAWFQQKPGKAPKLLIYSKSNRY TGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQYRSYPRTFGQGTKLEIKGGGS GGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIG YIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGY AWFIYWGQGTLVTVSSLGGGSGASTKGPSVFPLAPCSRSTSESTAALGCLVKDYF PEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHK PSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLYITREPEVTCWV DVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNG KEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCS VMHEALHNHYTQKSLSLSLG 68 DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRH TGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSG GGGQVQLVQSGAEVKKPGASVKVSCKASGYSFSTFFIHWVRQAPGQGLEWIGRI DPNSGATKYNEKFESRVTMTRDTSISTAYMELSRLRSDDTAVYYCARGEDLLIRT DALDYWGQGTSVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASWCLLNNFY PREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYAC EVTHQGLSSPVTKSFNRGEC 69 GGGSGGGG 70 LGGGSG 71 FNRGEC 72 VEPKSC 73 ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSWTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAP EFLGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSQEDPEVQFNWYVDGVEVHN AKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKG QPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPP VLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 74 ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSWTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAP EFLGGPSVFLFPPKPKDTLYITREPEVTCVWDVSQEDPEVQFNWYVDGVEVHNA KTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQ PREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 75 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGVE VHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTIS KAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNY KTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP G 76 EPKSCDKTHTCPPCP 77 RTVAAPSVFIFPPSDEQLKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQES VTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 78 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVWDVSHEDPEVKFNWYVDGVE VHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTIS KAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNY KTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP GK 79 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGVE VHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTIS KAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNY KTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP G 80 MDDSTEREQSRLTSCLKKREEMKLKECVSILPRKESPSVRSSKDGKLLAATLLLA

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LLSCCLTWSFYQVAALQGDLASLRAELQGHHAEKLPAGAPKAGLEEAPAVT AGLKIFEPPAPGEGNSSQNSRNKRAVQGPEETVTQDCLQLIADSETPTIQKGSYTF 81 MLGSRAVMLLLLLPWTAQGRAVPGGSSPAWTQCQQLSQKLCTLAWSAHPLVGH MDLREEGDEETTNNDVPHIQCGDGCDPQGLRDNSQFCLQRIHQGLIFYEKLLGSDIF TGEPSLLPDSPVGQLHASLLGLSQLLQPEGHHWETQQIPSLSPSQPWQRLLLRFKI LRSLQAFVAVAARVFAHGAATLSP 82 GGCGGGEVAACEKEVAALEKEVAALEKEVAALEK 83 GGCGGGKVAACKEKVAALKEKVAALKEKVAALKE 84 QVQLVQSGAEVKKPGSSVKVSCKAPDHIFSIHWMQWVRQAPGQGLEWMGEIFP GSGTTDYNEKFKGKVTITVDKSTSTAYMELSSLRSEDTAVYYCASGAFDYWGQG TTVTVSS 85 DIQMTQSPSSLSASVGDRVTITCRASQDIGNRLSWLQQEPGKAPKRLIYATSSLDS GVPSRFSGSRSGTEFTLTISSLQPEDFVTYYCLQYASSPFTFGQGTKLEIK 86 QVQLVQSGAEVKKPGSSVKVSCKAPDHIFSIHWMQWVRQAPGQGLEWMGEIFP GSGTTDYNEKFKGKVTITVDKSTSTAYMELSSLRSEDTAVYYCASGAFDYWGQG