Anti-human p40 protein domain antibodies and uses thereof

Abstract

To provide antibodies for treating or preventing autoimmune diseases.SOLUTION: Provided is an in vivo or in vitro method including: administering a cell including an antibody or antigen-binding fragment thereof, an antibody conjugate, a multispecific antibody, a fusion protein, or a pharmaceutical composition that specifically binds to human IL-12 / IL-23p40, the antibody or antigen-binding fragment thereof blocking the function of an interleukin IL-12 / IL-23p40 protein domain and including heavy and light chain variable regions that include specific amino acid sequences; and administering to a subject in need thereof an effective amount of any one of the disclosed antibodies or antigen-binding fragments thereof, antibody conjugates, multispecific antibodies, fusion proteins, or pharmaceutical compositions.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention is in the field of medicine and in particular relates to monoclonal antibodies and their use for blocking the function of the interleukin IL-12 / IL-23p40 protein domain.

[0002] background Interleukin-12 (IL-12), also known as cytotoxic lymphocyte maturation factor (CLMF) or natural killer cell-stimulating factor (NKSF), is a member of the interleukin family. IL-12 has a unique heterodimeric structure: a glycosylated peptide chain (with a relative molecular weight of 75,000 daltons) formed by covalently linking two protein domains, p40 and p35, via disulfide bonds. IL-12 has a heavy chain (p40) consisting of 306 amino acids, containing 10 cysteine residues and four potential glycosylation sites, and a light chain (p35) consisting of 197 amino acids, containing 7 cysteine residues and three potential glycosylation sites.

[0003] Interleukin-23 (IL-23), a new member of the interleukin-12 (IL-12) cytokine family, was discovered by Oppmann et al. in 2000. It is a biologically active complex cytokine formed by covalently linking the p19 and p40 protein domains of IL-12 via disulfide bonds. IL-23 can be secreted by activated antigen-presenting cells and dendritic cells. IL-23 binds to its receptors, IL-23R and IL-12Rβ1, but not IL-12Rβ2 (Oppmann B et al., Immunity, 2000, 13(5): 715-725). Although IL-23 and IL-12 share the same signaling pathways, including the Janus kinases Tyk2, Jak2, and STAT, each affects different T cells. IL-23 inhibits memory T cell CD4 +IL-12 induces acute proliferation of virgin CD4 + Like IL-12, human IL-23 stimulates the proliferation of PHA-activated T cells and CD45RO + Stimulates IFN-γ production and proliferation in T cells. By antagonizing the common subunit p40 of IL-12 and IL-23, it is possible to effectively antagonize both the IL-12 and IL-23 pathways simultaneously.

[0004] Interleukin-12 (IL-12) is primarily produced by dendritic cells, macrophages, B lymphocytes, and some other antigen-presenting cells (APCs). It enhances the cytotoxicity of natural killer cells (NK cells) and cytotoxic T cells (Tc cells), stimulates resting or activated T cells and NK cells to produce interferon-γ (IFN-γ), and promotes Th0-to-Th1 differentiation. IL-12 also promotes Th1 cells to secrete IFN-γ and IL2 to mediate cellular immune responses, induces T cells and NK cells to produce IFN-γ, and provides extramedullary hematopoiesis. IL-12 plays a crucial role in both the early nonspecific immune response and the subsequent antigen-specific adaptive immune process in the body, and is a multifunctional immunoregulator (Manetti R et al., Journal of Experimental Medicine, 1993, 177(4): 1199-1204). The role of IL-12 in inducing autoimmunity is (1) to promote the differentiation and proliferation of antigen-specific Th1 cells, resulting in the production of multiple cytokines and enhancing Th1-type immune responses, and (2) to stimulate monocyte macrophages to produce various active mediators, enhance the cytotoxicity of immunocompetent cells, and cause damage to self-tissues; IL-12 is also involved in antibody-mediated autoimmunity.

[0005] IL-23 plays an important role in various immune diseases and is thought to be involved in the pathogenesis of psoriasis. Abnormal accumulation of IL-2 and TNF-α in psoriatic skin lesions promotes the onset and progression of psoriasis, and the secretion of multiple cytokines by lymphocytes in the skin lesions, forming a large cellular network primarily containing Th1 cytokines, is thought to be an important pathological basis of the disease. Th1 cytokines secreted by DCs are thought to be an initiating factor in the development of psoriasis, and mature DCs can increase the expression of IFN-α and IL-12. The specific mechanisms of action of psychological, genetic, and infectious factors in the development of the disease are unknown. Saint-Mezard et al. found that psychological stress promotes the aggregation of skin DCs and enhances delayed allergic responses to haptens. IL-23 has also been experimentally confirmed to affect DC-induced inflammatory responses, but it is unclear whether it is a direct inflammatory factor. IL-12 induces the onset of psoriasis by mediating T cells to the skin surface via cutaneous lymphocyte antigen, and Rosmarin et al. analyzed the structure, receptor, and function of IL-12 and suggested that psoriasis could be treated by altering IL-12 levels (Rosmarin D et al., Journal of Drugs in Dermatology, 2005, 4(3): 318-325).

[0006] Systemic lupus erythematosus (SLE) is a complex systemic autoimmune disease. Currently, there is no effective treatment available in clinical practice in China, and conventional treatments mainly involve hormones and immunosuppressants. STELARA® (ustekinumab), a product of the well-known pharmaceutical company Johnson & Johnson, is a fully human IL-12 and IL-23 antagonist approved by the US FDA for the treatment of SLE, and has demonstrated the efficacy of anti-IL-12p40 antibodies in the treatment of SLE (Janssen R&D's STELARA (ustekinumab) has shown positive results in the treatment of systemic lupus erythematosus in Phase II trials).

[0007] Therefore, IL-12 and IL-23 are crucial in inducing autoimmunity and maintaining immune responses, and as a result, any moiety that blocks the production or signaling of IL-12 and IL-23 can prevent or suppress the onset and progression of autoimmune diseases.

[0008] Structurally, IL-12 and IL-23 both have a p40 protein domain, and monoclonal antibodies that specifically bind to the p40 protein domain can be used as blockers of the IL-12 and IL-23 pathways, thereby representing new drugs for treating autoimmune diseases (such as psoriasis vulgaris and systemic lupus erythematosus).

[0009] Ulcerative colitis is an IBD disease characterized primarily by immune dysfunction. It is an immunological disease with a complex disease pathology characterized by complex clinicopathological changes, a prolonged disease course, and repeated attacks. More than half of patients undergo conventional or biological therapy without relief. Ulcerative colitis is the result of the combined action of exogenous and host factors in certain genetic conditions. Lesions are usually located in the sigmoid colon and rectum and may extend to the descending colon or the entire colon. The disease course is long, and repeated attacks are common. The disease can occur at any age, but is most common in people in their 20s and 30s. Given the characteristics of ulcerative colitis, there is a need to identify better antibody drugs for treating patients with ulcerative colitis.

[0010] overview Based on the crystal structure of IL-12 / IL-23p40, the inventors used artificial intelligence to design monoclonal antibody technology, develop antibody sequences, and pre-screen the antibodies by methods such as ELISA, and finally selected antibodies with higher potency as candidate antibodies for subsequent pharmacodynamic studies.

[0011] Additionally, the present inventors have prepared humanized antibodies against the human IL-12 / IL-23p40 protein domain (e.g., the humanized antibodies designated H5L9, H5L10, H5L11, H5L12, H5L14, and H8L15). The present inventors have surprisingly found that the antibody can specifically bind to the human IL-12 / IL-23p40 protein domain and exhibit good binding activity.

[0012] Furthermore, the inventors have surprisingly found that the antibodies disclosed herein are capable of effectively blocking the binding of the human IL-12 / IL-23p40 protein domain to the cell surface receptors IL-12Rβ1 and IL-23R, and inhibiting IL-23-induced IL-17A secretion by human peripheral blood mononuclear lymphocytes.

[0013] The present inventors have also surprisingly found that the antibodies disclosed herein can effectively bind to the human IL-12 / IL-23p40 protein domain, block the binding of the human IL-12 / IL-23p40 protein domain to its ligands IL-12Rβ1 and IL-23R, and inhibit activation of the downstream signaling pathway of IL-12 / IL-23. The antibodies may be used to prepare medicaments for the prevention and treatment of autoimmune diseases (e.g., psoriasis vulgaris or systemic lupus erythematosus) and ulcerative colitis (e.g., refractory or recurrent).

[0014] The amino acid sequences of the CDR regions of the above-mentioned antibodies are analyzed by technical means well known to those skilled in the art, for example, using the VBASE2 database.

[0015] It will be understood by those skilled in the art that the CDR regions of an antibody are responsible for the binding specificity of the antibody to the antigen. Given the known sequences of the heavy and light chain variable regions of an antibody, there are several methods for determining the CDR regions of an antibody, including the Kabat, IMGT, Chothia, and AbM numbering systems.

[0016] Specifically, the AbM numbering system: the AbM method for defining CDRs is derived from Martin's related work (Martin ACR, Cheetham JC, Rees AR (1989) Modeling antibody hypervariable loops: A combined algorithm. Proc Natl Acad Sci USA 86: 9268-9272), and this method integrates partial definitions of the Kabat method and the Chothia method.

[0017] Kabat numbering system: See, e.g., Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md., 1991. Chothia numbering system: See, e.g., Chothia & Lesk (1987) J. Mol. Biol. 196: 901-917; Chothia et al. (1989), Nature 342: 878-883. IMGT numbering system: See, e.g., Lefranc et al., Dev. Comparat. Immunol. 27: 55-77, 2003.

[0018] However, application of all definitions of CDR to an antibody or variants thereof is intended to be within the scope of the term as defined and used herein. If the amino acid sequence of the variable region of the antibody is known, one of skill in the art can generally determine which residues comprise a particular CDR without relying on any experimental data other than the sequence itself. The appropriate amino acid residues of CDRs as defined by the Kabat and Chothia CDR numbering system are listed below for comparison. The exact number of residues that comprise a particular CDR will vary depending on the sequence and size of that CDR.

[0019] Preferably, the antibodies H5L9, H5L10, H5L11, H5L12 and H5L14 disclosed herein have the same HCDRs 1 to 3, The amino acid sequences of the three HCDR regions of the heavy chain variable region are as follows: HCDR1: GYSFTTYW (SEQ ID NO: 3) HCDR2: IMSPVDSDI (SEQ ID NO: 4) HCDR3: ARRRPGQGYFDF (SEQ ID NO: 5) The antibody H5L9 disclosed herein has LCDRs 1 to 3, The amino acid sequences of the three CDR regions of the light chain variable region are as follows: LCDR1:QNVGSW (SEQ ID NO: 8) LCDR2:ASS (SEQ ID NO: 9) LCDR3: QQYDIYPFT (SEQ ID NO: 10) The antibody H5L10 disclosed herein has LCDRs 1 to 3, The amino acid sequences of the three CDR regions of the light chain variable region are as follows: LCDR1:QSVGSW (SEQ ID NO: 19) LCDR2:ASN (SEQ ID NO: 21) LCDR3: QQYNIYPYT (SEQ ID NO: 22) The antibodies H5L11 and H5L12 disclosed herein have the same LCDRs 1 to 3, The amino acid sequences of the three CDR regions of the light chain variable region are as follows: LCDR1: QSVSSW (SEQ ID NO: 20) LCDR2:ASN (SEQ ID NO: 21) LCDR3: QQYNIYPYT (SEQ ID NO: 22) The antibody H5L14 disclosed herein has LCDRs 1-3: The amino acid sequences of the three CDR regions of the light chain variable region are as follows: LCDR1: QSVSSW (SEQ ID NO: 20) LCDR2:ASN (SEQ ID NO: 21) LCDR3: QQYNIYPFT (SEQ ID NO: 23) The antibody H8L15 disclosed herein has HCDRs 1-3 and LCDRs 1-3, The amino acid sequences of the three HCDR regions of the heavy chain variable region are as follows: HCDR1: GYTFTSYW (SEQ ID NO: 26) HCDR2: MSPVDSDI (SEQ ID NO: 4) HCDR3: ARRRPGQGYFDF (SEQ ID NO: 5) The amino acid sequences of the three CDR regions of the light chain variable region are as follows: LCDR1:QSVGTW (SEQ ID NO: 27) LCDR2: AAS (SEQ ID NO: 28) LCDR3: QQYNIYPYT (sequence number 22).

[0020] The present invention is described in detail below. One aspect of the present invention relates to an antibody or antigen-binding fragment thereof that preferably specifically binds to human IL-12 / IL-23p40, wherein: (1) The antibody comprising HCDR1, HCDR2 and HCDR3 contained in the heavy chain variable region set forth in SEQ ID NO: 1, (wherein preferably said HCDR1 comprises or consists of the sequence set forth in SEQ ID NO: 3, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to said sequence, or an amino acid sequence having one or more (preferably 1, 2 or 3) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence; said HCDR2 comprises or consists of the sequence set forth in SEQ ID NO: 4, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto, or an amino acid sequence having one or more (preferably 1, 2 or 3) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence, and said HCDR3 comprises or consists of the sequence set forth in SEQ ID NO: 5, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto, or an amino acid sequence having one or more (preferably 1, 2 or 3) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence; and the antibody further comprises: comprising LCDR1, LCDR2 and LCDR3 contained in the light chain variable region set forth in SEQ ID NO: 6, (wherein preferably said LCDR1 comprises or consists of the amino acid sequence set forth in SEQ ID NO: 8, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to said sequence, or an amino acid sequence having one or more (preferably 1, 2 or 3) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence; said LCDR2 comprises or consists of the amino acid sequence set forth in SEQ ID NO: 9, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto, or an amino acid sequence having one or more (preferably 1, 2 or 3) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence; and said LCDR3 comprises or consists of the sequence set forth in SEQ ID NO: 10, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to said sequence, or an amino acid sequence having one or more (preferably 1, 2 or 3) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence; (2) The antibody comprising HCDR1, HCDR2 and HCDR3 contained in the heavy chain variable region set forth in SEQ ID NO: 1, (wherein preferably said HCDR1 comprises or consists of the sequence set forth in SEQ ID NO: 3, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to said sequence, or an amino acid sequence having one or more (preferably 1, 2 or 3) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence; said HCDR2 comprises or consists of the sequence set forth in SEQ ID NO: 4, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto, or an amino acid sequence having one or more (preferably 1, 2 or 3) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence, and said HCDR3 comprises or consists of the sequence set forth in SEQ ID NO: 5, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto, or an amino acid sequence having one or more (preferably 1, 2 or 3) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence; The antibody further comprises: comprising LCDR1, LCDR2 and LCDR3 contained in the light chain variable region set forth in SEQ ID NO: 11, (wherein preferably said LCDR1 comprises or consists of the amino acid sequence set forth in SEQ ID NO: 19, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to said sequence, or an amino acid sequence having one or more (preferably 1, 2 or 3) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence; said LCDR2 comprises or consists of the amino acid sequence set forth in SEQ ID NO: 21, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto, or an amino acid sequence having one or more (preferably 1, 2 or 3) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence; and said LCDR3 comprises or consists of the sequence set forth in SEQ ID NO: 22, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto, or an amino acid sequence having one or more (preferably 1, 2 or 3) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence; (3) The antibody comprising HCDR1, HCDR2 and HCDR3 contained in the heavy chain variable region set forth in SEQ ID NO: 1, (wherein preferably said HCDR1 comprises or consists of the sequence set forth in SEQ ID NO: 3, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to said sequence, or an amino acid sequence having one or more (preferably 1, 2 or 3) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence; said HCDR2 comprises or consists of the sequence set forth in SEQ ID NO: 4, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto, or an amino acid sequence having one or more (preferably 1, 2 or 3) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence, and said HCDR3 comprises or consists of the sequence set forth in SEQ ID NO: 5, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto, or an amino acid sequence having one or more (preferably 1, 2 or 3) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence; The antibody further comprises: comprising LCDR1, LCDR2 and LCDR3 contained in the light chain variable region set forth in SEQ ID NOs: 13 and 15, (wherein preferably said LCDR1 comprises or consists of the amino acid sequence set forth in SEQ ID NO: 20, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to said sequence, or an amino acid sequence having one or more (preferably 1, 2 or 3) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence; said LCDR2 comprises or consists of the amino acid sequence set forth in SEQ ID NO: 21, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto, or an amino acid sequence having one or more (preferably 1, 2 or 3) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence; and said LCDR3 comprises or consists of the sequence set forth in SEQ ID NO: 22, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to said sequence, or an amino acid sequence having one or more (preferably 1, 2 or 3) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence; (4) The antibody is comprising HCDR1, HCDR2 and HCDR3 contained in the heavy chain variable region set forth in SEQ ID NO: 1, (wherein preferably said HCDR1 comprises or consists of the sequence set forth in SEQ ID NO: 3, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to said sequence, or an amino acid sequence having one or more (preferably 1, 2 or 3) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence; said HCDR2 comprises or consists of the sequence set forth in SEQ ID NO: 4, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto, or an amino acid sequence having one or more (preferably 1, 2 or 3) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence, and said HCDR3 comprises or consists of the sequence set forth in SEQ ID NO: 5, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto, or an amino acid sequence having one or more (preferably 1, 2 or 3) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence; The antibody further comprises: comprising LCDR1, LCDR2 and LCDR3 contained in the light chain variable region set forth in SEQ ID NO: 17, (wherein preferably said LCDR1 comprises or consists of the amino acid sequence set forth in SEQ ID NO: 20, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to said sequence, or an amino acid sequence having one or more (preferably 1, 2 or 3) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence; said LCDR2 comprises or consists of the amino acid sequence set forth in SEQ ID NO: 21, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto, or an amino acid sequence having one or more (preferably 1, 2 or 3) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence; and said LCDR3 comprises or consists of the sequence set forth in SEQ ID NO: 23, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to said sequence, or an amino acid sequence having one or more (preferably 1, 2 or 3) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence; (5) The antibody is: comprising HCDR1, HCDR2 and HCDR3 contained in a heavy chain variable region as set forth in SEQ ID NO: 24; (wherein preferably said HCDR1 comprises or consists of the sequence as set forth in SEQ ID NO: 26, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to said sequence, or an amino acid sequence having one or more (preferably 1, 2 or 3) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence; said HCDR2 comprises or consists of the sequence set forth in SEQ ID NO: 4, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto, or an amino acid sequence having one or more (preferably 1, 2 or 3) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence, and said HCDR3 comprises or consists of the sequence set forth in SEQ ID NO: 5, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto, or an amino acid sequence having one or more (preferably 1, 2 or 3) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence; The antibody further comprises: comprising LCDR1, LCDR2 and LCDR3 contained in the light chain variable region set forth in SEQ ID NO: 25 (wherein preferably said LCDR1 comprises or consists of the amino acid sequence set forth in SEQ ID NO: 27, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to said sequence, or an amino acid sequence having one or more (preferably 1, 2 or 3) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence; said LCDR2 comprises or consists of the amino acid sequence set forth in SEQ ID NO: 28, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto, or an amino acid sequence having one or more (preferably 1, 2 or 3) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence; and Said LCDR3 comprises or consists of the sequence set forth in SEQ ID NO: 22, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to said sequence, or an amino acid sequence having one or more (preferably 1, 2 or 3) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence).

