Anti-human P40 protein domain antibodies and their use

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] This invention belongs to the field of pharmaceuticals, and more particularly to monoclonal antibodies for blocking the function of the interleukin IL-12 / IL-23p40 protein domain and their use.

[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 heterodimer structure and is 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 consists of a heavy chain (p40) of 306 amino acids containing 10 cysteine ​​residues and 4 potential glycosylation sites, and a light chain (p35) of 197 amino acids containing 7 cysteine ​​residues and 3 potential glycosylation sites.

[0003] Interleukin-23 (IL-23), a new member of the interleukin-12 (IL-12) cytokine family discovered by Oppmann et al. in 2000, is a biologically active complex cytokine formed by the covalent linking of 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 to IL-12Rβ2 (Oppmann B et al., Immunity, 2000, 13(5): 715-725). IL-23 and IL-12 share the same signaling pathways, including the Janus kinases Tyk2, Jak2, and STAT, but each affects different T cells. IL-23 affects memory T cells (CD4). +induces acute proliferation, but IL-12 stimulates the proliferation of virgin CD4 + T cells. Similar to IL-12, human IL-23 stimulates the production and proliferation of IFN-γ in PHA-activated T cells and CD45RO + T cells. By antagonizing the common subunit p40 of IL-12 and IL-23, the pathways of IL-12 and IL-23 can be effectively antagonized simultaneously.

[0004] Interleukin 12 is mainly produced by dendritic cells, macrophages, B lymphocytes and some other antigen-presenting cells (APCs), 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 can promote the differentiation from Th0 to Th1. In addition, IL-12 promotes Thl cells to secrete IFN-γ and IL2, mediates the cellular immune response, induces T cells and NK cells to produce IFN-γ, and provides extramedullary hematopoietic function. IL-12 plays a very important role in both the initial non-specific immunity and the subsequent antigen-specific adaptive immune process in the body, and IL-12 is a multifunctional immunomodulatory factor (Manetti R et al., Journal of Experimental Medicine, 1993, 177(4): 1199-1204). The role of IL-12 in autoimmunity induction is (1) promoting the differentiation and proliferation of antigen-specific Thl cells to produce multiple cytokines and enhancing the Th1-type immune response, and (2) stimulating mononuclear macrophages to produce various active mediators, enhancing the cytotoxicity of immune cells and causing 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 onset of psoriasis. The abnormal accumulation of IL-2 and TNF-a in the skin lesions of psoriasis promotes the onset and progression of psoriasis. It is considered that the formation of a huge cell network mainly containing Th1 cytokines by lymphocytes secreting multiple cytokines in the skin lesions is an important pathological basis of the disease. Th1 cytokines secreted by DCs are thought to be the initiating factors in the onset of psoriasis, and mature DCs can increase the expression of IFN-α and IL-12. The specific mechanisms of action of mental, genetic, and infectious factors in the onset of the disease are unknown. Saint-Mezard et al. found that mental stress promotes the aggregation of skin DCs and enhances the delayed allergic reaction of the skin to haptens. Although it has been experimentally confirmed that IL-23 also acts on DC-induced inflammatory reactions, it is not clear whether it is a direct inflammatory factor. IL-12 induces the onset of psoriasis by mediating T cells to the skin surface via skin lymphocyte antigen, and Rosmarin et al. analyzed the structure, receptor, and function of IL-12 and suggested that psoriasis can be treated by changing the IL-12 level (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 are no effective treatment means for clinical treatment in China, and traditional treatments mainly include hormones and immunosuppressants. STELARA(R) (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 as a treatment drug for SLE, indicating the effectiveness of anti-IL-12p40 antibody in the treatment of SLE (STELARA (ustekinumab) of Janssen R&D shows positive results in the treatment of systemic lupus erythematosus in a phase II trial).

[0007] Therefore, IL-12 and IL-23 are extremely important in inducing autoimmunity and maintaining the immune response, and consequently, any part 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, both IL-12 and IL-23 possess 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 a new drug for treating autoimmune diseases (such as psoriasis vulgaris and systemic lupus erythematosus).

[0009] Ulcerative colitis is an IBD disease primarily characterized by immune dysfunction. It is an immune disorder with a complex pathogenesis, characterized by complex clinicopathological changes, a long disease course, and recurrent attacks. More than half of patients receive conventional or biological therapies without relief. Ulcerative colitis is the result of a combination of exogenous and host factors in a specific genetic context. Lesions are usually located in the sigmoid colon and rectum, but can extend to the descending colon or even the entire colon. The disease has a long course, and recurrent attacks are common. While the disease can occur at any age, it is most common in those in their 20s and 30s. Given the characteristics of ulcerative colitis, there is a need to find better antibody drugs to treat 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, developed antibody sequences, performed preliminary antibody screening using methods such as ELISA, and ultimately selected antibodies with higher efficacy as candidate antibodies for subsequent pharmacodynamic studies.

[0011] Furthermore, the inventors prepared humanized antibodies against the human IL-12 / IL-23p40 protein domain (e.g., humanized antibodies called H5L9, H5L10, H5L11, H5L12, H5L14, and H8L15). To our surprise, the inventors discovered that the antibody specifically binds to the human IL-12 / IL-23p40 protein domain and exhibits excellent binding activity.

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

[0013] The inventors have also found, surprisingly, 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 ligands IL-12Rβ1 and IL-23R, and inhibit the activation of downstream signaling pathways of IL-12 / IL-23. These antibodies may be used to prepare agents for the prevention and treatment of autoimmune diseases (e.g., psoriasis vulgaris or systemic lupus erythematosus) and ulcerative colitis (e.g., refractory or relapsing).

[0014] The amino acid sequence of the CDR region of the above antibody is analyzed by technical means well known to those skilled in the art, for example, by the VBASE2 database.

[0015] Those skilled in the art will understand that the CDR region of an antibody is responsible for the antibody's binding specificity to an antigen. Given the known sequences of the heavy chain and light chain variable regions of antibodies, there are several methods for determining the CDR region of an antibody, including the Kabat, IMGT, Chothia, and AbM numbering systems.

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

[0017] Kabat numbering system: See, for example, 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, for example, Chothia & Lesk (1987) J. Mol. Biol. 196: 901-917; Chothia et al. (1989), Nature 342: 878-883. IMGT numbering system: See, for example, Lefranc et al., Dev. Comparat. Immunol. 27: 55-77, 2003.

[0018] However, the application of all definitions of CDR to antibodies or their variants shall be within the scope of the terms defined and used herein. If the amino acid sequence of the variable region of the antibody is known, a person skilled in the art can generally determine which residues contain a particular CDR without relying on any experimental data other than the sequence itself. For comparison, appropriate amino acid residues of CDRs as defined by the Kabat and Chothia CDR numbering systems are listed below. The exact number of residues containing a specific CDR varies depending on the sequence and size of that CDR.

[0019] Preferably, the antibodies H5L9, H5L10, H5L11, H5L12, and H5L14 disclosed herein have the same HCDR1-3, The amino acid sequences of the three HCDR regions in the heavy chain variable region are as follows: HCDR1:GYSFTTYW (Sequence ID 3) HCDR2:IMSPVDSDI (Sequence ID 4) HCDR3:ARRRPGQGYFDF (Sequence ID 5) The antibody H5L9 disclosed herein has LCDR1-3, The amino acid sequences of the three CDR regions in the light chain variable region are as follows: LCDR1:QNVGSW (Sequence ID 8) LCDR2:ASS (Sequence ID 9) LCDR3:QQYDIYPFT(Sequence ID 10) The antibody H5L10 disclosed herein has LCDR1-3, The amino acid sequences of the three CDR regions in the light chain variable region are as follows: LCDR1:QSVGSW (Sequence ID 19) LCDR2:ASN (Sequence ID 21) LCDR3:QQYNIYPYT(Sequence ID 22) The antibodies H5L11 and H5L12 disclosed herein have the same LCDR1-3, The amino acid sequences of the three CDR regions in the light chain variable region are as follows: LCDR1:QSVSSW (Sequence ID 20) LCDR2:ASN (Sequence ID 21) LCDR3:QQYNIYPYT(Sequence ID 22) The antibody H5L14 disclosed herein has LCDR1-3: The amino acid sequences of the three CDR regions in the light chain variable region are as follows: LCDR1:QSVSSW (Sequence ID 20) LCDR2:ASN (Sequence ID 21) LCDR3:QQYNIYPFT (Sequence ID 23) The antibody H8L15 disclosed herein has HCDR1-3 and LCDR1-3, The amino acid sequences of the three HCDR regions in the heavy chain variable region are as follows: HCDR1:GYTFTSYW (Sequence ID 26) HCDR2:MSPVDSDI (Sequence ID 4) HCDR3:ARRRPGQGYFDF (Sequence ID 5) The amino acid sequences of the three CDR regions in the light chain variable region are as follows: LCDR1:QSVGTW (Sequence ID 27) LCDR2:A AS (Sequence ID 28) LCDR3:QQYNIYPYT (Sequence ID 22).