TTVTVSS 87 DIQMTQSPSSLSASVGDRVTITCRASQDIGNRLNWYQQKPGKAPKRLIYATSSLDS GVPSRFSGSRSGTEFTLTISSLQPEDFVTYYCLQYASSPFTFGQGTKLEIK 88 QVQLVQSGAEVKKPGASVKVSCKASGYSFSTFFIHWVRQRPGQGLEWIGRIDPNS GATKYNEKFESKVTLTRDTSISTAYMELSRLRSDDTAVYYCARGEDLLIRTDALD YWGQGTSVTVSS 89 DIQMTQSPSSLSASVGDRVSITCKASQNAGIDVAWFQQKPGKAPKLLIYSKSNRY TGVPSRFSGSGSGTDFTLTISSLQPEDFADYYCLQYRSYPRTFGGGTKLEIK 90 QVQLVQSGAEVKKPGASVKVSCKASGYSFSTFFIHWVRQAPGQGLEWIGRIDPNS GATKYNEKFESRVTMTRDTSISTAYMELSRLRSDDTAVYYCARGEDLLIRTDALD YWGQGTSVTVSS 91 DIQMTQSPSSLSASVGDRVSITCKASQNAGIDVAWFQQKPGKAPKLLIYSKSNRY TGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQYRSYPRTFGQGTKLEIK 92 GGTFNNAIN 93 GIIPMFGTAKYSQNFQG 94 SRDLLLFPHHALSP 95 QGDSLRSYYAS 96 GKNNRPS 97 SSRDSSGNHWV 98 GYTFTDQTIH 99 YIYPRDDSPKYNENFKG 100 PDRSGYAWFIY 101 KASRDVAIAVA 102 WASTRHT 103 HQYSSYPFT 104 DHIFSIHWMQ 105 EIFPGSGTTDYNEKFKG 106 GAFDY 107 RASQDIGNLS 108 ATSSLDS 109 LQYASSPFT BY DHIFSIHWMQ 111 EIFPGSGTTDYNEKFKG 112 GAFDY 113 RASQDIGNRLN 114 ATSSLDS 115 GYSFTFFIH 116 RIDPNSGATKYNEKFES 117 GEDLLIRTDALDY 118 KASQNAGIDVA 119 SKSNRYT 120 LQYRSYPRT 121 GYSFTFFIH

- 62 040031

122 GRIDPNSGATKYNEKFES 123 GEDLLIRTDALDY 124 KASQNAGIDVA 125 SKSNRYT 126 LQYRSYPRT 127 ESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLYITREPEVTCVWDVSQEDPEVQF NWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKG LPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWES NGQVMPENNYKTTCSVLDSDGSFFLYSRLTVSLVKH 128 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVWDVSHEDPEVKFNWYVDGVE VHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTIS KAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNY KTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP G 129 LEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVWDVSHED PEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCK VSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIA VEWESNGQVMPENNYKTGFSGNCSVLDSDGSFFLYSKWQVDK 130 LEPKSSDKTHTCPPCP 131 GCKWDLLIKQWVCDPLGSGSATGGSGSTASSGSGSATHMLPGCKWDLLIKQWV CDPLGGGGGVDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVWD VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGK EYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV MHEALHNHYTQKSLSLSPGK 132 QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHS GSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARGYYDILTGYYYY FDYWGQGTLVTVSS 133 EIVLTQSPATLSLSPGERATLSCRASQSVSRYLAWYQQKPGQAPRLLIYDASNRAT GIPARFSGSGSGTDSTLTISSLEPEDFAVYYCQQRSNWPRTFGQGTKVEIK 134 ESKYGPPCPPCP 135 GGCGGG 136 LGGCGGGS

Other embodiments of the invention.

All described features disclosed in this specification may be combined in any combination. Each property disclosed in this specification may be replaced by an alternative property that serves the same equivalent or similar purpose. Thus, unless otherwise indicated, each property is disclosed in this invention only to exemplify a general range of equivalent or similar properties.

From the above description, a person skilled in the art can easily determine the essential features of this disclosure and without departing from the spirit and scope of this invention can make various changes and modifications to the invention in order to adapt it to various applications and conditions. Thus, other embodiments of the invention are also within the scope of the claims.