[0021] In some embodiments of the invention, the antibody comprises: (1) the amino acid sequence set forth in SEQ ID NO: 1; a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the sequence set forth in SEQ ID NO: 1; a heavy chain variable region comprising or consisting of an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to the sequence set forth in SEQ ID NO:1; and the amino acid sequence set forth in SEQ ID NO: 6; a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the sequence set forth in SEQ ID NO: 6; or a light chain variable region comprising or consisting of an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to the sequence set forth in SEQ ID NO: 6; (2) the amino acid sequence set forth in SEQ ID NO: 1; a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the sequence set forth in SEQ ID NO: 1; a heavy chain variable region comprising or consisting of an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to the sequence set forth in SEQ ID NO:1; and the amino acid sequence set forth in SEQ ID NO: 11; a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the sequence set forth in SEQ ID NO: 11; a light chain variable region comprising or consisting of an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to the sequence set forth in SEQ ID NO: 11; (3) the amino acid sequence set forth in SEQ ID NO: 1; a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the sequence set forth in SEQ ID NO: 1; a heavy chain variable region comprising or consisting of an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to the sequence set forth in SEQ ID NO:1; and the amino acid sequence set forth in SEQ ID NO: 13; a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the sequence set forth in SEQ ID NO: 13, or a light chain variable region comprising or consisting of an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to the sequence set forth in SEQ ID NO: 13; (4) the amino acid sequence set forth in SEQ ID NO: 1; a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the sequence set forth in SEQ ID NO: 1; a heavy chain variable region comprising or consisting of an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to the sequence set forth in SEQ ID NO:1; and the amino acid sequence set forth in SEQ ID NO: 15; a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the sequence set forth in SEQ ID NO: 15; or a light chain variable region comprising or consisting of an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to the sequence set forth in SEQ ID NO: 15; (5) the amino acid sequence set forth in SEQ ID NO: 1; a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the sequence set forth in SEQ ID NO: 1; a heavy chain variable region comprising or consisting of an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to the sequence set forth in SEQ ID NO:1; and the amino acid sequence set forth in SEQ ID NO: 17; a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the sequence set forth in SEQ ID NO: 17, or a light chain variable region comprising or consisting of an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to the sequence set forth in SEQ ID NO: 17; (6) the amino acid sequence set forth in SEQ ID NO: 24; a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the sequence set forth in SEQ ID NO: 24, or a heavy chain variable region comprising or consisting of an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to the sequence set forth in SEQ ID NO: 24; and the amino acid sequence set forth in SEQ ID NO: 25; a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the sequence set forth in SEQ ID NO: 25; or a light chain variable region comprising or consisting of an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to the sequence set forth in SEQ ID NO: 25; Includes:

[0022] In one embodiment of the invention, the antibody further comprises framework regions (FRs) in the heavy chain variable region, preferably FRs comprising FR-H1, FR-H2, FR-H3 and FR-H4 (wherein the FR-H1 has an amino acid sequence set forth in SEQ ID NO: 29, a sequence which is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96% relative to the sequence set forth in SEQ ID NO: 29). , 97%, 98% or 99% sequence identity to the amino acid sequence set forth in SEQ ID NO: 29, or an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to the amino acid sequence set forth in SEQ ID NO: 30; , 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity, or an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to the amino acid sequence set forth in SEQ ID NO: 30; FR-H3 comprises or consists of the amino acid sequence set forth in SEQ ID NO: 31, or consisting of an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to said amino acid sequence as set forth in SEQ ID NO: 31, or an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said amino acid sequence as set forth in SEQ ID NO: 31;FR-H4 comprises or consists of the amino acid sequence set forth in SEQ ID NO: 32, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to said sequence set forth in SEQ ID NO: 32, or an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said amino acid sequence set forth in SEQ ID NO: 32);

[0023] In one embodiment of the invention, the antibody further comprises framework regions (FRs) in the light chain variable region, preferably FRs comprising FR-L1, FR-L2, FR-L3 and FR-L4 (wherein FR-L1 is the amino acid sequence set forth in SEQ ID NO: 33, at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, or 97%, relative to the sequence set forth in SEQ ID NO: 33, FR-L2 comprises or consists of an amino acid sequence having 97%, 98% or 99% sequence identity or an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to the amino acid sequence set forth in SEQ ID NO: 33; FR-L3 comprises or consists of an amino acid sequence set forth in SEQ ID NO: 34, an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 1109%, 1110%, 112%, 113%, 114%, 115%, 116%, 117%, 118%, 119%, 120%, 121%, 122%, 123%, 124%, 125%, 126%, 127%, 128%, 129%, 130%, 131%, 132%, 133%, 134%, 135%, 136%, 137%, 138%, 139%, 140%, 141%, 142%, 143%, 144%, 145%, 146%, 147%, 148%, 149%, 150%, 151%, 152%, 153%, 154%, 155%, 156%, 15 FR-L3 comprises or consists of an amino acid sequence having 9% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence set forth in SEQ ID NO: 34, or an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to the amino acid sequence set forth in SEQ ID NO: 35; FR-L4 comprises or consists of an amino acid sequence set forth in SEQ ID NO: 36, comprising or consisting of an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity, or having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said amino acid sequence set forth in SEQ ID NO: 35;FR-L4 comprises or consists of the amino acid sequence set forth in SEQ ID NO: 36, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to said sequence set forth in SEQ ID NO: 36, or an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said amino acid sequence set forth in SEQ ID NO: 36);

[0024] In one embodiment of the invention, the antibody further comprises framework regions (FRs) in the light chain variable region, preferably FRs comprising FR-L1, FR-L2, FR-L3 and FR-L4 (wherein said FR-L1 has an amino acid sequence set forth in SEQ ID NO: 41, a sequence similar to said sequence set forth in SEQ ID NO: 41, at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 1109%, 1111%, 112%, 113%, 114%, 115%, 116%, 117%, 118%, 119%, 120%, 121%, 122%, 123%, 124%, 125%, 126%, 127%, 128%, 129%, 130%, 131%, 132%, 133%, 134%, 135%, 136%, 137%, 138%, 139%, 140%, 141%, 142%, 143%, 144%, 145%, 146%, 147%, 148%, 149%, 150%, 151, 152%, 153%, 154%, 155%, 156%, 157%, 158%, 159%, 160%, 161%, 162%, 163%, 164%, 1 FR-L2 comprises or consists of an amino acid sequence having 7%, 98% or 99% sequence identity to the amino acid sequence set forth in SEQ ID NO: 41, or an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to the amino acid sequence set forth in SEQ ID NO: 41; said FR-L2 comprises or consists of an amino acid sequence set forth in SEQ ID NO: 42, an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 1109%, 1110%, 112%, 113%, 114%, 115%, 116%, 117%, 118%, 119%, 120%, 121%, 122%, 123%, 124%, 125%, 126%, 127%, 128%, 129%, 130%, 131%, 132%, 133%, 134%, 135%, 136%, 137%, 138%, 139%, 140%, 141%, 142%, 143%, 144%, 145%, 146%, 147%, 148%, 149%, 150%, 151%, 152%, 153%, FR-L3 comprises or consists of an amino acid sequence having 9% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity, or an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to the amino acid sequence set forth in SEQ ID NO: 42; said FR-L3 comprises or consists of an amino acid sequence set forth in SEQ ID NO: 43, or consisting of an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to said amino acid sequence as set forth in SEQ ID NO: 43, or having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said amino acid sequence as set forth in SEQ ID NO: 43;The FR-L4 comprises or consists of the amino acid sequence set forth in SEQ ID NO: 44, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the sequence set forth in SEQ ID NO: 44, or an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to the amino acid sequence set forth in SEQ ID NO: 44;

[0025] In one embodiment of the invention, the antibody further comprises framework regions (FRs) in the light chain variable region, preferably FRs comprising FR-L1, FR-L2, FR-L3 and FR-L4, wherein said FR-L1 has the amino acid sequence set forth in SEQ ID NO: 33, a sequence similar to that of SEQ ID NO: 33, at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96% , 97%, 98% or 99% sequence identity to the amino acid sequence set forth in SEQ ID NO: 33, or an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to the amino acid sequence set forth in SEQ ID NO: 33; said FR-L2 comprises or consists of the amino acid sequence set forth in SEQ ID NO: 45, an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, said FR-L3 comprises or consists of an amino acid sequence having 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to said amino acid sequence set forth in SEQ ID NO: 45, or an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said amino acid sequence set forth in SEQ ID NO: 46; said FR-L3 comprises or consists of an amino acid sequence set forth in SEQ ID NO: 46, or consisting of an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to said amino acid sequence as set forth in SEQ ID NO: 46, or having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said amino acid sequence as set forth in SEQ ID NO: 46;The FR-L4 comprises or consists of the amino acid sequence set forth in SEQ ID NO: 36, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the sequence set forth in SEQ ID NO: 36, or an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to the amino acid sequence set forth in SEQ ID NO: 36;

[0026] In one embodiment of the invention, the antibody further comprises framework regions (FRs) in the light chain variable region, preferably FRs comprising FR-L1, FR-L2, FR-L3 and FR-L4 (wherein said FR-L1 has an amino acid sequence set forth in SEQ ID NO: 33, a sequence which is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96% relative to said sequence set forth in SEQ ID NO: 33). , 97%, 98% or 99% sequence identity to the amino acid sequence set forth in SEQ ID NO: 33, or an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to the amino acid sequence set forth in SEQ ID NO: 33; said FR-L2 comprises or consists of the amino acid sequence set forth in SEQ ID NO: 47, an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88% sequence identity to the amino acid sequence set forth in SEQ ID NO: 47 , 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity, or an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to the amino acid sequence set forth in SEQ ID NO: 47; FR-L3 comprises or consists of the amino acid sequence set forth in SEQ ID NO: 46, or consisting of an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to said amino acid sequence as set forth in SEQ ID NO: 46, or an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said amino acid sequence as set forth in SEQ ID NO: 46;The FR-L4 comprises or consists of the amino acid sequence set forth in SEQ ID NO: 36, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the sequence set forth in SEQ ID NO: 36, or an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to the amino acid sequence set forth in SEQ ID NO: 36;

[0027] In one embodiment of the invention, the antibody further comprises framework regions (FRs) in the light chain variable region, preferably FRs comprising FR-L1, FR-L2, FR-L3 and FR-L4 (wherein the FR-L1 has an amino acid sequence as set forth in SEQ ID NO: 33, a sequence similar to the sequence set forth in SEQ ID NO: 33, at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, said FR-L2 comprises or consists of an amino acid sequence having 97%, 98% or 99% sequence identity, or an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said amino acid sequence set forth in SEQ ID NO: 33; said FR-L2 comprises or consists of an amino acid sequence set forth in SEQ ID NO: 45, an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, said FR-L3 comprises or consists of an amino acid sequence having 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to said amino acid sequence set forth in SEQ ID NO: 45, or an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said amino acid sequence set forth in SEQ ID NO: 48; said FR-L3 comprises or consists of an amino acid sequence set forth in SEQ ID NO: 48, or consisting of an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to said amino acid sequence as set forth in SEQ ID NO: 48, or having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said amino acid sequence as set forth in SEQ ID NO: 48;The FR-L4 comprises or consists of the amino acid sequence set forth in SEQ ID NO: 36, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the sequence set forth in SEQ ID NO: 36, or an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to the amino acid sequence set forth in SEQ ID NO: 36;

[0028] In one embodiment of the invention, the antibody comprises or consists of a heavy chain set forth in SEQ ID NO:39 and a light chain set forth in SEQ ID NO:40.

[0029] In one embodiment of the invention, the antibody comprises or consists of a heavy chain set forth in SEQ ID NO:49 and a light chain set forth in SEQ ID NO:50.

[0030] In one embodiment of the invention, the antibody comprises or consists of a heavy chain set forth in SEQ ID NO:49 and a light chain set forth in SEQ ID NO:51.

[0031] In one embodiment of the invention, the antibody comprises or consists of a heavy chain set forth in SEQ ID NO:49 and a light chain set forth in SEQ ID NO:52.

[0032] In one embodiment of the invention, the antibody comprises or consists of a heavy chain set forth in SEQ ID NO:49 and a light chain set forth in SEQ ID NO:53.

[0033] In one embodiment of the invention, the antibody comprises or consists of a heavy chain set forth in SEQ ID NO:49 and a light chain set forth in SEQ ID NO:54.

[0034] One aspect of the present invention relates to an isolated polypeptide comprising the sequence set forth in SEQ ID NOs: 3, 4 and 5, wherein said polypeptide specifically binds to a human IL-12 / IL-23p40 protein domain as part of an anti-human IL-12 / IL-23p40 protein domain antibody, said antibody further comprising the sequence set forth in SEQ ID NOs: 8, 9 and 10.

[0035] One aspect of the present invention relates to an isolated polypeptide comprising the sequence set forth in SEQ ID NOs: 8, 9 and 10, wherein said polypeptide specifically binds to a human IL-12 / IL-23p40 protein domain as part of an anti-human IL-12 / IL-23p40 protein domain antibody, said antibody further comprising the sequence set forth in SEQ ID NOs: 3, 4 and 5.

[0036] One aspect of the present invention relates to an isolated polypeptide comprising the sequence set forth in SEQ ID NOs: 3, 4 and 5, wherein said polypeptide specifically binds to a human IL-12 / IL-23p40 protein domain as part of an anti-human IL-12 / IL-23p40 protein domain antibody, said antibody further comprising the sequence set forth in SEQ ID NOs: 19, 21 and 22.

[0037] One aspect of the present invention relates to an isolated polypeptide comprising the sequence set forth in SEQ ID NOs: 19, 21 and 22, wherein said polypeptide specifically binds to a human IL-12 / IL-23p40 protein domain as part of an anti-human IL-12 / IL-23p40 protein domain antibody, said antibody further comprising the sequence set forth in SEQ ID NOs: 3, 4 and 5.

[0038] One aspect of the present invention relates to an isolated polypeptide comprising the sequence set forth in SEQ ID NOs: 3, 4 and 5, wherein said polypeptide specifically binds to a human IL-12 / IL-23p40 protein domain as part of an anti-human IL-12 / IL-23p40 protein domain antibody, said antibody further comprising the sequence set forth in SEQ ID NOs: 20, 21 and 22.

[0039] One aspect of the present invention relates to an isolated polypeptide comprising the sequence set forth in SEQ ID NOs: 20, 21 and 22, wherein said polypeptide specifically binds to a human IL-12 / IL-23p40 protein domain as part of an anti-human IL-12 / IL-23p40 protein domain antibody, said antibody further comprising the sequence set forth in SEQ ID NOs: 3, 4 and 5.

[0040] One aspect of the present invention relates to an isolated polypeptide comprising the sequence set forth in SEQ ID NOs: 3, 4 and 5, wherein said polypeptide specifically binds to a human IL-12 / IL-23p40 protein domain as part of an anti-human IL-12 / IL-23p40 protein domain antibody, said antibody further comprising the sequence set forth in SEQ ID NOs: 20, 21 and 23.

[0041] One aspect of the present invention relates to an isolated polypeptide comprising the sequence set forth in SEQ ID NOs: 20, 21 and 23, wherein said polypeptide specifically binds to a human IL-12 / IL-23p40 protein domain as part of an anti-human IL-12 / IL-23p40 protein domain antibody, said antibody further comprising the sequence set forth in SEQ ID NOs: 3, 4 and 5.