[0020] The present invention will be described in detail below. One aspect of the present invention preferably relates to an antibody or an antigen-binding fragment thereof that specifically binds to human IL-12 / IL-23p40, wherein, (1) The antibody is The heavy chain variable region described in Sequence ID No. 1 includes HCDR1, HCDR2, and HCDR3, (wherein preferably, HCDR1 includes or consists of the sequence described in Sequence ID No. 3, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with respect to the sequence, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, 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 the sequence, The HCDR2 comprises or consists of the sequence described in Sequence ID No. 4, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with respect to the sequence, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, 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 the sequence, and The HCDR3 comprises or consists of the sequence described in Sequence 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 with respect to the aforementioned 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 the aforementioned sequence. Furthermore, the antibody further, The light chain variable region described in Sequence ID No. 6 includes LCDR1, LCDR2, and LCDR3, (wherein preferably, LCDR1 includes or consists of the amino acid sequence described in Sequence ID No. 8, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with respect to the sequence, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, 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 the sequence, The LCDR2 includes or comprises the amino acid sequence described in Sequence ID No. 9, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with respect to the aforementioned sequence, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, 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 the aforementioned sequence, and The LCDR3 comprises or consists of the sequence described in Sequence ID No. 10, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with respect to the sequence, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, 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 the sequence described above. (2) The antibody is The heavy chain variable region described in Sequence ID No. 1 includes HCDR1, HCDR2, and HCDR3, (wherein preferably, HCDR1 includes or consists of the sequence described in Sequence ID No. 3, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with respect to the sequence, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, 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 the sequence, The HCDR2 comprises or consists of the sequence described in Sequence ID No. 4, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with respect to the sequence, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, 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 the sequence, and The HCDR3 comprises or consists of the sequence described in Sequence 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 with respect to the aforementioned 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 the aforementioned sequence. The aforementioned antibody was further described as follows: The light chain variable region described in SEQ ID NO: 11 includes LCDR1, LCDR2, and LCDR3, (wherein preferably, LCDR1 includes or comprises the amino acid sequence described in SEQ ID NO: 19, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with the sequence, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, 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 the sequence, The LCDR2 includes or comprises the amino acid sequence described in Sequence ID No. 21, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with respect to the aforementioned sequence, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, 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 the aforementioned sequence, and The LCDR3 comprises or consists of the sequence described in Sequence ID No. 22, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with respect to the sequence, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, 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 the sequence described above. (3) The antibody is The heavy chain variable region described in Sequence ID No. 1 includes HCDR1, HCDR2, and HCDR3, (wherein preferably, HCDR1 includes or consists of the sequence described in Sequence ID No. 3, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with respect to the sequence, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, 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 the sequence, The HCDR2 comprises or consists of the sequence described in Sequence ID No. 4, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with respect to the sequence, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, 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 the sequence, and The HCDR3 comprises or consists of the sequence described in Sequence 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 with respect to the 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 the sequence described above. The aforementioned antibody was further described as follows: The light chain variable regions described in SEQ ID NOs. 13 and 15 include LCDR1, LCDR2 and LCDR3, (wherein preferably, LCDR1 includes or comprises the amino acid sequence described in SEQ ID NOs. 20, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with respect to the aforementioned sequence, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, 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 the aforementioned sequence. The LCDR2 includes or comprises the amino acid sequence described in Sequence ID No. 21, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with respect to the aforementioned sequence, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, 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 the aforementioned sequence, and The LCDR3 comprises, or consists of, the sequence described in Sequence ID No. 22, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with respect to the sequence described above, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, 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 the sequence described above. (4) The antibody is The heavy chain variable region described in Sequence ID No. 1 includes HCDR1, HCDR2, and HCDR3, (wherein preferably, HCDR1 includes or consists of the sequence described in Sequence ID No. 3, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with respect to the sequence, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, 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 the sequence, The HCDR2 comprises or consists of the sequence described in Sequence ID No. 4, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with respect to the sequence, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, 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 the sequence, and The HCDR3 comprises or consists of the sequence described in Sequence 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 with respect to the 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 the sequence described above. The aforementioned antibody was further described as follows: The light chain variable region described in Sequence ID No. 17 includes LCDR1, LCDR2, and LCDR3, (wherein preferably, LCDR1 includes or consists of the amino acid sequence described in Sequence ID No. 20, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with respect to the sequence, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, 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 the sequence, The LCDR2 comprises or consists of the amino acid sequence described in Sequence ID No. 21, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with respect to the sequence, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, 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 the sequence, and The aforementioned LCDR3 includes or comprises the sequence described in Sequence ID No. 23, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with respect to the aforementioned sequence, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, 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 the aforementioned sequence); (5) The antibody is: The heavy chain variable region described in Sequence ID No. 24 includes HCDR1, HCDR2, and HCDR3: (wherein preferably, HCDR1 includes or consists of the sequence described in Sequence ID No. 26, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with respect to the sequence, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, 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 the sequence, The HCDR2 comprises or consists of the sequence described in Sequence ID No. 4, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with respect to the sequence, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, 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 the sequence, and The HCDR3 comprises or consists of the sequence described in Sequence 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 with respect to the aforementioned 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 the aforementioned sequence. The aforementioned antibody was further described as follows: The light chain variable region described in SEQ ID NO: 25 includes LCDR1, LCDR2, and LCDR3 (wherein preferably, LCDR1 includes or consists of the amino acid sequence described in SEQ ID NO: 27, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with the sequence, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, 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 the sequence, The LCDR2 comprises or consists of the amino acid sequence described in Sequence ID No. 28, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with respect to the aforementioned sequence, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, 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 the aforementioned sequence, and The LCDR3 comprises or consists of the sequence described in Sequence ID No. 22, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with respect to the sequence, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, 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 the sequence.

[0021] In some embodiments of the present invention, the antibody is (1) The amino acid sequence described in Sequence ID No. 1, A sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with respect to the sequence described in Sequence ID No. 1, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity, 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 described in Sequence ID No. 1; and The amino acid sequence described in Sequence ID No. 6, A sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with respect to the sequence described in Sequence ID No. 6, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity, 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 described in Sequence ID No. 6; (2) The amino acid sequence described in Sequence ID No. 1, A sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with respect to the sequence described in Sequence ID No. 1, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity, 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 described in Sequence ID No. 1; and The amino acid sequence described in Sequence ID No. 11, A sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with respect to the sequence described in Sequence ID No. 11, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity, 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 described in Sequence ID No. 11; (3) The amino acid sequence described in Sequence ID No. 1, A sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with respect to the sequence described in Sequence ID No. 1, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity, 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 described in Sequence ID No. 1; and The amino acid sequence described in SEQ ID NO: 13, A sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with respect to the sequence described in Sequence ID No. 13, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity, 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 described in Sequence ID No. 13; (4) The amino acid sequence described in Sequence ID No. 1, A sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with respect to the sequence described in Sequence ID No. 1, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity, 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 described in Sequence ID No. 1; and The amino acid sequence described in Sequence ID No. 15, A sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with respect to the sequence described in Sequence ID No. 15, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity, 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 described in Sequence ID No. 15; (5) The amino acid sequence described in Sequence ID No. 1, A sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with respect to the sequence described in Sequence ID No. 1, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity, 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 described in Sequence ID No. 1; and The amino acid sequence described in Sequence ID No. 17, A sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with the sequence described in Sequence ID No. 17, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity, 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 described in Sequence ID No. 17; (6) The amino acid sequence described in Sequence ID No. 24, A sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with respect to the sequence described in Sequence ID No. 24, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity, 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 described in Sequence ID No. 24; and The amino acid sequence described in Sequence ID No. 25, A sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with respect to the sequence described in Sequence ID No. 25, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity, 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 described in SEQ ID NO: 25. Includes.

[0022] In one embodiment of the present invention, the antibody further comprises FRs in the heavy chain variable region, preferably FR-H1, FR-H2, FR-H3 and FR-H4 (wherein FR-H1 is the amino acid sequence described in SEQ ID NO: 29, or at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, or 96% of the sequence described in SEQ ID NO: 29). , a 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 described in SEQ ID NO: 29, or comprising or consisting thereof; the FR-H2 is the amino acid sequence described in SEQ ID NO: 30, or at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88% of the sequence described in SEQ ID NO: 30 , a sequence having 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 described in SEQ ID NO: 30; FR-H3 is the amino acid sequence described in SEQ ID NO: 31, compared to the sequence described in SEQ ID NO: 31 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, 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 described in Sequence ID No. 31, or comprising such an amino acid sequence;FR-H4 includes or consists of the amino acid sequence described in SEQ ID NO: 32, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with the sequence described in SEQ ID NO: 32, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, 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 described in SEQ ID NO: 32.

[0023] In one embodiment of the present invention, the antibody further comprises FRs in the light chain variable region, preferably FR-L1, FR-L2, FR-L3 and FR-L4 (wherein FR-L1 is the amino acid sequence described 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%, or 96%, relative to the sequence described in SEQ ID NO: 33. A 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 described in SEQ ID NO: 33; FR-L2 is an amino acid sequence described in SEQ ID NO: 34, with respect to the sequence described in SEQ ID NO: 34, with respect to at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 8 A 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 described in SEQ ID NO: 34; FR-L3 is the amino acid sequence described in SEQ ID NO: 35, compared to the sequence described in SEQ ID NO: 35 A sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, 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 described in Sequence ID No. 35, or comprising such an amino acid sequence;FR-L4 includes or consists of the amino acid sequence described in SEQ ID NO: 36, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with the sequence described in SEQ ID NO: 36, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, 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 described in SEQ ID NO: 36.

[0024] In one embodiment of the present invention, the antibody further comprises FRs in the light chain variable region, preferably FR-L1, FR-L2, FR-L3 and FR-L4 (wherein FR-L1 is the amino acid sequence described in SEQ ID NO: 41, at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% of the sequence described in SEQ ID NO: 41, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 9 A sequence having 7%, 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 described in SEQ ID NO: 41; or FR-L2 having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 8% of the amino acid sequence described in SEQ ID NO: 42. A 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 described in SEQ ID NO: 42; the FR-L3 is the amino acid sequence described in SEQ ID NO: 43, compared to the sequence described in SEQ ID NO: 43 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, 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 described in Sequence ID No. 43, or comprising such an amino acid sequence;The aforementioned FR-L4 includes or consists of the amino acid sequence described in SEQ ID NO: 44, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with the aforementioned sequence described in SEQ ID NO: 44, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, 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 aforementioned amino acid sequence described in SEQ ID NO: 44.

[0025] In one embodiment of the present invention, the antibody further comprises FRs in the light chain variable region, preferably FR-L1, FR-L2, FR-L3 and FR-L4, where FR-L1 is the amino acid sequence described 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%, or 96% of the sequence described in SEQ ID NO: 33. , a 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 described in SEQ ID NO: 33; the FR-L2 is the amino acid sequence described in SEQ ID NO: 45, with respect to at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, A sequence having 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 described in SEQ ID NO: 45; or FR-L3 having the amino acid sequence described in SEQ ID NO: 46, or the sequence described in SEQ ID NO: 46 The 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%, or the 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 described in Sequence ID No. 46, or comprising or consisting of such sequences;The aforementioned FR-L4 includes or consists of the amino acid sequence described in SEQ ID NO: 36, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with the aforementioned sequence described in SEQ ID NO: 36, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, 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 aforementioned amino acid sequence described in SEQ ID NO: 36.

[0026] In one embodiment of the present invention, the antibody further comprises FRs in the light chain variable region, preferably FR-L1, FR-L2, FR-L3 and FR-L4 (wherein FR-L1 is the amino acid sequence described in SEQ ID NO: 33, or at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90%, preferably at least 91%, 92%, 93%, 94%, 95%, or 96% of the sequence described in SEQ ID NO: 33). , a 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 described in SEQ ID NO: 33; the FR-L2 is the amino acid sequence described in SEQ ID NO: 47, or at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, or 88% of the sequence described in SEQ ID NO: 47 , a sequence having 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 described in SEQ ID NO: 47, or comprising or consisting of such; FR-L3 is the amino acid sequence described in SEQ ID NO: 46, or compared to the sequence described in SEQ ID NO: 46 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, 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 described in Sequence ID No. 46, or comprising such an amino acid sequence;The aforementioned FR-L4 includes or consists of the amino acid sequence described in SEQ ID NO: 36, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with the aforementioned sequence described in SEQ ID NO: 36, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, 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 aforementioned amino acid sequence described in SEQ ID NO: 36.

