WO2023051798A1 - 抗il23抗体融合蛋白及用途 - Google Patents

抗il23抗体融合蛋白及用途 Download PDF

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WO2023051798A1
WO2023051798A1 PCT/CN2022/123370 CN2022123370W WO2023051798A1 WO 2023051798 A1 WO2023051798 A1 WO 2023051798A1 CN 2022123370 W CN2022123370 W CN 2022123370W WO 2023051798 A1 WO2023051798 A1 WO 2023051798A1
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seq
amino acid
acid sequence
antibody
variable region
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PCT/CN2022/123370
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French (fr)
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毛浪勇
应华
薛艺
金薪盛
陶维康
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江苏恒瑞医药股份有限公司
上海恒瑞医药有限公司
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Priority to CN202280059438.XA priority Critical patent/CN117916273A/zh
Priority to CA3233261A priority patent/CA3233261A1/en
Publication of WO2023051798A1 publication Critical patent/WO2023051798A1/zh

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/24—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against cytokines, lymphokines or interferons
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/46—Hybrid immunoglobulins
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K19/00—Hybrid peptides, i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/62—DNA sequences coding for fusion proteins

Definitions

  • the present disclosure belongs to the field of biotechnology, and more specifically, the present disclosure relates to an anti-IL23 antibody and its fusion protein and applications.
  • IL-23 (also known as IL23) is mainly produced by activated dendritic cells, macrophages and monocytes, and is a member of the IL-12 heterodimer cytokine family, produced by p19 (also known as IL23 p19 subunit) and p40 (also known as IL23 p40 subunit) two subunits, wherein the p40 subunit is a subunit shared with IL-12 (J Immunol.2018Sep 15; 201(6):1605-1613. ).
  • IL-23 exerts biological functions by interacting with its receptors and activating downstream signaling pathways.
  • IL-23 receptor includes two subunits of IL-12 receptor ⁇ 1 and IL-23 receptor (J Immunol.2002 Jun 1; 168(11):5699-708.).
  • IL-23 can promote the differentiation of Th17 cells, play an important role in the proliferation and stability of Th17 cells, and can promote Th17 cells to produce cytokines such as IL-17A, IL-17F and IL-22. These inflammatory factors act on keratinocytes, Causes keratinocyte activation and hyperproliferation.
  • Activated keratinocytes recruit and activate immune cells such as T cells by producing a large number of cytokines, chemokines, and antimicrobial peptides, forming a cascade of immune responses, causing symptoms of psoriasis (Cutis.2018 Mar; 101( 3S): 5-9.).
  • SLE Systemic lupus erythematosus
  • the source is neutrophils and apoptotic cells. Neutrophils release net-like extracellular traps, capture antibacterial proteins, and stimulate pDC to secrete a large amount of type I interferon under the combined action of interferon. Apoptotic cells will release high-mobility proteins and nucleic acids in the nucleus as their own antigens to induce immune cells to produce nucleotide antibodies.
  • the immune complexes of nucleic acids and antibodies interact with receptors on the cell surface to activate TLR signals.
  • IL-12 and 23 are two cytokines known to activate T cell differentiation. After activation, T cells secrete a variety of cytokines. Cytokines such as IL-17 participate in the recruitment of inflammatory cells and cause tissue damage, and cytokines such as IL-21 participate in the co-stimulation of B cells. BAFF and APRIL are activating factors of B cells. They stimulate B cells to mature and secrete antibodies by interacting with receptors on the surface of B cells (Nat Rev Rheumatol. 2016 Nov 22; 12(12):716-730.).
  • TACI is a membrane-bound receptor with an extracellular domain containing two cysteine-rich pseudo-repeats, a transmembrane domain, and CAML (calcium modulator and cyclophilic The cytoplasmic region where protein ligands interact.
  • TACI is associated with a subset of B cells and T cells.
  • the TACI receptor binds to BAFF of the tumor necrosis factor ligand family.
  • BAFF is a B cell activating factor belonging to the TNF family.
  • BAFF is mainly expressed on the surface of the bone marrow cell membrane and exists in the form of a trimer. The BAFF on the surface of the cell membrane will be hydrolyzed by protease to form soluble BAFF and enter the blood circulation system.
  • BAFF Characterized by multimerization up to 60-mers can be formed.
  • BAFF can also interact with another protein of the same family, APRIL, to form a heterologous trimer.
  • APRIL another protein of the same family
  • BAFF receptors there are three BAFF receptors on the surface of B cells, namely BAFF-R, BCMA and TACI. BAFF interacts with these three receptors and participates in the differentiation, maturation, survival and regulation of B cells.
  • APRIL and BAFF have two common receptors, namely BCMA and TACI, and APRIL interacts with these two receptors to participate in the survival and regulation of B cells (Samy, E., et al., Int Rev Immunol, 2017.36: p. 3-19; Kamal, A. and M.
  • BAFF is important for maintaining B cell homeostasis, and overactivation of BAFF signaling leads to the survival of self-reactive B cells and the production of autoantibodies to promote autoimmune responses (Cancro, M.P., D.P.D'Cruz, and M.A. Khamashta, J Clin Invest , 2009.119: p.1066-73).
  • the present disclosure constructs an anti-IL23 antibody fusion protein, which comprises an anti-IL23 antibody and a TACI polypeptide, wherein the anti-IL23 antibody specifically binds to human IL23 p19 subunit.
  • the anti-IL23 antibody fusion protein comprises or consists of an anti-IL23 antibody fused with a TACI polypeptide; preferably, the TACI polypeptide is contained in the anti-IL23 antibody
  • the N segment and/or C terminus of the heavy chain are fused with the anti-IL23 antibody.
  • the anti-IL23 antibody in the anti-IL23 antibody fusion protein comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3, wherein The light chain variable region comprises LCDR1, LCDR2 and LCDR3, wherein,
  • HCDR1, HCDR2 and HCDR3 of the heavy chain variable region respectively comprise the amino acid sequences of HCDR1, HCDR2 and HCDR3 in SEQ ID NO: 22, 21, 20, 19, 18, 17 or 1
  • the light LCDR1, LCDR2 and LCDR3 of the chain variable region comprise the amino acid sequence of LCDR1, LCDR2 and LCDR3 in SEQ ID NO: 26, 25, 24, 23 or 2, respectively;
  • HCDR1, HCDR2 and HCDR3 of the heavy chain variable region comprise the amino acid sequences of HCDR1, HCDR2 and HCDR3 in SEQ ID NO: 29, 30, 31, 32 or 3, respectively, and the light chain variable region
  • the LCDR1, LCDR2 and LCDR3 comprise the amino acid sequence of LCDR1, LCDR2 and LCDR3 in SEQ ID NO: 36, 35, 34, 33 or 4, respectively.
  • the HCDR1, HCDR2 and HCDR3 of the heavy chain variable region of the anti-IL23 antibody and the LCDR1, LCDR2 and LCDR3 of the light chain variable region are based on Defined by a numbering scheme selected from Kabat, IMGT, Chothia, AbM, and Contact.
  • HCDR1, HCDR2, and HCDR3 of the heavy chain variable region and LCDR1, LCDR2, and LCDR3 of the light chain variable region are defined according to the Kabat numbering convention; in some embodiments, the heavy chain can be HCDR1, HCDR2 and HCDR3 of the variable region and LCDR1, LCDR2 and LCDR3 of the light chain variable region are defined according to the IMGT numbering convention; in some embodiments, the HCDR1, HCDR2 and HCDR3 of the heavy chain variable region and the light chain LCDR1, LCDR2, and LCDR3 of the variable region are defined according to the Chothia numbering convention; defined by the AbM numbering convention; in some embodiments, the HCDR1, HCDR2, and HCDR3 of the heavy chain variable region and the LCDR1, LCDR2, and LCDR3 of the light chain variable region are defined according to the Contact numbering convention.
  • the anti-IL23 antibody comprises a heavy chain variable region and a light chain variable region, wherein:
  • HCDR1 of said heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 27, 28 or 5
  • HCDR2 comprises the amino acid sequence of SEQ ID NO: 6
  • HCDR3 comprises the amino acid sequence of SEQ ID NO: 7
  • LCDR1 of the light chain variable region comprises the amino acid sequence of SEQ ID NO: 8
  • LCDR2 comprises the amino acid sequence of SEQ ID NO: 9
  • LCDR3 comprises the amino acid sequence of SEQ ID NO: 10;
  • HCDR1 of the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 11
  • HCDR2 comprises the amino acid sequence of SEQ ID NO: 12
  • HCDR3 comprises the amino acid sequence of SEQ ID NO: 13
  • the light chain LCDR1 of the variable region comprises the amino acid sequence of SEQ ID NO: 14
  • LCDR2 comprises the amino acid sequence of SEQ ID NO: 37 or 15, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 16;
  • the anti-IL23 antibody comprises a heavy chain variable region and a light chain variable region, wherein
  • HCDR1 of the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 27
  • HCDR2 comprises the amino acid sequence of SEQ ID NO: 6
  • HCDR3 comprises the amino acid sequence of SEQ ID NO: 7
  • the light chain LCDR1 of the variable region comprises the amino acid sequence of SEQ ID NO: 8
  • LCDR2 comprises the amino acid sequence of SEQ ID NO: 9
  • LCDR3 comprises the amino acid sequence of SEQ ID NO: 10;
  • HCDR1 of the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 28
  • HCDR2 comprises the amino acid sequence of SEQ ID NO: 6
  • HCDR3 comprises the amino acid sequence of SEQ ID NO: 7
  • the light chain LCDR1 of the variable region comprises the amino acid sequence of SEQ ID NO: 8
  • LCDR2 comprises the amino acid sequence of SEQ ID NO: 9
  • LCDR3 comprises the amino acid sequence of SEQ ID NO: 10;
  • HCDR1 of the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 11
  • HCDR2 comprises the amino acid sequence of SEQ ID NO: 12
  • HCDR3 comprises the amino acid sequence of SEQ ID NO: 13
  • the light chain LCDR1 of the variable region comprises the amino acid sequence of SEQ ID NO:14
  • LCDR2 comprises the amino acid sequence of SEQ ID NO:37
  • LCDR3 comprises the amino acid sequence of SEQ ID NO:16.
  • the anti-IL23 antibody is a murine antibody, a chimeric antibody or a humanized antibody. In some embodiments, the antibody is a humanized antibody.
  • the anti-IL23 antibody comprises a heavy chain variable region and a light chain variable region, wherein:
  • said heavy chain variable region comprises any of SEQ ID NO: 22, 21, 20, 19, 18 or 17 or at least 90% thereof (e.g. at least 90%, 95%, 96%, 97% , 98% or 99%) sequence identity of amino acid sequences
  • said light chain variable region comprises any of SEQ ID NO: 26, 25, 24 or 23 or has at least 90% thereof (for example at least 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence identity; or
  • said heavy chain variable region comprises any of SEQ ID NO: 29, 30, 31 or 32 or at least 90% thereof (e.g. at least 90%, 95%, 96%, 97%, 98% or 99%) sequence identity amino acid sequence
  • said light chain variable region comprises any of SEQ ID NO: 36, 35, 34 or 33 or has at least 90% (such as at least 90%, 95%, 96%) therewith %, 97%, 98% or 99%) sequence identity of amino acid sequences; or
  • said heavy chain variable region comprises SEQ ID NO: 1 or an amino acid sequence having at least 90% (e.g., at least 90%, 95%, 96%, 97%, 98% or 99%) sequence identity thereto, and said light chain variable region comprising SEQ ID NO: 2 or an amino acid sequence having at least 90% (e.g., at least 90%, 95%, 96%, 97%, 98% or 99%) sequence identity thereto; or
  • said heavy chain variable region comprises SEQ ID NO: 3 or an amino acid sequence having at least 90% (e.g., at least 90%, 95%, 96%, 97%, 98% or 99%) sequence identity thereto, and said light chain variable region comprising SEQ ID NO: 4 or an amino acid sequence having at least 90% (eg, at least 90%, 95%, 96%, 97%, 98% or 99%) sequence identity thereto.
  • the framework region of the heavy chain variable region of the anti-IL23 antibody comprises One or more amino acid mutations in positions 89 and 93 (numbering according to the Kabat numbering system), and/or a frame region of the light chain variable region comprising positions selected from positions 4, 17, 36, 58, 60 and 68 ( One or more amino acid mutations in numbering according to the Kabat numbering system).
  • the HCDR1 of the heavy chain variable region of the anti-IL23 antibody comprises the amino acid sequence of SEQ ID NO: 27, 28 or 5, and the HCDR2 comprises the amino acid sequence of SEQ ID NO: 6 Amino acid sequence, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 7, and the LCDR1 of said light chain variable region comprises the amino acid sequence of SEQ ID NO: 8, LCDR2 comprises the amino acid sequence of SEQ ID NO: 9, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 9, and LCDR3 comprises the amino acid sequence of SEQ ID NO: The amino acid sequence of ID NO: 10; the framework region of the heavy chain variable region of the antibody comprises one selected from the group consisting of 1E, 30T, 37V, 44G, 49G, 73N, 89R and 93V (numbering according to the Kabat numbering system) or more amino acid mutations, and/or the framework region of the light chain variable region comprises one or more amino acids selected from
  • the framework region of the heavy chain variable region of the anti-IL23 antibody comprises a One or more amino acid mutations in (numbering according to the Kabat numbering system), and/or inclusions selected from positions 4, 36, 39, 71 and 100 (numbering according to the Kabat numbering system) on the framework region of the light chain variable region One or more amino acid mutations in .
  • the HCDR1 of the heavy chain variable region of the anti-IL23 antibody comprises the amino acid sequence of SEQ ID NO: 11
  • the HCDR2 comprises the amino acid sequence of SEQ ID NO: 12
  • HCDR3 comprises the amino acid sequence of SEQ ID NO: 13
  • LCDR1 of the light chain variable region comprises the amino acid sequence of SEQ ID NO: 14
  • LCDR2 comprises the amino acid sequence of SEQ ID NO: 37 or 15, and LCDR3 comprises the amino acid sequence of SEQ ID NO : the amino acid sequence of 16
  • the framework region of the heavy chain variable region of the antibody comprises one or more amino acid mutations selected from the group consisting of 1E, 2F, 28S, 44S, 71V and 73Q (numbering according to the Kabat numbering system)
  • the framework region of the light chain variable region comprises one or more amino acid mutations selected from 4I, 36F, 39R, 71L and 100S (numbered according to the Kabat numbering system).