[0042] One aspect of the present invention relates to an isolated polypeptide comprising the sequence set forth in SEQ ID NOs: 26, 4 and 5, wherein said polypeptide specifically binds to a human IL-12 / IL-23p40 protein domain as part of an anti-human IL-12 / IL-23p40 protein domain antibody, said antibody further comprising the sequence set forth in SEQ ID NOs: 27, 28 and 22.

[0043] One aspect of the present invention relates to an isolated polypeptide comprising the sequence set forth in SEQ ID NOs: 27, 28 and 22, wherein said polypeptide specifically binds to a human IL-12 / IL-23p40 protein domain as part of an anti-human IL-12 / IL-23p40 protein domain antibody, said antibody further comprising the sequence set forth in SEQ ID NOs: 26, 4 and 5.

[0044] One aspect of the present invention is a method for producing a medicament for the treatment of a pulmonary arthritis. 2. An isolated polypeptide comprising the sequence set forth in SEQ ID NO: 1 or 24, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, sequence identity to said sequence, or an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence, wherein said polypeptide is used as part of an anti-human IL-12 / IL-23p40 protein domain antibody. specifically binds to a human IL-12 / IL-23p40 protein domain, said antibody further comprising a sequence as set forth in SEQ ID NO: 6, 11, 13, 15, 17 or 25, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, sequence identity to said sequence, or an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence; or 2. An isolated polypeptide comprising a sequence as set forth in SEQ ID NO: 6, 11, 13, 15, 17 or 25, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to said sequence, or an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence, wherein said polypeptide is an antibody to an anti-human IL-12 / IL-23p40 protein. and specifically binds to the human IL-12 / IL-23p40 protein domain as part of a protein domain antibody, said antibody further comprising the sequence set forth in SEQ ID NO: 1 or 24, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to said sequence, or an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence. Regarding.

[0045] In one embodiment of the invention, the antigen-binding fragment is selected from the group consisting of Fab, Fab', F(ab')2, Fd, Fv, dAb, Fab / c, complementarity-determining region (CDR) fragments, single-chain antibodies (e.g., scFv), bivalent antibodies, and domain antibodies.

[0046] In one embodiment of the invention, the antibody is a humanized antibody, a chimeric antibody or a multispecific antibody (eg a bispecific antibody).

[0047] In one embodiment of the invention, the antibody is -5 Less than m, e.g., about 10 -6 M, 10 -7 M, 10 -8 M, 10 -9 Less than M or 10 -10 K below MD Preferably, the K D is measured by the Fortebio molecular interaction analyzer.

[0048] In one embodiment of the invention, the antibody has an EC50 of less than about 100 nM, for example, about 10 nM, 1 nM, 0.9 nM, 0.8 nM, 0.7 nM, 0.6 nM, 0.5 nM, 0.4 nM, 0.3 nM, 0.2 nM, 0.1 nM or less. 50 Specifically, the EC 50 is measured by indirect ELISA.

[0049] In one embodiment of the invention, the antibody comprises a constant region, and the constant region is derived from a species other than mouse, such as a human antibody, preferably human IgG, more preferably IgG1.

[0050] In one embodiment of the present invention, the constant region of the antibody is humanized, for example, the heavy chain constant region is an Igγ-1 chain C region, preferably the Igγ-1 chain C region of GenBank Accession No. P01857; and the light chain constant region is an Igκ chain C region, preferably the Igκ chain C region of GenBank Accession No. P01834.

[0051] Another aspect of the present invention relates to an isolated polynucleotide encoding a polypeptide comprising the sequence set forth in SEQ ID NOs: 3, 4 and 5, wherein said polypeptide specifically binds to a human IL-12 / IL-23p40 protein domain as part of an anti-human IL-12 / IL-23p40 protein domain antibody, said antibody further comprising the sequence set forth in SEQ ID NOs: 8, 9 and 10.

[0052] One aspect of the present invention relates to an isolated polynucleotide encoding a polypeptide comprising the sequence set forth in SEQ ID NOs: 8, 9 and 10, wherein said polypeptide specifically binds to a human IL-12 / IL-23p40 protein domain as part of an anti-human IL-12 / IL-23p40 protein domain antibody, said antibody further comprising the sequence set forth in SEQ ID NOs: 3, 4 and 5.

[0053] Another aspect of the present invention relates to an isolated polynucleotide encoding a polypeptide comprising the sequence set forth in SEQ ID NOs: 3, 4 and 5, wherein said polypeptide specifically binds to a human IL-12 / IL-23p40 protein domain as part of an anti-human IL-12 / IL-23p40 protein domain antibody, said antibody further comprising the sequence set forth in SEQ ID NOs: 19, 21 and 22.

[0054] One aspect of the present invention relates to an isolated polynucleotide encoding a polypeptide comprising the sequence set forth in SEQ ID NOs: 19, 21 and 22, wherein said polypeptide specifically binds to a human IL-12 / IL-23p40 protein domain as part of an anti-human IL-12 / IL-23p40 protein domain antibody, said antibody further comprising the sequence set forth in SEQ ID NOs: 3, 4 and 5.

[0055] Another aspect of the present invention relates to an isolated polynucleotide encoding a polypeptide comprising the sequence set forth in SEQ ID NOs: 3, 4 and 5, wherein said polypeptide specifically binds to a human IL-12 / IL-23p40 protein domain as part of an anti-human IL-12 / IL-23p40 protein domain antibody, said antibody further comprising the sequence set forth in SEQ ID NOs: 20, 21 and 22.

[0056] One aspect of the present invention relates to an isolated polynucleotide encoding a polypeptide comprising the sequence set forth in SEQ ID NOs: 20, 21 and 22, wherein said polypeptide specifically binds to a human IL-12 / IL-23p40 protein domain as part of an anti-human IL-12 / IL-23p40 protein domain antibody, said antibody further comprising the sequence set forth in SEQ ID NOs: 3, 4 and 5.

[0057] Another aspect of the present invention relates to an isolated polynucleotide encoding a polypeptide comprising the sequence set forth in SEQ ID NOs: 3, 4 and 5, wherein said polypeptide specifically binds to a human IL-12 / IL-23p40 protein domain as part of an anti-human IL-12 / IL-23p40 protein domain antibody, said antibody further comprising the sequence set forth in SEQ ID NOs: 20, 21 and 23.

[0058] One aspect of the present invention relates to an isolated polynucleotide encoding a polypeptide comprising the sequence set forth in SEQ ID NOs: 20, 21 and 23, wherein said polypeptide specifically binds to a human IL-12 / IL-23p40 protein domain as part of an anti-human IL-12 / IL-23p40 protein domain antibody, said antibody further comprising the sequence set forth in SEQ ID NOs: 3, 4 and 5.

[0059] Another aspect of the present invention relates to an isolated polynucleotide encoding a polypeptide comprising the sequence set forth in SEQ ID NOs: 26, 4 and 5, wherein said polypeptide specifically binds to a human IL-12 / IL-23p40 protein domain as part of an anti-human IL-12 / IL-23p40 protein domain antibody, said antibody further comprising the sequence set forth in SEQ ID NOs: 27, 28 and 22.

[0060] One aspect of the present invention relates to an isolated polynucleotide encoding a polypeptide comprising the sequence set forth in SEQ ID NOs: 27, 28 and 22, wherein said polypeptide specifically binds to a human IL-12 / IL-23p40 protein domain as part of an anti-human IL-12 / IL-23p40 protein domain antibody, said antibody further comprising the sequence set forth in SEQ ID NOs: 26, 4 and 5.

[0061] One aspect of the present invention relates to an isolated polynucleotide encoding a polypeptide comprising the sequence set forth in SEQ ID NO: 1 or 24, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, sequence identity to said sequence, or an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence, wherein said polypeptide is an anti-human IL-12 / IL-23p40 specifically binds to the human IL-12 / IL-23p40 protein domain as part of a protein domain antibody, said antibody further comprising the sequence set forth in SEQ ID NO: 6, 11, 13, 15, 17 or 25, an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto, or having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared thereto; or 2. An isolated polynucleotide encoding a polypeptide comprising the sequence set forth in SEQ ID NO: 6, 11, 13, 15, 17 or 25, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, sequence identity to said sequence, or an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence, wherein said polypeptide is an anti-human IL-12 / IL-12 antibody. specifically binds to the human IL-12 / IL-23p40 protein domain as part of an IL-23p40 protein domain antibody, said antibody further comprising the sequence set forth in SEQ ID NO: 1 or 24, an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to said sequence, or having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence Regarding.

[0062] In particular, the polynucleotide comprises or consists of the nucleotide sequence set forth in SEQ ID NO: 2 or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto.

[0063] In particular, the polynucleotide comprises or consists of the nucleotide sequence set forth in SEQ ID NO: 7 or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto.

[0064] In particular, the polynucleotide comprises or consists of the nucleotide sequence set forth in SEQ ID NO: 12, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto.

[0065] In particular, the polynucleotide comprises or consists of the nucleotide sequence set forth in SEQ ID NO: 14, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto.

[0066] In particular, the polynucleotide comprises or consists of the nucleotide sequence set forth in SEQ ID NO: 16, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto.

[0067] In particular, the polynucleotide comprises or consists of the nucleotide sequence set forth in SEQ ID NO: 18, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto.

[0068] In particular, the polynucleotide comprises or consists of the nucleotide sequence set forth in SEQ ID NO: 37, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto.

[0069] In particular, the polynucleotide comprises or consists of the nucleotide sequence set forth in SEQ ID NO: 38, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto.

[0070] Yet another aspect of the present invention relates to a vector comprising any one of the polynucleotide molecules disclosed herein as described above.

[0071] Yet another aspect of the present invention relates to a host cell comprising any one of the polynucleotide molecules disclosed herein or the vectors disclosed herein, as described above.

[0072] Yet another aspect of the present invention relates to a method of preparing any one of the antibodies or antigen-binding fragments thereof disclosed herein as described above, culturing under suitable conditions the host cells disclosed herein, and isolating the antibody or antigen-binding fragment thereof from the cell culture.

[0073] One aspect of the present invention further provides an antibody conjugate comprising the anti-human IL-12 / IL-23p40 protein domain antibody or antigen-binding fragment thereof and a conjugate moiety attached thereto, wherein the conjugate moiety is a purification tag (e.g., His tag), a cytotoxic agent, or a detectable label. Preferably, the conjugate moiety is a radioisotope, a luminescent substance, a colored substance, an enzyme, or polyethylene glycol.

[0074] One aspect of the present invention further provides a fusion protein comprising any one of the anti-human IL-12 / IL-23p40 protein domain antibodies or antigen-binding fragments thereof as described above.

[0075] One aspect of the present invention provides a multispecific antibody, preferably a bispecific antibody, comprising an antibody or antigen-binding fragment thereof described herein.

[0076] One aspect of the present invention further provides a kit comprising any one of the antibodies or antigen-binding fragments thereof disclosed herein, or an antibody conjugate, fusion protein, or multispecific antibody disclosed herein, as described above.

[0077] Preferably, the kit further comprises a second antibody that specifically identifies the antibody or antigen-binding fragment thereof; optionally, the second antibody further comprises a detectable label such as a radioisotope, a luminescent substance, a colored substance, an enzyme, or polyethylene glycol.

[0078] Yet another aspect of the present invention relates to the use of any one of the antibodies or antigen-binding fragments thereof, antibody conjugates, fusion proteins or multispecific antibodies disclosed herein as described above in the preparation of a kit for detecting the presence or level of human IL-12 / IL-23p40 in a sample.

[0079] Yet another aspect of the present invention relates to a pharmaceutical composition comprising any one of the antibodies or antigen-binding fragments thereof, said antibody conjugates, said multispecific antibodies or said fusion proteins disclosed herein as described above, and optionally a pharmaceutically acceptable carrier and / or excipient.

[0080] Yet another aspect of the present invention is a method for producing a semiconductor device comprising: an agent that blocks the binding of human IL-12 / IL-23p40 to human IL-12Rβ1 or human IL-23R; an agent for blocking the activity or downregulating the levels of human IL-12 / IL-23p40, or an agent for blocking a cellular response mediated by binding of human IL-12Rβ1 or human IL-23R to p40; (Preferably, the ligand for human IL-12 / IL-23p40 is human IL-12Rβ1 or human IL-23R) The present invention relates to the use of any one of the antibodies or antigen-binding fragments thereof, or the antibody conjugates, the multispecific antibodies, the fusion proteins, or the pharmaceutical compositions as described above in the preparation of a therapeutic agent for the treatment of a disease.

[0081] One aspect of the present invention relates to the use of any one of the antibodies or antigen-binding fragments thereof, the antibody conjugates, the multispecific antibodies, the fusion proteins, or the pharmaceutical compositions as described above in the preparation of a medicament for treating a disease selected from the group consisting of an autoimmune disease (e.g., psoriasis vulgaris or systemic lupus erythematosus) and ulcerative colitis (e.g., refractory or recurrent).

[0082] Yet another aspect of the present invention relates to an in vivo or in vitro method comprising administering to a subject in need thereof a cell comprising an antibody or antigen-binding fragment thereof, antibody conjugate, multispecific antibody, fusion protein or pharmaceutical composition as described herein, or administering to a subject in need thereof an effective amount of any one of the antibodies or antigen-binding fragments thereof, antibody conjugate, multispecific antibody, fusion protein or pharmaceutical composition as disclosed herein as above, said method comprising: A method for blocking the binding of human IL-12 / IL-23p40 to its ligand IL-12Rβ1 or IL-23R; Methods for downregulating the activity or levels of human IL-12 / IL-23p40, and A method for blocking a cellular response mediated by binding of human IL-12Rβ1 or human IL-23R to p40; (Preferably, the ligand for IL-12 / IL-23p40 is IL-12Rβ1 or IL-23R) is selected from the group consisting of:

[0083] In one embodiment of the invention, the in vitro method is non-therapeutic and / or non-diagnostic.

[0084] Yet another aspect of the present invention relates to the use of any one of the antibodies or antigen-binding fragments thereof, antibody conjugates, multispecific antibodies, fusion proteins or pharmaceutical compositions disclosed herein as described above in the preparation of a medicament for the prevention, treatment, adjuvant treatment and / or diagnosis of an autoimmune disease (e.g., psoriasis vulgaris or systemic lupus erythematosus) or ulcerative colitis (e.g., refractory or recurrent), or their use for the prevention, treatment, adjuvant treatment and / or diagnosis of an autoimmune disease (e.g., psoriasis vulgaris or systemic lupus erythematosus) or ulcerative colitis (e.g., refractory or recurrent).

[0085] In one embodiment of the invention, the medicament is in a form suitable for administration by subcutaneous, intradermal, intravenous, intramuscular or intralesional injection.

[0086] Yet another aspect of the present invention relates to a method for the prevention, treatment, adjuvant treatment and / or diagnosis of an autoimmune disease (e.g., psoriasis vulgaris or systemic lupus erythematosus) or ulcerative colitis (e.g., refractory or recurrent) comprising administering to a subject in need thereof any one of the antibodies or antigen-binding fragments thereof, antibody conjugates, multispecific antibodies, fusion proteins or pharmaceutical compositions disclosed herein as described above.

[0087] The present invention also provides a method of treating a patient suffering from ulcerative colitis, comprising: (i) measuring the level of human IL-12 / IL-23p40 in a sample from the patient, wherein the patient is positive for human IL-12 / IL-23p40; and (ii) administering to the patient a therapeutically effective amount of an anti-human IL-12 / IL-23p40 antibody, or antigen-binding portion thereof.

[0088] The ulcerative colitis described herein can be refractory and recurrent. For example, in some patients, the ulcerative colitis is recurrent; in some patients, the ulcerative colitis is refractory.

[0089] In some aspects of the invention, the patient is undergoing conventional treatment or is inadequately responsive, non-responsive, or intolerant to a biologic agent, hi some specific embodiments, the patient's undergoing conventional treatment or inadequately responsive, non-responsive, or intolerant to a biologic agent results in a failure to achieve a complete or partial response.

[0090] As used herein, the H8L15H1L1 is an anti-human IL-12 / IL-23p40 monoclonal antibody, and its sequence and structure can be found in Patent CN201910706137.1. In the H8L15H1L1 monoclonal antibody, the HCDR1 comprises the sequence GYTFTSYW (SEQ ID NO: 3), the HCDR2 comprises the sequence MSPVDSDI (SEQ ID NO: 4), the HCDR3 comprises the sequence ARRRPGQGYFDF (SEQ ID NO: 5), the LCDR1 comprises the sequence QSVGTW (SEQ ID NO: 6), the LCDR2 comprises the sequence AAS (SEQ ID NO: 7), and the LCDR3 comprises the sequence QQYNIYPYT (SEQ ID NO: 8).

[0091] In the present invention, unless otherwise specified, scientific and technical terms used herein have the meanings commonly understood by those skilled in the art. Furthermore, the experimental procedures of cell culture, molecular genetics, nucleic acid chemistry, and immunology used in the present invention are routine procedures widely used in the corresponding fields. In order to better understand the present invention, definitions and explanations of relevant terms are provided below.