[0027] In one embodiment of the present invention, the antibody further comprises FRs in the light chain variable region, preferably FR-L1, FR-L2, FR-L3 and FR-L4 (wherein FR-L1 is the amino acid sequence described 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%, or 96%, relative to the sequence described in SEQ ID NO: 33. A 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 described in SEQ ID NO: 33; the FR-L2 is the amino acid sequence described in SEQ ID NO: 45, with respect to the sequence described in SEQ ID NO: 45, with respect to at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88% A sequence having 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 described in SEQ ID NO: 45; or FR-L3 having the amino acid sequence described in SEQ ID NO: 48, or the sequence described in SEQ ID NO: 48 The 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%, or the 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 described in Sequence ID No. 48, or comprising or consisting of such sequences;The aforementioned FR-L4 includes or consists of the amino acid sequence described in SEQ ID NO: 36, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with the aforementioned sequence described in SEQ ID NO: 36, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, 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 aforementioned amino acid sequence described in SEQ ID NO: 36.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

[0044] One aspect of the present invention is, An isolated polypeptide comprising the sequence described in SEQ ID NO: 1 or 24, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with the sequence, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, 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 sequence (where the polypeptide is part of an anti-human IL-12 / IL-23p40 protein domain antibody). The antibody specifically binds to the human IL-12 / IL-23p40 protein domain, and further comprises the sequence described in SEQ ID NOs. 6, 11, 13, 15, 17, or 25, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with the sequence, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, 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 sequence; or An isolated polypeptide comprising the 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 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 specifically binds to the human IL-12 / IL-23p40 protein domain as part of an anti-human IL-12 / IL-23p40 protein domain antibody, and said antibody further comprises the sequence according to 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) relates to

[0045] In one embodiment of the invention, said 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, said antibody is a humanized antibody, a chimeric antibody or a multispecific antibody (e.g., a bispecific antibody).

[0047] In one embodiment of the invention, said antibody has a K -5 less than about 10 -6 M, e.g., about 10 -7 M, 10 -8 M, 10 -9 less than M, or 10 -10 M or lessD It binds to the human IL-12 / IL-23p40 protein domain having the K. Preferably, the K D This is measured using the Fortebio molecular interaction analyzer.

[0048] In one embodiment of the present invention, 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 It binds to the human IL-12 / IL-23p40 protein domain having the EC. Specifically, the EC 50 It is measured by the indirect ELISA method.

[0049] In one embodiment of the present invention, the antibody includes a constant region, and the constant region is derived from a species other than mouse, for example, 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 an Igγ-1 chain C region of GenBank accession number P01857; and the light chain constant region is an Igκ chain C region, preferably an Igκ chain C region of GenBank accession number P01834.

[0051] Another aspect of the present invention relates to an isolated polynucleotide encoding a polypeptide comprising the sequences described in SEQ ID NOs: 3, 4, and 5 (wherein the polypeptide specifically conjugates to the human IL-12 / IL-23p40 protein domain as part of an anti-human IL-12 / IL-23p40 protein domain antibody, the antibody further comprising the sequences described 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 sequences described in SEQ ID NOs: 8, 9, and 10 (wherein the polypeptide specifically conjugates to the human IL-12 / IL-23p40 protein domain as part of an anti-human IL-12 / IL-23p40 protein domain antibody, the antibody further comprising the sequences described 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 sequences described in SEQ ID NOs: 3, 4, and 5 (wherein the polypeptide specifically conjugates to the human IL-12 / IL-23p40 protein domain as part of an anti-human IL-12 / IL-23p40 protein domain antibody, the antibody further comprising the sequences described 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 sequences described in SEQ ID NOs: 19, 21, and 22 (wherein the polypeptide specifically conjugates to the human IL-12 / IL-23p40 protein domain as part of an anti-human IL-12 / IL-23p40 protein domain antibody, the antibody further comprising the sequences described 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 sequences described in SEQ ID NOs: 3, 4, and 5 (wherein the polypeptide specifically conjugates to the human IL-12 / IL-23p40 protein domain as part of an anti-human IL-12 / IL-23p40 protein domain antibody, the antibody further comprising the sequences described 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 sequences described in SEQ ID NOs: 20, 21, and 22 (wherein the polypeptide specifically conjugates to the human IL-12 / IL-23p40 protein domain as part of an anti-human IL-12 / IL-23p40 protein domain antibody, the antibody further comprising the sequences described 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 sequences described in SEQ ID NOs: 3, 4, and 5 (wherein the polypeptide specifically conjugates to the human IL-12 / IL-23p40 protein domain as part of an anti-human IL-12 / IL-23p40 protein domain antibody, the antibody further comprising the sequences described 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 sequences described in SEQ ID NOs: 20, 21, and 23 (wherein the polypeptide specifically conjugates to the human IL-12 / IL-23p40 protein domain as part of an anti-human IL-12 / IL-23p40 protein domain antibody, the antibody further comprising the sequences described 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 sequences described in SEQ ID NOs: 26, 4, and 5 (wherein the polypeptide specifically conjugates to the human IL-12 / IL-23p40 protein domain as part of an anti-human IL-12 / IL-23p40 protein domain antibody, the antibody further comprising the sequences described 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 sequences described in SEQ ID NOs: 27, 28, and 22 (wherein the polypeptide specifically conjugates to the human IL-12 / IL-23p40 protein domain as part of an anti-human IL-12 / IL-23p40 protein domain antibody, the antibody further comprising the sequences described 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 described in SEQ ID NO: 1 or 24, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with the sequence, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, or an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) conserved amino acid mutations (preferably substitutions, insertions, or deletions) compared to the sequence (wherein the polypeptide is anti-human IL-12 / IL-23p40 A protein domain antibody that specifically binds to the human IL-12 / IL-23p40 protein domain as part of a protein domain antibody, wherein the antibody further comprises the sequence described in SEQ ID NOs. 6, 11, 13, 15, 17, or 25, sequence identity of 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%, with respect to the sequence, or an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) conserved amino acid mutations (preferably substitutions, insertions, or deletions) compared to the sequence; or An isolated polynucleotide encoding a polypeptide comprising the sequence described in SEQ ID NOs. 6, 11, 13, 15, 17, or 25, a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% sequence identity with the sequence, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, or an amino acid sequence having one or more (preferably 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) conserved amino acid mutations (preferably substitutions, insertions, or deletions) compared to the sequence (wherein the polypeptide is anti-human IL-12 / As part of an IL-23p40 protein domain antibody, the antibody specifically binds to the human IL-12 / IL-23p40 protein domain, and further comprises the sequence described in SEQ ID NO: 1 or 24, sequence identity of at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90% of the sequence, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, 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 sequence. Regarding.

[0062] Specifically, the polynucleotide includes or consists of the nucleotide sequence described in Sequence ID No. 2, or sequences 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 with respect to the aforementioned sequence.

[0063] Specifically, the polynucleotide includes or consists of the nucleotide sequence described in Sequence ID No. 7, or sequences 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 with respect to the aforementioned sequence.

[0064] Specifically, the polynucleotide includes or consists of a nucleotide sequence described in Sequence 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 with respect to the aforementioned sequence.

[0065] Specifically, the polynucleotide includes or consists of the nucleotide sequence described in Sequence 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 with respect to the aforementioned sequence.

[0066] Specifically, the polynucleotide includes or consists of the nucleotide sequence described in Sequence ID No. 16, or sequences 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 with respect to said sequence.

[0067] Specifically, the polynucleotide includes or consists of a nucleotide sequence described in Sequence 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 with respect to the aforementioned sequence.

[0068] Specifically, the polynucleotide includes or consists of a nucleotide sequence described in Sequence 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 with respect to the aforementioned sequence.

[0069] Specifically, the polynucleotide includes or consists of a nucleotide sequence described in Sequence 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 with respect to the aforementioned sequence.

[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 either a polynucleotide molecule or a vector disclosed herein as described above.

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

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

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

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

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

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

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

[0079] A further aspect of the present invention relates to a pharmaceutical composition comprising one of the antibodies or antigen-binding fragments thereof disclosed herein as described above, the antibody conjugate, the multispecific antibody, or the fusion protein, and optionally comprising a pharmaceutically acceptable carrier and / or excipient.

[0080] Another aspect of the present invention is, Drugs that block the binding of human IL-12 / IL-23p40 to human IL-12Rβ1 or human IL-23R, A drug for blocking the activity of human IL-12 / IL-23p40 or for downregulating its levels, Agents for blocking cellular responses mediated by the binding of human IL-12Rβ1 or human IL-23R to p40; (Preferably, the ligand for the human IL-12 / IL-23p40 is human IL-12Rβ1 or human IL-23R.) This relates to the use of any one of the following in the preparation of an antibody or its antigen-binding fragment or antibody conjugate, the multispecific antibody, the fusion protein, or the pharmaceutical composition.

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

[0082] A further aspect of the present invention relates to an in vivo or in vitro method comprising administering cells containing the antibodies described herein or their antigen-binding fragments, antibody conjugates, multispecific antibodies, fusion proteins, or pharmaceutical compositions, or administering to a subject requiring an effective amount of any one of the antibodies disclosed herein as described above or their antigen-binding fragments, antibody conjugates, multispecific antibodies, fusion proteins, or pharmaceutical compositions. The method is as follows: A method for blocking the binding of human IL-12 / IL-23p40 to ligand IL-12Rβ1 or IL-23R. A method for downregulating the activity or level of human IL-12 / IL-23p40, and A method for blocking cellular responses mediated by the 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.) Selected from the group consisting of

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

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

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

[0086] A further aspect of the present invention relates to a method for the prevention, treatment, adjuvant therapy and / or diagnosis of autoimmune diseases (e.g., psoriasis vulgaris or systemic lupus erythematosus) or ulcerative colitis (e.g., refractory or recurrent), comprising administering to a subject requiring 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 for treating a patient suffering from ulcerative colitis, comprising (i) measuring the level of human IL-12 / IL-23p40 in a sample of the patient (where 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 its antigen-binding moiety.