  • the anti-IL23 antibody comprises a heavy chain variable region and a light chain variable region, wherein,
  • said heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 22, 21, 20, 19, 18 or 17, and said light chain variable region comprises SEQ ID NO: 26, 25, 24 or 23 the amino acid sequence of; or
  • said heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 29, 30, 31 or 32, and said light chain variable region comprises the amino acid sequence of SEQ ID NO: 36, 35, 34 or 33; or
  • said heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 1, and said light chain variable region comprises the amino acid sequence of SEQ ID NO: 2;
  • said heavy chain variable region comprises the amino acid sequence of SEQ ID NO:3
  • said light chain variable region comprises the amino acid sequence of SEQ ID NO:4.
  • the anti-IL23 antibody comprises a heavy chain variable region and a light chain variable region, wherein,
  • said heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 22, and said light chain variable region comprises the amino acid sequence of SEQ ID NO: 26;
  • said heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 21, and said light chain variable region comprises the amino acid sequence of SEQ ID NO: 25;
  • said heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 29 and said light chain variable region comprises the amino acid sequence of SEQ ID NO: 36.
  • the anti-IL23 antibody further comprises an antibody heavy chain constant region and a light chain constant region.
  • the heavy chain constant region is a human IgG heavy chain constant region; in some embodiments, the heavy chain constant region is selected from human IgG1, IgG2, IgG3 and IgG4 constant regions; in some embodiments, The light chain constant region is selected from a human antibody ⁇ or ⁇ chain constant region; in some embodiments, the heavy chain constant region is a human IgG1 heavy chain constant region, and the light chain constant region is a human ⁇ light chain constant region.
  • the Fc region of the heavy chain constant region has one or more amino acid substitutions that reduce binding of the Fc region to an Fc receptor.
  • the Fc region has L234A, L235A mutations, and/or S228P mutations, and/or YTE mutations (M252Y, S254T, and T256E), and the numbering of the mutations is based on the EU index.
  • the heavy chain constant region comprises the amino acid sequence of SEQ ID NO: 38
  • the light chain constant region comprises the amino acid sequence of SEQ ID NO: 39.
  • the anti-IL23 antibody comprises a heavy chain and a light chain, wherein,
  • the heavy chain of said anti-IL23 antibody comprises an amino acid sequence having at least 90% (e.g., at least 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO: 42, and said light chain comprises an amino acid sequence having at least 90% (e.g., at least 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO: 43; or
  • the heavy chain of said anti-IL23 antibody comprises an amino acid sequence having at least 90% (e.g., at least 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO: 40, and said light chain comprises an amino acid sequence having at least 90% (e.g., at least 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO: 41; or
  • the heavy chain of the anti-IL23 antibody comprises an amino acid sequence having at least 90% (e.g., at least 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO: 44
  • said light chain comprises an amino acid sequence having at least 90% (eg, at least 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO: 45.
  • the anti-IL23 antibody comprises a heavy chain and a light chain, wherein,
  • the heavy chain of the anti-IL23 antibody comprises the amino acid sequence of SEQ ID NO: 42, and the light chain comprises the amino acid sequence of SEQ ID NO: 43;
  • the heavy chain of the anti-IL23 antibody comprises the amino acid sequence of SEQ ID NO: 40, and the light chain comprises the amino acid sequence of SEQ ID NO: 41;
  • the heavy chain of the anti-IL23 antibody comprises the amino acid sequence of SEQ ID NO:44, and the light chain comprises the amino acid sequence of SEQ ID NO:45.
  • the TACI polypeptide has a better function of preventing fragmentation.
  • the TACI polypeptide is a polypeptide comprising the 48th to 85th amino acid residues of SEQ ID NO: 58 or a variant thereof; wherein, The variant has amino acid substitutions at one or more positions selected from positions 49, 52, 53, 57, 65, 82 and 83, and the position of the amino acid substitution is relative to the sequence SEQ ID NO : 58 natural sequence numbered amino acid residue positions.
  • the variant of the TACI polypeptide has one or more amino acid substitutions selected from the group consisting of 49T or 49R, 52S, 53E or 53Q, 57E, 65T or 65A, 82A or 82R, and 83Y , the amino acid replacement site is the amino acid residue site numbered relative to the natural sequence of the sequence SEQ ID NO:58.
  • the TACI polypeptide is shown in SEQ ID NO: 58 or a truncated fragment of SEQ ID NO: 58 or a variant thereof; wherein The truncated fragment comprises the 48th to the 85th amino acid residues of SEQ ID NO: 58, and the variant has an amino acid residue selected from the group consisting of 49, 52, 53, 57, 65 on SEQ ID NO: 58 or a truncated fragment thereof. , one or more amino acid substitutions in positions 82 and 83, wherein the position of the amino acid substitution is an amino acid residue position numbered relative to the natural sequence of the sequence SEQ ID NO:58.
  • the truncated fragment of the TACI polypeptide comprises: the 48th to the 86th amino acid residues of SEQ ID NO: 58; SEQ ID NO: The 48th to the 87th amino acid residue of 58; or the 48th to the 88th amino acid residue of SEQ ID NO:58.
  • the sequence of the TACI polypeptide is shown in any one of SEQ ID NO: 60-63.
  • the sequence of the TACI polypeptide is a variant of SEQ ID NO: 58 or a truncated fragment of SEQ ID NO: 58 (such as SEQ ID NO: 60.
  • the TACI polypeptide is in SEQ ID NO: 58, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62 or SEQ ID NO: 63 sequence has one or more amino acid substitutions selected from the group consisting of 49T or 49R, 52S, 53E or 53Q, 57E, 65T or 65A, 82A or 82R, and 83Y (for example, 1, 2 , 3, 4, 5, 6 or 7 amino acid substitutions), wherein the amino acid substitution position is the amino acid residue position numbered relative to the natural sequence of the sequence SEQ ID NO:58.
  • the TACI polypeptide is: in SEQ ID NO: 58, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62 or On the basis of the sequence of SEQ ID NO: 63, there is any amino acid substitution selected from 49T, 52S, 53E, 53Q, 57E and 82A; in SEQ ID NO: 58, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 61, SEQ ID NO: ID NO: 62 or SEQ ID NO: 63 based on the sequence has 49R and 65T amino acid substitutions; in SEQ ID NO: 58, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62 or SEQ ID NO: 45R and 65A amino acid substitutions on the basis of the 63 sequence; 49R, 65T on the basis of the sequence of SEQ ID NO: 58, SEQ ID NO: 60,
  • SEQ ID NO: 61, SEQ ID NO: 62 or SEQ ID NO: 63 there are 49T and 82A amino acid substitutions; in SEQ ID NO: 58, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 61, SEQ ID NO: 63 ID NO: 62 or SEQ ID NO: 63 sequence based on 49T and 83Y amino acid substitutions; in SEQ ID NO: 58, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62 or SEQ ID NO: 49T, 82A and 83Y amino acid substitutions on the basis of the 63 sequence; or on the basis of the sequence of SEQ ID NO: 58, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62 or SEQ ID NO: 63 49T, 53E, 57E and 82A amino acid substitutions.
  • the aforementioned amino acid replacement site is the amino acid residue site numbered relative to the natural sequence
  • the amino acid sequence of the TACI polypeptide is as shown in any one of SEQ ID NO:51 to 83.
  • the amino acid sequence of the TACI polypeptide is as shown in any one of SEQ ID NO: 60-63 and SEQ ID NO: 66-83.
  • the amino acid sequence of the TACI polypeptide is shown in SEQ ID NO: 83.
  • the anti-IL23 antibody fusion protein as described in any one of the above comprises:
  • Second chain light chain of anti-IL23 antibody
  • the TACI polypeptide 1 and the TACI polypeptide 2 are the same or not, optionally, the TACI polypeptide 1 and the TACI polypeptide 2 can be independently selected from any TACI polypeptide described herein; the linker 1 and linker 2 are the same or different; in some embodiments, the linker is selected from (G x S) y linker, wherein, x is selected from an integer of 1-5, and y is selected from 0-6 In some embodiments, the linker is selected from (G x S) y linker, wherein, x is selected from the integer of 1-5, y is selected from the integer of 1-6, in some embodiments , the linker is GGGGSGGGGSGGGGS (as shown in SEQ ID NO: 84) or GGGS (as shown in SEQ ID NO: 85).
  • the anti-IL23 antibody fusion protein has: a first chain comprising the amino acid sequence of SEQ ID NO: 47, and comprising SEQ ID NO: 43
  • the second chain of the amino acid sequence; or the anti-IL23 antibody fusion protein has: the first chain comprising the amino acid sequence of SEQ ID NO: 46, and the second chain comprising the amino acid sequence of SEQ ID NO: 41.
  • the anti-IL23 antibody fusion protein has 2 first chains comprising the amino acid sequence of SEQ ID NO: 47, and 2 second chains comprising the amino acid sequence of SEQ ID NO: 43; in some In one embodiment, the anti-IL23 antibody fusion protein has two first chains comprising the amino acid sequence of SEQ ID NO: 46, and two second chains comprising the amino acid sequence of SEQ ID NO: 41.
  • the present disclosure provides an anti-IL23 antibody, wherein the anti-IL23 antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3, the The light chain variable region comprises LCDR1, LCDR2 and LCDR3, wherein,
  • HCDR1, HCDR2 and HCDR3 of the heavy chain variable region respectively comprise the amino acid sequences of HCDR1, HCDR2 and HCDR3 in SEQ ID NO: 22, 21, 20, 19, 18, 17 or 1
  • the light LCDR1, LCDR2 and LCDR3 of the chain variable region comprise the amino acid sequence of LCDR1, LCDR2 and LCDR3 in SEQ ID NO: 26, 25, 24, 23 or 2, respectively;
  • HCDR1, HCDR2 and HCDR3 of the heavy chain variable region comprise the amino acid sequences of HCDR1, HCDR2 and HCDR3 in SEQ ID NO: 29, 30, 31, 32 or 3, respectively, and the light chain variable region
  • the LCDR1, LCDR2 and LCDR3 comprise the amino acid sequence of LCDR1, LCDR2 and LCDR3 in SEQ ID NO: 36, 35, 34, 33 or 4, respectively.
  • HCDR1, HCDR2 and HCDR3 of the heavy chain variable region and the LCDR1, LCDR2 and LCDR3 of the light chain variable region are selected from Kabat, IMGT , Chothia, AbM and Contact numbering rules defined.
  • HCDR1, HCDR2, and HCDR3 of the heavy chain variable region and LCDR1, LCDR2, and LCDR3 of the light chain variable region are defined according to the Kabat numbering convention; in some embodiments, the heavy chain can be HCDR1, HCDR2 and HCDR3 of the variable region and LCDR1, LCDR2 and LCDR3 of the light chain variable region are defined according to the IMGT numbering convention; in some embodiments, the HCDR1, HCDR2 and HCDR3 of the heavy chain variable region and the light chain LCDR1, LCDR2, and LCDR3 of the variable region are defined according to the Chothia numbering convention; defined by the AbM numbering convention; in some embodiments, the HCDR1, HCDR2, and HCDR3 of the heavy chain variable region and the LCDR1, LCDR2, and LCDR3 of the light chain variable region are defined according to the Contact numbering convention.
  • HCDR1 of said heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 27, 28 or 5
  • HCDR2 comprises the amino acid sequence of SEQ ID NO: 6
  • HCDR3 comprises the amino acid sequence of SEQ ID NO: 7
  • LCDR1 of the light chain variable region comprises the amino acid sequence of SEQ ID NO: 8
  • LCDR2 comprises the amino acid sequence of SEQ ID NO: 9
  • LCDR3 comprises the amino acid sequence of SEQ ID NO: 10;
  • HCDR1 of the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 11
  • HCDR2 comprises the amino acid sequence of SEQ ID NO: 12
  • HCDR3 comprises the amino acid sequence of SEQ ID NO: 13
  • the light chain LCDR1 of the variable region comprises the amino acid sequence of SEQ ID NO: 14
  • LCDR2 comprises the amino acid sequence of SEQ ID NO: 37 or 15, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 16;
  • HCDR1 of the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 27, and HCDR2 comprises the amino acid sequence of SEQ ID NO: 6, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 7, and LCDR1 of the light chain variable region comprises the amino acid sequence of SEQ ID NO: 8, LCDR2 comprises the amino acid sequence of SEQ ID NO: 9, and LCDR3 comprises the amino acid sequence of SEQ ID NO: the amino acid sequence of 10; or
  • HCDR1 of the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 28
  • HCDR2 comprises the amino acid sequence of SEQ ID NO: 6
  • HCDR3 comprises the amino acid sequence of SEQ ID NO: 7
  • the light chain LCDR1 of the variable region comprises the amino acid sequence of SEQ ID NO: 8
  • LCDR2 comprises the amino acid sequence of SEQ ID NO: 9
  • LCDR3 comprises the amino acid sequence of SEQ ID NO: 10;
  • HCDR1 of the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 11
  • HCDR2 comprises the amino acid sequence of SEQ ID NO: 12
  • HCDR3 comprises the amino acid sequence of SEQ ID NO: 13
  • the light chain LCDR1 of the variable region comprises the amino acid sequence of SEQ ID NO:14
  • LCDR2 comprises the amino acid sequence of SEQ ID NO:37
  • LCDR3 comprises the amino acid sequence of SEQ ID NO:16.
  • the anti-IL23 antibody is a murine antibody, a chimeric antibody or a humanized antibody. In some embodiments, the antibody is a humanized antibody.
  • the anti-IL23 antibody comprises a heavy chain variable region and a light chain variable region, wherein:
  • said heavy chain variable region comprises any of SEQ ID NO: 22, 21, 20, 19, 18 or 17 or at least 90% thereof (e.g. at least 90%, 95%, 96%, 97% , 98% or 99%) sequence identity of amino acid sequences
  • said light chain variable region comprises any of SEQ ID NO: 26, 25, 24 or 23 or has at least 90% thereof (for example at least 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence identity; or
  • said heavy chain variable region comprises any of SEQ ID NO: 29, 30, 31 or 32 or at least 90% thereof (e.g. at least 90%, 95%, 96%, 97%, 98% or 99%) sequence identity amino acid sequence
  • said light chain variable region comprises any of SEQ ID NO: 36, 35, 34 or 33 or has at least 90% (such as at least 90%, 95%, 96%) therewith %, 97%, 98% or 99%) sequence identity of amino acid sequences; or
  • said heavy chain variable region comprises SEQ ID NO: 1 or an amino acid sequence having at least 90% (e.g., at least 90%, 95%, 96%, 97%, 98% or 99%) sequence identity thereto, and said light chain variable region comprising SEQ ID NO: 2 or an amino acid sequence having at least 90% (e.g., at least 90%, 95%, 96%, 97%, 98% or 99%) sequence identity thereto; or
  • said heavy chain variable region comprises SEQ ID NO: 3 or an amino acid sequence having at least 90% (e.g., at least 90%, 95%, 96%, 97%, 98% or 99%) sequence identity thereto, and said light chain variable region comprising SEQ ID NO: 4 or an amino acid sequence having at least 90% (eg, at least 90%, 95%, 96%, 97%, 98% or 99%) sequence identity thereto.