[0092] As used herein, the term "antigen-binding region" refers to a protein or a portion of a protein that specifically binds to a given antigen. For example, a portion of an antibody that contains amino acid residues that interact with an antigen and confer specificity and affinity to the antibody for the antigen is called the "antigen-binding region." The antigen-binding region generally contains one or more complementarity-determining regions (CDRs). Some antigen-binding regions further contain one or more "framework" regions (FRs). CDRs are amino acid sequences that contribute to antigen-binding specificity and affinity.

[0093] As used herein, the term "antibody" refers to an intact immunoglobulin of any isotype or antigen-binding fragment thereof that can compete with the intact antibody for specific binding to a target antigen, and includes, for example, chimeric, humanized, fully human, and bispecific antibodies or antigen-binding fragments thereof. Such "antibodies" are antigen-binding proteins.

[0094] An intact antibody generally comprises at least two full-length heavy chains and two full-length light chains, but may optionally comprise fewer chains, such as antibodies naturally occurring in camelids, which may comprise only heavy chains. An antibody or antigen-binding fragment thereof may be derived from only a single source or may be "chimeric," i.e., different portions of the antibody may be derived from two different sources, as described further below. Antibodies or antigen-binding fragments thereof may be produced in hybridomas by recombinant DNA techniques or by enzymatic or chemical cleavage of intact antibodies. Unless otherwise specified, the term "antibody" includes antibodies comprising two full-length heavy chains and two full-length light chains, as well as derivatives, variants, and fragments thereof.

[0095] As used herein, the term "antigen-binding fragment" (or abbreviated "fragment") of an "antibody" or "immunoglobulin chain" (heavy or light chain) includes a portion of an antibody (whether obtained or synthetic) that lacks at least some of the amino acid residues present in the full-length antibody, but is capable of specifically binding to its antigen. Such fragments are biologically active because they specifically bind to the target antigen and can compete with other antibodies or antigen-binding fragments thereof for specific binding to a given epitope. In one aspect, such a fragment retains at least one CDR present in a full-length light or heavy chain of the antibody, and in some embodiments, such a fragment comprises a single heavy and / or light chain or portion thereof. Such biologically active fragments can be produced by recombinant DNA techniques or by, for example, enzymatic or chemical cleavage of intact antibodies.

[0096] Immunologically functional immunoglobulin fragments include, but are not limited to, Fab, Fab', F(ab')2, Fv, Fd, dAb, Fab / c, complementarity-determining region (CDR) fragments, single-chain antibodies (e.g., scFv), bivalent antibodies, and domain antibodies, and can be derived from any mammal, including, but not limited to, human, mouse, rat, camelid, or rabbit. It is further contemplated that functional portions of the antibodies disclosed herein, such as one or more CDRs, can be covalently linked to a second protein or small molecule to generate therapeutic agents directed to specific targets in the body, thereby having bifunctional therapeutic properties or extended serum half-life, such as fusion proteins.

[0097] As used herein, the terms "full antibody chain," "full length antibody," "intact antibody," and "whole antibody" are used interchangeably herein and refer to an antibody having a structure substantially similar to a native antibody structure as defined herein, or an antibody having a heavy chain in the Fc region.

[0098] The term "light chain" includes full-length light chains and fragments thereof containing sufficient variable region sequence to confer binding specificity. L and constant region domain C L The light chain variable region domain is at the amino terminus of the polypeptide. The light chain includes kappa chains and lambda chains.

[0099] The term "heavy chain" includes full-length heavy chains and fragments thereof containing sufficient variable region sequence to confer binding specificity. H and three constant region domains C H1 , C H2 and C H3 The V H The domain is at the amino terminus of the polypeptide and H The C domain is at its carboxyl terminus. H3 is closest to the carboxyl terminus of the polypeptide. The heavy chain may be of any isotype, including IgG (including subtypes IgG1, IgG2, IgG3, and IgG4), IgA (including subtypes IgA1 and IgA2), IgM, and IgE.

[0100] As used herein, the term "Fab fragment" refers to a fragment containing one light chain, C H1 and one heavy chain variable region. The heavy chain of a Fab molecule cannot form disulfide bonds with another heavy chain molecule.

[0101] As used herein, the term "Fc" region refers to the C region of an antibody. H1 Domain and the C H2 The two heavy chain fragments comprise two heavy chain fragments containing the C domain, the two heavy chain fragments being connected by two or more disulfide bonds and the C H3 The domains are held together by hydrophobic interactions.

[0102] As used herein, the term "Fab' fragment" refers to a fragment that contains one light chain and a portion of one heavy chain (the V H Domain, C H1domain, and the C H1 Domain and C H2 The Fab' fragments contain a portion of the region between the Fab' domains, resulting in the formation of an interchain disulfide bond between the two heavy chains of the two Fab' fragments to give rise to an F(ab')2 molecule.

[0103] As used herein, the term "F(ab')2 fragment" refers to the C H1 Domain and C H2 It contains two light chains and two heavy chains, including portions of the constant regions between the domains, such that interchain disulfide bonds are formed between the two heavy chains. Thus, the F(ab')2 fragment consists of two Fab' fragments held together by disulfide bonds between the two heavy chains.

[0104] As used herein, the term "Fv region" comprises the variable regions from the heavy and light chains, but lacks the constant regions.

[0105] As used herein, the term "Fd" fragment refers to a V H Domain and C H1 It refers to an antibody fragment consisting of domains (Ward et al., Nature, 341:544-546 (1989)).

[0106] As used herein, the term "dAb" fragment refers to a V H It consists of domains (Ward et al., Nature 341:544-546 (1989)).

[0107] As used herein, the term "Fab'-SH" is the designation herein for Fab' in which one or more cysteine residues of the constant domains contain a free thiol group.

[0108] As used herein, the term "Fab / c" fragment refers to an intermediate formed by pepsin digestion of immunoglobulins, which combines the advantages of the Fab and Fc regions, i.e., high diffusibility in vivo and low metabolic clearance, while retaining high affinity (Liu Jianjun, Chinese Journal of Cellular and Molecular Immunology, 1989(4):29-29).

[0109] As used herein, the term "single-chain antibody" refers to an Fv molecule in which the heavy and light chain variable regions are linked by a flexible linker to form a single polypeptide chain (which forms the antigen-binding region) (see, e.g., Bird et al., Science, 242:423-426 (1988), and Huston et al., Proc. Natl. Acad. Sci. USA, 90:5879-5883 (1988)). Single-chain antibodies are described in detail in International Patent Publication No. 88 / 01649 and U.S. Patent Nos. 4,946,778 and 5,260,203, the disclosures of which are incorporated herein by reference.

[0110] As used herein, the term "domain antibody" refers to an immunologically functional immunoglobulin fragment that contains only the variable region of the heavy chain or the light chain. In some cases, two or more V H The domains are covalently linked by peptide linkers to form multivalent domain antibodies (particularly bivalent domain antibodies). H The regions may target the same or different antigens.

[0111] As used herein, the term "bivalent antigen-binding protein" or "bivalent antibody" comprises two antigen-binding sites. In some cases, the two binding sites have the same antigen specificity. The bivalent antibody may be bispecific.

[0112] As used herein, the term "multispecific antigen-binding protein" or "multispecific antibody" is an antigen-binding protein or antibody that targets multiple antigens or epitopes.

[0113] As used herein, the terms "bispecific," "dual-specific," or "bifunctional" antigen-binding protein or antibody refer to a hybrid antigen-binding protein or antibody, each having two different antigen-binding sites. Bispecific antibodies are multispecific antigen-binding proteins or antibodies and can be produced by a variety of methods, including, but not limited to, fusion of hybridomas or linking of Fab' fragments. See, for example, Songsivilai and Lachmann, 1990, Clin. Exp. Immunol. 79:315-321; Kostelny et al., 1992, J. Immunol. 148:1547-1553. The two binding sites of a bispecific antigen-binding protein or antibody bind to two different epitopes present on the same or different protein targets.

[0114] As used herein, the terms "mAb" and "monoclonal antibody" refer to an antibody or antibody fragment derived from a population of highly homologous antibodies, i.e., derived from a population of identical antibody molecules except for naturally occurring mutations that may occur spontaneously. Monoclonal antibodies are highly specific to a single epitope on an antigen. Compared to monoclonal antibodies, polyclonal antibodies generally contain at least two or more different antibodies that typically identify different epitopes on an antigen. Monoclonal antibodies can generally be obtained by the hybridoma technique first described by Kohler et al. (Nature, 256:495, 1975), and can also be obtained by recombinant DNA technology (see, e.g., U.S. Pat. No. 4,816,567).

[0115] As used herein, the term "humanized antibody" refers to an antibody or antibody fragment obtained when all or part of the CDR regions of a human immunoglobulin (receptor antibody) are replaced by the CDR regions of a non-human antibody (donor antibody), where the donor antibody may be a non-human (e.g., mouse, rat, or rabbit) antibody having the desired specificity, affinity, or reactivity. Furthermore, some amino acid residues in the framework regions (FRs) of the receptor antibody can also be replaced by corresponding amino acid residues of a non-human antibody or by amino acid residues of other antibodies to further improve or optimize the performance of the antibody. For further details on humanized antibodies, see, e.g., Jones et al., Nature, 321:522-525 (1986); Reichmann et al., Nature, 332:323-329 (1988); Presta, Curr. Op. Struct. Biol., 2:593-596 (1992); and Clark, Immunol. Today 21:397-402 (2000).

[0116] As used herein, the term "epitope" refers to a site on an antigen to which an immunoglobulin or antibody specifically binds. In this field, "epitope" is also referred to as "antigenic determinant." The epitope or antigenic determinant generally consists of a chemically active surface grouping of molecules such as amino acids, carbohydrates, or sugar side chains, and usually has specific three-dimensional structural and charge characteristics. For example, the epitope generally includes at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 consecutive or non-consecutive amino acids in a unique spatial conformation, which can be "linear" or "conformational." See, for example, Epitope Mapping Protocols in Methods in Molecular Biology, Vol. 66, GE Morris, Ed. (1996). In a linear epitope, all interaction sites between a protein and an interacting molecule (e.g., an antibody) are located linearly along the primary amino acid sequence of the protein, whereas in a conformational epitope, the interaction sites are located across amino acid residues of the protein that are separated from each other.

[0117] The terms "polypeptide" or "protein" as used herein are used to refer to a polymer of amino acid residues.

[0118] The terms are also used to refer to amino acid polymers in which one or more amino acid residues are analogs or mimetics of naturally occurring amino acids, and to naturally occurring amino acid polymers. The terms can also include amino acid polymers that have been modified, for example, by the addition of sugar residues to form glycoproteins, or phosphorylated amino acid polymers. Polypeptides and proteins can be produced by naturally occurring cells and non-recombinant cells, or they can be produced by genetically engineered or recombinant cells, and include molecules with the amino acid sequence of a naturally occurring protein or molecules with deletions, insertions, and / or substitutions of one or more amino acids of the naturally occurring sequence.

[0119] Specifically, the terms "polypeptide" and "protein" include antibodies, such as anti-human p40 antibodies (also called p40 antibodies), p40 binding proteins, and antigen-binding proteins or sequences having deletions, insertions, and / or substitutions in one or more amino acids.

[0120] The term "polypeptide fragment" refers to a polypeptide that has an amino-terminal deletion, a carboxyl-terminal deletion, and / or an internal deletion compared to the full-length protein. Such fragments may also contain modified amino acids compared to the full-length protein. In certain embodiments, such fragments are about 5 to 500 amino acids in length. For example, fragments can be at least 5, 6, 8, 10, 14, 20, 50, 70, 100, 110, 150, 200, 250, 300, 350, 400, or 450 amino acids in length. Useful polypeptide fragments include immunologically functional fragments of antibodies that contain a binding domain. In the case of human p40 antibodies, useful fragments include, but are not limited to, CDR regions, heavy or light chain variable domains, portions of antibody chains, and variable domains that contain exactly two CDRs.

[0121] A "derivative" of a polypeptide is a polypeptide (e.g., an antigen-binding protein or antibody) that is chemically modified in a way other than by insertion, deletion, or substitution, e.g., by conjugation with another chemical moiety (e.g., a PEG-conjugated polypeptide).

[0122] As used herein, the term "isolated" refers to something obtained by artificial means from a natural state. When a particular "isolated" substance or component exists in nature, it may be altered in its natural environment, separated from its natural environment, or both. For example, a particular non-isolated polynucleotide or polypeptide naturally exists in a particular living animal, and the same polynucleotide or polypeptide, isolated in high purity from such a natural state, is referred to as an isolated polynucleotide or polypeptide. The term "isolated" does not exclude the presence of artificial or synthetic substances or other impurities that do not affect the activity of the substance.

[0123] As used herein, the term "vector" refers to a nucleic acid vehicle into which a polynucleotide can be inserted. If the vector allows for the expression of a protein encoded by the inserted polynucleotide, the vector is called an expression vector. A vector can be introduced into a host cell by transformation, transduction, or transfection, so that the genetic material elements carried by the vector can be expressed in the host cell. Vectors include, but are not limited to, plasmids; phagemids; cosmids; artificial chromosomes such as yeast artificial chromosomes (YACs), bacterial artificial chromosomes (BACs), or P1-derived artificial chromosomes (PACs); phages such as lambda phage or M13 phage; and animal viruses, which are well known to those skilled in the art. Animal viruses that can be used as vectors include, but are not limited to, retroviruses (including lentiviruses), adenoviruses, adeno-associated viruses, herpes viruses (such as herpes simplex viruses), poxviruses, baculoviruses, papilloma viruses, and papovaviruses (such as SV40). Vectors may contain various elements that control expression, including, but not limited to, promoter sequences, transcription initiation sequences, enhancer sequences, selection elements, and reporter genes. Furthermore, vectors may further contain a replication origin.

[0124] As used herein, the term "host cell" refers to a cell that can be introduced with a vector, including, but not limited to, prokaryotic cells such as E. coli or Bacillus subtilis, fungal cells such as yeast cells or Aspergillus, insect cells such as S2 Drosophila cells or Sf9, or animal cells such as fibroblasts, CHO cells, COS cells, NSO cells, HeLa cells, BHK cells, HEK293 cells or human cells.

[0125] As used herein, the term "specifically binds" refers to a non-random binding reaction between two molecules, such as the reaction between an antibody and its target antigen. In some embodiments, an antibody that specifically binds to an antigen (or is specific for an antigen) has a binding affinity of about 10 -5 Less than M, e.g., about 10 -6 M, 10 -7 M, 10 -8 M, 10 -9 M, less than or equal to 10 -10 Affinity (K D ) refers to an antibody that binds to an antigen.

[0126] As used herein, the term "K D " refers to the dissociation equilibrium constant for a specific antibody-antigen interaction, which is used to describe the binding affinity between an antibody and an antigen. Among several parameters measured by molecular binding kinetics, K DThe KD value is the dissociation equilibrium constant. In antibody drug research, the KD value is a parameter that characterizes the strength of the affinity effect between a target antibody and a target antigen molecule, and is expressed by the formula: K D =k dis / k on A smaller equilibrium dissociation constant indicates stronger antibody-antigen binding and higher affinity between the antibody and the antigen. k on (association rate constant) is the rate of formation of the antigen-antibody complex, and is the smaller of k on indicates a faster binding rate of the antibody to the antigen. dis (dissociation rate constant) is the rate at which an antibody dissociates from an antigen-antibody complex, and is the smaller of k dis This suggests that the antibody dissociates from the antigen more slowly and that the binding between the antibody and the antigen is stronger. Generally, antibodies have a binding affinity of about 10, as measured, for example, by a Fortebio molecular interaction analyzer using biolayer interferometry (BLI) technology. -5 Less than m, e.g., about 10 -6 M, 10 -7 M, 10 -8 M, 10 -9 Less than M or 10 -10 The dissociation equilibrium constant (K D ) and binds to an antigen (e.g., L1 protein).

[0127] As used herein, the term "EC 50 " refers to the effective concentration that is 50% of the maximal antibody response.

[0128] As used herein, the terms "monoclonal antibody" and "McAb" have the same meaning and are used interchangeably; the terms "polyclonal antibody" and "PcAb" have the same meaning and are used interchangeably. Additionally, amino acids are generally represented herein by one-letter and three-letter abbreviations known in the art. For example, alanine can be represented by A or Ala.

[0129] As used herein, the terms "percent sequence identity" and "percent sequence homology" are used interchangeably.

[0130] As used herein, the terms "similarity," "sequence similarity," and "identity" refer to the correlation between the sequences of two or more protein or polypeptide molecules, as determined by aligning and comparing the sequences. "Percent identity" refers to the percentage of identical amino acid residues in the compared molecules and can be calculated based on the size of the smallest molecule being compared. For such calculations, gaps in the alignment, if any, must be addressed by a particular mathematical model or computer program (i.e., "algorithm").

[0131] When used in reference to polypeptides, the term "substantial identity" means that two peptide sequences have at least 70%, 75% or 80% sequence identity, at least 90% or 95% sequence identity, or at least 97%, 98% or 99% sequence identity when optimally aligned using, for example, the programs GAP or BESTFIT using the default gap weights provided by the programs.

[0132] In some cases, residue positions that are not identical differ by conservative amino acid substitutions. A "conservative amino acid substitution" is one in which an amino acid residue is substituted with another amino acid residue having a side chain R group that possesses similar chemical properties (e.g., charge, hydrophilicity, or hydrophobicity). Generally, conservative amino acid substitutions substantially preserve the function and properties of the protein.