[0088] The ulcerative colitis described herein may 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 present invention, the patient is receiving conventional treatment or is insufficiently responsive, unresponsive, or intolerant to biological agents. In some specific embodiments, the patient's receiving conventional treatment or insufficient responsiveness, unresponsiveness, or intolerance to biological agents results in failure of complete or partial response.

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

[0091] In this invention, unless otherwise specified, the scientific and technical terms used herein have meanings that are generally understood by those skilled in the art. Furthermore, the experimental procedures of cell culture, molecular genetics, nucleic acid chemistry, and immunology used in this invention are routine procedures widely used in their respective fields. For further understanding of this invention, definitions and explanations of relevant terms are provided below.

[0092] As used herein, the term “antigen-binding region” means 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 that antigen to the antibody is called an “antigen-binding region.” Such an antigen-binding region generally includes one or more complementarity-determining regions (CDRs). Some antigen-binding regions further include one or more “framework” regions (FRs). A CDR is an amino acid sequence that contributes to the specificity and affinity of antigen binding.

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

[0094] Intact antibodies generally contain at least two full-length heavy chains and two full-length light chains, but may contain fewer chains, such as antibodies naturally occurring in camelids that may contain only heavy chains. Antibodies or their antigen-binding fragments may originate from a single source or be "chimeric," meaning that different parts of an antibody may originate from two different sources, as further described below. Antibodies or their antigen-binding fragments may be produced in hybridomas by recombinant DNA technology or by enzymatic or chemical cleavage of intact antibodies. Unless otherwise stated, the term "antibody" includes not only antibodies containing two full-length heavy chains and two full-length light chains, but also their derivatives, variants, and fragments.

[0095] As used herein, the terms “antibody” or “immunoglobulin chain” (heavy or light chain) “antigen-binding fragment” (or “fragment”) include a portion of an antibody (which may be acquired or synthesized) that lacks at least some of the amino acid residues present in the full-length antibody but is capable of specifically binding to that antigen. Such fragments are biologically active because they specifically bind to a target antigen and can specifically bind to a given epitope in competition with other antibodies or their antigen-binding fragments. In one embodiment, such a fragment retains at least one CDR present in the full-length light or heavy chain of the antibody, and in several embodiments, such a fragment includes a single heavy and / or light chain or a portion thereof. Such biologically active fragments can be produced by recombinant DNA technology or, for example, by enzymatic or chemical cleavage of an intact antibody.

[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 may be derived from any mammal, including, but not limited to, humans, mice, rats, camelids, or rabbits. One or more functional portions of antibodies disclosed herein, such as CDRs, can be covalently bound to a second protein or small molecule to generate therapeutic agents directed at specific targets in the body, thereby having bifunctional therapeutic properties or, further intended, having an extended serum half-life, such as a fusion protein.

[0097] As used herein, the terms “full-length antibody chain,” “full-length antibody,” “intact antibody,” and “whole antibody” are interchangeable herein and refer to antibodies having a structure substantially similar to a natural antibody structure as defined herein, or antibodies having a heavy chain in the Fc region.

[0098] The term "light chain" includes full-length light chains and their fragments having a variable region sequence sufficient to confer binding specificity. The full-length light chain has a variable region domain V L and steady-state domain C L It includes the variable region domain of the light chain, which is located at the amino terminus of its polypeptide. The light chain includes a κ chain and a λ chain.

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

[0100] As used herein, the term “Fab fragment” means a single light chain of C H1 and consists of a variable region of one heavy chain. The heavy chain of the Fab molecule cannot form a disulfide bond 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 It comprises two heavy chain fragments containing a domain. The two heavy chain fragments are connected by two or more disulfide bonds and the C H3 They are retained together due to the hydrophobic interactions between the domains.

[0102] As used herein, the term “Fab' fragment” means a portion of one light chain and one heavy chain (the V H Domain, the C H1Domain, and the C H1 Domain and C H2 This includes a portion of the region between the domains, resulting in the formation of an interchain disulfide bond between the two heavy chains of the two Fab' fragments, thus yielding the F(ab')2 molecule.

[0103] As used herein, the term “F(ab')2 fragment” means the C H1 Domain and C H2 It comprises two light chains and two heavy chains, including a portion of the constant region between the domains, and as a result, an interchain disulfide bond is formed between the two heavy chains. Thus, the F(ab')2 fragment consists of two Fab' fragments held together by a disulfide bond between the two heavy chains.

[0104] As used herein, the term "Fv region" includes the variable region derived from the heavy and light chains, but lacks the steady region.

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

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

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

[0108] As used herein, the term "Fab / c" fragment refers to an intermediate formed by the pepsin digestion of immunoglobulins, which combines the advantages of both Fab and Fc regions—namely, strong in vivo diffusibility 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-chain variable region and the light-chain variable region are linked by a flexible linker to form a single polypeptide chain (which forms an 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. Patents 4,946,778 and 5,260,203, which are incorporated herein by reference.

[0110] As used herein, the term “domain antibody” is an immunofunctional immunoglobulin fragment comprising only the variable region of the heavy chain or the light chain. In some cases, two or more V H The regions are covalently linked by peptide linkers to form a multivalent domain antibody (especially a bivalent domain antibody). The two Vs of the bivalent domain antibody H The region may target the same or different antigens.

[0111] As used herein, the terms “bivalent antigen-binding protein” or “bivalent antibody” include 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 terms “multispecific antigen-binding protein” or “multispecific antibody” refer to an antigen-binding protein or antibody that targets multiple antigens or epitopes.

[0113] As used herein, the terms “bispecific,” “bispecific,” or “bifunctional” antigen-binding proteins or antibodies refer to hybrid antigen-binding proteins or antibodies having two distinct antigen-binding sites. Bispecific antibodies are multispecific antigen-binding proteins or multispecific antibodies that can be produced by a variety of methods, including but not limited to hybridoma fusion or Fab' fragment linking. 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 group of highly homologous antibodies, i.e., from a group of identical antibody molecules, excluding spontaneously occurring natural mutations. The monoclonal antibody is highly specific to a single epitope on an antigen. The polyclonal antibody generally contains at least two different antibodies that generally identify different epitopes on an antigen, compared to the monoclonal antibody. Monoclonal antibodies can generally be obtained by hybridoma technology, first reported by Kohler et al. (Nature, 256:495, 1975), and also by recombinant DNA technology (see, e.g., U.S. Patent 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 region of a human immunoglobulin (receptor antibody) is replaced by the CDR region 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 may also be replaced by amino acid residues of the corresponding 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, for example, Jonesetal, 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 the same art, “epitope” is also called an “antigenicity determinant.” The epitope or antigenicity determinant generally consists of a chemically active surface group of a molecule, such as an amino acid, carbohydrate, or sugar side chain, and usually has specific three-dimensional structural properties and specific charge properties. For example, the epitope generally contains at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 consecutive or discontinuous amino acids in a unique spatial conformation, which can be “linear” or “conformation.” 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 arranged linearly along the primary amino acid sequence of the protein. In a conformational epitope, the interaction sites are arranged across amino acid residues of proteins that are separated from each other.

[0117] The terms “polypeptide” or “protein” are used herein to refer to polymers of amino acid residues.

[0118] The terms described above are also used to refer to amino acid polymers, and naturally occurring amino acid polymers, in which one or more amino acid residues are analogs or mimetic versions of naturally occurring amino acids. The terms may also include, for example, amino acid polymers modified 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 polypeptides and proteins may be produced by genetically modified cells or recombinant cells, and include molecules having the amino acid sequence of a natural protein, or molecules having deletions, insertions and / or substitutions in one or more amino acids of the natural sequence.

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

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

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

[0122] As used herein, the term “isolated” means obtained from a natural state by artificial means. Where a particular “isolated” substance or component exists in nature, it may undergo changes in its natural environment, be isolated from its natural environment, or both. For example, a particular unisolated polynucleotide or polypeptide may exist naturally in a particular living animal, and a high-purity version of the same polynucleotide or polypeptide isolated from such a natural state is called 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 polynucleotides can be inserted. If a vector enables 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, thereby enabling the expression of elements of the genetic material carried by the vector in the host cell. Examples of 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 phages or M13 phages; 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, herpesviruses (such as herpes simplex virus), poxviruses, baculoviruses, papillomaviruses, and papovaviruses (such as SV40). A vector may include, but is not limited to, a promoter sequence, a transcription start sequence, an enhancer sequence, a selection element, and a reporter gene, as well as various other elements that control expression. Furthermore, a vector may further include a replication start site.

[0124] As used herein, the term “host cell” refers to a cell that can be introduced using a vector, including, but not limited to, prokaryotic cells such as Escherichia coli or Bacillus subtilis, fungal cells such as yeast cells or Aspergillus, insect cells such as Drosophila S2 cells or Sf9 cells, 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 the antigen it targets. In some embodiments, an antibody that specifically binds to an antigen (or an antibody that is specific to an antigen) is about 10 -5 Less than M, for example, about 10 -6 M, 10 -7 M, 10 -8 M, 10 -9 M, less than or 10 -10 Affinity (K) below M D ) refers to an antibody that binds to an antigen.

[0126] When used in this specification, the term "K D K is a dissociation equilibrium constant for a specific antibody-antigen interaction, used to describe the binding affinity between an antibody and an antigen. Of several parameters measured by molecular binding kinetics, K is a dissociation equilibrium constant for a specific antibody-antigen interaction. DThe 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 the target antibody and the target antigen molecule, and is expressed by the formula: K D =k dis / k on It is calculated by k. A smaller equilibrium dissociation constant indicates stronger antibody-antigen binding and higher affinity between the antibody and antigen. on (Association rate constant) is the rate of antigen-antibody complex formation, and a smaller k on This suggests that the antibody binding rate to the antigen is faster. dis The dissociation rate constant (k) is the rate at which an antibody dissociates from an antigen-antibody complex, and a smaller k is a constant. dis This suggests that the rate at which the antibody dissociates from the antigen is slower, and that the binding between the antibody and antigen is stronger. Generally, antibodies are measured, for example, by the Fortebio molecular interaction analyzer using biolayer interferometry (BLI) technology, at approximately 10 -5 Less than M, for example, about 10 -6 M, 10 -7 M, 10 -8 M, 10 -9 Less than M or 10 -10 Dissociation equilibrium constants (K) less than or equal to M D It binds to the antigen (for example, L1 protein) via [a specific mechanism].

[0127] As used herein, the term "EC 50 " refers to the effective concentration, which is 50% of the antibody's maximum response.