  • the framework region of the heavy chain variable region of the anti-IL23 antibody comprises One or more amino acid mutations in position 93 (numbering according to Kabat numbering system), and/or inclusions selected from positions 4, 17, 36, 58, 60 and 68 (according to Kabat numbering system) on the framework region of the light chain variable region One or more amino acid mutations in numbering system numbering).
  • HCDR1 of the heavy chain variable region of the anti-IL23 antibody comprises the amino acid sequence of SEQ ID NO: 27, 28 or 5
  • HCDR2 comprises the amino acid sequence of SEQ ID NO: 6
  • HCDR3 comprises the amino acid sequence of SEQ ID NO: 6
  • the amino acid sequence of NO: 7 and the LCDR1 of the light chain variable region comprises the amino acid sequence of SEQ ID NO: 8
  • the LCDR2 comprises the amino acid sequence of SEQ ID NO: 9
  • the LCDR3 comprises the amino acid sequence of SEQ ID NO: 10; comprising one or more amino acid mutations selected from the group consisting of 1E, 30T, 37V, 44G, 49G, 73N, 89R and 93V (numbered according to the Kabat numbering system) on the framework region of the heavy chain variable region of the antibody, and /or the framework region of the light chain variable region comprises one or more amino acid mutations selected from 4L, 17Q, 36F, 58I, 60A and 68R (numbering according to the
  • the framework region of the heavy chain variable region of the anti-IL23 antibody comprises positions selected from positions 1, 2, 28, 44, 71 and 73 (according to One or more amino acid mutations in the Kabat numbering system numbering), and/or the framework region of the light chain variable region contains an amino acid selected from positions 4, 36, 39, 71 and 100 (numbering according to the Kabat numbering system) One or more amino acid mutations.
  • HCDR1 of the heavy chain variable region of the anti-IL23 antibody comprises the amino acid sequence of SEQ ID NO: 11
  • HCDR2 comprises the amino acid sequence of SEQ ID NO: 12
  • HCDR3 comprises the amino acid sequence of SEQ ID NO: 13
  • Amino acid sequence and the LCDR1 of the light chain variable region comprises the amino acid sequence of SEQ ID NO: 14, the LCDR2 comprises the amino acid sequence of SEQ ID NO: 37 or 15, and the LCDR3 comprises the amino acid sequence of SEQ ID NO: 16;
  • the heavy chain variable region of the antibody comprises one or more amino acid mutations selected from the group consisting of 1E, 2F, 28S, 44S, 71V and 73Q (numbered according to the Kabat numbering system), and/or the light chain
  • the framework region of the variable region comprises one or more amino acid mutations selected from 4I, 36F, 39R, 71L and 100S (numbered according to the Kabat numbering system).
  • the anti-IL23 antibody of any one of the above comprises a heavy chain variable region and a light chain variable region, wherein,
  • said heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 22, 21, 20, 19, 18 or 17, and said light chain variable region comprises SEQ ID NO: 26, 25, 24 or 23 the amino acid sequence of; or
  • said heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 29, 30, 31 or 32, and said light chain variable region comprises the amino acid sequence of SEQ ID NO: 36, 35, 34 or 33; or
  • said heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 1, and said light chain variable region comprises the amino acid sequence of SEQ ID NO: 2;
  • said heavy chain variable region comprises the amino acid sequence of SEQ ID NO:3
  • said light chain variable region comprises the amino acid sequence of SEQ ID NO:4.
  • the anti-IL23 antibody of any one of the above comprises a heavy chain variable region and a light chain variable region, wherein,
  • said heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 22, and said light chain variable region comprises the amino acid sequence of SEQ ID NO: 26;
  • said heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 21, and said light chain variable region comprises the amino acid sequence of SEQ ID NO: 25;
  • said heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 29 and said light chain variable region comprises the amino acid sequence of SEQ ID NO: 36.
  • the anti-IL23 antibody further comprises an antibody heavy chain constant region and a light chain constant region.
  • the heavy chain constant region is a human IgG heavy chain constant region; in some embodiments, the heavy chain constant region is selected from human IgG1, IgG2, IgG3 and IgG4 constant regions; in some embodiments, The light chain constant region is selected from a human antibody ⁇ or ⁇ chain constant region; in some embodiments, the heavy chain constant region is a human IgG1 heavy chain constant region, and the light chain constant region is a human ⁇ light chain constant region.
  • the Fc region of the heavy chain constant region has one or more amino acid substitutions that reduce binding of the Fc region to an Fc receptor.
  • the Fc region has L234A, L235A mutations, and/or S228P mutations, and/or YTE mutations (M252Y, S254T, and T256E), and the numbering of the mutations is based on the EU index.
  • the heavy chain constant region comprises the amino acid sequence of SEQ ID NO: 38
  • the light chain constant region comprises the amino acid sequence of SEQ ID NO: 39.
  • the anti-IL23 antibody of any of the above comprises a heavy chain and a light chain, wherein,
  • the heavy chain of said anti-IL23 antibody comprises an amino acid sequence having at least 90% (e.g., at least 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO: 42, and a light chain comprising an amino acid sequence having at least 90% (e.g., at least 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO: 43; or
  • the heavy chain of said anti-IL23 antibody comprises an amino acid sequence having at least 90% (e.g., at least 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO: 40, and a light chain comprising an amino acid sequence having at least 90% (e.g., at least 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO: 41; or
  • the heavy chain of the anti-IL23 antibody comprises an amino acid sequence having at least 90% (e.g., at least 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO: 44, and the light chain comprise an amino acid sequence having at least 90% (eg, at least 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO: 45.
  • the anti-IL23 antibody of any of the above comprises a heavy chain and a light chain, wherein
  • the heavy chain of the anti-IL23 antibody comprises the amino acid sequence of SEQ ID NO: 42, and the light chain comprises the amino acid sequence of SEQ ID NO: 43;
  • the heavy chain of the anti-IL23 antibody comprises the amino acid sequence of SEQ ID NO: 40, and the light chain comprises the amino acid sequence of SEQ ID NO: 41;
  • the heavy chain of the anti-IL23 antibody comprises the amino acid sequence of SEQ ID NO: 44, and the light chain comprises the amino acid sequence of SEQ ID NO: 45.
  • the present disclosure also provides an isolated anti-IL23 antibody or anti-IL23 antibody fusion protein, which competes with the anti-IL23 antibody or anti-IL23 antibody fusion protein described in any one of the foregoing for binding to human IL23.
  • the anti-IL23 antibody or anti-IL23 antibody fusion protein according to any one of the foregoing has one or more of the following properties:
  • KD value and human IL- 23 p19 binding and/or with a KD value less than 9.00E-11M (for example, less than 9.00E-11M, less than 8.00E-11M, less than 7.00E-11M, less than 6.00E-11M or less) with cynomolgus monkey IL-23 p19 binding, said KD value is measured by surface plasmon resonance assay, e.g. by Measured by surface plasmon resonance assay; in some embodiments, the KD value is detected by the method of Test 6 of the present disclosure;
  • the B. have blocking IL-23/IL-23R binding activity;
  • the IC value of blocking human IL-23/IL-23R binding is less than 0.6nM (for example, less than 0.6nM, less than 0.5nM, less than 0.4nM , less than 0.3nM or less), the IC 50 value is detected by Elisa method; in some embodiments, the IC 50 value is detected by the test 2 method of the present disclosure;
  • the D. have blocking BAFF/BCMA binding activity;
  • the IC value of blocking BAFF/BCMA binding is less than 4.50nM (for example, less than 4.50nM, less than 3.00nM, less than 2.00nM, less than 1.00nM or less),
  • the IC50 value is detected by an Elisa method; in some embodiments, the IC50 value is detected by a test 2 method of the present disclosure;
  • the IC value of blocking BAFF/TACI binding is less than 6.00nM (for example, less than 6.00nM, less than 5.00nM, less than 4.00nM, less than 3.00nM, less than 2.00nM, less than 1.00nM or less), said IC 50 value is detected by Elisa method; in some embodiments, said IC 50 value is detected by the test 2 method of the present disclosure;
  • the F. have blocking APRIL/BCMA binding activity;
  • the IC value of blocking APRIL/BCMA binding is less than 1.00nM (for example, less than 1.00nM, less than 0.5nM, less than 0.4nM, less than 0.1nM or less),
  • the IC50 value is detected by an Elisa method; in some embodiments, the IC50 value is detected by a test 2 method of the present disclosure;
  • the G. have blocking APRIL/TACI binding activity;
  • the IC 50 value of blocking APRIL/TACI binding is less than 3.00nM (for example, less than 3.00nM, less than 2.00nM, less than 1.50nM or less), said IC 50 The value is detected by the Elisa method; in some embodiments, the IC50 value is detected by the test 2 method of the present disclosure;
  • H. has the activity of suppressing IL-17 secretion; Preferably, with less than 0.03nM (for example, less than 0.03nM, less than 0.02nM nM, less than 0.01nM or less) IC50 value inhibits IL-17 secretion; In some embodiments Among them, the IC50 value is detected by the test 4 method of the present disclosure;
  • the value inhibits the proliferation of BaF3-IL-23R cells; the IC 50 value is detected by PerkinElmer; in some embodiments, the IC 50 value is detected by the disclosed test 3 method;
  • TNF ⁇ , IL-22 and IgA cytokines have the activity of inhibiting the secretion of TNF ⁇ , IL-22 and IgA cytokines; in some embodiments, said TNF ⁇ , IL-22 and IgA are detected by the method of test 9 of the present disclosure; or
  • K. has the activity of inhibiting B cell proliferation; in some embodiments, said TNF ⁇ , IL-22 and IgA are detected by the assay 5 method of the present disclosure.
  • the present disclosure also provides a pharmaceutical composition, which comprises the anti-IL23 antibody fusion protein or anti-IL23 antibody described in any one of the foregoing, and one or more pharmaceutically acceptable carriers, diluents or excipients.
  • the present disclosure provides a nucleic acid molecule encoding the anti-IL23 antibody fusion protein or anti-IL23 antibody of any one of the foregoing.
  • the present disclosure provides an expression vector comprising the aforementioned nucleic acid molecule.
  • the present disclosure provides a host cell comprising the aforementioned nucleic acid molecule.
  • the present disclosure provides a host cell comprising the aforementioned expression vector.
  • the present disclosure provides a method for treating or improving B cell disorders or autoimmune diseases, the method comprising administering a therapeutically effective amount of any one of the anti-IL23 described above to a subject in need Steps of antibody fusion protein or anti-IL23 antibody or pharmaceutical composition.
  • the present disclosure provides the use of the anti-IL23 antibody fusion protein or anti-IL23 antibody, nucleic acid molecule or pharmaceutical composition described in any one of the foregoing in the preparation of medicines for treating or preventing diseases.
  • the present disclosure also provides the anti-IL23 antibody fusion protein or anti-IL23 antibody, nucleic acid molecule or composition described in any one of the foregoing for use as a medicament.
  • the medicament is used to treat a B cell disorder or an autoimmune disease.
  • the disease of any one of the preceding is a disease or disorder associated with IL23 expression.
  • the autoimmune disease is selected from the group consisting of: systemic lupus erythematosus, myasthenia gravis, multiple sclerosis, insulin-dependent diabetes mellitus, Crohn's disease, rheumatoid arthritis, polyarticular juvenile rheumatoid Arthritis and psoriatic arthritis; said B-cell disorder is selected from the group consisting of neoplasms, chronic leukaemia, multiple myeloma, non-Hodgkin's lymphoma, post-transplantation lymphoproliferative disease, and light chain gammopathies .
  • the disease is systemic lupus erythematosus.
  • the treatment of any of the preceding further comprises administering to the subject an additional therapeutic agent.
  • FIG. 1 Schematic diagram of the structure of Hu29-19T and Hu29-24T;
  • Figure 2 Experimental results of the erythema severity score of animals with imiquimod-induced psoriasis
  • Figure 3 Experimental results of the skin desquamation severity score of animals with imiquimod-induced psoriasis
  • Figure 4 The comprehensive scoring results of erythema and skin desquamation in animals with psoriasis induced by imiquimod;
  • Figure 5 The results of the right ear thickness experiment of psoriasis animals induced by human IL-23;
  • Figure 7 Experimental results of the weight of the right ear of psoriasis animals induced by human IL-23 (in the accompanying drawings, compared with the negative control, **P ⁇ 0.01, ***P ⁇ 0.001);
  • Figure 8 The results of the anti-IL23 antibody fusion protein inhibiting the secretion of IgA
  • Figure 10 The results of the anti-IL23 antibody fusion protein inhibiting the secretion of IL-22;
  • Figure 11 Renal pathology scoring results in animal efficacy experiments for systemic lupus erythematosus
  • Figure 12 Results of skin damage scores in animal drug efficacy experiments for systemic lupus erythematosus
  • cytokine is a general term for proteins released by a population of cells that act as intercellular mediators on other cells.
  • cytokines include lymphokines, monokines, chemokines and traditional polypeptide hormones.
  • exemplary cytokines include: IL-2, IFN- ⁇ , IL-6, TNF ⁇ , IL-17, and IL-5.
  • TACI described in the present disclosure is a membrane-bound receptor
  • wild-type human TACI comprises two extracellular regions rich in cysteine-rich pseudo-repeats (cysteine-rich pseudo-repeats), a transmembrane region and Cytoplasmic region that interacts with CAML (Calcium Modulator and Cyclophilin Ligand).
  • Wild-type human TACI extracellular region (positions 1-165) refers to SEQ ID NO: 51 of the present disclosure.
  • "TACI extracellular domain" and "TACI extracellular region” can be replaced with each other.
  • IL-23 (also known as IL23) is mainly produced by activated dendritic cells, macrophages and monocytes, and is a member of the IL-12 heterodimer cytokine family, produced by p19 (also known as IL23 p19 subunit) and p40 (also known as IL23 p40 subunit) two subunits, in which the p40 subunit is a subunit shared with IL-12 (J Immunol.2018 Sep 15; 201(6):1605-1613 .).
  • amino acid refers to naturally occurring and synthetic amino acids, as well as amino acid analogs and amino acid mimetics that function in a manner similar to naturally occurring amino acids.