[0133] When two or more amino acid sequences differ from each other by conservative substitutions, the percent sequence identity may be increased to correct for the conservative nature of the substitutions. Methods for making this adjustment are well known to those of skill in the art. See, e.g., Pearson, Methods Mol. Biol., 243:307-31 (1994).

[0134] Examples of groups of amino acids with side chains possessing similar chemical properties include: 1) aliphatic hydroxyl side chains: glycine, alanine, valine, leucine, and isoleucine, 2) aliphatic hydroxyl side chains: serine and threonine, 3) amide-containing side chains: asparagine and glutamine, 4) aromatic side chains: phenylalanine, tyrosine, and tryptophan, 5) basic side chains: lysine, arginine, and histidine, 6) acidic side chains: aspartic acid and glutamic acid, and 7) sulfur-containing side chains: cysteine and methionine. For example, conservative amino acid substitution groups are valine-leucine-isoleucine-alanine-glycine, phenylalanine-tyrosine, lysine-arginine, threonine-serine, glutamic acid-aspartic acid, and asparagine-glutamine.

[0135] Optionally, a conservative substitution is any change that has a positive value in the PAM250 log-likelihood matrix disclosed in Gonnet et al., Science, 256:1443-45 (1992), which is incorporated herein by reference. A "reasonably conservative" substitution is any change that has a non-negative value in the PAM250 log-likelihood matrix.

[0136] Polypeptide sequence identity is typically measured by sequence analysis software, which matches sequences using measures of similarity assigned to different substitutions, deletions, and other modifications (including conservative amino acid substitutions).

[0137] For example, GCG, including programs such as "Gap" and "Bestfit" (using default parameters specified by the program), can be used to determine sequence homology or sequence identity between closely related polypeptides (e.g., homologous polypeptides from different species) or between a wild-type protein and its mutant protein. See, e.g., GCG Version 6.1 (University of Wisconsin, WI). Polypeptide sequences can also be compared using FASTA with default or recommended parameters. See GCG Version 6.10 FASTA (e.g., FASTA2 and FASTA3), which provides alignments for the regions of optimal overlap between the challenge and query sequences (Pearson, Methods Enzymol. 183:63-98 (1990); Pearson, Methods Mol. Biol., 132:185-219 (2000)).

[0138] Another preferred algorithm for use in comparing sequences to databases containing large sequences from different organisms is the computer program BLAST, particularly blasp or blasn (using the default parameters provided by the program). See, e.g., Altschul et al., Mol. Biol., 215:403-410 (1990); Altschul et al., Nucleic Acids Res., 25:3389-402 (1997).

[0139] The term "treatment" or "treating" generally refers to an operation to obtain a desired pharmacological and / or physiological effect. The effect may be prophylactic, in that a disease or its symptoms are completely or partially prevented; and / or the effect may be therapeutic, in that a disease and / or its side effects are partially or completely stabilized or cured.

[0140] As used herein, "treatment" or "treating" encompasses any treatment for a disease in a patient, including (a) preventing a disease or a symptom of a disease from occurring in a patient who may be susceptible to, but has not yet been diagnosed as having, the disease; (b) suppressing the symptoms of the disease, i.e., arresting its progression; or (c) alleviating the symptoms of the disease, i.e., causing regression of the disease or symptom.

[0141] As used herein, the term "general treatment" refers to a treatment in which a drug substance is transported through the bloodstream to reach and affect cells throughout the body.

[0142] As used herein, the term "systemic chemotherapy" refers to general chemotherapy excluding chemotherapy for locally advanced disease as part of a multimodal treatment, where said chemotherapy for locally advanced disease includes induction chemotherapy, chemotherapy with concurrent radiation therapy, and adjuvant chemotherapy.

[0143] As used herein, the term "subject" (sometimes referred to herein as "patient") refers to mammals such as rodents, cats, dogs, and primates. Preferably, the subject of the present invention is a human.

[0144] "Administer," "administration," or "administering" refers to the physical introduction of a composition containing a therapeutic agent into a subject using any of a variety of methods and delivery systems known to those skilled in the art. Routes of administration of inhibitors of autoimmune disease-associated factors (e.g., anti-human IL-12 / IL-23p40 antibodies) include intravenous, intramuscular, subcutaneous, intraperitoneal, spinal, or other parenteral routes of administration, for example, by injection or infusion. As used herein, the phrase "parenteral administration" refers to modes of administration other than enteral and topical administration, typically by injection, and includes, but is not limited to, intravenous, intramuscular, intraarterial, intrathecal, intralymphatic, intralesional, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intra-articular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injection and infusion, and in vivo electroporation.

[0145] In certain embodiments, the inhibitor of an autoimmune disease-associated factor (e.g., an anti-human IL-12 / IL-23p40 antibody) is administered parenterally, and in some embodiments, the inhibitor is administered orally. Other parenteral routes include topical, epidermal, or mucosal routes of administration, such as intranasal, vaginal, rectal, sublingual, or topical administration.

[0146] Administration may be carried out, for example, once, multiple times, and / or over one or more extended periods of time.

[0147] A "subject" includes either a human or a non-human animal. The term "non-human animal" includes, but is not limited to, vertebrates such as non-human primates, sheep, dogs, and rodents such as mice, rats, and guinea pigs. In certain embodiments, the subject is a human. The terms "subject" and "patient" are used interchangeably herein.

[0148] The terms "about," "approximately," or "substantially comprising" refer to a value or composition that is within an acceptable error range for a particular value or composition as determined by one of ordinary skill in the art, which will depend in part on how the value or composition is measured or determined, i.e., the limitations of the measurement system. For example, "about," "approximately," or "substantially comprising" may refer to being within one or more standard deviations as practiced in the art. Alternatively, "about" or "substantially comprising" may refer to a range that differs by up to 10% or 20% (i.e., ±10% or ±20%) from the parameter or value modified thereby. For example, about 3 mg may include any number between 2.7 mg and 3.3 mg (for 10%) or between 2.4 mg and 3.6 mg (for 20%).

[0149] Mode of administration The following is not intended to limit the mode of administration of the agents disclosed herein.

[0150] In one embodiment, the agents disclosed herein can be formulated into pharmaceutical compositions suitable for single or multiple administration.

[0151] The agents disclosed herein may be administered by a variety of suitable routes, including, but not limited to, oral or parenteral administration (by intravenous, intramuscular, topical or subcutaneous routes).

[0152] In some embodiments, the agents disclosed herein are administered orally or by injection, for example, intravenously or intraperitoneally.

[0153] Suitable dosage forms of the agents disclosed herein include, but are not limited to, tablets, troches, pills, capsules (e.g., hard capsules, soft capsules, enteric-coated capsules and microcapsules), elixirs, granules, syrups, injections (intramuscular, intravenous, intraperitoneal), granules, emulsions, suspensions, solutions, powders, and sustained release formulations for oral or parenteral administration.

[0154] The medicaments disclosed herein include pharmaceutically acceptable carriers and / or excipients.

[0155] Beneficial effects of the present invention: The humanized antibody against the human IL-12 / IL-23p40 protein domain can specifically bind to the IL-12 / IL-23p40 protein domain, effectively block the binding of the IL-12 / IL-23p40 protein domain to the cell surface receptors IL-12Rβ1 and IL-23R, inhibit the activation of the downstream signal pathway of the human IL-12 / IL-23p40 protein domain, and inhibit IL-23-induced IL-17A secretion.

[0156] The binding activity of the antibodies disclosed herein to human p40 is significantly better than that of the control antibodies ustekinumab and Ab123FR1.

[0157] The antibodies disclosed herein have the potential to be used to prepare drugs for preventing and treating autoimmune diseases (e.g., psoriasis vulgaris or systemic lupus erythematosus) and ulcerative colitis (e.g., refractory or recurrent), while having good applicability and market value. [Brief explanation of the drawings]

[0158] [Figure 1] Results of detecting the binding activity of H5L9 and Ab123FR1 to the human IL-12 / IL-23p40 protein domain. [Figure 2] Results of detecting the binding activity of H5L10 and Ab123FR1 to the human IL-12 / IL-23p40 protein domain. [Figure 3] Results of detecting the binding activity of H5L11, H5L12, H5L14, and Ab123FR1 to the human IL-12 / IL-23p40 protein domain. [Figure 4] Results of detecting the activity of H8L15 and Ab123FR1 against the human IL-12 / IL-23p40 protein domain. [Figure 5] Results of determining the affinity constant of H5L9 for the human IL-12 / IL-23p40 protein domain. The antibody concentrations corresponding to the top and bottom curve pairs are 5 nM, 2.5 nM, 1.25 nM, 0.75 nM, and 0.31 nM, respectively. [Figure 6] Results of determining the affinity constant of H5L10 for the human IL-12 / IL-23p40 protein domain. The antibody concentrations corresponding to the top and bottom curve pairs are 5 nM, 2.5 nM, 1.25 nM, 0.75 nM, and 0.31 nM, respectively. [Figure 7] Results of determining the affinity constant of H5L11 for the human IL-12 / IL-23p40 protein domain. The antibody concentrations corresponding to the top and bottom curve pairs are 5 nM, 2.5 nM, 1.25 nM, 0.75 nM, and 0.31 nM, respectively. [Figure 8] Results of determining the affinity constant of H5L12 for the human IL-12 / IL-23p40 protein domain. The antibody concentrations corresponding to the top and bottom curve pairs are 5 nM, 2.5 nM, 1.25 nM, 0.75 nM, and 0.31 nM, respectively. [Figure 9] Results of determining the affinity constant of H5L14 for the human IL-12 / IL-23p40 protein domain. The antibody concentrations corresponding to the top and bottom curve pairs are 5 nM, 2.5 nM, 1.25 nM, 0.75 nM, and 0.31 nM, respectively. [Figure 10] Results of determining the affinity constant of Ab123FR1 for the human IL-12 / IL-23p40 protein domain. The antibody concentrations corresponding to the top and bottom curve pairs are 5 nM, 2.5 nM, 1.25 nM, 0.75 nM, and 0.31 nM, respectively. [Figure 11] Results of determining the affinity constant of H8L15 for the human IL-12 / IL-23p40 protein domain. The antibody concentrations corresponding to the top and bottom curve pairs are 5 nM, 2.5 nM, 1.25 nM, 0.75 nM, and 0.31 nM, respectively. [Figure 12]Figure 1 shows the affinity constant of ustekinumab for the human IL-12 / IL-23p40 protein domain. The antibody concentrations corresponding to the top and bottom curve pairs are 5 nM, 2.5 nM, 1.25 nM, 0.75 nM, and 0.31 nM, respectively. [Figure 13] Antibodies H5L9, H5L10, H5L11, H5L12, H5L14 and ustekinumab competitively block the binding of human IL-12 to 293T-IL-12Rβ1 and IL-23R cells (FACS). [Figure 14] Antibodies H5L9, H5L10, H5L11, H5L12, H5L14 and ustekinumab competitively block binding of human IL-23 to 293T-IL-12Rβ1 and IL-23R cells (FACS). [Figure 15] H8L15 significantly inhibits IL-23-induced IL-17A secretion by splenocytes from mice with spontaneous systemic lupus erythematosus. [Figure 16] H8L15 significantly improves skin damage in psoriasis model mice. [Figure 17] Statistical results of pathological scores of ulcerative colitis in each experimental group. H8L15 significantly reduces pathological changes and clinical symptoms in ulcerative colitis model mice. [Figure 18] Changes in the weight of experimental mice. [Figure 19] Demonstration of skin histopathological observations on H8L15 treatment of mice with DSS-induced colitis in combination with recombinant human IL-23. [Figure 20] The antibodies Ab123FR1, H8L15, and ustekinumab competitively block the binding of human IL-12 to 293T-IL-12Rβ1 and IL-23R cells (FACS). [Figure 21] The antibodies Ab123FR1, H8L15, and ustekinumab competitively block the binding of human IL-23 to 293T-IL-12Rβ1 and IL-23R cells (FACS).

[0159] Detailed Description Hereinafter, the embodiments of the present invention will be described in detail with reference to examples. Those skilled in the art will understand that the following examples are used only to illustrate the present invention and should not be considered to limit the scope of the present invention. If techniques or conditions are not specified, the examples were carried out according to the techniques or conditions described in the literature of the art (see, for example, Molecular Cloning: A Laboratory Manual, authored by J. Sambrook et al., and translated by Huang Peitang et al., Third Edition, Science Press) or product manuals. If the manufacturer of the reagents or equipment used is not specified, they are conventional products available on the market.

[0160] In the following examples of the present invention, C5BL / 6 mice were purchased from Guangdong Medical Laboratory Animal Center.

[0161] In the following examples of the present invention, IL-12 (human IL12 (HisTag)) was purchased from Sino Biological (catalog number: CT-050-H08H-20, lot number: LC11MC2805).

[0162] In the following examples of the present invention, the anti-IL-12 / IL-23p40 antibody Ab123FR1 was used as a control antibody, and its preparation method can be referred to Chinese Patent No. CN103275222B, which was produced by Akeso Biopharma, Inc., and its sequence is shown as positions 20-468 of SEQ ID NO: 3 and positions 20-233 of SEQ ID NO: 4 in Chinese Patent No. CN103275222B.

[0163] In the following examples of the present invention, ustekinumab (trade name Stelara), an anti-p40 antibody against the same target, was purchased from Johnson & Johnson as a control antibody.

[0164] In the following examples of the present invention, the cell lines 293T-IL-12Rβ1 and IL-23R used were produced by Akeso Biopharma, Inc. The cell lines 293T-IL-12Rβ1 and IL-23R were produced using a third-generation lentivirus system (e.g., A Third Generation Lentivirus Vector with a Conditional Packaging System, Dull T, Zufferey R, Kelly M, Mandel RJ, Nguyen M, Trono D, and Naldini L., J Virol. 1998). 72(11):8463-8471), where the lentiviral expression vectors used were pLenti-IL-12Rβ1-BSD (IL-12Rβ1, GenBank accession number 3549; vector pLenti-BSD purchased from Invitrogen, catalog number: K497000) and pCDH-IL-23R-puro (IL-23R, GenBank accession number 149233; vector pCDH-CMV-MCS-EF1-Puro purchased from Yubio, product number VT1480).

[0165] The isotype control antibody was human anti-hen egg lysozyme IgG (anti-HEL, i.e., human IgG (abbreviated as hIgG1)), the sequence of which was taken from "Affinity maturation increases the stability and plasticity of the Fv domain of anti-protein antibodies" (Acierno et al., J Mol Biol., 2007, 374(1):130-46, where the amino acid sequence of the heavy chain is set forth in SEQ ID NO: 21 and the amino acid sequence of the light chain is set forth in SEQ ID NO: 22). The isotype control antibody was prepared in the laboratory of Akeso Biopharma.

[0166] The hIL-23 recombinant protein (IL-23, GeneBank accession number 51561) was prepared in the laboratory of Akeso Biopharma, Inc.

[0167] The following examples further illustrate the present invention and are not intended to limit it.

[0168] Example 1. Design, expression, and purification of the heavy and light chain sequences of anti-human p40 antibody H8L15

[0169] 1. Antibody Design To prepare the anti-human IL-12 / IL-23p40 antibody H8L15, the inventors determined the amino acid sequence of the CDR region of the antibody that binds to the antigen using quantum simulation calculations based on the structure of the IL-12 / IL-23p40 protein domain and a structural biology-based three-dimensional spatial structure simulation technique for antigen-antibody binding and the interaction between the CDR region of the antibody and the antigen. Meanwhile, they correspondingly optimized the framework region of the antibody without affecting the three-dimensional structure of the CDR region, ultimately obtaining antibodies H5L9, H5L10, H5L11, H5L12, H5L14, and H8L15 that specifically bind to human IL-12 / IL-23p40.

[0170] The amino acid sequences of the heavy and light chain variable regions of the antibody and the DNA sequences encoding them are as follows: H5L9: The amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO:1, and the DNA sequence encoding it is set forth in SEQ ID NO:2. H5L9: The amino acid sequence of the light chain variable region is set forth in SEQ ID NO:6, and the DNA sequence encoding it is set forth in SEQ ID NO:7. H5L10: The amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO:1, and the DNA sequence encoding it is set forth in SEQ ID NO:2. H5L10: The amino acid sequence of the light chain variable region is set forth in SEQ ID NO:11, and the DNA sequence encoding it is set forth in SEQ ID NO:12. H5L11: The amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO:1, and the DNA sequence encoding it is set forth in SEQ ID NO:2. H5L11: The amino acid sequence of the light chain variable region is set forth in SEQ ID NO:13, and the encoding DNA sequence is set forth in SEQ ID NO:14. H5L12: The amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO:1, and the DNA sequence encoding it is set forth in SEQ ID NO:2. H5L12: The amino acid sequence of the light chain variable region is set forth in SEQ ID NO:15, and the encoding DNA sequence is set forth in SEQ ID NO:16. H5L14: The amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO:1, and the DNA sequence encoding it is set forth in SEQ ID NO:2. H5L14: The amino acid sequence of the light chain variable region is set forth in SEQ ID NO:17, and the DNA sequence encoding it is set forth in SEQ ID NO:18. H8L15: The amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO:24, and the DNA sequence encoding it is set forth in SEQ ID NO:37. H8L15: The amino acid sequence of the light chain variable region is set forth in SEQ ID NO:25, and the DNA sequence encoding it is set forth in SEQ ID NO:38.