[0128] As used herein, the terms “monoclonal antibody” and “McAb” have the same meaning and are interchangeable; the terms “polyclonal antibody” and “PcAb” have the same meaning and are interchangeable. In addition, in this specification, amino acids are generally represented by one- and three-letter abbreviations known in the art. For example, alanine can be represented as A or Ala.

[0129] As used herein, the terms “percent sequence identity” and “percent sequence homology” are interchangeable.

[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, determined by aligning and comparing their sequences. “Percent identity” refers to the percentage of identical amino acid residues in the molecules being compared, which can be calculated based on the size of the smallest molecule being compared. For such calculations, any alignment gaps must be addressed by a specific mathematical model or computer program (i.e., an “algorithm”).

[0131] When used for polypeptides, the term “substantial identity” means, for example, that two peptide sequences, when optimally aligned using the default gap weights provided by the program and using the program GAP or BESTFIT, 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.

[0132] In some cases, the positions of non-identical residues differ due to conservative amino acid substitutions. A "conservative amino acid substitution" is when an amino acid residue is replaced by another amino acid residue having a side-chain R group with similar chemical properties (e.g., charge, hydrophilicity, or hydrophobicity). Generally, conservative amino acid substitutions substantially preserve the function and properties of the protein.

[0133] If two or more amino acid sequences differ from each other due to conservative substitutions, the percentage sequence identity may be increased to compensate for the conservative nature of the substitutions. Methods for making this adjustment are well known to those skilled in the art. See, for example, Pearson, Methods Mol. Biol., 243:307-31 (1994).

[0134] Examples of amino acid groups 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 substituents are valine-leucine-isoleucine-alanine-glycine, phenylalanine-tyrosine, lysine-arginine, threonine-serine, glutamic acid-aspartic acid, and asparagine-glutamine.

[0135] In some cases, a conservative permutation 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 “moderately conservative” permutation 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. This software matches sequences using similarity measures assigned to different substitutions, deletions, and other modifications (including conserved 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, for example, 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 alignment for the best overlap region 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 used when comparing sequences with databases containing large sequences from different biological sources is the computer program BLAST, particularly blasp or blasn (using the default parameters provided by the program). See, for example, Altschul et al., Mol. Biol., 215:403-410 (1990); Altschul et al., Nucleic Acids Res., 25:3389-402 (1997).

[0139] The term “treatment” or “to treat” usually refers to an operation to obtain a desired pharmacological and / or physiological effect. Such effect may be prophylactic in that it completely or partially prevents a disease or its symptoms; and / or may be therapeutic in that it partially or completely stabilizes or treats a disease and / or its side effects.

[0140] As used herein, “treatment” or “to treat” encompasses any treatment of a disease in a patient, including (a) preventing the occurrence of the disease or symptoms of the disease in a patient who is potentially susceptible to the disease but has not yet been diagnosed with the disease; (b) suppressing the symptoms of the disease, i.e., halting its progression; or (c) alleviating the symptoms of the disease, i.e., causing regression of the disease or symptoms.

[0141] As used herein, the term “general treatment” refers to a treatment in which a drug substance is transported through the bloodstream, reaches cells throughout the body, and exerts an effect.

[0142] As used herein, the term “systemic chemotherapy” refers to general chemotherapy, excluding chemotherapy for locally progressive disease as part of a multimodal treatment, wherein such chemotherapy for locally progressive disease includes induction chemotherapy, chemotherapy combined with radiotherapy, and adjuvant chemotherapy.

[0143] As used herein, the term “subject” (which may also be referred to herein as “patient”) refers to mammals such as rodents, cats, dogs, and primates. Preferably, the subject of the present invention is human.

[0144] "Administer," "administer," or "to administer" means physically introducing a composition containing a therapeutic agent into a subject using any of the various methods and delivery systems known to those skilled in the art. Routes of administration for inhibitors for autoimmune disease-related factors (e.g., anti-human IL-12 / IL-23p40 antibodies) include intravenous, intramuscular, subcutaneous, intraperitoneal, spinal, or other parenteral routes of administration, such as by injection or infusion. As used herein, the term "parenteral administration" refers to administration methods other than enteral and topical administration, typically by injection, and includes, but is not limited to, intravenous, intramuscular, intra-arterial, intrathecal, intralymphatic, intrafocal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subepidermal, intra-articular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injections and infusions and in vivo electroporation.

[0145] In certain embodiments, the inhibitor against autoimmune disease-related factors (e.g., anti-human IL-12 / IL-23p40 antibody) is administered via parenteral routes, and in some embodiments, the inhibitor is administered orally. Other parenteral routes include, for example, topical, epidermal, or mucosal administration routes such as intranasal, vaginal, rectal, sublingual, or local administration.

[0146] The administration may be carried out, for example, once, multiple times, and / or over one or more extension periods.

[0147] The term "subject" includes either a human or a non-human animal. The term "non-human animal" includes, but is not limited to, non-human primates, vertebrates such as sheep and 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” refer to values ​​or compositions within an acceptable margin of error for a particular value or composition as determined by those skilled in the art, and they depend in part on how such value or composition is measured or determined, i.e., on the limitations of the measuring system. For example, “about,” “approximately,” or “substantially” may mean being within one or more standard deviations practiced in the art. Alternatively, “about” or “substantially” may mean a range that differs by up to 10% or 20% (i.e., ±10% or ±20%) for the parameter or value it modifies. For example, about 3 mg may include any number between 2.7 mg and 3.3 mg (10%) or between 2.4 mg and 3.6 mg (20%).

[0149] Method of administration The following is not intended to limit the methods of administration of the drugs disclosed herein.

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

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

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

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

[0154] The agents 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 blocking the binding of the IL-12 / IL-23p40 protein domain to the cell surface receptors IL-12Rβ1 and IL-23R, inhibiting the activation of downstream signaling pathways of the human IL-12 / IL-23p40 protein domain, and inhibiting 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 may be used to prepare agents for the prevention and treatment of autoimmune diseases (e.g., psoriasis vulgaris or systemic lupus erythematosus) and ulcerative colitis (e.g., refractory or relapsing). Meanwhile, the antibodies possess good applicability and market value. [Brief explanation of the drawing]

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

[0159] Detailed explanation Embodiments of the present invention will be described in detail below with reference to examples. Those skilled in the art will understand that the following examples are for illustrative purposes only and should not be considered to limit the scope of the invention. Where the art or conditions are not specified, the examples were carried out in accordance with the art 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. The reagents or apparatus used are commercially available conventional products unless their manufacturer is specified.

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

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

[0162] In the following embodiments of the present invention, Ab123FR1, an anti-IL-12 / IL-23p40 antibody, is used as a control antibody, and its preparation method can be referred to in Chinese Registered Patent No. CN103275222B. This antibody was manufactured 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 embodiments of the present invention, ustekinumab (trademark Stellara), 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 manufactured by AkesoBiopharma, Inc. The cell lines 293T-IL-12Rβ1 and IL-23R are third-generation lentivirus systems (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). The cells were prepared by viral infection of 293 T cells using 72(11):8463-8471, where the lentiviral expression vectors used were pLenti-IL-12Rβ1-BSD (IL-12Rβ1, GenBank registry number 3549; vector pLenti-BSD, catalog number: K497000, purchased from Invitrogen) and pCDH-IL-23R-puro (IL-23R, GenBank registry number 149233; vector pCDH-CMV-MCS-EF1-Puro, product number VT1480, purchased from Yubio).

[0165] The control antibody for the aforementioned isotype is human anti-hen egg lysozyme IgG (anti-HEL, i.e., human IgG (abbreviated as hIgG1)), and its sequence is derived from "Affinity maturation enhances 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 described in SEQ ID NO: 21 and the amino acid sequence of the light chain is described in SEQ ID NO: 22). The control antibody for the aforementioned isotype was prepared in the laboratory of Akeso Biopharma.

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

[0167] The following examples are further illustrative of 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 the 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 to which the antibody binds to the antigen by quantum simulation calculations based on the structure of the IL-12 / IL-23p40 protein domain and three-dimensional spatial structure simulation techniques based on structural biology for antigen-antibody binding and interaction between the antibody's CDR region and the antigen. Meanwhile, without affecting the three-dimensional structure of the CDR region, the framework portion of the antibody was optimized accordingly, ultimately yielding antibodies H5L9, H5L10, H5L11, H5L12, H5L14, and H8L15 that specifically bind to human IL-12 / IL-23p40.

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

[0171] 1. Antibody expression and purification The nucleotide sequences encoding the heavy chain variable region (described in SEQ ID NO: 37; constant region: Igγ-1 chain C region; accession number P01857) and the light chain variable region (described in SEQ ID NO: 38; constant region: Igκ chain C region; accession number P01834) of the above antibody, such as H8L15, were cloned into the vector pUC Simple (provided by Genscript), respectively, to obtain pUC57 Simple-H8L15H containing the full-length heavy chain nucleotides of H8L15 and pUC57 Simple-H8L15L containing the full-length light chain nucleotides of H8L15, respectively.

[0172] Plasmids pUC57 simple-H8L15H and pUC57 simple-H8L15L were digested (HindIII & EcoRI), and the heavy and light chain nucleotide sequences isolated by electrophoresis were subcloned into the vector pcDNA3.1. Recombinant plasmids were then extracted and cotransfected into 293F cells.

[0173] After culturing transfected 293F cells for 7 days, the culture medium was centrifuged at high speed, the supernatant was concentrated, and packed into a HiTrap MabSelect SuRe column. The target sample was isolated by eluting the protein in a single step using an eluent. 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 the binding activity of antibodies H5L9, H5L10, H5L11, H5L12, H5L14, H8L15, Ab123FR1, and ustekinumab against 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 1% BSA solution in PBS. After blocking was complete, the plates were washed with PBST, tapped dry. Antibodies diluted in a concentration gradient with PBST solution were added to the plate wells, and the antibody dilution gradients are detailed in Table 1. Plates containing the test antibodies were incubated at 37°C for 30 minutes, then washed with PBST, tapped dry. A 1:5000 diluted HRP-labeled goat anti-human IgG(H+L) (purchased from Jackson ImmunoResearch Inc., catalog number: 109-035-088) secondary antibody action solution was added, and the resulting mixture was incubated at 37°C for 30 minutes. After incubation, the plates were washed with PBST and lightly tapped to dry. TMB (Neogen, 308177) was added and allowed to develop color in the absence of light for 5 minutes, then the color reaction was stopped by adding stop solution. The plates were then immediately placed in a plate reader, and the OD values ​​of each well in the plate were read at 450 nm. This data was analyzed using SoftMax Pro 6.2.1.