  • Naturally occurring amino acids are those encoded by the genetic code, as well as those amino acids that are later modified, eg, hydroxyproline, gamma-carboxyglutamic acid, and O-phosphoserine.
  • Amino acid analogs are compounds that have the same basic chemical structure (i.e., the alpha carbon bonded to a hydrogen, carboxyl, amino group, and R group) as a naturally occurring amino acid, such as homoserine, norleucine, methionine sulfoxide , Methylsulfonium methionine.
  • Such analogs have modified R groups (eg, norleucine) or modified peptide backbones, but retain the same basic chemical structure as a naturally occurring amino acid.
  • An amino acid mimetic refers to a chemical compound that has a structure that differs from the general chemical structure of an amino acid, but functions in a manner similar to a naturally occurring amino acid.
  • amino acid mutation includes amino acid substitutions (also called amino acid substitutions), deletions, insertions and modifications. Any combination of substitutions, deletions, insertions and modifications can be made to achieve the final construct so long as the final construct possesses the desired properties, such as reduced or binding to Fc receptors.
  • Amino acid sequence deletions and insertions include deletions and insertions at the amino and/or carboxyl termini of the polypeptide chain.
  • Specific amino acid mutations may be amino acid substitutions.
  • the amino acid mutation is a non-conservative amino acid substitution, that is, replacing one amino acid with another amino acid having different structural and/or chemical properties.
  • Amino acid substitutions include substitutions with non-naturally occurring amino acids or with derivatives of the 20 natural amino acids (e.g., 4-hydroxyproline, 3-methylhistidine, ornithine, homoserine, 5-hydroxylysine) .
  • Amino acid mutations can be generated using genetic or chemical methods well known in the art. Genetic methods can include site-directed mutagenesis, PCR, gene synthesis, and the like. It is anticipated that methods other than genetic engineering to alter amino acid side chain groups, such as chemical modification, may also be available. Various names may be used herein to refer to the same amino acid mutation.
  • amino acid residue at a specific position can be expressed in the form of position + amino acid residue, for example, 366W means that the amino acid residue at position 366 is W. T366W means that the amino acid residue at the 366th position is mutated from the original T to W.
  • antibody is used in the broadest sense and encompasses various antibody structures including, but not limited to, monoclonal antibodies, polyclonal antibodies; monospecific antibodies, multispecific antibodies (e.g., bispecific antibodies); full-length antibodies and antibody Fragments (or antigen-binding fragments, or antigen-binding portions) as long as they exhibit the desired antigen-binding activity.
  • Native antibody refers to a naturally occurring immunoglobulin molecule. For example, native IgG antibodies are heterotetrameric glycoproteins of approximately 150,000 Daltons, composed of two identical light chains and two identical heavy chains joined by disulfide bonds.
  • each heavy chain has a variable region (VH), also known as variable heavy domain, heavy chain variable region, followed by a heavy chain constant region, the natural IgG heavy chain constant region usually contains three Constant domains (CH1, CH2 and CH3).
  • VH variable region
  • VL variable light domain, or light chain variable domain, followed by a constant light domain (light chain constant region, CL ).
  • full-length antibody “intact antibody” and “whole antibody” are used interchangeably herein to refer to an antibody having a heavy chain structure substantially similar to that of a native antibody or having an Fc region as defined herein.
  • Natural complete antibody light chain includes light chain variable region VL and constant region CL, VL is at the amino terminal of light chain, light chain constant region includes ⁇ chain and ⁇ chain; heavy chain includes variable region VH and constant region (CH1, CH2 and CH3), the VH is at the amino-terminus of the heavy chain, the constant region is at the carboxy-terminus, wherein CH3 is closest to the carboxy-terminus of the polypeptide, and the heavy chain can belong to any isotype, including IgG (including IgG1, IgG2, IgG3 and IgG4 subtypes) , IgA (including IgA1 and IgA2 subtypes), IgM and IgE.
  • IgG including IgG1, IgG2, IgG3 and IgG4 subtypes
  • IgA including IgA1 and IgA2 subtypes
  • IgM and IgE IgE.
  • variable region or “variable domain” of an antibody refers to the domain of an antibody's heavy or light chain that is involved in binding the antibody to antigen.
  • the antibody heavy chain variable region (VH) and light chain variable region (VL) each comprise four conserved framework regions (FR) and three complementarity determining regions (CDR).
  • FR conserved framework regions
  • CDR complementarity determining region
  • VH contains 3 CDR regions: HCDR1, HCDR2 and HCDR3
  • VL contains 3 CDR regions: LCDR1, LCDR2 and LCDR3.
  • Each VH and VL consists of three CDRs and four FRs arranged in the following order from the amino terminus (also known as the N terminus) to the carboxyl terminus (also known as the C terminus): FR1, CDR1, FR2, CDR2, FR3, CDR3 , FR4.
  • amino acid sequence boundaries of CDRs can be determined by various known schemes, for example: “Kabat” numbering convention (see Kabat et al. (1991), “Sequences of Proteins of Immunological Interest", 5th Edition, Public Health Service, National Institutes of Health , Bethesda, MD), “Chothia” numbering sequence, “ABM” numbering sequence, "contact” numbering sequence (see Martin, ACR. Protein Sequence and Structure Analysis of Antibody Variable Domains [J].
  • antibody fragment refers to a molecule other than an intact antibody that comprises the portion of an intact antibody that binds to the antigen to which the intact antibody binds.
  • antibody fragments include, but are not limited to, Fv, Fab, Fab', Fab'-SH, F(ab') 2 , single domain antibody, single chain Fab (scFab), diabody, linear antibody, single chain antibody molecule (e.g. scFv); and multispecific antibodies formed from antibody fragments.
  • Fc region or “fragment crystallizable region” is used to define the C-terminal region of an antibody heavy chain, including native and engineered Fc regions.
  • the Fc region comprises the same or different two subunits.
  • the Fc region of a human IgG heavy chain is defined as extending from the amino acid residue at position Cys226 or from Pro230 to its carboxyl terminus.
  • Suitable Fc regions for use in the antibodies described herein include the Fc regions of human IgGl, IgG2 (IgG2A, IgG2B), IgG3 and IgG4.
  • the boundaries of the Fc region can also be varied, such as deletion of the C-terminal lysine of the Fc region (residue 447 according to the EU numbering system) or deletion of the C-terminal glycine and lysine of the Fc region (residue 447 according to the EU numbering system). system residues 446 and 447).
  • the numbering convention for the Fc region is the EU numbering system, also known as the EU index.
  • chimeric antibody refers to an antibody in which a portion of the heavy and/or light chains is derived from a particular source or species, while the remaining portion of the heavy and/or light chains is derived from another, different source or species.
  • humanized antibody is an antibody that retains the reactivity of a non-human antibody while being less immunogenic in humans. This can be achieved, for example, by retaining the non-human CDR regions and replacing the remainder of the antibody with their human counterparts (ie, the constant regions and the framework portion of the variable regions).
  • human antibody “human antibody”, “fully human antibody”, and “fully human antibody” are used interchangeably to refer to antibodies whose variable and constant regions are human sequences.
  • the term encompasses antibodies that are derived from human genes but have, for example, altered sequences that reduce potential immunogenicity, increase affinity, eliminate cysteines or glycosylation sites that might cause undesired folding.
  • the term encompasses such antibodies produced recombinantly in non-human cells which may confer glycosylation not characteristic of human cells.
  • the term also encompasses antibodies that have been raised in transgenic mice containing some or all of the immunoglobulin heavy and light chain loci.
  • the meaning of human antibody expressly excludes humanized antibodies comprising non-human antigen-binding residues.
  • affinity refers to the overall strength of the non-covalent interaction between a single binding site of a molecule (eg, an antibody) and its binding partner (eg, an antigen). As used herein, unless otherwise indicated, binding “affinity” refers to internal binding affinity, which reflects a 1:1 interaction between members of a binding pair (eg, antibody and antigen).
  • the affinity of a molecule X for its ligand Y can generally be expressed by a dissociation constant (KD). Affinity can be measured by routine methods known in the art, including those described herein.
  • the term “kassoc” or “ka” refers to the on-rate of a particular antibody-antigen interaction and the term “kdis” or “kd” refers to the dissociation rate of a particular antibody-antigen interaction.
  • KD refers to the dissociation constant, which is obtained from the ratio of kd to ka (ie, kd/ka) and is expressed as molarity (M).
  • M molarity
  • the KD value of an antibody can be determined using methods well known in the art. For example, using biosensing systems such as systems measuring surface plasmon resonance (eg Biacore), or measuring affinity in solution by solution equilibrium titration (SET).
  • effector function refers to those biological activities attributable to an antibody Fc region (either native sequence Fc region or amino acid sequence mutated Fc region) and which vary with the antibody isotype.
  • antibody effector functions include, but are not limited to: C1q binding and complement-dependent cytotoxicity, Fc receptor binding, antibody-dependent cell-mediated cytotoxicity (ADCC), phagocytosis, cell surface receptors (e.g., B cell receptors, body) downregulation; and B cell activation.
  • the term "monoclonal antibody” refers to a population of substantially homogeneous antibodies, i.e., the antibody molecules comprised in the population are identical in amino acid sequence, except for natural mutations that may be present in minor amounts.
  • polyclonal antibody preparations typically comprise multiple different antibodies with different amino acid sequences in their variable domains, often specific for different epitopes.
  • “Monoclonal” denotes the characteristics of an antibody obtained from a substantially homogeneous population of antibodies and should not be construed as requiring that the antibody be produced by any particular method.
  • the antibodies provided by the present disclosure are monoclonal antibodies.
  • antigen refers to a molecule or portion of a molecule capable of being bound by a selective binding agent such as an antigen binding protein (including, for example, an antibody), and which can additionally be used in an animal to generate antibodies capable of binding the antigen.
  • a selective binding agent such as an antigen binding protein (including, for example, an antibody)
  • An antigen may have one or more epitopes capable of interacting with different antigen binding proteins (eg antibodies).
  • epitope refers to an area (area or region) on an antigen capable of specifically binding to an antibody or antigen-binding fragment thereof.
  • Epitopes may be formed from contiguous strings of amino acids (linear epitopes) or comprise non-contiguous amino acids (conformational epitopes), for example brought into spatial proximity by folding of the antigen, ie by tertiary folding of the proteinaceous antigen.
  • the difference between a conformational epitope and a linear epitope is that antibody binding to a conformational epitope is lost in the presence of denaturing solvents.
  • An epitope comprises at least 3, at least 4, at least 5, at least 6, at least 7, or 8-10 amino acids in a unique spatial conformation.
  • Screening for antibodies that bind a particular epitope can be performed using methods routine in the art, such as, but not limited to, alanine scanning, peptide blotting, peptide cleavage analysis, epitope excision, epitope extraction, Chemical modification of antigens (see Prot. Sci. 9 (2000) 487-496), and cross-blocking.
  • the term “capable of specifically binding”, “specifically binds” or “binds” means that an antibody is capable of binding to a certain antigen or an epitope within the antigen with a higher affinity than to other antigens or epitopes.
  • the antibody is prepared in an amount of about 1 ⁇ 10 -7 M or less (e.g., about 1 ⁇ 10 -8 M, 1 ⁇ 10 -9 M, 1 ⁇ 10 -10 M, 1 ⁇ 10 -11 M or less).
  • KD equilibrium dissociation constant
  • the antibody binds an antigen with a KD that is 10% or less (eg, 1%) of the antibody's KD for binding to a non-specific antigen (eg, BSA, casein).
  • KD can be measured using known methods, for example by measured by surface plasmon resonance.
  • antibodies that specifically bind to an antigen or an epitope within an antigen may have cross-reactivity to other related antigens, e.g. (cynomolgus, cyno), chimpanzee (Pan troglodytes) (chimpanzee, chimp)) or marmoset (Callithrix jacchus) (commonmarmoset, marmoset) are cross-reactive.
  • anti-IL23 antibody and "IL23-binding antibody” refer to antibodies capable of binding IL23 with sufficient affinity.
  • the antibody binds to an unrelated, non-IL23 protein to an extent less than about 10% of the antibody's binding to IL23 and IL12 by Surface plasmon resonance measurements.
  • the antibody that binds to the IL23 protein has a dissociation constant (KD) of ⁇ about 1 ⁇ M, ⁇ about 100 nM, ⁇ about 10 nM, ⁇ about 1 nM, ⁇ about 0.1 nM, ⁇ about 0.01 nM, or ⁇ about 0.001 nM.
  • KD dissociation constant
  • the anti-IL23 antibody binds to an epitope of the p19 subunit of human or cynomolgus IL23.
  • linker refers to a connecting unit connecting two polypeptide fragments, usually with a certain degree of flexibility, and the use of the linker will not lose the original function of the protein domain.
  • linkers appearing in the same structure may be the same or different.
  • the linker may be a peptide linker comprising one or more amino acids, typically about 1-30, 2-24 or 3-15 amino acids.
  • the linkers used herein may be the same or different.
  • antibody-dependent cellular cytotoxicity refers to a mechanism of inducing cell death that relies on antibody coating of target cells with lytically active effector cells ( Cells such as natural killer (NK), monocytes, macrophages and neutrophils) interact via Fc ⁇ receptors (Fc ⁇ Rs) expressed on effector cells.
  • NK cells express FcyRIIIa
  • monocytes express FcyRI, FcyRII, and FcyRIIIa.
  • the ADCC activity of the antibodies provided herein can be assessed using an in vitro assay using antigen-expressing cells as target cells and NK cells as effector cells. Cell lysis is detected based on labels released from lysed cells, such as radioactive substrates, fluorescent dyes, or native intracellular proteins.
  • ADCP antibody-dependent cellular phagocytosis
  • complement-dependent cytotoxicity refers to a cell death-inducing mechanism in which the Fc effector domain of a target-binding antibody binds and activates the complement component C1q, which in turn activates the complement cascade, resulting in target cell death.
  • Activation of complement can also result in the deposition of complement components on the surface of target cells that promote CDC by binding to complement receptors (eg, CR3) on leukocytes.
  • complement receptors eg, CR3
  • nucleic acid is used herein interchangeably with the term “polynucleotide” and refers to deoxyribonucleotides or ribonucleotides and polymers thereof in single- or double-stranded form.
  • the term encompasses nucleic acids containing known nucleotide analogs or modified backbone residues or linkages, synthetic, naturally occurring and non-naturally occurring, having similar binding properties to the reference nucleic acid, and defined in Metabolized in a manner similar to the reference nucleotide.
  • nucleic acid refers to a nucleic acid molecule that has been separated from components of its natural environment.