[0171] 1. Antibody Expression and Purification The nucleotide sequence encoding the heavy chain variable region (described in SEQ ID NO: 37; constant region: Ig γ-1 chain C region; accession number P01857) and the nucleotide sequence encoding the light chain variable region (described in SEQ ID NO: 38; constant region: Ig κ chain C region; accession number P01834) of the above-mentioned antibody, such as H8L15, were each cloned into the vector pUCsimple (provided by Genscript) to obtain pUC57simple-H8L15H containing the full-length heavy chain nucleotides of H8L15 and pUC57simple-H8L15L containing the full-length light chain nucleotides of H8L15, respectively.

[0172] The plasmids pUC57simple-H8L15H and pUC57simple-H8L15L were digested (HindIII & EcoRI), and the heavy and light chain nucleotide sequences isolated by electrophoresis were subcloned into the vector pcDNA3.1, and the recombinant plasmids were extracted and co-transfected into 293F cells.

[0173] After culturing the transfected 293F cells for 7 days, the medium was centrifuged at high speed, and the resulting supernatant was concentrated and loaded onto a HiTrap MabSelect SuRe column. The protein was eluted in a single step with an eluent to isolate the target sample. The antibody sample was stored in PBS buffer.

[0174] Other antibodies were prepared in the same manner. The antibodies H5L9, H5L10, H5L11, H5L12, H5L14, and H8L15 prepared in this example were used in Examples 2 to 4 below.

[0175] Example 2. Detection of binding activity of antibodies H5L9, H5L10, H5L11, H5L12, H5L14, H8L15, Ab123FR1, and ustekinumab to human IL-12 / IL-23p40 by ELISA

[0176] Microplates were coated with human p40-His (Akeso Biopharma, Inc.; gene p40: GeneBank accession number NM002187). After incubation at 4°C for over 12 hours, the plates were washed with PBST, tapped dry, and blocked with a solution of 1% BSA in PBS. After blocking was complete, the plates were washed with PBST and tapped dry. Gradient dilutions of antibodies in PBST were added to the plate wells; the antibody dilution gradient is detailed in Table 1. The plates containing the test antibodies were incubated at 37°C for 30 minutes, then washed with PBST and tapped dry. A 1:5000 dilution of HRP-labeled goat anti-human IgG (H+L) (purchased from Jackson ImmunoResearch Inc., catalog number: 109-035-088) secondary antibody working solution was added, and the resulting mixture was incubated at 37°C for 30 minutes. After incubation, the plate was washed with PBST and gently tapped dry. TMB (Neogen, 308177) was added to allow color development for 5 minutes in the absence of light, and then a stop solution was added to stop the color reaction. The plate was then immediately placed in a plate reader, and the OD value of each well in the plate was read at 450 nm. The data was analyzed using SoftMax Pro 6.2.1.

[0177] The results of detecting the binding of antibodies H5L9 and Ab123FR1 to the antigen human p40-His are shown in Figure 1. The OD values for all doses are shown in Table 1. The antibody binding EC 50 The results are shown in Table 1.

[0178] The results of detecting the binding of antibodies H5L10 and Ab123FR1 to the antigen human p40-His are shown in Figure 2. The OD values for all doses are shown in Table 2. The antibody binding EC 50 was calculated, and the results are shown in Table 2.

[0179] The results of detecting the binding of antibodies H5L11, H5L12, H5L14, and Ab123FR1 to the antigen human p40-His are shown in Figure 3. The OD values for all doses are shown in Table 3. The binding EC values of the antibodies were calculated by curve fitting with the antibody concentration on the horizontal axis and the absorbance value on the vertical axis. 50 was calculated and the results are shown in Table 3.

[0180] The results of detecting the binding of antibodies H8L15 and Ab123FR1 to the antigen human p40-His are shown in Figure 4. The OD values for all doses are shown in Table 4. The antibody binding EC 50 was calculated and the results are shown in Table 4.

[0181] The results show that the binding efficiencies of H5L9, H5L10, H5L11, H5L12, H5L14 and H8L15 to the antigen human p40-His are dose-dependent.

[0182] As shown in Figure 1 and Table 1, H5L9 binds human p40-His with an EC 50 Ab123FR1 binds to human p40-His with an EC 50 The binding efficiency of H5L9 was comparable to that of Ab123FR1.

[0183] As shown in Figure 2 and Table 2, H5L10 binds human p40-His with an EC 50 Ab123FR1 binds to human p40-His with an EC 50 The binding efficiency of H5L10 was comparable to that of Ab123FR1.

[0184] As shown in Figure 3 and Table 3, H5L11, H5L12, H5L14, and Ab123FR1 had EC of 0.082 nM, 0.082 nM, 0.107 nM, and 1.181 nM, respectively. 50 The binding efficiencies of H5L11, H5L12, and H5L14 were clearly better than that of Ab123FR1.

[0185] As shown in Figure 4 and Table 4, H8L15, Ab123FR1, and ustekinumab had EC values of 0.059 nM, 0.074 nM, and 0.077 nM, respectively. 50 In terms of binding efficiency, Ab123FR1 is comparable to ustekinumab, while H8L15 is significantly better than the other two.

[0186] [Table 1]

[0187] [Table 2]

[0188] [Table 3]

[0189] [Table 4]

[0190] Example 3. Determination of affinity constants of H5L9, H5L10, H5L11, H5L12, H5L14, H8L15, Ab123FR1, and ustekinumab for human IL-12 / IL-23p40 antigen by Fortebio

[0191] The sample dilution buffer for H5L9, H5L10, H5L11, H5L12, H5L14, H8L15, Ab123FR1, and ustekinumab was PBS (0.02% Tween-20, 0.1% BSA, pH 7.4). p40-His was immobilized on a HIS1K (manufacturer: Fortebio, catalog number: 18-5120) sensor at a concentration of 1 μg / mL for 40 seconds. The sensor was equilibrated in buffer for 60 seconds, and the immobilized p40-His was allowed to bind to the antibody at concentrations of 5–0.31 nM (two-fold dilutions) for 120 seconds. The protein was then dissociated in buffer for 300 seconds. The sensor was refreshed with 10 mM glycine solution (pH = 1.5). The detection temperature was 37°C, the detection frequency was 0.3 Hz, and the sample plate shaking speed was 500 rpm. The data were analyzed by 1:1 model fitting to obtain affinity constants.

[0192] The results of measuring the affinity constants of the humanized antibodies H5L9, H5L10, H5L11, H5L12, H5L14, Ab123FR1, H8L15, and ustekinumab (as a control antibody) for human p40-His are shown in Table 5, and the detection results are shown in Figures 5, 6, 7, 8, 9, 10, 11, and 12.

[0193] The results show that the affinity constant of H5L9 for human p40-His is 8.49E-10 M, the affinity constant of H5L10 for human p40-His is 1.21E-10 M, the affinity constant of H5L11 for human p40-His is 1.36E-10 M, the affinity constant of H5L12 for human p40-His is 9.05E-11 M, the affinity constant of H5L14 for human p40-His is 6.20E-11 M, the affinity constant of Ab123FR1 for human p40-His is 7.40E-11 M, the affinity constant of H8L15 for human p40-His is 6.09E-11 M, and the affinity constant of ustekinumab for human p40-His is 8.64E-11 M.

[0194] In terms of affinity, they were ranked from strongest to weakest: H8L15, H5L14, Ab123FR1, ustekinumab, H5L12, H5L10, H5L11 and H5L9. H8L15 and H5L14 showed stronger affinity for human p40-His than Ab123FR1 and ustekinumab.

[0195] [Table 5]

[0196] Example 4. Flow cytometric detection of anti-human IL-12 / IL-23p40 antibodies that competitively block the binding of human IL-12 and IL-23 to 293T-IL-12Rβ1 and IL-23R cells

[0197] 1.1. Flow Cytometric Detection of Antibodies H5L9, H5L10, H5L11, H5L12, H5L14, and Ustekinumab, Which Competitively Block the Binding of Human IL-12 to 293T-IL-12Rβ1 and IL-23R Cells 293T-IL-12Rβ1 and IL-23R cells were digested using conventional methods and divided into several samples containing 300,000 cells each. 200 μL of 1% PBSA was added to each sample, and the mixture was centrifuged at 700 × g for 5 minutes. The supernatant was discarded. According to the experimental design, the corresponding diluted antibody (maximum final concentration of 30 μg / mL, 3-fold dilutions, a total of 8 concentrations) and human IL-12 (Sino Biological, catalog number: CT-050-H08H-20) (final concentration of 20 nM) were mixed at a 1:1 ratio to set up a blank control. The antibody and human IL-12 mixture was incubated on ice for 30 minutes, and then 100 μL per sample was added to the cell pellet. The resulting mixture was mixed thoroughly and incubated on ice for 60 minutes. 200 μL of 1% PBSA was added, and the mixture was centrifuged at 700 × g for 5 minutes. The supernatant was discarded, and the sample was then washed twice. Alexa Fluor® 488 anti-His tag antibody (Biolegend, catalog number: 652509) was diluted 1:400 and added to each tube at 100 μL. The mixture was mixed thoroughly and incubated on ice for 40 minutes in the absence of light. 200 μL of 1% PBSA was added, and the mixture was centrifuged at 700 × g for 5 minutes. The supernatant was discarded and then washed twice. 200 μL of 1% PBSA was added per tube, and the cells were resuspended and transferred to flow cytometry tubes for testing. The results of antibodies H5L9, H5L10, H5L11, H5L12, H5L14, and ustekinumab competitively blocking human IL-12 binding to 293T-IL-12Rβ1 and IL-23R cells, as detected by FACS, are shown in Table 6 and Figure 13.

[0198] According to the results shown in Table 6 and Figure 13, H5L9, H5L10, H5L11, H5L12, H5L14, and ustekinumab can all competitively block the binding of IL-12 to IL-12Rβ1 on the cell membrane surface of 293T-IL-12Rβ1 and IL-23R, where the competitive binding activity of H5L9 was not significant, and the EC 50were 0.3312 μg / mL, 0.414 μg / mL, 0.3172 μg / mL, 0.5320 μg / mL, and 0.3770 μg / mL, respectively.

[0199] In terms of potency in competitively blocking IL-12 binding to IL-12Rβ1 on the cell membrane surface of 293T-IL-12Rβ1 and IL-23R, the antibodies were ranked from strongest to weakest: H5L12, H5L10, ustekinumab, H5L11, H5L14, and H5L9.

[0200] The above results indicate that the competitive binding activity of H5L12 and H5L10 to competitively block the binding of IL-12 to IL-12Rβ1 on the cell membrane surface is better than that of ustekinumab.

[0201] [Table 6]

[0202] 1.2. Flow cytometric detection of antibodies Ab123FR1, H8L15, and ustekinumab that competitively block human IL-12 binding to 293T-IL-12Rβ1 and IL-23R cells 293T-IL-12Rβ1 and IL-23R cells were digested using conventional methods and divided into several samples containing 300,000 cells each. 200 μL of 1% PBSA was added to each sample, and the mixture was centrifuged at 1200 rpm for 5 minutes. The supernatant was discarded. According to the experimental design, the corresponding diluted antibody (maximum final concentration 60 μg / mL, 3-fold dilutions, a total of 8 concentrations) and IL12-His (Sino Biological, catalog number: CT-050-H08H-20) (40 nM) were mixed at a 1:1 ratio to set up a blank control. The antibody and IL12 mixture was incubated on ice for 30 minutes, and then 100 μL per sample was added to the cell pellet. The resulting mixture was mixed thoroughly and incubated on ice for 60 minutes. 200 μL of 1% PBSA was added, and the mixture was centrifuged at 1200 rpm for 5 minutes. The supernatant was discarded, followed by two washes. The THE™ His-tag antibody (FITC) (Genscript, catalog number: A01620) was diluted 1:500 and added to each tube at 100 μL. The mixture was mixed thoroughly and incubated on ice for 40 minutes in the absence of light. 200 μL of 1% PBSA was added, and the mixture was centrifuged at 1200 rpm for 5 minutes. The supernatant was discarded and then washed twice. 200 μL of 1% PBSA was added per tube, and the cells were resuspended and transferred to flow cytometry tubes for testing. The results of antibodies Ab123FR1, H8L15, and ustekinumab competitively blocking the binding of human IL-12 to 293T-IL-12Rβ1 and IL-23R cells, as detected by FACS, are shown in Table 7 and Figure 20. According to the results shown in Table 7 and Figure 20, Ab123FR1, H8L15, and ustekinumab were all able to competitively block the binding of IL-12 to IL-12Rβ1 on the cell membrane surface of 293T-IL-12Rβ1 and IL-23R, and the binding EC 50 The values are 1.99 μg / mL, 1.46 μg / mL and 1.58 μg / mL, respectively.

[0203] The above results indicate that the competitive binding activity of H8L15, which competitively blocks the binding of IL-12 to IL-12Rβ1 on the cell membrane surface, is better than that of Ab123FR1 and ustekinumab.

[0204] [Table 7]

[0205] 2.1. Flow Cytometric Detection of Antibodies H5L9, H5L10, H5L11, H5L12, H5L14, and Ustekinumab, Which Competitively Block the Binding of Human IL-23 to 293T-IL-12Rβ1 and IL-23R Cells 293T-IL-12Rβ1 and IL-23R cells were digested using the standard method and divided into several samples containing 300,000 cells each. 200 μL of 1% PBSA was added to each sample, and the mixture was centrifuged at 700 × g for 5 minutes. The supernatant was discarded. According to the experimental design, the corresponding diluted antibody (maximum final concentration 30 μg / mL, 3-fold dilutions, a total of 8 concentrations) and human IL-23 (IL-23-His-Biotin (Akesobio, Lot No. 20161209) (final concentration 2 μg / mL) were mixed at a 1:1 ratio to set up a blank control. The antibody and human IL-23 mixture was incubated on ice for 30 minutes, and then 100 μL per sample was added to the cell pellet. The resulting mixture was thoroughly mixed and incubated on ice for 60 minutes. 200 μL of 1% PBSA was added to each sample. PBSA was added, the mixture was centrifuged at 700 × g for 5 minutes, the supernatant was discarded, and the cells were washed twice. FITC-steptavidin (Biolegend, catalog number: 405202) was diluted 1:500 and added to each tube at 100 μL. The mixture was mixed thoroughly and incubated on ice for 40 minutes in the absence of light. 200 μL of 1% PBSA was added, the mixture was centrifuged at 700 × g for 5 minutes, the supernatant was discarded, and the cells were washed twice. 200 μL of 1% PBSA was added per tube, and the cells were resuspended and transferred to flow cytometry tubes for testing. The results of antibodies H5L9, H5L10, H5L11, H5L12, H5L14, and ustekinumab competitively blocking human IL-23 binding to 293T-IL-12Rβ1 and IL-23R cells, as detected by FACS, are shown in Table 8 and Figure 14.

[0206] According to the results shown in Table 8 and Figure 14, H5L9, H5L10, H5L11, H5L12, H5L14, and ustekinumab can competitively block the binding of IL-23 to the IL-23 receptor complex on the cell membrane surface of 293T-IL-12Rβ1 and IL-23R, and the competitive binding EC 50 The values are 4.252 μg / mL, 0.6995 μg / mL, 0.8643 μg / mL, 0.7748 μg / mL, 0.8806 μg / mL and 1.158 μg / mL, respectively.

[0207] In terms of potency in competitively blocking IL-23 binding to the IL-23 receptor complex on the cell membrane surface of 293T-IL-12Rβ1 and IL-23R, the antibodies were ranked from strongest to weakest: H5L10, H5L12, H5L11, H5L14, ustekinumab, and H5L9.

[0208] The above results indicate that the competitive binding activity of H5L10, H5L12, H5L11, and H5L14 to competitively block the binding of IL-23 to the IL-23 receptor complex on the cell membrane surface of 293T-IL-12Rβ1 and IL-23R is better than that of ustekinumab.

[0209] [Table 8]

[0210] 2.2. Flow Cytometric Detection of Antibodies Ab123FR1, H8L15, and Ustekinumab, Which Competitively Block Human IL-23 Binding to 293T-IL-12Rβ1 and IL-23R Cells 293T-IL-12Rβ1 and IL-23R cells were digested using the standard method and divided into several samples containing 300,000 cells each. 200 μL of 1% PBSA was added to each sample, and the mixture was centrifuged at 1200 rpm for 5 minutes. The supernatant was discarded. According to the experimental design, the corresponding diluted antibody (maximum concentration 60 μg / mL, 3-fold dilutions, total 8 concentrations) and human IL23-His-biotin (Akeso Biopharma, Inc., Lot No.: 20161209) (4 μg / mL) were mixed at a 1:1 ratio to set up a blank control. The antibody and IL-His-biotin mixture was incubated on ice for 30 minutes and then added to the cell pellet at 100 μL per sample. The resulting mixture was mixed thoroughly and incubated on ice for 60 minutes. 200 μL of 1% PBSA was added, the mixture was centrifuged at 1200 rpm for 5 minutes, the supernatant was discarded, and the cells were washed twice. FITC-steptavidin (Biolegend, catalog number: 405202) was diluted 1:500 and added to each tube at 100 μL. The mixture was mixed thoroughly and incubated on ice for 40 minutes in the absence of light. 200 μL of 1% PBSA was added, the mixture was centrifuged at 1200 rpm for 5 minutes, the supernatant was discarded, and the cells were washed twice. 200 μL of 1% PBSA was added per tube, and the cells were resuspended and transferred to flow cytometry tubes for testing. The results of FACS detection of antibodies Ab123FR1, H8L15, and ustekinumab, which competitively block human IL-23 binding to 293T-IL-12Rβ1 and IL-23R cells, are shown in Table 9 and Figure 21.