Claims (39)

CLAIM 1. A compound that specifically binds FAFL (BAFF) and IL-23A, containing a first polypeptide and a second polypeptide, or containing two of said first polypeptides and two of said second polypeptides, where said two first polypeptides are linked together via at least one disulfide bond, and : (A) said first polypeptide contains: (i) a first immunoglobulin (VL1) light chain variable domain specific for the first target protein; (ii) a second immunoglobulin heavy chain variable domain (VH2) specific for a second target protein; and (iii) a hinge region, a heavy chain constant region 2 (CH2), and a heavy chain constant region 3 (CH3); and (B) said second polypeptide contains: (i) a second immunoglobulin (VL2) light chain variable domain specific for said second target protein; (ii) a heavy chain variable domain of the first immunoglobulin (VH1) specific for the indicated - 63 040031 to the first target protein, moreover: a) said VL1 and VH1 combine to form an attachment site that binds said first target protein; b) said VL2 and VH2 combine to form an attachment site that binds said second target protein; c) said heavy chain constant region 2 (CH2) contains tyrosine at position 252, threonine at position 254 and glutamic acid at position 256, numbered according to the EC index, as in Kabat; And d) said first target protein is FALV (BAFF) and said second target protein is IL-23A, or said first target protein is IL-23A and said second target protein is FALV (BAFF), and wherein: (i) said VL1 contains SEQ ID NO: 2, said VH1 contains SEQ ID NO: 1, said VL2 contains SEQ ID NO: 4, and said VH2 contains SEQ ID NO: 3; or (ii) said VL1 contains SEQ ID NO: 4, said VH1 contains SEQ ID NO: 3, said VL2 contains SEQ ID NO: 2, and said VH2 contains SEQ ID NO: 1; or (iii) said VL1 contains SEQ ID NO: 89, said VH1 contains SEQ ID NO: 88, said VL2 contains SEQ ID NO: 4, and said VH2 contains SEQ ID NO: 3; or (iv) said VL1 contains SEQ ID NO: 4, said VH1 contains SEQ ID NO: 3, said VL2 contains SEQ ID NO: 89, and said VH2 contains SEQ ID NO: 88; or (v) said VL1 contains SEQ ID NO: 91, said VH1 contains SEQ ID NO: 90, said VL2 contains SEQ ID NO: 4, and said VH2 contains SEQ ID NO: 3; or (vi) said VL1 contains SEQ ID NO: 4, said VH1 contains SEQ ID NO: 3, said VL2 contains SEQ ID NO: 91, and said VH2 contains SEQ ID NO: 90. 2. The compound of claim 1, wherein said first polypeptide further comprises a first linker between said VL1 and said VH2, and said second polypeptide further comprises a second linker between said VL2 and said VH1. 3. The compound according to claim 2, characterized in that said first linker or said second linker contains the amino acid sequence GGGSGGG (SEQ ID NO: 69). 4. The compound according to claim 2, characterized in that said first linker and said second linker contain the amino acid sequence GGGSGGG (SEQ ID NO: 69). 5. The compound of claim 1, wherein said first polypeptide further comprises a heavy chain constant region 1 (CH1) domain and said second polypeptide further comprises a light chain constant region (CL) domain, wherein said CL and said CH1 are joined via a disulfide bond in order to form the C1 domain. 6. The compound of claim 5, wherein said first polypeptide further comprises a third linker between said VH2 and said CH1 and said second polypeptide further comprises a fourth linker between said VH1 and said CL. 7. The compound according to claim 6, characterized in that said third linker or said fourth linker contains the amino acid sequence LGGGSG (SEQ ID NO: 70). 8. The compound according to claim 6, characterized in that said third linker and said fourth linker contain the amino acid sequence LGGGSG (SEQ ID NO: 70). 9. A compound according to any one of the preceding claims, characterized in that said heavy chain constant region 2 (CH2) contains alanine at position 234 and alanine at position 235, numbered according to the EC index as in Kabat. 10. A compound according to any one of the preceding claims, wherein the amino acid sequence of said heavy chain constant region 2 (CH2) or said heavy chain constant region 3 (CH3) is derived from IgG1 or from IgG4. 11. A compound according to any one of the preceding claims, characterized in that said hinge region contains the amino acid sequence EPKSCDKTHTCPPCP (SEQ ID NO: 76), the amino acid sequence LEPKSSDKTHTCPPCP (SEQ ID NO: 130) or the amino acid sequence ESKYGPPCPPCP (SEQ ID NO: 134). 