  • An isolated nucleic acid includes a nucleic acid molecule contained in a cell that normally contains the nucleic acid molecule, but which is present extrachromosomally or at a chromosomal location other than its natural chromosomal location.
  • An isolated nucleic acid encoding a polypeptide or fusion protein refers to one or more nucleic acid molecules encoding a polypeptide or fusion protein, including such one or more nucleic acid molecules in a single vector or in separate vectors, and present in a host cell Such one or more nucleic acid molecules at one or more positions.
  • a particular nucleic acid sequence also implicitly encompasses conservatively modified variants thereof (eg, degenerate codon substitutions) and complementary sequences as well as the explicitly indicated sequence.
  • degenerate codon substitutions can be obtained by generating sequences in which the third position of one or more selected (or all) codons is mixed with bases and/or deoxygenated Inosine residue substitution.
  • polypeptide and "protein” are used interchangeably herein to refer to a polymer of amino acid residues.
  • the term applies to amino acid polymers in which one or more amino acid residues are an artificial chemical mimetic of the corresponding naturally occurring amino acid, and to both naturally occurring amino acid polymers and non-naturally occurring amino acid polymers. Unless otherwise stated, a particular polypeptide sequence also implicitly encompasses conservatively modified variants thereof.
  • sequence identity means that when two sequences are optimally aligned, gaps are introduced as necessary to obtain the maximum percent sequence identity and any conservative substitutions are not considered part of the sequence identity, two The degree (percentage) to which amino acids/nucleic acids of a sequence are identical at equivalent positions.
  • alignment can be achieved by techniques known in the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN, ALIGN-2 or Megalign (DNASTAR) software. Those skilled in the art can determine suitable parameters for measuring alignment, including any algorithms needed to achieve maximal alignment over the full length of the sequences being compared.
  • the position relative to the XX sequence means that the sequence to be tested is optimally aligned with the XX sequence to obtain the highest percentage identity.
  • the positions corresponding to the sequence to be tested and the XX sequence are two The relative position of the sequence.
  • the natural sequence amino acid residue positions of SEQ ID NO: 51 and SEQ ID NO: 60 relative to SEQ ID NO: 58 in the extracellular region of TACI are shown in Table 2:
  • the 2nd (natural sequence) residue site of SEQ ID NO: 60 and the 49th (natural sequence) residue site of the sequence SEQ ID NO: 58 are corresponding sites.
  • fused or “linked” refer to the joining of components (eg, TACI polypeptide and antibody heavy chain) by a covalent bond, either directly or via one or more linkers.
  • linker is a peptide linker
  • the covalent bond is a peptide bond.
  • anti-IL23 antibody fusion protein refers to a protein in which an anti-IL23 antibody is fused to an active protein.
  • a TACI polypeptide is fused to the heavy chain terminus of an anti-IL23 antibody to form a protein.
  • vector means a polynucleotide molecule capable of transporting another polynucleotide to which it has been linked.
  • plasmid refers to a circular double-stranded DNA loop into which additional DNA segments can be ligated.
  • viral vector such as an adeno-associated viral vector (AAV or AAV2), in which additional DNA segments can be ligated into the viral genome.
  • AAV adeno-associated viral vector
  • Certain vectors are capable of autonomous replication in the host cells into which they are introduced (eg, bacterial vectors and episomal mammalian vectors with a bacterial origin of replication).
  • vectors can integrate into the genome of the host cell after introduction into the host cell, thereby replicating along with the host genome.
  • expression vector or "expression construct” refers to a vector that can transform a host cell and contains a vector that directs and/or controls (along with the host cell) the expression of one or more heterologous coding regions operably linked thereto.
  • Expression constructs may include, but are not limited to, sequences that affect or control transcription, translation, and, when an intron is present, RNA splicing of the coding region to which it is operably linked.
  • host cell refers to a cell into which exogenous nucleic acid has been introduced, including the progeny of such cells.
  • Host cells include “transformants” and “transformed cells,” which include the primary transformed cell and progeny derived therefrom, regardless of the number of passages. Progeny may not be identical to the parental cell in nucleic acid content, but may contain mutations. Mutant progeny having the same function or biological activity as screened or selected for in the originally transformed cell are included herein.
  • Host cells include prokaryotic and eukaryotic host cells, where eukaryotic host cells include, but are not limited to, mammalian cells, insect cell lines, plant cells, and fungal cells.
  • Mammalian host cells include human, mouse, rat, dog, monkey, pig, goat, cow, horse, and hamster cells, including but not limited to Chinese hamster ovary (CHO) cells, NSO, SP2 cells, HeLa cells, baby hamster cells Kidney (BHK) cells, monkey kidney cells (COS), human hepatocellular carcinoma cells (eg, Hep G2), A549 cells, 3T3 cells, and HEK-293 cells.
  • Fungal cells include yeast and filamentous fungal cells including, for example, Pichia pastoris, Pichia finlandica, Pichia trehalophila, Pichia koclamae, Pichia membranaefaciens, Pichia minuta (Ogataea minuta, Pichia lindneri), Pichia puntiae, Pichia thermotolerans, Pichia willow salictaria), Pichia guercuum, Pichia pijperi, Pichia stiptis, Pichia methanolica, Pichia, Saccharomycescerevisiae, Saccharomyces cerevisiae , Hansenula polymorpha, Kluyveromyces, Kluyveromyces lactis, Candida albicans, Aspergillus nidulans, Aspergillus niger, Aspergillus oryzae, Trichoderma reesei, Chrysosporium lucknowense, Fus
  • Pichia any Saccharomyces, Hansenula polymorpha, any Kluyveromyces, Candida albicans, any Aspergillus, Trichoderma reesei, Luke Mold (Chrysosporium lucknowense), any Fusarium species, Yarrowia lipolytica, and Neurospora crassa.
  • the expressions "cell”, “cell line” and “cell culture” are used interchangeably and all such designations include progeny.
  • the words “transformants” and “transformed cells” include primary subject cells and cultures derived therefrom, regardless of the number of passages. It should also be understood that not all progeny will have the exact same DNA content due to deliberate or unintentional mutations. Mutant progeny having the same function or biological activity as the original transformed cell from which they were screened are included.
  • composition refers to a mixture containing one or more anti-IL23 antibody fusion proteins described herein and other chemical components, such as physiological/pharmaceutically acceptable carriers and excipients.
  • pharmaceutically acceptable carrier refers to an ingredient in a pharmaceutical formulation that is different from the active ingredient and is nontoxic to the subject.
  • Pharmaceutically acceptable carriers include, but are not limited to, buffers, excipients, stabilizers or preservatives.
  • subject or “individual” includes humans and non-human animals.
  • Non-human animals include all vertebrates (eg, mammals and non-mammals) such as non-human primates (eg, cynomolgus monkeys), sheep, dogs, cows, chickens, amphibians, and reptiles.
  • patient or “subject” are used interchangeably herein unless otherwise indicated.
  • cyno or “cynomolgus” refers to Macaca fascicularis.
  • the individual or subject is a human.
  • administering when applied to an animal, human, experimental subject, cell, tissue, organ or biological fluid, refers to the interaction of an exogenous drug, therapeutic agent, diagnostic agent or composition with an animal, human , subjects, cells, tissues, organs or biological fluids.
  • sample refers to a collection of similar fluids, cells, or tissues isolated from a subject, as well as fluids, cells, or tissues present in a subject.
  • exemplary samples are biological fluids such as blood, serum and serosal fluids, plasma, lymph, urine, saliva, cystic fluid, tears, faeces, sputum, mucous membrane secretions of secretory tissues and organs, vaginal secretions, ascites , pleura, pericardium, peritoneum, peritoneal and other body cavity fluids, fluid collected from bronchial lavage, synovial fluid, liquid solutions in contact with subjects or biological sources, such as cell and organ culture media (including cell or organ condition culture medium), lavage fluid, etc., tissue biopsy samples, fine needle aspirations, surgically resected tissues, organ cultures, or cell cultures.
  • biological fluids such as blood, serum and serosal fluids, plasma, lymph, urine, saliva, cystic fluid, tears, faeces, sputum, mucous membrane
  • Treatment refers to clinical intervention that attempts to alter the natural course of the individual being treated, and may be performed for prophylaxis or during the course of clinical pathology. Desired effects of treatment include, but are not limited to, prevention of occurrence or recurrence of disease, alleviation of symptoms, alleviation/reduction of any direct or indirect pathological consequences of disease, prevention of metastasis, reduction of rate of disease progression, amelioration or palliation of disease state, and regression or amelioration of prognosis.
  • the antibodies of the disclosure are used to delay the development of a disease or slow the progression of a disease.
  • an “effective amount” is generally sufficient to reduce the severity and/or frequency of symptoms, eliminate these symptoms and/or underlying causes, prevent the occurrence of symptoms and/or their underlying causes, and/or ameliorate or ameliorate the impairment caused by or associated with the disease state (e.g. lung disease).
  • the effective amount is a therapeutically or prophylactically effective amount.
  • a “therapeutically effective amount” is sufficient to treat a disease state or symptom, especially a state or symptom associated with the disease state, or otherwise prevent, hinder, delay or reverse the disease state or any other adverse effect in any way related to the disease state. The amount of progression of the desired symptoms.
  • a “prophylactically effective amount” is an amount that, when administered to a subject, will have a predetermined prophylactic effect, such as preventing or delaying the onset (or recurrence) of the disease state, or reducing the likelihood of the onset (or recurrence) of the disease state or associated symptoms .
  • Complete therapeutic or prophylactic effect does not necessarily occur after administration of one dose, but may occur after administration of a series of doses.
  • a therapeutically or prophylactically effective amount may be administered in one or more administrations.
  • “Therapeutically effective amount” and “prophylactically effective amount” can vary depending on factors such as the disease state, age, sex and weight of the individual, and the ability of the therapeutic agent or combination of therapeutic agents to elicit a desired response in the individual.
  • Exemplary indicators of an effective therapeutic agent or combination of therapeutic agents include, for example, improved health status of a patient.
  • Anti-IL23 antibodies of the present disclosure are provided.
  • the present disclosure contemplates a novel anti-IL23 antibody and fusion protein thereof. It has one or more of the following properties:
  • KD value and human IL- 23 p19 binding and/or with a KD value less than 9.00E-11M (for example, less than 9.00E-11M, less than 8.00E-11M, less than 7.00E-11M, less than 6.00E-11M or less) with cynomolgus monkey IL-23 p19 binding, said KD value is measured by surface plasmon resonance assay, e.g. by as measured by surface plasmon resonance assay; in some embodiments, the detection
  • the B. have blocking IL-23/IL-23R binding activity;
  • the IC value of blocking human IL-23/IL-23R binding is less than 0.6nM (for example, less than 0.6nM, less than 0.5nM, less than 0.4nM , less than 0.3nM or less), the IC50 value is detected by Elisa method;
  • C. have the activity of inhibiting IL-17 secretion;
  • IC 50 value inhibits IL-17 secretion; Said IC 50 The value is detected by Elisa;
  • D It has the activity of inhibiting the proliferation of BaF3-IL-23R cells; preferably, with an IC of less than 0.5nM (for example, less than 0.5nM, less than 0.4nM nM, less than 0.3nM, less than 0.2nM, less than 0.1nM or less)
  • the value inhibits the proliferation of BaF3-IL-23R cells; the IC50 value is detected by PerkinElmer;
  • E. have the activity of inhibiting the secretion of TNF ⁇ and/or IL-22;
  • the present disclosure provides an anti-IL23 antibody, wherein the anti-IL23 antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3, and the light chain variable region comprises HCDR1, HCDR2 and HCDR3, and the light chain variable region comprises The chain variable region comprises LCDR1, LCDR2 and LCDR3, wherein,
  • the HCDR1, HCDR2 and HCDR3 of the heavy chain variable region and the LCDR1, LCDR2 and LCDR3 of the light chain variable region of the anti-IL23 antibody according to any one of the above are defined according to the numbering convention selected from Kabat of which:
  • HCDR1 of the heavy chain variable region is shown in SEQ ID NO: 27, 28 or 5
  • HCDR2 is shown in SEQ ID NO: 6
  • HCDR3 is shown in SEQ ID NO: 7
  • the light LCDR1 of the chain variable region is set forth in SEQ ID NO: 8
  • LCDR2 is set forth in SEQ ID NO: 9
  • LCDR3 is set forth in SEQ ID NO: 10; or
  • HCDR1 of the heavy chain variable region is shown in SEQ ID NO: 11
  • HCDR2 is shown in SEQ ID NO: 12
  • HCDR3 is shown in SEQ ID NO: 13
  • the light chain variable region LCDR1 is set forth in SEQ ID NO: 14
  • LCDR2 is set forth in SEQ ID NO: 37 or 15
  • LCDR3 is set forth in SEQ ID NO: 16.
  • the anti-IL23 antibody of any one of the above comprises a heavy chain variable region and a light chain variable region, wherein
  • HCDR1 of the heavy chain variable region is shown in SEQ ID NO: 27, HCDR2 is shown in SEQ ID NO: 6, and HCDR3 is shown in SEQ ID NO: 7, and the light chain variable region LCDR1 is set forth in SEQ ID NO: 8, LCDR2 is set forth in SEQ ID NO: 9, and LCDR3 is set forth in SEQ ID NO: 10; or
  • HCDR1 of the heavy chain variable region is shown in SEQ ID NO: 28
  • HCDR2 is shown in SEQ ID NO: 6
  • HCDR3 is shown in SEQ ID NO: 7
  • the light chain variable region LCDR1 is set forth in SEQ ID NO: 8
  • LCDR2 is set forth in SEQ ID NO: 9
  • LCDR3 is set forth in SEQ ID NO: 10; or
  • HCDR1 of the heavy chain variable region is shown in SEQ ID NO: 11
  • HCDR2 is shown in SEQ ID NO: 12
  • HCDR3 is shown in SEQ ID NO: 13
  • the light chain variable region LCDR1 is set forth in SEQ ID NO: 14
  • LCDR2 is set forth in SEQ ID NO: 37
  • LCDR3 is set forth in SEQ ID NO: 16.
  • the anti-IL23 antibody of any of the preceding is murine, chimeric or humanized. In some embodiments, the anti-IL23 antibody is humanized. In some embodiments, the heavy chain variable region of the anti-IL23 antibody has FR1, FR2, FR3 derived from IGHV2-26*01 and FR4 derived from IGHJ6*01, and it is unsubstituted or has selected One or more amino acid substitutions from the group consisting of 1E, 30T, 37V, 44G, 49G, 73N, 89R and 93V; and/or the light chain variable region has FR1, FR2 derived from IGKV4-1*01 , FR3 and FR4 derived from IGKJ4*01 and which are unsubstituted or have one or more amino acid substitutions selected from the group consisting of 4L, 17Q, 36F, 58I, 60A and 68R.