[0177] Figure 1 shows the detection results for the binding of antibodies H5L9 and Ab123FR1 to the human p40-His antigen. Table 1 shows the OD values ​​for all doses. Curve fitting with antibody concentration on the x-axis and absorbance on the y-axis was performed to determine the antibody binding EC. 50 The calculation is performed, and the results are shown in Table 1.

[0178] Figure 2 shows the detection results for the binding of antibodies H5L10 and Ab123FR1 to the human p40-His antigen. Table 2 shows the OD values ​​for all doses. Curve fitting with antibody concentration on the x-axis and absorbance on the y-axis was performed to determine the antibody binding EC. 50 The result is calculated and shown in Table 2.

[0179] Figure 3 shows the detection results of the binding of antibodies H5L11, H5L12, H5L14, and Ab123FR1 to the human p40-His antigen. Table 3 shows the OD values ​​for all doses. Curve fitting with antibody concentration on the x-axis and absorbance on the y-axis was performed to determine the antibody binding EC. 50 The result is calculated and shown in Table 3.

[0180] Figure 4 shows the detection results for the binding of antibodies H8L15 and Ab123FR1 to the human p40-His antigen. Table 4 shows the OD values ​​for all doses. Curve fitting with antibody concentration on the x-axis and absorbance on the y-axis was performed to determine the antibody binding EC. 50 The result is calculated and shown in Table 4.

[0181] The results above indicate that the binding efficiency of H5L9, H5L10, H5L11, H5L12, H5L14, and H8L15 to the human p40-His antigen is dose-dependent.

[0182] As shown in Figure 1 and Table 1, H5L9 has an EC of 0.079 nM in human p40-His. 50 It binds to Ab123FR1, and Ab123FR1 has an EC of 0.063 nM to human p40-His. 50 The binding occurs via this method. The binding efficiency of H5L9 was similar to that of Ab123FR1.

[0183] As shown in Figure 2 and Table 2, H5L10 has an EC of 0.057 nM in human p40-His. 50 It binds to Ab123FR1, and Ab123FR1 has an EC of 0.051 nM to human p40-His. 50 The binding occurs via this method. The binding efficiency of H5L10 was similar to that of Ab123FR1.

[0184] As shown in Figure 3 and Table 3, H5L11, H5L12, H5L14, and Ab123FR1 have EC values ​​of 0.082 nM, 0.082 nM, 0.107 nM, and 1.181 nM, respectively. 50 It binds to human p40-His at a specific value. The binding efficiencies of H5L11, H5L12, and H5L14 were clearly better than those of Ab123FR1.

[0185] As shown in Figure 4 and Table 4, H8L15, Ab123FR1, and ustekinumab have EC values ​​of 0.059 nM, 0.074 nM, and 0.077 nM, respectively. 50 It binds to human p40-His at a certain value. 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. Fortebio measurement of affinity constants of H5L9, H5L10, H5L11, H5L12, H5L14, H8L15, Ab123FR1 and ustekinumab for human IL-12 / IL-23p40 antigen.

[0191] The buffer used for diluting samples of 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 on the sensor was bound to the antibody at a concentration of 5-0.31 nM (2-fold dilution) for 120 seconds, after which the protein was 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 was analyzed by 1:1 model fitting to obtain affinity constants.

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

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

[0194] In terms of affinity, they were ranked from strongest to weakest as follows: 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 4.293 Detection of anti-human IL-12 / IL-23p40 antibodies that competitively block the binding of human IL-12 and IL-23 to T-IL-12Rβ1 and IL-23R cells by flow cytometry.

[0197] 1.1.293 Flow cytometry detection of antibodies H5L9, H5L10, H5L11, H5L12, H5L14, and ustekinumab that competitively block the binding of human IL-12 to T-IL-12Rβ1 and IL-23R cells. 293T-IL-12Rβ1 and IL-23R cells were digested using a standard method and divided into several samples, each containing 300,000 cells. 200 μL of 1% PBSA was added to each sample, and the mixture was centrifuged at 700 × g for 5 minutes, discarding the supernatant. According to the experimental design, a blank control was established by mixing the corresponding diluted antibody (maximum final concentration of 30 μg / mL, 3-fold dilution, total of 8 concentrations) and human IL-12 (Sino Biological, catalog number: CT-050-H08H-20) (final concentration of 20 nM) in a 1:1 ratio. The antibody and human IL-12 mixture was incubated on ice for 30 minutes, and then added to the cell precipitate at 100 μL / sample. The resulting mixture was thoroughly mixed 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, discarding the supernatant, and then washed twice. Alexa Fluor(R)488 anti-His tag antibody (Biolegend, catalog number: 652509) was diluted to a ratio of 1:400 and added to each tube in 100 μL. The mixture was thoroughly mixed 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 to resuspend the cells, which were then transferred to flow cytometry tubes for testing. The results for antibodies H5L9, H5L10, H5L11, H5L12, H5L14, and ustekinumab, which competitively blocked the binding of human IL-12 to 293T-IL-12Rβ1 and IL-23R cells detected by FACS, are shown in Table 6 and Figure 13.

[0198] As shown in Table 6 and Figure 13, H5L9, H5L10, H5L11, H5L12, H5L14, 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. Here, no significant competitive binding activity was observed for H5L9, and the EC of H5L10, H5L11, H5L12, H5L14, and ustekinumab was also shown. 50They were 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 the strength of competitively blocking the binding of IL-12 to IL-12Rβ1 on the cell membrane surface of 293T-IL-12Rβ1 and IL-23R, the antibodies were ranked from strong to weak as follows: H5L12, H5L10, ustekinumab, H5L11, H5L14, and H5L9.

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

[0201]

Table 6

[0202] 1.2. Detection by flow cytometry of antibodies Ab123FR1, H8L15, and ustekinumab that competitively block the binding of human IL-12 to 293T-IL-12Rβ1 and IL-23R cells The 293T-IL-12Rβ1 and IL-23R cells were digested in the usual way and divided into several samples each having 300,000 cells. 200 μL of 1% PBSA was added to each sample, and the mixture was centrifuged at 1200 rpm for 5 minutes to discard the supernatant. According to the experimental plan, the corresponding diluted antibodies (highest final concentration 60 μg / mL, 3-fold dilution, a total of 8 concentrations) and IL12-His (Sino Biological, catalog number: CT-050-H08H-20) (40 nM) were mixed at a ratio of 1:1, and a blank control was set. The mixture of the antibody and IL12 was incubated on ice for 30 minutes and then added to the cell pellet at 100 μL / sample. The resulting mixture was thoroughly mixed 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 to discard the supernatant and then washed twice. The THE™ His-tag antibody (FITC) (Genscript, catalog number: A01620) was diluted at a ratio of 1:500 and added to each tube at 100 μL. The mixture was thoroughly mixed 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 to discard the supernatant and then washed twice. 200 μL / tube of 1% PBSA was added to resuspend the cells, which were then transferred to flow cytometry tubes for testing. The results of the antibodies Ab123FR1, H8L15 and ustekinumab that competitively block the binding of human IL-12 to 293T-IL-12Rβ1 and IL-23R cells are shown in Table 7 and Figure 20. According to the results shown in Table 7 and Figure 20, all of Ab123FR1, H8L15 and ustekinumab can 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 values of Ab123FR1, H8L15 and ustekinumab are 1.99 μg / mL, 1.46 μg / mL and 1.58 μg / mL, respectively.

[0203] The results above indicate that the competitive binding activity of H8L15 in competitively blocking 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] 2.1.293 Detection by flow cytometry of antibodies H5L9, H5L10, H5L11, H5L12, H5L14, and ustekinumab that competitively block the binding of human IL-23 to T-IL-12Rβ1 and IL-23R cells. 293T-IL-12Rβ1 and IL-23R cells were digested using a standard method and divided into several samples, each containing 300,000 cells. 200 μL of 1% PBSA was added to each sample, and the mixture was centrifuged at 700 × g for 5 minutes, with the supernatant discarded. According to the experimental design, the corresponding diluted antibodies (maximum final concentration 30 μg / mL, 3-fold dilution, 8 concentrations in total) and human IL-23 (IL-23-His-biotin (Biotin) (Akesobio, lot number: 20161209)) (final concentration 2 μg / mL) were mixed in a 1:1 ratio to establish a blank control. The antibody and human IL23 mixtures were incubated on ice for 30 minutes, and then added to the cell precipitate at a rate of 100 μL / sample. The resulting mixtures were thoroughly mixed and incubated on ice for 60 minutes. 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. FITC steptavidin (Biolegend, catalog number: 405202) was diluted to a ratio of 1:500 and added to each tube in 100 μL. The mixture was thoroughly mixed 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. 1% PBSA was added at a rate of 200 μL / tube, the cells were resuspended, and transferred to flow cytometry tubes for testing. The results for 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 detected by FACS, are shown in Table 8 and Figure 14.

[0206] As shown in Table 8 and Figure 14, H5L9, H5L10, H5L11, H5L12, H5L14, and ustekinumab were all able 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, and the competitive binding EC of these antibodies 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 competitive blocking of 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 as follows: H5L10, H5L12, H5L11, H5L14, ustekinumab, and H5L9.

[0208] The results above indicate that the competitive binding activity of H5L10, H5L12, H5L11, and H5L14 in blocking 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] 2.2.293 Detection of antibodies Ab123FR1, H8L15, and ustekinumab that competitively block the binding of human IL-23 to T-IL-12Rβ1 and IL-23R cells by flow cytometry 293T-IL-12Rβ1 and IL-23R cells were digested using a standard method and divided into several samples, each containing 300,000 cells. 200 μL of 1% PBSA was added to each sample, and the mixture was centrifuged at 1200 rpm for 5 minutes, after which the supernatant was discarded. According to the experimental design, a blank control was established by mixing the corresponding diluted antibody (maximum concentration 60 μg / mL, 3-fold dilution, total of 8 concentrations) and human IL23-His-biotin (Akeso Biopharma, Inc., Lot No.: 20161209) (4 μg / mL) in a 1:1 ratio. The antibody and IL-His-biotin mixtures were incubated on ice for 30 minutes and then added to the cell precipitate at a rate of 100 μL / sample. The resulting mixtures were thoroughly mixed 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 stepavidin (Biolegend, catalog number: 405202) was diluted to a ratio of 1:500 and added to each tube in 100 μL. The mixtures were thoroughly mixed 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. 1% PBSA was added at a rate of 200 μL / tube to resuspend the cells, which were then transferred to flow cytometry tubes for testing. The results of FACS detection of antibodies Ab123FR1, H8L15, and ustekinumab, which competitively block the binding of human IL-23 to 293T-IL-12Rβ1 and IL-23R cells, are shown in Table 9 and Figure 21.