[0211] According to the results shown in Table 9 and Figure 21, Ab123FR1, H8L15, and ustekinumab can all competitively block the binding of IL-23 to IL-23R on the cell membrane surface of 293T-IL-12Rβ1 and IL-23R. The binding EC of Ab123FR1, H8L15, and ustekinumab 50 The values are 1.41 μg / mL, 0.8942 μg / mL and 1.434 μg / mL, respectively.

[0212] The above results indicate that the competitive binding activity of H8L15, which competitively blocks the binding of IL-23 to IL-23R on the cell membrane surface of 293T-IL-12Rβ1 and IL-23R, is better than that of Ab123FR1 and ustekinumab.

[0213] [Table 9]

[0214] Example 5. H8L15 effectively inhibits IL-17A secretion by splenocytes from mice with spontaneous systemic lupus erythematosus

[0215] A spontaneous systemic lupus erythematosus model (Jeltsch-David H. Autoimmun Rev. 2014;13(9):963-973) mice (MRL / lpr mice purchased from Shanghai SLAC Laboratory Animal Co., Ltd.) were anesthetized with chloral hydrate, disinfected by immersion in 75% ethyl alcohol, and transferred to a biosafety cabinet. The spleens were then dissected and harvested. The spleens were washed in a dish containing 1640 ml of complete medium to remove fat and fascia. The washed mouse spleens were placed on a 70 μm cell strainer and gently crushed with a syringe plunger. The resulting suspension of spleen cells was washed repeatedly with culture medium. The filtrate was collected and centrifuged at 170 × g for 5 minutes, and the supernatant was discarded. 7 mL of red blood cell lysate was added to resuspend the cell pellet. The mixture was mixed gently and thoroughly, left on ice for 8 minutes, and then an equal volume of complete medium was added to stop lysis. The mixture was centrifuged at 170 × g for 5 minutes, and the supernatant was discarded. The cell pellet was centrifuged and washed twice with 1640 complete medium. The cell pellet was then resuspended in 1640 complete medium, counted, and adjusted for cell density and plated in a 96-well plate (1 × 10 6Cells were seeded in 100 μL of IL-17A / 100 μL. According to the experimental design, 50 μL of antibody was pre-incubated with 50 μL of IL-23 (20 ng / mL final concentration) for 1 hour, and then 50 μL of IL-2 (100 U / mL final concentration) was added. The mixture was incubated at 37°C / 5% CO2 in an incubator for 6 days. After 6 days, the cell supernatant was collected by centrifugation, and the concentration of IL-17A in the supernatant was detected by ELISA.

[0216] As shown in Figure 15, IL-23 effectively promotes IL-17A secretion by splenocytes from spontaneous systemic lupus erythematosus model mice. Addition of H8L15 during the action of IL-23 significantly inhibited IL-17A secretion, and the inhibitory activity was significantly dose-dependent, with pharmacodynamic activity significantly superior to that of AB123FR1 (also referred to as Ab123FR1).

[0217] Example 6. H8L5 effectively improves skin damage in psoriasis model mice

[0218] After shaving, C57BL / 6 mice (purchased from the Animal Center of Guangdong Medical Research Institute) were randomly divided into normal, model, positive control, and H8L15 groups, with 10 mice per group. One day before the first injection of recombinant human IL-23, the model group received a subcutaneous injection of an isotype control antibody (i.e., human anti-chicken egg lysozyme), the H8L15-treated group received a corresponding concentration of H8L15, and the normal group received an equal volume of saline subcutaneous injection. One day after administration, the C57BL / 6 mice were anesthetized with an intraperitoneal injection of 3.5% chloral hydrate at a dose of 7.5 mL / kg. The normal group received an intradermal injection of 25 μL saline per mouse, while the remaining mice received an intradermal injection of 10 μg / 25 μL recombinant human IL-23 per mouse. Injections were performed once daily for six consecutive days. Two days after the final intradermal injection of recombinant human IL-23, mice from each group were subjected to cervical dislocation, and small pieces of cervical skin (approximately 0.5 cm x 0.5 cm) were excised and fixed in formalin tissue fixative. Pathological sections of the mouse skin were prepared 24 hours later for six mice from each group. After the pathological sections were prepared, one representative field was selected under a 100x microscope, and six areas within the original image were randomly selected to measure the epidermal thickness of the mice's skin. Data are presented as mean ± standard error, and results were evaluated by one-way analysis of variance after intergroup comparisons processed using GraphPad software. P<0.05 indicates significant differences, and P<0.01 indicates ultra-significant differences.

[0219] The results are shown in Figure 16. Compared with the normal group, the thickness of the skin epidermis of the mice in the model group was obviously increased (P<0.01). After administration, H8L15 was effective in inhibiting acanthosis in psoriasis mice (P<0.01).

[0220] Example 7. Treatment of colitis with anti-IL-12 / IL-23p40 antibodies

[0221] Anti-IL-12 / IL-23p40 antibodies, such as H8L15, were found to be effective in reducing pathological changes and clinical symptoms in a mouse model of ulcerative colitis. A colitis model was established in C57BL / 6 mice by inducing colitis with DSS (dextran sodium sulfate). Experimental mice were divided into a normal group (3 mice) and each of the other groups (6 mice). The following groups were established: a positive control group (DSS group), an isotype control antibody group (anti-HEL), a high-dose H8L15 group (120 mg / kg), and a low-dose H8L15 group (40 mg / kg). The drugs were injected subcutaneously on days 0, 3, and 6. In the normal group, the animal model was established by providing sterile water through a drinking bottle; in the DSS (MP Bio, Cat. No. Q1723) group, a 1% DSS solution (prepared by adding 2.5 g of DSS to 250 mL of sterile water) was provided through a drinking bottle for 9 consecutive days to create the animal model; in the antibody-treated experimental group, a 1% DSS solution (prepared by adding 2.5 g of DSS to 250 mL of sterile water) was provided through a drinking bottle and hIL-23 recombinant protein (200 μL / 100 μg / mouse) was intraperitoneally injected daily (D1-D5, D7-D9).

[0222] The use and welfare of laboratory animals was carried out in accordance with the regulations of the Association for the Assessment and Accreditation of Laboratory Animal Care International (AAALAC). Animal health and mortality were monitored daily, and routine examinations included observation of the effects of the test substance or drug on the animals' daily behavior, including behavioral activity, weight change, and appearance.

[0223] The experimental parameters were used to examine the effects of the drugs on colitis, and specific parameters were based on the pathological score table of mouse colitis shown in Table 10.

[0224] [Table 10]

[0225] [Table 11]

[0226] Regarding the experimental results, Table 10 shows the criteria for pathological scoring of mouse colitis, Table 11 shows the mouse model establishment method and antibody administration scheme for each experimental group, and the pathological scoring results of colitis are shown in Figure 17. A demonstration of skin histopathological observations for the treatment of H8L15 in mice with colitis induced by DSS in combination with recombinant human IL-23 (HE staining × 100) (a: normal group; b: DSS group; c: DSS + hIL-23 + hIgG1 (120 mg / kg) group; d: DSS + hIL-23 + H8L15 (120 mg / kg) group; e: DSS + hIL-23 + H8L15 (40 mg / kg) group) is shown in Figure 19.

[0227] Conclusion: According to the weight of mice during the experiment period shown in Figure 18, the weight of mice in the model group always decreased, while the weight of mice in the treatment group did not decrease, which is obviously different from that of the model group.

[0228] According to the colitis pathology scores shown in Figure 17, the model group showed clear characteristics of colitis, while the high-dose treatment group and the low-dose treatment group showed statistically significant differences from the model group, indicating that antibody H8L15 is effective in treating ulcerative colitis.

[0229] The sequence information is as follows: The amino acid sequences of the heavy chain variable regions of H5L9, H5L10, H5L11, H5L12 and H5L14 are set forth in SEQ ID NO:1. [ka]

[0230] The nucleotide sequences of the heavy chain variable regions of H5L9, H5L10, H5L11, H5L12 and H5L14 are set forth in SEQ ID NO:2. [ka]

[0231] For H5L9, H5L10, H5L11, H5L12 and H5L14, HCDR1 is set forth in SEQ ID NO:3, HCDR2 is set forth in SEQ ID NO:4 and HCDR3 is set forth in SEQ ID NO:5. HCDR1: GYSFTTYW (SEQ ID NO: 3) HCDR2: MSPVDSDI (SEQ ID NO: 4) HCDR3: ARRRPGQGYFDF (SEQ ID NO: 5)

[0232] The amino acid sequence of the light chain variable region of H5L9 is set forth in SEQ ID NO:6. [ka]

[0233] The nucleotide sequence of the light chain variable region of H5L9 is set forth in SEQ ID NO:7. [ka]

[0234] For H5L9, LCDR1 is set forth in SEQ ID NO:8, LCDR2 is set forth in SEQ ID NO:9, and LCDR3 is set forth in SEQ ID NO:10. LCD 1: QNVGSW (SEQ ID NO: 8) LCDR2:ASS (SEQ ID NO: 9) LCDR3: QQYDIYPFT (SEQ ID NO: 10)

[0235] The amino acid sequence of the light chain variable region of H5L10 is set forth in SEQ ID NO:11. [ka]

[0236] The nucleotide sequence of the light chain variable region of H5L10 is set forth in SEQ ID NO:12. [ka]

[0237] The amino acid sequence of the light chain variable region of H5L11 is set forth in SEQ ID NO:13. [ka]

[0238] The nucleotide sequence of the light chain variable region of H5L11 is set forth in SEQ ID NO:14. [ka]

[0239] The amino acid sequence of the light chain variable region of H5L12 is set forth in SEQ ID NO:15. [ka]

[0240] The nucleotide sequence of the light chain variable region of H5L12 is set forth in SEQ ID NO:16. [ka]

[0241] The amino acid sequence of the light chain variable region of H5L14 is set forth in SEQ ID NO:17. [ka]

[0242] The nucleotide sequence of the light chain variable region of H5L14 is set forth in SEQ ID NO:18. [ka]

[0243] The LCDR1 of L10 is set forth in SEQ ID NO:19. QSVGSW The LCDR1 of L11, L12 and L14 is set forth in SEQ ID NO:20. QSVSSW The LCDR2 of L10, L11, L12 and L14 is set forth in SEQ ID NO:21. ASN The LCDR3 of L10, L11 and L12 is set forth in SEQ ID NO:22. QQYNIYPYT The LCDR3 of L14 is set forth in SEQ ID NO:23. QQYNIYPFT

[0244] The amino acid sequence of the heavy chain variable region of H8L15 is set forth in SEQ ID NO:24. [ka]

[0245] The amino acid sequence of the light chain variable region of H8L15 is set forth in SEQ ID NO:25. [ka]

[0246] For H8L15, HCDR1 is set forth in SEQ ID NO:26, HCDR2 is set forth in SEQ ID NO:4, HCDR3 is set forth in SEQ ID NO:5, LCDR1 is set forth in SEQ ID NO:27, LCDR2 is set forth in SEQ ID NO:28, LCDR3 is set forth in SEQ ID NO:22, FR-H1 is set forth in SEQ ID NO:29, FR-H2 is set forth in SEQ ID NO:30, FR-H3 is set forth in SEQ ID NO:31, FR-H4 is set forth in SEQ ID NO:32, FR-L1 is set forth in SEQ ID NO:33, FR-L2 is set forth in SEQ ID NO:34, FR-L3 is set forth in SEQ ID NO:35, and FR-L4 is set forth in SEQ ID NO:36. HCDR1: GYTFTSYW (SEQ ID NO: 26) HCDR2: MSPVDSDI (SEQ ID NO: 4) HCDR3: ARRRPGQGYFDF (SEQ ID NO: 5) LCDR1:QSVGTW (SEQ ID NO: 27) LCDR2: AAS (SEQ ID NO: 28) LCDR3: QQYNIYPYT (SEQ ID NO: 22) FR-H1:EVQLVQSGAEVKKPGESLKISCQSS (SEQ ID NO: 29) FR-H2:IGWVRQMPGQGLEWIGI (SEQ ID NO: 30) FR-H3: RYNPMFRGQVTMSVDKSSSTAYLQWSSLKASDTAMYYC (SEQ ID NO: 31) FR-H4:WGQGTMVTVSS (SEQ ID NO: 32) FR-L1:EIVLTQSPATLSASPGERATISCRAS (SEQ ID NO: 33) FR-L2: VAWYQQKPGQAPRSLIY (SEQ ID NO: 34) FR-L3:NLQSGIPARFSGSGSGTDFTLTISSLEPEDFAVYYC (SEQ ID NO: 35) FR-L4: FGQGTRLEIK (SEQ ID NO: 36)

[0247] The nucleotide sequence of the heavy chain variable region of H8L15 is set forth in SEQ ID NO:37. [ka]

[0248] The nucleotide sequence of the light chain variable region of H8L15 is set forth in SEQ ID NO:38. [ka]

[0249] The heavy chain amino acid sequence of H8L15 is set forth in SEQ ID NO:39. [ka]

[0250] The light chain amino acid sequence of H8L15 is set forth in SEQ ID NO:40. [ka]

[0251] For H5L9, the sequences of FR-H1, FR-H2, FR-H3, and FR-H4 are the same as those of H8L15, the sequence of FR-L1 is set forth in SEQ ID NO: 41, the sequence of FR-L2 is set forth in SEQ ID NO: 42, the sequence of FR-L3 is set forth in SEQ ID NO: 43, and the sequence of FR-L4 is set forth in SEQ ID NO: 44. FR-L1:DIQMTQSPSSLSASVGDRVTITCKAS (SEQ ID NO: 41) FR-L2: LAWYQQKPGKAPKSLIYS (SEQ ID NO: 42) FR-L3: RQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYC (SEQ ID NO: 43) FR-L4: FGQGTKLEIK (SEQ ID NO: 44)

[0252] For H5L10, the sequences of FR-H1, FR-H2, FR-H3, and FR-H4 are the same as those of H8L15, the sequence of FR-L1 is set forth in SEQ ID NO: 33, the sequence of FR-L2 is set forth in SEQ ID NO: 45, the sequence of FR-L3 is set forth in SEQ ID NO: 46, and the sequence of FR-L4 is set forth in SEQ ID NO: 36. FR-L1:EIVLTQSPATLSASPGERATISCRAS (SEQ ID NO: 33) FR-L2: LAWYQQKPGQAPRSLIYA (SEQ ID NO: 45) FR-L3: LQSGIPARFSGSGSGTDFTLTISSLEPEDFAVYYC (SEQ ID NO: 46) FR-L4: FGQGTRLEIK (SEQ ID NO: 36)

[0253] For H5L11, the sequences of FR-H1, FR-H2, FR-H3, and FR-H4 are the same as those of H8L15, the sequence of FR-L1 is set forth in SEQ ID NO: 33, the sequence of FR-L2 is set forth in SEQ ID NO: 47, the sequence of FR-L3 is set forth in SEQ ID NO: 46, and the sequence of FR-L4 is set forth in SEQ ID NO: 36. FR-L1:EIVLTQSPATLSASPGERATISCRAS (SEQ ID NO: 33) FR-L2: LAWYQQKPGQAPRSLIYS (SEQ ID NO: 47) FR-L3: LQSGIPARFSGSGSGTDFTLTISSLEPEDFAVYYC (SEQ ID NO: 46) FR-L4: FGQGTRLEIK (SEQ ID NO: 36)

[0254] For H5L12, the sequences of FR-H1, FR-H2, FR-H3, and FR-H4 are the same as those of H8L15, the sequence of FR-L1 is set forth in SEQ ID NO: 33, the sequence of FR-L2 is set forth in SEQ ID NO: 45, the sequence of FR-L3 is set forth in SEQ ID NO: 48, and the sequence of FR-L4 is set forth in SEQ ID NO: 36. FR-L1:EIVLTQSPATLSASPGERATISCRAS (SEQ ID NO: 33) FR-L2: LAWYQQKPGQAPRSLIYA (SEQ ID NO: 45) FR-L3: RQSGIPARFSGSGSGTDFTLTISSLEPEDFAVYYC (SEQ ID NO: 48) FR-L4: FGQGTRLEIK (SEQ ID NO: 36)

[0255] For H5L14, the sequences of FR-H1, FR-H2, FR-H3, and FR-H4 are the same as those of H8L15, the sequence of FR-L1 is set forth in SEQ ID NO: 33, the sequence of FR-L2 is set forth in SEQ ID NO: 45, the sequence of FR-L3 is set forth in SEQ ID NO: 46, and the sequence of FR-L4 is set forth in SEQ ID NO: 36. FR-L1:EIVLTQSPATLSASPGERATISCRAS (SEQ ID NO: 33) FR-L2: LAWYQQKPGQAPRSLIYA (SEQ ID NO: 45) FR-L3: LQSGIPARFSGSGSGTDFTLTISSLEPEDFAVYYC (SEQ ID NO: 46) FR-L4: FGQGTRLEIK (SEQ ID NO: 36)