  • the HCDR1 of the heavy chain variable region is as shown in SEQ ID NO: 27, 28 or 5
  • HCDR2 is as shown in SEQ ID NO: 6
  • HCDR3 is as shown in SEQ ID NO: 7
  • LCDR1 of the light chain variable region is shown in SEQ ID NO: 8
  • LCDR2 is shown in SEQ ID NO: 9
  • LCDR3 is shown in SEQ ID NO: 10.
  • the anti-IL23 antibody as described in any one of the preceding is a humanized antibody
  • the heavy chain variable region of the anti-IL23 antibody has FR1 derived from IGHV1-3*01, FR2, FR3 and FR4 derived from IGHJ1*01 and which are unsubstituted or have one or more amino acid substitutions selected from the group consisting of 1E, 2F, 28S, 44S, 71V and 73Q; and/or all
  • the light chain variable region has FR1, FR2, FR3 derived from IGKV2-40*01 and FR4 derived from IGKJ2*01, and it is unsubstituted or has a composition selected from 4I, 36F, 39R, 71L and 100S One or more amino acid substitutions in the group.
  • the heavy chain variable region of the anti-IL23 antibody has HCDR1 as shown in SEQ ID NO: 11, HCDR2 as shown in SEQ ID NO: 12, and HCDR3 as shown in SEQ ID NO: 13, and LCDR1 of the light chain variable region is shown in SEQ ID NO: 14, LCDR2 is shown in SEQ ID NO: 37 or 15, and LCDR3 is shown in SEQ ID NO: 16.
  • the above variable regions and CDRs are defined according to the Kabat numbering convention.
  • the anti-IL23 antibody of any one of the above comprises a heavy chain variable region and a light chain variable region, wherein,
  • said heavy chain variable region is as set forth in SEQ ID NO: 22, 21, 20, 19, 18 or 17, and said light chain variable region is as set forth in SEQ ID NO: 26, 25, 24 or 23 show; or
  • said heavy chain variable region is set forth in SEQ ID NO: 29, 30, 31 or 32, and said light chain variable region is set forth in SEQ ID NO: 36, 35, 34 or 33; or
  • said heavy chain variable region is as shown in SEQ ID NO: 1, and said light chain variable region is as shown in SEQ ID NO: 2; or
  • the heavy chain variable region is as shown in SEQ ID NO:3, and the light chain variable region is as shown in SEQ ID NO:4.
  • the anti-IL23 antibody of any one of the above comprises a heavy chain variable region and a light chain variable region, wherein,
  • said heavy chain variable region is as shown in SEQ ID NO: 22, and said light chain variable region is as shown in SEQ ID NO: 26;
  • said heavy chain variable region is as shown in SEQ ID NO: 21, and said light chain variable region is as shown in SEQ ID NO: 25; or
  • said heavy chain variable region is as shown in SEQ ID NO: 29
  • said light chain variable region is as shown in SEQ ID NO: 36.
  • the anti-IL23 antibody of any one of the above further comprises an antibody heavy chain constant region and a light chain constant region.
  • the heavy chain constant region is a human IgG heavy chain constant region; in some embodiments, the heavy chain constant region is selected from human IgG1, IgG2, IgG3 and IgG4 constant regions; in some embodiments, The light chain constant region is selected from a human antibody ⁇ or ⁇ chain constant region; in some embodiments, the heavy chain constant region is a human IgG1 heavy chain constant region, and the light chain constant region is a human ⁇ light chain constant region.
  • the Fc region of the heavy chain constant region has one or more amino acid substitutions that reduce binding of the Fc region to an Fc receptor.
  • the Fc region has L234A, L235A mutations, and/or S228P mutations, and/or YTE mutations (M252Y, S254T, and T256E), and the numbering of the mutations is based on the EU index. In some embodiments, the Fc region has L234A, L235A mutations. In some embodiments, the heavy chain constant region comprises the amino acid sequence of SEQ ID NO: 38, and the light chain constant region comprises the amino acid sequence of SEQ ID NO: 39.
  • the anti-IL23 antibody of any one of the above comprises a heavy chain and a light chain, wherein,
  • the heavy chain of said anti-IL23 antibody comprises an amino acid sequence having at least 90% (e.g., at least 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO: 42, and a light chain comprising an amino acid sequence having at least 90% (e.g., at least 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO: 43; or
  • the heavy chain of said anti-IL23 antibody comprises an amino acid sequence having at least 90% (e.g., at least 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO: 40, and a light chain comprising an amino acid sequence having at least 90% (e.g., at least 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO: 41; or
  • the heavy chain of the anti-IL23 antibody comprises an amino acid sequence having at least 90% (e.g., at least 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO: 44, and the light chain comprise an amino acid sequence having at least 90% (eg, at least 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO: 45.
  • the anti-IL23 antibody of any of the above comprises a heavy chain and a light chain, wherein
  • Anti-IL23 antibody fusion protein of the present disclosure is provided.
  • an anti-IL23 antibody fusion protein which comprises an anti-IL23 antibody and a TACI polypeptide, wherein the anti-IL23 antibody specifically binds to human IL23, preferably specifically binds to the p19 subunit of human IL23.
  • the anti-IL23 antibody or anti-IL23 antibody fusion protein according to any one of the foregoing has one or more of the following characteristics:
  • the B. have blocking IL-23/IL-23R binding activity;
  • the IC value of blocking human IL-23/IL-23R binding is less than 0.6nM (for example, less than 0.6nM, less than 0.5nM, less than 0.4nM , less than 0.3nM or less), the IC50 value is detected by Elisa method;
  • the D. have blocking BAFF/BCMA binding activity;
  • the IC value of blocking BAFF/BCMA binding is less than 4.50nM (for example, less than 4.50nM, less than 3.00nM, less than 2.00nM, less than 1.00nM or less), The IC50 value is detected by Elisa method;
  • the IC value of blocking BAFF/TACI binding is less than 6.00nM (for example, less than 6.00nM, less than 5.00nM, less than 4.00nM, less than 3.00nM, less than 2.00nM, Less than 1.00nM or less), the IC50 value is detected by Elisa method;
  • the F. have blocking APRIL/BCMA binding activity;
  • the IC value of blocking APRIL/BCMA binding is less than 1.00nM (for example, less than 1.00nM, less than 0.5nM, less than 0.4nM, less than 0.1nM or less), The IC50 value is detected by Elisa method;
  • the G. have blocking APRIL/TACI binding activity;
  • the IC 50 value of blocking APRIL/TACI binding is less than 3.00nM (for example, less than 3.00nM, less than 2.00nM, less than 1.50nM or less), said IC 50 The value is detected by Elisa method;
  • H. has the activity of inhibiting IL-17 secretion; Preferably, with less than 0.03nM (for example, less than 0.03nM, less than 0.02nM nM, less than 0.01nM or less) IC 50 value inhibits IL-17 secretion; Said IC 50 The value is detected by Elisa;
  • It has the activity of inhibiting the proliferation of BaF3-IL-23R cells; preferably, with an IC of less than 0.5nM (for example, less than 0.5nM, less than 0.4nM nM, less than 0.3nM, less than 0.2nM, less than 0.1nM or less)
  • the value inhibits the proliferation of BaF3-IL-23R cells; the IC50 value is detected by PerkinElmer;
  • K. has the activity of inhibiting B cell proliferation.
  • the anti-IL23 antibody comprises a heavy chain variable region and a light chain variable region
  • the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3
  • the The light chain variable region comprises LCDR1, LCDR2 and LCDR3, wherein the HCDR1, HCDR2 and HCDR3 of the heavy chain variable region and the LCDR1, LCDR2 and LCDR3 of the light chain variable region are defined according to the numbering rules selected from Kabat, in:
  • HCDR1 of the heavy chain variable region is shown in SEQ ID NO: 27, 28 or 5
  • HCDR2 is shown in SEQ ID NO: 6
  • HCDR3 is shown in SEQ ID NO: 7
  • the light LCDR1 of the chain variable region is set forth in SEQ ID NO: 8
  • LCDR2 is set forth in SEQ ID NO: 9
  • LCDR3 is set forth in SEQ ID NO: 10; or
  • HCDR1 of the heavy chain variable region is shown in SEQ ID NO: 11
  • HCDR2 is shown in SEQ ID NO: 12
  • HCDR3 is shown in SEQ ID NO: 13
  • the light chain variable region LCDR1 is set forth in SEQ ID NO: 14
  • LCDR2 is set forth in SEQ ID NO: 37 or 15
  • LCDR3 is set forth in SEQ ID NO: 16.
  • HCDR1 of the heavy chain variable region is shown in SEQ ID NO: 27, HCDR2 is shown in SEQ ID NO: 6, and HCDR3 is shown in SEQ ID NO: 7, and the light chain variable region LCDR1 is set forth in SEQ ID NO: 8, LCDR2 is set forth in SEQ ID NO: 9, and LCDR3 is set forth in SEQ ID NO: 10; or
  • HCDR1 of the heavy chain variable region is shown in SEQ ID NO: 28
  • HCDR2 is shown in SEQ ID NO: 6
  • HCDR3 is shown in SEQ ID NO: 7
  • the light chain variable region LCDR1 is set forth in SEQ ID NO: 8
  • LCDR2 is set forth in SEQ ID NO: 9
  • LCDR3 is set forth in SEQ ID NO: 10; or
  • HCDR1 of the heavy chain variable region is shown in SEQ ID NO: 11
  • HCDR2 is shown in SEQ ID NO: 12
  • HCDR3 is shown in SEQ ID NO: 13
  • the light chain variable region LCDR1 is set forth in SEQ ID NO: 14
  • LCDR2 is set forth in SEQ ID NO: 37
  • LCDR3 is set forth in SEQ ID NO: 16.
  • the anti-IL23 antibody is a murine antibody, a chimeric antibody or a humanized antibody. In some embodiments, the anti-IL23 antibody is a humanized antibody.
  • the heavy chain variable region of the anti-IL23 antibody has FR1, FR2, FR3 derived from IGHV2-26*01 and FR4 derived from IGHJ6*01, and it is unsubstituted or has selected One or more amino acid substitutions from the group consisting of 1E, 30T, 37V, 44G, 49G, 73N, 89R and 93V; and/or the light chain variable region has FR1, FR2 derived from IGKV4-1*01 , FR3 and FR4 derived from IGKJ4*01 and which are unsubstituted or have one or more amino acid substitutions selected from the group consisting of 4L, 17Q, 36F, 58I, 60A and 68R.
  • the heavy chain variable region of the anti-IL23 antibody has HCDR1 as shown in SEQ ID NO: 27, 28 or 5, HCDR2 as shown in SEQ ID NO: 6, and HCDR3 as shown in SEQ ID NO: 7 As shown, and LCDR1 of the light chain variable region is shown in SEQ ID NO: 8, LCDR2 is shown in SEQ ID NO: 9, and LCDR3 is shown in SEQ ID NO: 10.
  • the above variable regions and CDRs are defined according to the Kabat numbering convention.
  • the anti-IL23 antibody is a humanized antibody
  • the heavy chain variable region of the anti-IL23 antibody has a protein derived from IGHV1-3*01 and / Or the light chain variable region has FR1, FR2, FR3 derived from IGKV2-40*01 and FR4 derived from IGKJ2*01, and it is unsubstituted or has a group selected from 4I, 36F, 39R, 71L and One or more amino acid substitutions in the group consisting of 100S.
  • the heavy chain variable region of the anti-IL23 antibody has HCDR1 as shown in SEQ ID NO: 11, HCDR2 as shown in SEQ ID NO: 12, and HCDR3 as shown in SEQ ID NO: 13, and LCDR1 of the light chain variable region is shown in SEQ ID NO: 14, LCDR2 is shown in SEQ ID NO: 37 or 15, and LCDR3 is shown in SEQ ID NO: 16.
  • the above variable regions and CDRs are defined according to the Kabat numbering convention.
  • the anti-IL23 antibody comprises a heavy chain variable region and a light chain variable region, wherein,
  • said heavy chain variable region is as set forth in SEQ ID NO: 22, 21, 20, 19, 18 or 17, and said light chain variable region is as set forth in SEQ ID NO: 26, 25, 24 or 23 show; or
  • said heavy chain variable region is set forth in SEQ ID NO: 29, 30, 31 or 32, and said light chain variable region is set forth in SEQ ID NO: 36, 35, 34 or 33; or
  • said heavy chain variable region is as shown in SEQ ID NO: 1, and said light chain variable region is as shown in SEQ ID NO: 2; or
  • the heavy chain variable region is as shown in SEQ ID NO:3, and the light chain variable region is as shown in SEQ ID NO:4.
  • the anti-IL23 antibody comprises a heavy chain variable region and a light chain variable region, wherein,
  • said heavy chain variable region is as shown in SEQ ID NO: 22, and said light chain variable region is as shown in SEQ ID NO: 26; or
  • said heavy chain variable region is as shown in SEQ ID NO: 21, and said light chain variable region is as shown in SEQ ID NO: 25; or
  • said heavy chain variable region is as shown in SEQ ID NO: 29
  • said light chain variable region is as shown in SEQ ID NO: 36.
  • the anti-IL23 antibody further comprises an antibody heavy chain constant region and a light chain constant region.
  • the heavy chain constant region is a human IgG heavy chain constant region; in some embodiments, the heavy chain constant region is selected from human IgG1, IgG2, IgG3 and IgG4 constant regions; in some embodiments, The light chain constant region is selected from a human antibody ⁇ or ⁇ chain constant region; in some embodiments, the heavy chain constant region is a human IgG1 heavy chain constant region, and the light chain constant region is a human ⁇ light chain constant region.
  • the Fc region of the heavy chain constant region has one or more amino acid substitutions that reduce binding of the Fc region to an Fc receptor.
  • the Fc region has L234A, L235A mutations, and/or S228P mutations, and/or YTE mutations (M252Y, S254T, and T256E), and the numbering of the mutations is based on the EU index.
  • the heavy chain constant region comprises the amino acid sequence of SEQ ID NO: 38
  • the light chain constant region comprises the amino acid sequence of SEQ ID NO: 39.