[0211] As 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. 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 results above indicate that the competitive binding activity of H8L15 in competitively blocking 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 spleen cells in mice with spontaneous systemic lupus erythematosus.

[0215] A model of spontaneous systemic lupus erythematosus (Jeltsch-David H. Autoimmun Rev. 2014;13(9):963-973.) mouse (MRL / lpr mouse purchased from Shanghai SLAC Laboratory Animal Co., Ltd.) was anesthetized with chloral hydrate, disinfected by immersion in 75% ethyl alcohol, transferred to a biosafety cabinet, and then dissected to collect the spleen. The spleen was washed in a dish containing 1640 complete medium to remove fat and fascial tissue. The washed mouse spleen was placed in a 70 μm cell strainer and gently crushed with a syringe plunger, and the spleen cell suspension was repeatedly washed with culture medium. The filtrate was collected and centrifuged at 170 × g for 5 minutes, and the supernatant was discarded. 7 mL of erythrocyte lysate was added to resuspend the cell precipitate. The mixture was gently and thoroughly mixed, left on ice for 8 minutes, and then an equal volume of complete medium was added to stop the dissolution. The mixture was centrifuged at 170 × g for 5 minutes, and the supernatant was discarded. The cell precipitate was centrifuged, washed twice with 1640 complete medium, then resuspended in 1640 complete medium, counted to adjust the cell density, and plated in a 96-well plate (1 × 10⁶). 6Seeds were seeded in 100 μL of solution. According to the experimental plan, 50 μL of antibody was pre-incubated with 50 μL of IL-23 (final concentration 20 ng / mL) for 1 hour, then 50 μL of IL-2 (final concentration 100 U / mL) was added, and the mixture was incubated in an incubator at 37°C / 5% CO2 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 is effective in promoting IL-17A secretion by spleen cells in a spontaneous systemic lupus erythematosus model mouse. When H8L15 is added while IL-23 is acting, IL-17A secretion is significantly inhibited, and this inhibitory activity is significantly dose-dependent, with pharmacodynamic activity being remarkably superior to that of AB123FR1 (sometimes called 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 Institute of Medicine) were randomly divided into a normal group, a model group, a positive control group, and an H8L15 group with 10 mice per group. One day before the first injection of recombinant human IL-23, an isotype control antibody (i.e., human anti-chicken egg lysozyme) was subcutaneously injected into the model group, the corresponding concentration of H8L15 was injected into the H8L15 administration group, and an equal volume of physiological saline was subcutaneously injected into the normal group. One day after administration, 3.5% chloral hydrate was intraperitoneally injected at a dose of 7.5 mL / kg to anesthetize C57BL / 6 mice. 25 μL / mouse of physiological saline was intradermally injected into the mice in the normal group, and 10 μg / 25 μL / mouse of recombinant human IL-23 was intradermally injected into the remaining mice. The injection was performed once a day for 6 consecutive days. On the second day after the final intradermal injection of recombinant human IL-23, the mice in each group were sacrificed by cervical dislocation, a small piece of cervical skin (about 0.5 cm × 0.5 cm) was excised, and fixed with formalin tissue fixative. Pathological sections of mouse skin were prepared 24 hours later for 6 mice in each group. After preparing the pathological sections, one representative field was selected under a 100× microscope, and 6 sites in the original image were randomly selected to measure the thickness of the skin epidermis of the mice. The data were expressed as mean ± standard error, and the results were evaluated by one-way analysis of variance after comparison between groups processed with GraphPad software. P < 0.05 indicated a significant difference, and P < 0.01 indicated a highly significant difference.

[0219] The results are shown in Fig. 16. Compared with the normal group, the thickness of the skin epidermis of the mice in the model group was significantly increased (P < 0.01). After administration, H8L15 was effective in inhibiting epidermal hyperplasia in psoriatic mice (P < 0.01).

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

[0221] Anti-IL-12 / IL-23p40 antibodies such as H8L15 have been found to be effective in reducing pathological changes and clinical symptoms in ulcerative colitis model mice. A colitis model was established by inducing C57BL / 6 mice with DSS (dextran sulfate sodium). Experimental mice were divided into a normal group of 3 and each of the other groups of 6. 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) were established. The drugs were administered subcutaneously on days D0, D3, and D6. In the normal group, the animal model was established by administering sterile water through a drinking bottle; in the DSS (MP Bio, catalog number: Q1723) group, an animal model was created by administering a 1% DSS solution (prepared by adding 2.5 g of DSS to 250 mL of sterile water) through a drinking bottle for 9 consecutive days; in the antibody-based experimental group, an animal model was established by administering a 1% DSS solution (prepared by adding 2.5 g of DSS to 250 mL of sterile water) through a drinking bottle and by intraperitoneal injection of recombinant hIL-23 protein (200 μL / 100 μg / mouse) daily (D1-D5, D7-D9).

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

[0223] The experimental indicators were used to examine the effects of the aforementioned drugs on colitis, and the specific indicators were based on the pathological score table for 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 method for establishing the mouse model and the antibody administration scheme in each experimental group, and Figure 17 shows the pathological scoring results for colitis. Figure 19 demonstrates the cutaneous histopathological observation for H8L15 treatment of DSS-induced colitis mice combined with recombinant human IL-23 (HE stained ×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).

[0227] Conclusion: As shown in Figure 18, the body weight of mice in the model group consistently decreased, while the body weight of mice in the treatment group did not decrease, which is clearly different from that of the model group.

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

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

[0230] The nucleotide sequences of the heavy chain variable regions H5L9, H5L10, H5L11, H5L12, and H5L14 are described in Sequence ID No. 2. [ka]

[0231] For H5L9, H5L10, H5L11, H5L12, and H5L14, HCDR1 is described in sequence number 3, HCDR2 is described in sequence number 4, and HCDR3 is described in sequence number 5. HCDR1:GYSFTTYW (Sequence ID 3) HCDR2:MSPVDSDI (Sequence ID 4) HCDR3:ARRRPGQGYFDF (Sequence ID 5)

[0232] The amino acid sequence of the light chain variable region of H5L9 is described in Sequence ID No. 6. [ka]

[0233] The nucleotide sequence of the light chain variable region of H5L9 is described in Sequence ID No. 7. [ka]

[0234] For H5L9, LCDR1 is described in sequence number 8, LCDR2 is described in sequence number 9, and LCDR3 is described in sequence number 10. LCD 1: QNVGSW (Sequence ID 8) LCDR2:ASS (Sequence ID 9) LCDR3:QQYDIYPFT(Sequence ID 10)

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

[0236] The nucleotide sequence of the light chain variable region of H5L10 is described in Sequence ID No. 12. [ka]

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

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

[0239] The amino acid sequence of the light chain variable region of H5L12 is described in Sequence ID No. 15. [ka]

[0240] The nucleotide sequence of the light chain variable region of H5L12 is described in Sequence ID No. 16. [ka]

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

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

[0243] L10's LCDR1 is described in sequence number 19. QSVGSW LCDR1 for L11, L12, and L14 is described in Sequence ID No. 20. QSVSSW LCDR2 for L10, L11, L12, and L14 is described in Sequence ID No. 21. ASN LCDR3 for L10, L11, and L12 is described in Sequence ID No. 22. QQYNIYPYT L14's LCDR3 is described in sequence number 23. QQYNIYPFT

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

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

[0246] For H8L15, HCDR1 is listed in sequence number 26, HCDR2 in sequence number 4, HCDR3 in sequence number 5, LCDR1 in sequence number 27, LCDR2 in sequence number 28, LCDR3 in sequence number 22, FR-H1 in sequence number 29, FR-H2 in sequence number 30, FR-H3 in sequence number 31, FR-H4 in sequence number 32, FR-L1 in sequence number 33, FR-L2 in sequence number 34, FR-L3 in sequence number 35, and FR-L4 in sequence number 36. HCDR1:GYTFTSYW (Sequence ID 26) HCDR2:MSPVDSDI (Sequence ID 4) HCDR3:ARRRPGQGYFDF (Sequence ID 5) LCDR1:QSVGTW (Sequence ID 27) LCDR2:A AS (Sequence ID 28) LCDR3:QQYNIYPYT(Sequence ID 22) FR-H1:EVQLVQSGAEVKKPGESLKISCQSS (Sequence ID 29) FR-H2:IGWVRQMPGQGLEWIGI(Sequence ID 30) FR-H3:RYNPMFRGQVTMSVDKSSSTAYLQWSSLKASDTAMYYC (Sequence ID 31) FR-H4:WGQGTMVTVSS (Sequence ID 32) FR-L1:EIVLTQSPATLSASPGERATISCRAS(Sequence ID 33) FR-L2:VAWYQQKPGQAPRSLIY(Sequence ID 34) FR-L3:NLQSGIPARFSGSGSGTDFTLTISSLEPEDFAVYYC(Sequence ID 35) FR-L4:FGQGTRLEIK (Sequence ID 36)

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

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

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

[0250] The light chain amino acid sequence of H8L15 is described in Sequence ID No. 40. [ka]

[0251] For H5L9, the sequences of FR-H1, FR-H2, FR-H3, and FR-H4 are the same as the sequences of H8L15. The sequence of FR-L1 is described in sequence number 41, the sequence of FR-L2 is described in sequence number 42, the sequence of FR-L3 is described in sequence number 43, and the sequence of FR-L4 is described in sequence number 44. FR-L1:DIQMTQSPSSLSASVGDRVTITCKAS(Sequence ID 41) FR-L2:LAWYQQKPGKAPKSLIYS (Sequence ID 42) FR-L3:RQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYC(Sequence ID 43) FR-L4:FGQGTKLEIK (Sequence ID 44)