12. A compound according to any one of the preceding claims, characterized in that said compound contains two said first polypeptides and two said second polypeptides, said two first polypeptides being linked together via at least one disulfide bond. 13. The compound according to claim 1, characterized in that: (i) said first polypeptide contains the amino acid sequence of SEQ ID NO: 5, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 6; or (ii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 7, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 8; or (iii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 9, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 10; or (iv) said first polypeptide contains the amino acid sequence of SEQ ID NO: 11, - 64 040031 and the specified second polypeptide contains the amino acid sequence of SEQ ID NO: 12; or (v) said first polypeptide contains the amino acid sequence of SEQ ID NO: 13, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 14; or (vi) said first polypeptide contains the amino acid sequence of SEQ ID NO: 15, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 16; or (vii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 17, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 18; or (viii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 19, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 20; or (ix) said first polypeptide contains the amino acid sequence of SEQ ID NO: 37 and said second polypeptide contains the amino acid sequence of SEQ ID NO: 38; or (x) said first polypeptide contains the amino acid sequence of SEQ ID NO: 39 and said second polypeptide contains the amino acid sequence of SEQ ID NO: 40; or (xi) said first polypeptide contains the amino acid sequence of SEQ ID NO: 41 and said second polypeptide contains the amino acid sequence of SEQ ID NO: 42; or (xii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 43 and said second polypeptide contains the amino acid sequence of SEQ ID NO: 44; or (xiii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 45, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 46; or (xiv) said first polypeptide contains the amino acid sequence of SEQ ID NO: 47, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 48; or (xv) said first polypeptide contains the amino acid sequence of SEQ ID NO: 49 and said second polypeptide contains the amino acid sequence of SEQ ID NO: 50; or (xvi) said first polypeptide contains the amino acid sequence of SEQ ID NO: 51 and said second polypeptide contains the amino acid sequence of SEQ ID NO: 52; or (xvii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 61 and said second polypeptide contains the amino acid sequence of SEQ ID NO: 62; or (xviii) said first polypeptide contains the amino acid sequence of SEQ ID NO: 63 and said second polypeptide contains the amino acid sequence of SEQ ID NO: 64; or (xix) said first polypeptide contains the amino acid sequence of SEQ ID NO: 65, and said second polypeptide contains the amino acid sequence of SEQ ID NO: 66; or (xx) said first polypeptide contains the amino acid sequence of SEQ ID NO: 67 and said second polypeptide contains the amino acid sequence of SEQ ID NO: 68. 14. The compound according to claim 13, characterized in that said compound contains two of said first polypeptides and two of said second polypeptides, wherein said two first polypeptides are linked together via at least one disulfide bond, and wherein each said first polypeptide is linked to one said second polypeptide through at least one disulfide bond. 15. The compound according to claim 13 or 14, characterized in that said compound contains two of said first polypeptides and two of said second polypeptides, wherein each of said first polypeptides contains CH1, hinge, CH2 and CH3, and each of said second polypeptides contains CL wherein the hinge, CH2 and CH3 of one of the first polypeptides is combined with the hinge, CH2 and CH3 of another of the first polypeptides, and the CH1 of each of said first polypeptides is combined with the CL of one of said second polypeptides to form a tetravalent molecule. 