  • the anti-IL23 antibody comprises a heavy chain and a light chain, wherein,
  • the heavy chain of said anti-IL23 antibody comprises an amino acid sequence having at least 90% (e.g., at least 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO: 42, and a light chain comprising an amino acid sequence having at least 90% (e.g., at least 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO: 43; or
  • the heavy chain of said anti-IL23 antibody comprises an amino acid sequence having at least 90% (e.g., at least 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO: 40, and a light chain comprising an amino acid sequence having at least 90% (e.g., at least 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO: 41; or
  • the heavy chain of the anti-IL23 antibody comprises an amino acid sequence having at least 90% (e.g., at least 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO: 44, and the light chain comprise an amino acid sequence having at least 90% (eg, at least 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO: 45.
  • the anti-IL23 antibody comprises a heavy chain and a light chain, wherein
  • the TACI polypeptide is less likely to be broken than the wild-type TACI polypeptide (sequence such as SEQ ID NO: 51).
  • the TACI polypeptide is shown in SEQ ID NO: 58 or a truncated fragment of SEQ ID NO: 58 or a variant thereof; wherein the The truncated fragment comprises the 48th to the 85th amino acid residues of SEQ ID NO: 58, and the variant has a residue selected from the group consisting of 49, 52, 53, 57 on the basis of SEQ ID NO: 58 or a truncated fragment thereof. , one or more amino acid substitutions in positions 65, 82 and 83, wherein the position of the amino acid substitution is an amino acid residue position numbered relative to the natural sequence of the sequence SEQ ID NO:58.
  • the truncated fragment of the TACI polypeptide comprises: the 48th to the 86th amino acid residues of SEQ ID NO: 58; SEQ ID NO: The 48th to the 87th amino acid residue of 58; or the 48th to the 88th amino acid residue of SEQ ID NO:58.
  • the sequence of the TACI polypeptide is shown in any one of SEQ ID NO: 60-63.
  • the sequence of the TACI polypeptide is a variant of SEQ ID NO: 58 or a truncated fragment of SEQ ID NO: 58 (such as SEQ ID NO: 60 , SEQ ID NO: 61, SEQ ID NO: 62 or SEQ ID NO: 63), said variant has a sequence selected from 49, 52, Any 1, 2, 3, 4, 5, 6 or 7 amino acid substitutions in positions 53, 57, 65, 82 and 83, the position of the amino acid substitution is relative to the sequence SEQ ID NO: 58 amino acid residue positions numbered in natural order.
  • the TACI polypeptide is in SEQ ID NO: 58, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62 or SEQ ID NO: 63 sequence has one or more amino acid substitutions selected from the group consisting of 49T or 49R, 52S, 53E or 53Q, 57E, 65T or 65A, 82A or 82R, and 83Y (for example, 1, 2 , 3, 4, 5, 6 or 7 amino acid substitutions), wherein the amino acid substitution position is the amino acid residue position numbered relative to the natural sequence of the sequence SEQ ID NO:58.
  • the TACI polypeptide is in SEQ ID NO: 58, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62 or SEQ ID NO: 62 or SEQ ID NO: 63 sequence, there is any amino acid substitution selected from 49T, 52S, 53E, 53Q, 57E and 82A; in SEQ ID NO: 58, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID Based on the sequence of NO: 62 or SEQ ID NO: 63, there are 49R and 65T amino acid substitutions; in SEQ ID NO: 58, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62 or SEQ ID NO: 63 45R and 65A amino acid substitutions on the basis of the sequence; 49R, 65T and 82R amino acid substitution; 53E and 57E amino acid substitutions based on
  • SEQ ID NO: 60 SEQ ID NO: 61, SEQ ID NO: 62 or SEQ ID NO: 63
  • the sequence of the TACI polypeptide is shown in any one of SEQ ID NO: 60-63 and SEQ ID NO: 66-83. In some embodiments of the anti-IL23 antibody fusion protein according to any one of the above, the TACI polypeptide sequence is shown in SEQ ID NO: 83.
  • the anti-IL23 antibody fusion protein as described in any one of the above comprises:
  • the first chain from N-terminus to C-terminus, is: [TACI polypeptide 1]-[Linker 1]-[anti-IL23 antibody heavy chain]-[Linker 2]-[TACI polypeptide 2], and
  • the second chain which is an anti-IL23 antibody light chain
  • the TACI polypeptide 1 and the TACI polypeptide 2 can be the same or different, and the TACI polypeptide 1 and the TACI polypeptide 2 can be independently selected from any TACI polypeptide as described above; the linker 1 and the linker 2 may be the same or different; in some embodiments, the TACI polypeptide 1 and TACI polypeptide 2 are the same; in some embodiments, the linker is selected from (G x S) y linker, wherein X is selected from An integer of 1-5, Y selected from an integer of 0-6, in some embodiments, the linker is GGGGSGGGGSGGGGS (shown in SEQ ID NO: 84) or GGGS (shown in SEQ ID NO: 85).
  • the first chain of the anti-IL23 antibody fusion protein is shown in SEQ ID NO: 47, and the second chain is shown in SEQ ID NO: 43 or the first strand is as shown in SEQ ID NO:46, and the second strand is as shown in SEQ ID NO:41.
  • the anti-IL23 antibody fusion protein as described in any one of the above has 2 first chains such as SEQ ID NO: 47, and 2 second chains such as SEQ ID NO: 43; in some embodiments Among them, the anti-IL23 antibody fusion protein has two first chains such as SEQ ID NO: 46, and two second chains such as SEQ ID NO: 41.
  • amino acid sequence variants of the anti-IL23 antibodies provided herein, or fusion proteins thereof are contemplated.
  • Amino acid sequence variants of antibodies can be prepared by introducing appropriate modifications into the nucleotide sequence encoding the antibody, or by peptide synthesis. Such modifications include, for example, deletions, and/or insertions, and/or substitutions of residues within the amino acid sequence of the anti-IL23 antibody or its fusion protein. Any combination of deletions, insertions, and substitutions can be made to arrive at the final construct, so long as the final construct possesses the desired characteristics, such as antigen-binding properties.
  • antibody variants having one or more amino acid substitutions are provided.
  • Sites of interest for substitution mutagenesis include CDRs and FRs.
  • Conservative substitutions are shown in Table 3 under the heading "Preferred Substitutions”. More substantial changes are provided in Table 3 under the heading "Exemplary Substitutions" and are described further below with reference to amino acid side chain classes.
  • Amino acid substitutions can be introduced into an antibody of interest, and the products screened for desired activity, such as retained/improved antigen binding, reduced immunogenicity, or improved ADCC or CDC.
  • amino acids can be grouped as follows:
  • Non-conservative substitutions would entail replacing a member of one of these classes for a member of another class.
  • substitutional variant involves substituting one or more CDR residues of a parent antibody (eg, a humanized or human antibody).
  • a parent antibody eg. a humanized or human antibody
  • the resulting variant selected for further study will have an altered (e.g. improved) certain biological property (e.g. increased affinity, reduced immunogenicity) relative to the parent antibody, and/or will be substantially Some of the biological properties of the parental antibody are retained.
  • An exemplary substitution variant is an affinity matured antibody, which can be conveniently produced, for example, using phage display-based affinity maturation techniques such as those described herein. Briefly, one or more CDR residues are mutated, and the variant antibodies are displayed on phage and screened for specific biological activity (eg, binding affinity).
  • Alterations can be made to the CDRs, eg, to improve antibody affinity. Such changes can be made to CDR "hot spots", i.e. residues encoded by codons that undergo mutation at high frequency during the somatic maturation process, and/or residues that contact antigen, while making changes to the resulting variant VH or VL test for binding affinity.
  • affinity maturation diversity is introduced into the variable genes selected for maturation by any of a variety of methods, such as error-prone PCR, strand shuffling, or oligonucleotide-directed mutagenesis middle. Then, create secondary libraries. The library is then screened to identify any antibody variants with the desired affinity.
  • CDR residues involved in antigen binding can be specifically identified, for example, using alanine scanning mutagenesis or modeling.
  • HCDR3 and LCDR3 are frequently targeted.
  • substitutions, insertions or deletions may occur within one or more CDRs, so long as such changes do not substantially reduce the ability of the antibody to bind antigen.
  • conservative changes eg, conservative substitutions, as provided herein
  • Such changes may eg be outside antigen contacting residues in the CDRs.
  • each CDR is unchanged, or contains no more than 1, 2 or 3 amino acid substitutions.
  • alanine scanning mutagenesis One method that can be used to identify residues or regions of an antibody that can be targeted for mutagenesis is called "alanine scanning mutagenesis".
  • a residue or group of target residues e.g. charged residues such as Arg, Asp, His, Lys and Glu
  • neutral or negatively charged amino acids e.g. Ala or polyalanine
  • Further substitutions may be introduced at amino acid positions showing functional sensitivity to the initial substitution.
  • contact points between antibody and antigen can be identified by studying the crystal structure of the antigen-antibody complex. These contact residues and neighboring residues can be targeted or eliminated as candidates for substitution.
  • Variants can be screened to determine whether they contain desired properties.
  • Amino acid sequence insertions include amino and/or carboxyl terminal fusions ranging in length from 1 residue to polypeptides containing 100 or more residues, and intrasequence insertions of single or multiple amino acid residues.
  • terminal insertions include antibodies with an N-terminal methionyl residue.
  • Other insertional variants of antibody molecules include fusions of the N- or C-terminus of the antibody to enzymes or polypeptides that extend the serum half-life of the antibody.
  • the Fc region of an anti-IL23 antibody or anti-IL23 antibody fusion protein of the present disclosure comprises one or more amino acid substitutions that reduce its binding to an Fc receptor, such as its binding to an Fc ⁇ receptor binding and reduce or eliminate effector function.
  • a native IgG Fc region specifically an IgG 1 Fc region or an IgG 4 Fc region, may cause the fusion proteins of the present disclosure to target cells expressing Fc receptors, rather than cells expressing antigens.
  • an engineered Fc region of the present disclosure exhibits reduced binding affinity for an Fc receptor and/or reduced effector function.
  • the engineered Fc region has a binding affinity for Fc receptors that is reduced by more than 50%, 80%, 90%, or 95% compared to a native Fc region.
  • the Fc receptor is an Fc gamma receptor.
  • the Fc receptor is a human Fc ⁇ receptor, eg, Fc ⁇ RI, Fc ⁇ RIIa, Fc ⁇ RIIB, Fc ⁇ RIIIa.
  • the engineered Fc region also has reduced binding affinity for complement, such as C1q, compared to a native Fc region.
  • the engineered Fc region has no reduced binding affinity for neonatal Fc receptor (FcRn) compared to a native Fc region.
  • the engineered Fc region has reduced effector function, which may include, but is not limited to, one or more of the following: reduced complement-dependent cytotoxicity (CDC), reduced Antibody-dependent cell-mediated cytotoxicity (ADCC), decreased antibody-dependent cellular phagocytosis (ADCP), decreased cytokine secretion, decreased immune complex-mediated antigen uptake by antigen-presenting cells, decreased interaction with NK cells decreased binding to macrophages, decreased binding to monocytes, decreased binding to polymorphonuclear cells, decreased direct signaling-induced apoptosis, decreased dendritic cell maturation, or decreased T cells primed.
  • CDC complement-dependent cytotoxicity
  • ADCC Antibody-dependent cell-mediated cytotoxicity
  • ADCP antibody-dependent cellular phagocytosis
  • cytokine secretion decreased immune complex-mediated antigen uptake by antigen-presenting cells
  • decreased interaction with NK cells decreased binding to macrophages
  • monocytes decreased binding to monocytes
  • polymorphonuclear cells
  • amino acid residue substitutions at positions 238, 265, 269, 270, 297, 327, and 329 may reduce effector function.
  • the Fc region is a human IgG 1 Fc region, and the amino acid residues at positions 234 and 235 are A, and the numbering is based on the EU index.
  • amino acid residue substitutions at positions such as 228 may reduce effector function.
  • Anti-IL23 antibodies or anti-IL23 antibody fusion proteins may comprise different binding domains fused to the two subunits of the Fc region, thus potentially leading to undesired homodimerization.
  • the Fc region of the present disclosure comprises modifications according to the knob-into-hole (KIH) technique, which involves the introduction of a knob at the interface of the first subunit and the introduction of a knob at the interface of the second subunit.
  • KH knob-into-hole
  • the bulge structure is constructed by replacing small amino acid side chains from the interface of the first subunit with larger side chains such as tyrosine or tryptophan. Instead, the pore structure is created in the interface of the second subunit by replacing large amino acid side chains with smaller ones, such as alanine or threonine.
  • the protrusion structure and hole structure are prepared by changing the nucleic acid encoding the polypeptide, and the optional amino acid substitutions are shown in Table 4 below:
  • knob-and-hole technique In addition to the knob-and-hole technique, other techniques for modifying the CH3 domain of the heavy chain of a multispecific antibody to achieve heterodimerization are known in the art, for example WO96/27011, WO98/050431, EP1870459, WO2007/ 110205, WO 007/147901, WO2009/089004, WO2010/129304, WO2011/90754, WO2011/143545, WO2012/058768, WO2013/157954 and WO013/096291.
  • the C-terminus of the Fc region may be a complete C-terminus ending with the amino acid residue PGK; it may also be a shortened C-terminus in which, for example, one or two C-terminal amino acid residues have been removed.
  • the C-terminus of the heavy chain is a shortened C-terminus ending in PG.
  • a composition of intact antibodies can include a population of antibodies from which all K447 residues and/or G446+K447 residues have been removed.
  • a composition of intact antibodies can include a population of antibodies in which the K447 residue and/or the G446+K447 residues have not been removed.
  • the composition of whole antibodies has a population of antibodies with and without a K447 residue and/or a mixture of antibodies with G446+K447 residues.
  • Anti-IL23 antibodies or anti-IL23 antibody fusion proteins can be produced using recombinant methods. For these methods, one or more isolated nucleic acids encoding a polypeptide or fusion protein are provided.
  • the present disclosure provides an isolated nucleic acid encoding an anti-IL23 antibody or anti-IL23 antibody fusion protein as previously described. Such nucleic acid may be derived from an independent polypeptide chain encoding any of the foregoing.
  • the present disclosure provides one or more vectors (eg, expression vectors) comprising such nucleic acids.
  • the disclosure provides host cells comprising such nucleic acids.
  • a method of producing a polypeptide or fusion protein comprising, under conditions suitable for expression, culturing a host cell comprising a nucleic acid encoding said polypeptide or fusion protein, as provided above, and The anti-IL23 antibody or anti-IL23 antibody fusion protein is optionally recovered from the host cell (or host cell culture medium).
  • nucleic acid encoding the protein is isolated and inserted into one or more vectors for further cloning and/or expression in host cells.
  • nucleic acids can be readily isolated and sequenced using conventional procedures, or produced by recombinant methods or obtained by chemical synthesis.