[0252] For H5L10, the sequences of FR-H1, FR-H2, FR-H3, and FR-H4 are the same as the sequences of H8L15. The sequence of FR-L1 is described in sequence number 33, the sequence of FR-L2 is described in sequence number 45, the sequence of FR-L3 is described in sequence number 46, and the sequence of FR-L4 is described in sequence number 36. FR-L1:EIVLTQSPATLSASPGERATISCRAS(Sequence ID 33) FR-L2:LAWYQQKPGQAPRSLIYA (Sequence ID 45) FR-L3:LQSGIPARFSGSGSGTDFTLTISSLEPEDFAVYYC(Sequence ID 46) FR-L4:FGQGTRLEIK (Sequence ID 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 described in sequence number 33, the sequence of FR-L2 is described in sequence number 47, the sequence of FR-L3 is described in sequence number 46, and the sequence of FR-L4 is described in sequence number 36. FR-L1:EIVLTQSPATLSASPGERATISCRAS(Sequence ID 33) FR-L2:LAWYQQKPGQAPRSLIYS (Sequence ID 47) FR-L3:LQSGIPARFSGSGSGTDFTLTISSLEPEDFAVYYC(Sequence ID 46) FR-L4:FGQGTRLEIK (Sequence ID 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 described in sequence number 33, the sequence of FR-L2 is described in sequence number 45, the sequence of FR-L3 is described in sequence number 48, and the sequence of FR-L4 is described in sequence number 36. FR-L1:EIVLTQSPATLSASPGERATISCRAS(Sequence ID 33) FR-L2:LAWYQQKPGQAPRSLIYA (Sequence ID 45) FR-L3:RQSGIPARFSGSGSGTDFTLTISSLEPEDFAVYYC(Sequence ID 48) FR-L4:FGQGTRLEIK (Sequence ID 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 described in sequence number 33, the sequence of FR-L2 is described in sequence number 45, the sequence of FR-L3 is described in sequence number 46, and the sequence of FR-L4 is described in sequence number 36. FR-L1:EIVLTQSPATLSASPGERATISCRAS(Sequence ID 33) FR-L2:LAWYQQKPGQAPRSLIYA (Sequence ID 45) FR-L3:LQSGIPARFSGSGSGTDFTLTISSLEPEDFAVYYC(Sequence ID 46) FR-L4:FGQGTRLEIK (Sequence ID 36)

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

[0257] The light chain amino acid sequence of H5L9 is described in Sequence ID No. 50. [ka]

[0258] The light chain amino acid sequence of H5L10 is described in Sequence ID No. 51. [ka]

[0259] The light chain amino acid sequence of H5L11 is described in Sequence ID No. 52. [ka]

[0260] The light chain amino acid sequence of H5L12 is described in Sequence ID No. 53. [ka]

[0261] The light chain amino acid sequence of H5L14 is described in Sequence ID No. 54. [ka]

Claims

1. An antibody or an antigen-binding fragment thereof that specifically binds to human IL-12 / IL-23p40, wherein the antibody is: A heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity with SEQ ID NO: 1, comprising HCDR1 described in SEQ ID NO: 3, HCDR2 described in SEQ ID NO: 4, and HCDR3 described in SEQ ID NO: 5; and A light chain variable region comprising an amino acid sequence having at least 90% sequence identity with SEQ ID NO: 6, comprising LCDR1 described in SEQ ID NO: 8, LCDR2 described in SEQ ID NO: 9, and LCDR3 described in SEQ ID NO:

10. An antibody or its antigen-binding fragment, including an antibody.

2. The aforementioned antibody is: Heavy chain variable region containing the amino acid sequence described in Sequence ID No. 1; and Light chain variable region containing the amino acid sequence described in SEQ ID NO: 6 The antibody or antigen-binding fragment thereof according to claim 1, comprising:

3. The antibody or antigen-binding fragment thereof according to claim 1 or 2, further comprising a heavy chain constant region and a light chain constant region, wherein the constant regions are derived from a species other than mouse.

4. The antibody or antigen-binding fragment thereof according to claim 3, wherein the constant region is derived from human IgG.

5. The antibody or antigen-binding fragment thereof according to claim 3, wherein the constant region is derived from IgG1.

6. The antibody or antigen-binding fragment thereof according to claim 3, wherein the heavy chain constant region is the Igγ-1 chain C region; and the light chain constant region is the Igκ chain C region.

7. The antibody contains or consists of the heavy chain described in SEQ ID NO: 49 and the light chain described in SEQ ID NO:

50. The antibody or antigen-binding fragment thereof according to claim 1 or 2.

8. The antigen-binding fragment is Fab, Fab', F(ab') ２ An antibody or antigen-binding fragment thereof according to claim 1 or 2, selected from the group consisting of Fv, Fab / c, single-chain antibodies, and bivalent antibodies.

9. The antibody or antigen-binding fragment according to claim 1 or 2, wherein the antigen-binding fragment is scFv.

10. The antibody or antigen-binding fragment thereof according to claim 1 or 2, wherein the antibody is a humanized antibody, a chimeric antibody, or a multispecific antibody.

11. The antibody or antigen-binding fragment thereof according to claim 1 or 2, wherein the antibody is a bispecific antibody.

12. The aforementioned antibody is 10 －５ K less than M Ｄ An antibody or antigen-binding fragment thereof according to claim 1 or 2, which binds to the human IL-12 / IL-23p40 protein domain.

13. The antibody has an EC of less than 100 nM. ５０ An antibody or antigen-binding fragment thereof according to claim 1 or 2, which binds to the human IL-12 / IL-23p40 protein domain.

14. An isolated polynucleotide, wherein the polynucleotide includes or consists of the nucleotide sequence described in SEQ ID NO: 2 and the nucleotide sequence described in SEQ ID NO:

7. Polynucleotide.

15. A vector comprising the polynucleotide described in claim 14.

16. A host cell comprising the polynucleotide described in claim 14 or the vector described in claim 15.

17. A method for preparing an antibody or an antigen-binding fragment according to any one of claims 1 to 13, comprising culturing the host cells according to claim 16 under appropriate conditions, and isolating the antibody or an antigen-binding fragment thereof from the cell culture.

18. An antibody conjugate comprising an antibody or an antigen-binding fragment thereof according to any one of claims 1 to 13, and a conjugate portion bound thereto, wherein the conjugate portion is a purified tag, a cytotoxic drug, or a detectable label.

19. The antibody conjugate according to claim 18, wherein the conjugate portion is a His tag, a radioisotope, a luminescent substance, a coloring substance, an enzyme, or polyethylene glycol.

20. A fusion protein comprising an antibody or an antigen-binding fragment thereof according to any one of claims 1 to 13.

21. A multispecific antibody comprising the antibody or antigen-binding fragment thereof according to any one of claims 1 to 13.

22. A kit comprising an antibody or antigen-binding fragment thereof according to any one of claims 1 to 13, an antibody conjugate according to claim 18 or 19, a fusion protein according to claim 20, or a multispecific antibody according to claim 21.

23. The kit according to claim 22, further comprising a second antibody that specifically identifies the antibody or its antigen-binding fragment.

24. The kit according to claim 23, wherein the second antibody further comprises a detectable label.

25. The kit according to claim 24, wherein the detectable label is a radioisotope, a luminescent substance, a coloring substance, an enzyme, or polyethylene glycol.

26. Using an antibody or antigen-binding fragment thereof according to any one of claims 1 to 13, an antibody conjugate according to claim 18 or 19, a fusion protein according to claim 20, or a multispecific antibody according to claim 21 in the preparation of a kit for detecting the presence or level of human IL-12 / IL-23p40 protein domains in a sample.

27. A pharmaceutical composition comprising an antibody or antigen-binding fragment thereof according to any one of claims 1 to 13, an antibody conjugate according to claim 18 or 19, a fusion protein according to claim 20 or a multispecific antibody according to claim 21, and a pharmaceutically acceptable carrier and / or excipient.

28. The pharmaceutical composition according to claim 27, wherein the pharmaceutical composition is in a form suitable for administration by subcutaneous injection, intradermal injection, intravenous injection, intramuscular injection, or intralesional injection.

29. A drug that blocks the binding of the human IL-12 / IL-23p40 protein domain to human IL-12Rβ1 or human IL-23R. A drug for blocking the activity of the human IL-12 / IL-23p40 protein domain or for downregulating its level, Drugs for blocking cellular responses mediated by the binding of human IL-12Rβ1 or human IL-23R to the p40 protein domain. The preparation of the antibody or antigen-binding fragment thereof according to any one of claims 1 to 13, the antibody conjugate according to claim 18 or 19, the fusion protein according to claim 20, the multispecific antibody according to claim 21, or the pharmaceutical composition according to claim 27 or 28.

30. The use according to claim 29, wherein the ligand for the human IL-12 / IL-23p40 protein domain is human IL-12Rβ1 or human IL-23R.

31. Use of an antibody or antigen-binding fragment thereof according to any one of claims 1 to 13, an antibody conjugate according to claim 18 or 19, a fusion protein according to claim 20, a multispecific antibody according to claim 21, or a pharmaceutical composition according to claim 27 or 28 in the preparation of a drug for treating an autoimmune disease or ulcerative colitis.

32. The use according to claim 31, wherein the autoimmune disease is psoriasis vulgaris or systemic lupus erythematosus, or the ulcerative colitis is refractory or recurrent.

33. It blocks the binding of the human IL-12 / IL-23p40 protein domain to ligand IL-12Rβ1 or IL-23R. Downregulating the activity or level of the human IL-12 / IL-23p40 protein domain, and It blocks cellular responses mediated by the binding of human IL-12Rβ1 or human IL-23R to the p40 protein domain. A pharmaceutical composition according to claim 27 or 28 for the purpose of...

34. The pharmaceutical composition according to claim 33, wherein the ligand for IL-12 / IL-23p40 is IL-12Rβ1 or IL-23R.

35. An antibody or antigen-binding fragment thereof according to any one of claims 1 to 13, an antibody conjugate according to claim 18 or 19, a fusion protein according to claim 20, a multispecific antibody according to claim 21, or a pharmaceutical composition according to claim 27 or 28, for the prevention, treatment, adjuvant treatment and / or diagnosis of autoimmune diseases or ulcerative colitis.

36. The antibody or antigen-binding fragment thereof, antibody conjugate, fusion protein, multispecific antibody or pharmaceutical composition according to claim 35, wherein the autoimmune disease is psoriasis vulgaris or systemic lupus erythematosus, or the ulcerative colitis is refractory or recurrent.

37. An antibody or antigen-binding fragment thereof according to any one of claims 1 to 13, an antibody conjugate according to claim 18 or 19, a fusion protein according to claim 20, a multispecific antibody according to claim 21, or a pharmaceutical composition according to claim 27 or 28, for the prevention, treatment, adjuvant treatment and / or diagnosis of autoimmune disease or ulcerative colitis in subjects who have received conventional treatment or who are insufficiently responsive, unresponsive or intolerant to biological agents and have failed to achieve complete or partial response.

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