16. The compound of claim 1, comprising two first polypeptides and two second polypeptides, wherein said two first polypeptides are linked together via at least one disulfide bond, and wherein each said first polypeptide is linked to one said second polypeptide via at least one a disulfide bond, wherein each of said first polypeptides contains: (i) a first immunoglobulin (VL1) light chain variable domain specific for the first target protein; (ii) a second immunoglobulin heavy chain variable domain (VH2) specific for a second target protein; (iii) a heavy chain constant region 1 (CH1), a hinge region, a heavy chain constant region 2 (CH2), and a heavy chain constant region 3 (CH3), wherein said second polypeptides each comprise: (i) a second immunoglobulin (VL2) light chain variable domain specific for said second target protein; (ii) a first immunoglobulin (VH1) heavy chain variable domain specific for said first target protein; (iii) a light chain constant region (CL) domain, wherein the hinge, CH2 and CH3 of one of the first polypeptides combines with the hinge, CH2 and CH3 - 65 040031 of another of the first polypeptides, and CH1 of each of these first polypeptides combines with the CL of each of the second polypeptides to form a tetravalent molecule, while: a) said VL1 and VH1 combine to form an attachment site that binds said first target protein; b) said VL2 and VH2 combine to form an attachment site that binds said second target protein; c) said heavy chain constant region 2 (CH2) contains tyrosine at position 252, threonine at position 254 and glutamic acid at position 256, numbered according to the EC index, as in Kabat; And d) said first target protein is FALV (BAFF) and said second target protein is IL-23A, or said first target protein is IL-23A and said second target protein is FALV (BAFF), and wherein: (i) said VL1 contains SEQ ID NO: 2, said VH1 contains SEQ ID NO: 1, said VL2 contains SEQ ID NO: 4, and said VH2 contains SEQ ID NO: 3; or (ii) said VL1 contains SEQ ID NO: 4, said VH1 contains SEQ ID NO: 3, said VL2 contains SEQ ID NO: 2, and said VH2 contains SEQ ID NO: 1; or (iii) said VL1 contains SEQ ID NO: 89, said VH1 contains SEQ ID NO: 88, said VL2 contains SEQ ID NO: 4, and said VH2 contains SEQ ID NO: 3; or (iv) said VL1 contains SEQ ID NO: 4, said VH1 contains SEQ ID NO: 3, said VL2 contains SEQ ID NO: 89, and said VH2 contains SEQ ID NO: 88; or (v) said VL1 contains SEQ ID NO: 91, said VH1 contains SEQ ID NO: 90, said VL2 contains SEQ ID NO: 4, and said VH2 contains SEQ ID NO: 3; or (vi) said VL1 contains SEQ ID NO: 4, said VH1 contains SEQ ID NO: 3, said VL2 contains SEQ ID NO: 91, and said VH2 contains SEQ ID NO: 90. 17. The compound of claim 16, wherein each of said first polypeptides further comprises a first linker between said VL1 and said VH2, and each of said second polypeptides further comprises a second linker between said VL2 and said VH1. 18. The compound according to claim 17, characterized in that said first linker or said second linker contains the amino acid sequence GGGSGGGG (SEQ ID NO: 69). 19. The compound according to claim 17, characterized in that said first linker and said second linker contain the amino acid sequence GGGSGGGG (SEQ ID NO: 69). 20. The compound of claim 17, wherein each of said first polypeptides further comprises a third linker between said VH2 and said CH1, and each of said second polypeptides further comprises a fourth linker between said VH1 and said CL. 21. A compound according to claim 20, wherein said third linker or said fourth linker contains the amino acid sequence LGGGSG (SEQ ID NO: 70). 22. A compound according to claim 20, wherein said third linker and said fourth linker contain the amino acid sequence LGGGSG (SEQ ID NO: 70). 23. A compound according to claim 16, characterized in that said heavy chain constant region 2 (CH2) contains alanine at position 234 and alanine at position 235, numbered according to the EC index, as in Kabat. 24. The compound of claim 16 wherein the amino acid sequence of said hinge region, said heavy chain constant region 2 (CH2) or said heavy chain constant region 3 (CH3) is derived from IgG1 or from IgG4. 25. The compound according to claim 16, characterized in that said hinge region contains the amino acid sequence EPKSCDKTHTCPPCP (SEQ ID NO: 76), the amino acid sequence LEPKSSDKTHTCPPCP (SEQ ID NO: 130) or the amino acid sequence ESKYGPPCPPCP (SEQ ID NO: 134). 