[0256] The heavy chain amino acid sequences of H5L9, H5L10, H5L11, H5L12 and H5L14 are set forth in SEQ ID NO:49. [ka]

[0257] The light chain amino acid sequence of H5L9 is set forth in SEQ ID NO:50. [ka]

[0258] The light chain amino acid sequence of H5L10 is set forth in SEQ ID NO:51. [ka]

[0259] The light chain amino acid sequence of H5L11 is set forth in SEQ ID NO:52. [ka]

[0260] The light chain amino acid sequence of H5L12 is set forth in SEQ ID NO:53. [ka]

[0261] The light chain amino acid sequence of H5L14 is set forth in SEQ ID NO:54. [ka]

Claims

1. An antibody or antigen-binding fragment thereof that specifically binds to human IL-12 / IL-23p40, the antibody comprising: (1) HCDR1, HCDR2, and HCDR3 contained in the heavy chain variable region set forth in SEQ ID NO: 1 or SEQ ID NO: 24; and (2) LCDR1, LCDR2, and LCDR3 contained in a light chain variable region set forth in any one of SEQ ID NO: 6, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, and SEQ ID NO: 25 Including, Preferably, said HCDR1 comprises or consists of a sequence as set forth in SEQ ID NO: 3 or 26, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto, or an amino acid sequence having one or more (preferably 1, 2 or 3) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence; said HCDR2 comprises or consists of the sequence set forth in SEQ ID NO: 4, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto, or an amino acid sequence having one or more (preferably 1, 2 or 3) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence; and HCDR3 comprises or consists of the sequence set forth in SEQ ID NO:5, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto, or an amino acid sequence having one or more (preferably 1, 2 or 3) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence; and The antibody further comprises: LCDR1 comprising or consisting of a sequence according to SEQ ID NO: 8, 19, 20 or 27, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, sequence identity thereto, or an amino acid sequence having one or more (preferably 1, 2 or 3) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence, LCDR2 comprising or consisting of an amino acid sequence as set forth in SEQ ID NO: 9, 21 or 28, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, sequence identity thereto, or an amino acid sequence having one or more (preferably 1, 2 or 3) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence, and LCDR3 comprising or consisting of a sequence as set forth in SEQ ID NO: 10, 22 or 23, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, sequence identity thereto, or an amino acid sequence having one or more (preferably 1, 2 or 3) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence. An antibody or an antigen-binding fragment thereof comprising:

2. 2. The antibody or antigen-binding fragment thereof of claim 1, wherein the antibody comprises framework regions (FRs) in the heavy chain variable region and a FR in the light chain variable region, wherein: (1) The FR in the heavy chain variable region includes FR-H1, FR-H2, FR-H3, and FR-H4, wherein the FR-H1 is an amino acid sequence set forth in SEQ ID NO:29, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the sequence set forth in SEQ ID NO:29, or ... No. 29; said FR-H2 comprises or consists of an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said amino acid sequence set forth in SEQ ID NO: 30; said FR-H2 comprises or consists of an amino acid sequence set forth in SEQ ID NO: 31, a sequence which is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% similar to said sequence set forth in SEQ ID NO: 30; said FR-H3 comprises or consists of an amino acid sequence having 1%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to said amino acid sequence set forth in SEQ ID NO: 30, or having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said amino acid sequence set forth in SEQ ID NO: 31; said FR-H3 comprises or consists of an amino acid sequence set forth in SEQ ID NO: 32, an amino acid sequence having at least 80% sequence identity to said sequence set forth in SEQ ID NO: 32 , 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to said amino acid sequence according to SEQ ID NO: 31, or comprising or consisting of an amino acid sequence which has one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said amino acid sequence according to SEQ ID NO: 31;FR-H4 comprises or consists of the amino acid sequence set forth in SEQ ID NO:32, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to said sequence set forth in SEQ ID NO:32, or an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said amino acid sequence set forth in SEQ ID NO:32; (2) The FR in the light chain variable region includes FR-L1, FR-L2, FR-L3 and FR-L4 (wherein the FR-L1 is an amino acid sequence set forth in SEQ ID NO: 33 or 41, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the sequence set forth in SEQ ID NO: 33 or 41, or the amino acid sequence set forth in SEQ ID NO: 33 or 41). FR-L2 comprises or consists of an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence set forth in SEQ ID NO: 34, 42, 45 or 47; FR-L3 comprises or consists of an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, said FR-L3 comprises or consists of an amino acid sequence having 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity, an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said amino acid sequence set forth in SEQ ID NO: 34, 42, 45 or 47; said FR-L3 comprises or consists of an amino acid sequence set forth in SEQ ID NO: 35, 43, 46 or 48, an amino acid sequence having at least one conservative amino acid mutation (preferably substitutions, insertions or deletions) compared to said amino acid sequence set forth in SEQ ID NO: 35, 43, 46 or 48; or consisting of an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity, or having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said amino acid sequence as set forth in SEQ ID NO: 35, 43, 46 or 48;FR-L4 comprises or consists of an amino acid sequence as set forth in SEQ ID NO: 36 or 44, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to said sequence as set forth in SEQ ID NO: 36 or 44, or an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said amino acid sequence as set forth in SEQ ID NO: 36 or 44); An antibody or an antigen-binding fragment thereof.

3. The antibody or antigen-binding fragment thereof according to claim 1 or 2, (1) an amino acid sequence set forth in SEQ ID NO: 1 or 24; a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to said sequence as set forth in SEQ ID NO: 1 or 24, or an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence set forth in SEQ ID NO: 1 or 24; A heavy chain variable region comprising or consisting of: (2) an amino acid sequence set forth in SEQ ID NO: 6, 11, 13, 15, 17, or 25; a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 86% 96%, 97%, 98% or 99% sequence identity to the sequence set forth in SEQ ID NO: 6, 11, 13, 15, 17 or 25, or a light chain variable region comprising or consisting of an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to the amino acid sequence set forth in SEQ ID NO: 6, 11, 13, 15, 17 or 25; An antibody or an antigen-binding fragment thereof comprising:

4. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 3, The antibody further comprises a heavy chain variable region and a light chain variable region, and the constant region is derived from a species other than mouse, e.g., a human antibody, preferably from human IgG, more preferably from IgG1; preferably, the heavy chain constant region is an Igγ-1 chain C region (more preferably, the Igγ-1 chain C region of GenBank Accession No. P01857); and the light chain constant region is an Igκ chain C region (more preferably, the Igκ chain C region of GenBank Accession No. P01834). An antibody or an antigen-binding fragment thereof.

5. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 4, wherein the antibody comprises or consists of a heavy chain set forth in SEQ ID NO: 19 and a light chain set forth in SEQ ID NO:

20.

6. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 5, wherein the antigen-binding fragment is selected from the group consisting of Fab, Fab', F(ab') ２ , Fd, Fv, dAb, Fab / c, complementarity determining region (CDR) fragment, single chain antibody (e.g.: scFv), bivalent antibody and domain antibody.

7. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 6, wherein the antibody is a humanized antibody, a chimeric antibody or a multispecific antibody (e.g., a bispecific antibody).

8. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 7, wherein the antibody has a binding activity of about 10 －５ Less than M, for example, about 10 －６ M, 10 －７ M, 10 －８ M, 10 －９ M or 10 －１０ K below M Ｄ An antibody or an antigen-binding fragment thereof that binds to a human IL-12 / IL-23p40 protein domain at

9. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 7, wherein the antibody has an EC of less than about 100 nM, for example, about 10 nM, 1 nM, 0.9 nM, 0.8 nM, 0.7 nM, 0.6 nM, 0.5 nM, 0.4 nM, 0.3 nM, 0.2 nM, 0.1 nM or less. ５０ An antibody or an antigen-binding fragment thereof that binds to a human IL-12 / IL-23p40 protein domain at

10. (1) An isolated polypeptide comprising the sequences set forth in SEQ ID NOs:3, 4 and 5 or SEQ ID NOs:26, 4 and 5, said polypeptide specifically binding to a human IL-12 / IL-23p40 protein domain as part of an anti-human IL-12 / IL-23p40 protein domain antibody, said antibody further comprising the sequences set forth in SEQ ID NOs:8, 19 or 20, SEQ ID NOs:9 or 21, SEQ ID NOs:10, 22 or 23, or SEQ ID NOs:27, 28 and 22; (2) An isolated polypeptide comprising a sequence set forth in SEQ ID NO:8, 19 or 20, SEQ ID NO:9 or 21, SEQ ID NO:10, 22 or 23, or SEQ ID NOs:27, 28 and 22, said polypeptide specifically binds to a human IL-12 / IL-23p40 protein domain as part of an anti-human IL-12 / IL-23p40 protein domain antibody, said antibody further comprising a sequence set forth in SEQ ID NO:3, 4 and 5; (3) An isolated polypeptide comprising a sequence as set forth in SEQ ID NO: 1 or 24, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, sequence identity to said sequence, or an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence, wherein said polypeptide is an isolated polypeptide comprising a human IL-12 / IL-23p40 protein domain antibody as part of an anti-human IL-12 / IL-23p40 protein domain antibody. an isolated polypeptide that specifically binds to an IL-12 / IL-23p40 protein domain, said antibody further comprising a sequence according to SEQ ID NO: 6, 11, 13, 15, 17 or 25, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto, or an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence; and (4) An isolated polypeptide comprising a sequence set forth in SEQ ID NO: 6, 11, 13, 15, 17 or 25, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to said sequence, or an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence, wherein said polypeptide is an anti-human IL-12 / IL-23p 10. An isolated polypeptide which specifically binds to the human IL-12 / IL-23 p40 protein domain as part of a 40 protein domain antibody, said antibody comprising a sequence as set forth in SEQ ID NO: 1 or 24, a sequence having 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto, or further comprising one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence. An isolated polypeptide selected from the group consisting of:

11. (1) an isolated polynucleotide encoding a polypeptide comprising a sequence as set forth in SEQ ID NOs:3, 4 and 5, said polypeptide specifically binds to a human IL-12 / IL-23p40 protein domain as part of an anti-human IL-12 / IL-23p40 protein domain antibody, said antibody further comprising a sequence as set forth in SEQ ID NOs:8, 19 or 20, SEQ ID NOs:9 or 21, SEQ ID NOs:10, 22 or 23, or SEQ ID NOs:27, 28 and 22; (2) an isolated polynucleotide encoding a polypeptide comprising a sequence set forth in SEQ ID NO:8, 19 or 20, SEQ ID NO:9 or 21, SEQ ID NO:10, 22 or 23, or SEQ ID NOs:27, 28 and 22, said polypeptide specifically binding to a human IL-12 / IL-23p40 protein domain as part of an anti-human IL-12 / IL-23p40 protein domain antibody, said antibody further comprising a sequence set forth in SEQ ID NO:3, 4 and 5; (3) An isolated polynucleotide encoding a polynucleotide comprising a sequence set forth in SEQ ID NO: 1 or 24, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to said sequence, or an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence, wherein said polypeptide is a member of an anti-human IL-12 / IL-23p40 protein domain antibody. an isolated polynucleotide which specifically binds to a human IL-12 / IL-23p40 protein domain as part of said antibody, said antibody further comprising an amino acid sequence according to SEQ ID NO: 6, 11, 13, 15, 17 or 25, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto, or an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence; and (4) An isolated polynucleotide encoding a polypeptide comprising a sequence set forth in SEQ ID NO: 6, 11, 13, 15, 17 or 25, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to said sequence, or a sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence, wherein said polypeptide is an anti-human IL-12 / IL-23p 10. An isolated polynucleotide that specifically binds to the human IL-12 / IL-23p40 protein domain as part of a 40 protein domain antibody, said antibody comprising a sequence as set forth in SEQ ID NO: 1 or 24, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to said antibody, or further comprising one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid mutations (preferably substitutions, insertions or deletions) compared to said sequence. An isolated polynucleotide encoding a polypeptide selected from the group consisting of:

12. 12. A polynucleotide encoding a polypeptide according to claim 11, said polynucleotide comprising or having a nucleotide sequence as set forth in SEQ ID NO: 2 or 37, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto. or said polynucleotide comprises or consists of the nucleotide sequence set forth in SEQ ID NO: 7, 12, 14, 16, 18 or 38, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto.

13. A vector comprising the polynucleotide according to any one of claims 11 to 12.

14. A host cell comprising the polynucleotide according to any one of claims 11 to 12 or the vector according to claim 13.

15. A method for preparing the antibody or antigen-binding fragment thereof according to any one of claims 1 to 9, comprising culturing a host cell according to claim 14 under suitable conditions and isolating the antibody or antigen-binding fragment thereof from the cell culture.

16. An antibody conjugate comprising the antibody or antigen-binding fragment thereof according to any one of claims 1 to 9 and a conjugate moiety attached thereto, wherein the conjugate moiety is a purification tag (e.g., a His tag), a cytotoxic agent or a detectable label; preferably, the conjugate moiety is a radioisotope, a luminescent substance, a colored substance, an enzyme or polyethylene glycol.

17. A fusion protein comprising the antibody or antigen-binding fragment thereof according to any one of claims 1 to 9.

18. A multispecific antibody, preferably a bispecific antibody, comprising the antibody or antigen-binding fragment thereof according to any one of claims 1 to 9.

19. A kit comprising the antibody or antigen-binding fragment thereof according to any one of claims 1 to 9, the antibody conjugate according to claim 16, the fusion protein according to claim 17, or the multispecific antibody according to claim 18, preferably further comprising a second antibody which specifically identifies the antibody or antigen-binding fragment thereof, optionally wherein the second antibody further comprises a detectable label, such as a radioisotope, a luminescent substance, a colored substance, an enzyme or polyethylene glycol.

20. Use of an antibody or an antigen-binding fragment thereof according to any one of claims 1 to 9, an antibody conjugate according to claim 16, a fusion protein according to claim 17 or a multispecific antibody according to claim 18 in the preparation of a kit for detecting the presence or level of a human IL-12 / IL-23p40 protein domain in a sample.

21. A pharmaceutical composition comprising an antibody or an antigen-binding fragment thereof according to any one of claims 1 to 9, an antibody conjugate according to claim 16, a fusion protein according to claim 17 or a multispecific antibody according to claim 18, and optionally a pharma- ceutically acceptable carrier and / or excipient.

22. 22. The pharmaceutical composition of claim 21, wherein the pharmaceutical composition is in a form suitable for administration by subcutaneous, intradermal, intravenous, intramuscular or intralesional injection.

23. Agents that block the binding of human IL-12 / IL-23p40 protein domains to human IL-12Rβ1 or human IL-23R; Agents for blocking the activity or downregulating the levels of the human IL-12 / IL-23 p40 protein domain, or Agent for blocking a cellular response mediated by binding of human IL-12Rβ1 or human IL-23R to a p40 protein domain, preferably wherein the ligand of the human IL-12 / IL-23 p40 protein domain is human IL-12Rβ1 or human IL-23R. Use of the antibody or antigen-binding fragment thereof according to any one of claims 1 to 9, the antibody conjugate according to claim 16, the fusion protein according to claim 17, the multispecific antibody according to claim 18 or the pharmaceutical composition according to claim 21 in the preparation of

24. Use of an antibody or antigen-binding fragment thereof according to any one of claims 1 to 9, an antibody conjugate according to claim 16, a fusion protein according to claim 17, a multispecific antibody according to claim 18 or a pharmaceutical composition according to claim 21 in the preparation of a medicament for treating an autoimmune disease (e.g. psoriasis vulgaris or systemic lupus erythematosus) or ulcerative colitis (e.g. refractory or recurrent).

25. An in vivo or in vitro method comprising administering a cell comprising the antibody or antigen-binding fragment thereof according to any one of claims 1 to 9, the antibody conjugate of claim 16, the fusion protein of claim 17, the multispecific antibody of claim 18, or the pharmaceutical composition of claim 21, or administering to a subject in need thereof an effective amount of the antibody or antigen-binding fragment thereof according to any one of claims 1 to 9, the antibody conjugate of claim 16, the fusion protein of claim 17, the multispecific antibody of claim 18, or the pharmaceutical composition of claim 21, said method comprising: A method for blocking the binding of the human IL-12 / IL-23p40 protein domain to the ligand IL-12Rβ1 or IL-23R, Methods for downregulating the activity or levels of the human IL-12 / IL-23 p40 protein domain, and 1. A method for blocking a cellular response mediated by binding of human IL-12Rβ1 or human IL-23R to a p40 protein domain, comprising: Preferably, the ligand for IL-12 / IL-23q40 is IL-12Rβ1 or IL-23R. An in vivo or in vitro method selected from the group consisting of:

26. 20. A method for the prevention, treatment, adjuvant treatment and / or diagnosis of an autoimmune disease (e.g. psoriasis vulgaris or systemic lupus erythematosus) or ulcerative colitis (e.g. refractory or recurrent) comprising administering to a subject in need thereof an antibody or antigen-binding fragment thereof according to any one of claims 1 to 9, an antibody conjugate according to claim 16, a fusion protein according to claim 17, a multispecific antibody according to claim 18 or a pharmaceutical composition according to claim 21, preferably wherein said subject is undergoing conventional treatment or is insufficiently responsive, unresponsive or intolerant to biological agents and has failed to achieve a complete or partial response.

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