26. The compound according to claim 16, characterized in that: (i) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 5 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 6; or (ii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 7 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 8; or (iii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 13 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 14; or (iv) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 15 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 16; or (v) each of said first polypeptides contains the amino acid sequence - 66 040031 SEQ ID NO: 37 and each of these second polypeptides contains the amino acid sequence of SEQ ID NO: 38; or (vi) each of said first polypeptides contains the amino acid sequence SEQ ID NO: 41 and each of these second polypeptides contains the amino acid sequence of SEQ ID NO: 42; or (vii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 45 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 46; or (viii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 49 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 50; or (ix) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 61 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 62; or (x) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 63 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 64; or (xi) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 65 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 66; or (xii) each of said first polypeptides contains the amino acid sequence of SEQ ID NO: 67 and each of said second polypeptides contains the amino acid sequence of SEQ ID NO: 68. 27. Composition for binding FAVL (BAFF) and IL-23A, containing a compound according to any one of claims 1-26 and a pharmaceutically acceptable carrier. 28. A method of treating an autoimmune or inflammatory disease, comprising administering to a subject: (a) compounds according to any one of paragraphs.1-26; or (b) a pharmaceutical composition containing (i) said compound and (ii) a pharmaceutically acceptable carrier. 29. A compound according to any one of claims 1 to 26 which inhibits the function of BAFF and IL-23A for the treatment of an autoimmune or inflammatory disease. 30. Pharmaceutical composition containing: (i) a compound according to any one of claims 1 to 26; and (ii) a pharmaceutically acceptable carrier, wherein said composition is for use in the treatment of an autoimmune or inflammatory disease. 31. A nucleic acid containing a nucleotide sequence encoding a polypeptide according to any one of claims 1-26. 32. A vector containing a nucleic acid according to claim 31, which can be used to express a compound according to any one of claims 1-26. 33. The vector of claim 32, further comprising a promoter operably linked to said nucleic acid. 34. A cell containing a nucleic acid according to claim 31, or a vector according to claim 32 or 33. 35. A method for producing a polypeptide according to any one of claims 1 to 26, comprising obtaining a cell according to claim 34 and expressing said nucleic acid in said cell. 36. The method of claim 35, further comprising isolating and purifying said polypeptide. 37. The method according to claim 28, wherein the autoimmune or inflammatory disease includes systemic lupus erythematosus (SLE (SLE)), systemic lupus erythematosus with kidney damage/lupus nephritis (LN (LN)), ANCA-associated vasculitis ( AAV (AAV)), primary Sjögren's syndrome (PSS (pSS)), chronic graft-versus-host disease (CCVHD (cGVHD)), systemic sclerosis (SSc)), rheumatoid arthritis (PA (RA)), psoriasis (Ps ( Ps)), ankylosing spondylitis (AC (AS)) and psoriatic arthritis (PA (RA)). 38. The compound according to claim 29, characterized in that the autoimmune or inflammatory disease includes systemic lupus erythematosus (SLE (SLE)), systemic lupus erythematosus with kidney damage / lupus nephritis (LN (LN)), ANCA-associated vasculitis ( AAV (AAV)), primary Sjögren's syndrome (PSS (pSS)), chronic graft-versus-host disease (CCVHD (cGVHD)), systemic sclerosis (SSc)), rheumatoid arthritis (PA (RA)), psoriasis (Ps ( Ps)), ankylosing spondylitis (AC (AS)) and psoriatic arthritis (PA (RA)). 39. The pharmaceutical composition according to claim 30, characterized in that the autoimmune or inflammatory disease includes systemic lupus erythematosus (SLE (SLE)), systemic lupus erythematosus with kidney damage / lupus nephritis (LN (LN)), ANCA-associated vasculitis (AAB - 67 040031 (AAV)), primary Sjogren's syndrome (PSS (pSS)), chronic graft-versus-host disease (CCVHD (cGVHD)), systemic sclerosis (SSc)), rheumatoid arthritis (PA (RA)), psoriasis ( Ps (Ps)), ankylosing spondylitis (AC (AS)) and psoriatic arthritis (PA (RA)). Fig. 1A Fig. 1B - 68 040031 Fig. 2 Fig. 3 Fig. 4 * equals p less than 0.05 One-way analysis of variance followed by Dunnett's multiple comparison test versus hFAVLMk control Fig. 5 - 69 040031 * equals p less than 0.05 One-way analysis of variance followed by Dunnett's multiple comparison test versus rhIL-23 control