  • Suitable host cells for cloning or expressing vectors encoding anti-IL23 antibodies or anti-IL23 antibody fusion proteins include prokaryotic or eukaryotic cells described herein. For example, it can be produced in bacteria, especially when glycosylation and Fc effector functions are not required. After expression, it can be isolated from the bacterial cell paste in a soluble fraction and can be further purified.
  • eukaryotic microbes such as filamentous fungi or yeast are suitable cloning or expression hosts for fusion protein-encoding vectors, including fungal and yeast strains.
  • Suitable host cells for expression of fusion proteins may also be derived from multicellular organisms (invertebrates and vertebrates); examples of invertebrate cells include plant and insect cells.
  • a number of baculovirus strains have been identified for use in combination with insect cells, particularly for the transfection of Spodoptera frugiperda cells; plant cell cultures can also be used as hosts, e.g.
  • vertebrate cells can also be used as hosts, such as mammalian cell lines adapted for growth in suspension.
  • suitable mammalian host cell lines are the SV40-transformed monkey kidney CV1 line (COS-7); the human embryonic kidney line (293 or 293T cells); baby hamster kidney cells (BHK); Sertoli) cells (TM4 cells); monkey kidney cells (CV1); African green monkey kidney cells (VERO-76); human cervical cancer cells (HELA); canine kidney cells (MDCK); buffalo rat liver cells ( BRL3A); human lung cells (W138); human hepatocytes (Hep G2); mouse mammary tumor (MMT 060562); TRI cells; MRC 5 cells; and FS4 cells.
  • Suitable mammalian host cell lines include Chinese Hamster Ovary (CHO) cells, including DHFR-CHO cells; and myeloma cell lines, such as YO, NSO and Sp2/0.
  • CHO Chinese Hamster Ovary
  • myeloma cell lines such as YO, NSO and Sp2/0.
  • Anti-IL23 antibodies or anti-IL23 antibody fusion proteins provided herein can be identified, screened or characterized for their physical/chemical characteristics and/or biological activity by various assays known in the art.
  • the anti-IL23 antibody or anti-IL23 antibody fusion protein of the present disclosure is tested for activity, eg, by known methods such as ELISA, Western blot, and the like.
  • any of the anti-IL23 antibodies or anti-IL23 antibody fusion proteins provided herein can be used in methods of treatment.
  • the present disclosure provides the use of an anti-IL23 antibody or an anti-IL23 antibody fusion protein in the manufacture or preparation of a medicament.
  • the B cell disorder or autoimmune disease is an IL23-associated disease or condition.
  • the autoimmune disease is selected from the group consisting of: systemic lupus erythematosus, myasthenia gravis, multiple sclerosis, insulin-dependent diabetes mellitus, Crohn's disease, rheumatoid arthritis, polyarticular juvenile rheumatoid Arthritis and psoriatic arthritis; said B-cell disorder is selected from the group consisting of neoplasms, chronic leukaemia, multiple myeloma, non-Hodgkin's lymphoma, post-transplantation lymphoproliferative disease, and light chain gammopathies .
  • the autoimmune disease is systemic lupus erythematosus.
  • the use further comprises administering to the subject an effective amount of at least one additional therapeutic agent (e.g., one, two, three, four, five, or six additional therapeutic agents agent).
  • additional therapeutic agent e.g., one, two, three, four, five, or six additional therapeutic agents agent.
  • a "subject" according to any of the above embodiments may be a human.
  • a pharmaceutical composition comprising the anti-IL23 antibody or anti-IL23 antibody fusion protein, for example, for any of the above pharmaceutical uses or methods of treatment.
  • a pharmaceutical composition comprises any of the polypeptides or fusion proteins provided herein and a pharmaceutically acceptable carrier.
  • the pharmaceutical composition further comprises at least one additional therapeutic agent.
  • the anti-IL23 antibody or anti-IL23 antibody fusion protein of the present disclosure can be used alone or in combination with other agents for therapy.
  • an anti-IL23 antibody or anti-IL23 antibody fusion protein of the present disclosure can be co-administered with at least one additional therapeutic agent.
  • Anti-IL23 antibodies or anti-IL23 antibody fusion proteins of the present disclosure can be administered by any suitable means, including parenteral, intrapulmonary, and intranasal, and if desired for local treatment, intralesional administration.
  • Parenteral infusions include intramuscular, intravenous, intraarterial, intraperitoneal or subcutaneous administration. Administration may be by any suitable route, eg, by injection, such as intravenous or subcutaneous injection, depending in part on whether the administration is short-term or chronic.
  • a variety of dosing schedules are contemplated herein, including, but not limited to, single or multiple administrations at multiple time points, bolus administration, and pulse infusion.
  • the anti-IL23 antibodies or anti-IL23 antibody fusion proteins of the present disclosure will be formulated, dosed, and administered in a manner consistent with good medical practice.
  • Factors considered in this context include the particular condition being treated, the particular mammal being treated, the clinical condition of the individual patient, the cause of the condition, the site of delivery of the agent, the method of administration, the timing of administration, and others known to the medical practitioner.
  • a polypeptide or fusion protein may or may not be formulated with one or more agents currently used to prevent or treat the disorder. The effective amount of such other agents depends on the amount present in the pharmaceutical composition, the type of disorder or treatment, and other factors. These are generally used at the same dosages and routes of administration as described herein, or at about 1 to 99% of the dosages described herein, or at other dosages, and any route empirically/clinically determined to be appropriate.
  • the appropriate dose of the anti-IL23 antibody or anti-IL23 antibody fusion protein of the present disclosure (when used alone or in combination with one or more other additional therapeutic agents) will depend on the disease to be treated type of drug, type of therapeutic molecule, severity and course of the disease, whether administered for prophylactic or therapeutic purposes, previous therapy, patient's clinical history and response to the therapeutic molecule, and the judgment of the attending physician.
  • the therapeutic molecule is suitably administered to the patient at one time or over a series of treatments.
  • an article of manufacture comprising materials useful for the treatment, prevention and/or diagnosis of the disorders described above.
  • the article comprises a container and a label or package insert on or associated with the container.
  • Suitable containers include, for example, bottles, vials, syringes, IV solution bags, and the like.
  • Containers can be formed from various materials such as glass or plastic.
  • the container contains a composition effective, alone or in combination with another composition, for the treatment, prophylaxis and/or diagnosis of a condition, and may have a sterile access opening (e.g., the container may have a stopper pierceable by a hypodermic needle). IV solution bag or vial).
  • At least one active agent in the composition is an anti-IL23 antibody or anti-IL23 antibody fusion protein of the present disclosure.
  • the label or package insert indicates that the composition is used to treat the condition of choice.
  • the article of manufacture may comprise: (a) a first container having a composition therein, wherein the composition comprises an anti-IL23 antibody or an anti-IL23 antibody fusion protein of the present disclosure; and (b) a second container having the composition therein A container, wherein the composition comprises an additional cytotoxic or other therapeutic agent.
  • the article of manufacture of this embodiment of the present disclosure may further comprise a package insert indicating that the composition may be used to treat a particular condition.
  • the article of manufacture may further comprise a second (or third) container comprising a pharmaceutically acceptable buffer. It may further comprise other materials as desired from a commercial and user standpoint, including other buffers, diluents, filters, needles and syringes.
  • Embodiment 1 Preparation of mouse anti-human IL-23 p19 monoclonal antibody
  • mice were immunized with human IL-23 protein (Sino Biological, CT048-H08H). After 3 times of immunization, blood was taken to measure the titer of the antibody in the serum, and the mice with high antibody titer in the serum and the titer tended to plateau were selected for splenocyte fusion, and the fused hybridoma cells were plated in a 96-well cell culture plate , placed in a 37°C, 5% CO 2 incubator for cultivation. The cell culture supernatant was taken for detection by enzyme-linked immunosorbent assay (ELISA). The screened positive clones were expanded, cryopreserved and subcloned two to three times until single-cell clones were obtained.
  • ELISA enzyme-linked immunosorbent assay
  • the selected hybridoma clones were further prepared and purified by serum-free cell culture method.
  • the resulting hybridoma antibody is detected by ELISA for the binding of the antibody to the human IL-23 protein and the blocking of the receptor (see test example 1 and test example 2 of this disclosure for the method), and the hybridization with good binding activity and blocking activity is selected. tumor cell lines.
  • Sequences of monoclonal antibodies cloned from monoclonal hybridoma cell lines mAb29 and mAb38 were selected. The process is as follows: hybridoma cells in logarithmic growth phase were collected, RNA was extracted with Trizol (Invitrogen, Cat#15596-018), and reverse transcribed into cDNA. Use cDNA as a template for PCR amplification and send it to the sequencing company for sequencing.
  • the amino acid sequence of the variable region of the antibody corresponding to the obtained DNA sequence is as follows:
  • variable region sequences of the above mAb29 and mAb38 candidate molecules were respectively amplified by PCR to amplify the VH/VK sequences, and then homologously recombined with the expression vector pHr (with signal peptide and hIgG1/hkappa constant region gene (CH1-Fc/CL) fragment) ,
  • the human heavy chain IgG1 constant region sequence is shown in SEQ ID NO: 38
  • the human light chain ⁇ constant region sequence is shown in SEQ ID NO: 39
  • the recombinant chimeric antibody full-length expression plasmid VH-CH1- Fc-pHr/VL-CL-pHr and then obtain its chimeric antibodies Ch29 and Ch38.
  • the heavy and light chain variable region germline genes with high homology were selected as templates, and the CDRs of the mouse antibody were grafted (Grafted) into the corresponding In the human source template, a variable region sequence in the order of FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4 is formed, and then the variable region sequence is fused with a human constant region sequence to obtain a humanized antibody.
  • the humanization of mAb29 and mAb38 murine antibodies is exemplarily described below, and the CDR amino acid residues of the antibodies in the examples are determined and annotated by the Kabat numbering system.
  • the heavy and light chain variable region germline genes with high homology were selected as templates.
  • IGKV4-1*01 and IGKJ4*01 were selected as templates for the humanized light chain of the mouse antibody mAb29, that is, FR1, FR2, FR3 of the human germline light chain IGKV4-1*01, and the JK4 region of IGKJ4*01 were selected (as FR4) is used as the framework region of the humanized antibody light chain;
  • the humanized heavy chain templates are IGHV2-26*01 and IGHJ6*01, that is, FR1, FR2, FR3, and IGHJ6 of the human germline heavy chain IGHV2-26*01 are selected *
  • the JH6 region of 01 was used as the humanized antibody light chain framework region.
  • the CDRs of the murine antibody mAb29 were grafted into the corresponding human templates selected to replace the CDR regions of the human templates; then, the 4th, 17th, 36th, and 58th , 60 and/or 68 amino acid residues (numbering according to the Kabat numbering system) are mutated, and the 1st, 30th, 37th, 44, 49, 73, 89 and/or 93 (according to the Kabat numbering system) of the heavy chain variable region are mutated Number) amino acid residues were mutated, in addition, the first amino acid residue of HCDR1: SYAIS (SEQ ID NO: 5) in the heavy chain variable region was mutated to N or Q, and a new HCDR1: NYAIS (SEQ ID NO: 5) was obtained NO: 27) or QYAIS (SEQ ID NO: 28), the humanized antibody variable region sequence of mAb29 is as follows:
  • sequence order is FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4, the underlined part in the sequence is the CDR region (CDR amino acid residues are determined by the Kabat numbering system), and the rest is the FR region.
  • IGKV2-40*01 and IGKJ2*01 were selected as templates for the humanized light chain of the mouse antibody mAb38, that is, FR1, FR2, FR3 of the human germline light chain IGKV2-40*01, and the JK2 region of IGKJ2*01 were selected (as FR4) is used as the framework region of the humanized antibody light chain; the humanized heavy chain templates are IGHV1-3*01 and IGHJ1*01, that is, FR1, FR2, FR3, and IGHJ1 of the human germline heavy chain IGHV1-3*01 are selected * The JH1 region of 01 (as FR4) was used as the heavy chain framework region of the humanized antibody.
  • the CDRs of the murine antibody mAb38 were transplanted into corresponding selected human templates to replace the CDR regions of the human templates.
  • the 4th, 36th, 39th, and 39th regions of the light chain variable regions of the humanized antibody were Amino acid residues 71 and/or 100 (numbered according to the Kabat numbering system) are mutated, and amino acid residues 1, 2, 28, 44, 71 and/or 73 (numbered according to the Kabat numbering system) of the heavy chain variable region
  • the third amino acid residue of LCDR2: LMSTRAS (SEQ ID NO: 15) in the light chain variable region was mutated to Q to obtain a new LCDR2: LMQTRAS (SEQ ID NO: 37), mAb38
  • the sequence of the variable region of the humanized antibody is as follows:
  • sequence order is FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4, the underlined part in the sequence is the CDR region (CDR amino acid residues are determined by the Kabat numbering system), and the rest is the FR region.
  • the expression vectors of antibody light chain and heavy chain were respectively constructed, and the humanized antibody light and heavy chains were cross-paired and combined, and the culture supernatant was collected and purified after transfection into 293E cells to obtain the humanized full-length antibody.
  • the constant region of the heavy chain of the humanized antibody can be selected from constant regions of IgG1, IgG2, IgG3, and IgG4.
  • the constant region of the human heavy chain IgG1 is used
  • the constant region of the light chain of the humanized antibody can be selected from the group consisting of human ⁇ , Lambda chain constant region, the sequence of the constant region of an exemplary antibody is as follows:
  • the humanized antibody heavy chain variable region of the aforementioned mAb29 is fused with the human heavy chain IgG1 constant region (sequence shown in SEQ ID NO: 38) to form an antibody full-length heavy chain, and the humanized antibody light chain
  • the variable region was fused with the human light chain ⁇ constant region (sequence shown in SEQ ID NO: 39) to form the full-length light chain of the antibody, and the mAb29 humanized antibody shown in Table 6 below was obtained:
  • the heavy chain variable region of the humanized antibody derived from mAb38 is fused with the human heavy chain IgG1 constant region (sequence shown in SEQ ID NO: 38) to form the full-length heavy chain of the antibody, and the light chain of the humanized antibody can be
  • the variable region was fused with the human light chain ⁇ constant region (sequence shown in SEQ ID NO: 39) to form the full-length light chain of the antibody, and the mAb38 humanized antibody shown in Table 7 below was obtained:
  • Embodiment 3 Construction of anti-IL23 antibody fusion protein
  • the aforementioned anti-IL23 antibody was fused with the TACI polypeptide to construct an anti-IL23 antibody fusion protein.
  • the anti-IL23 antibody is Hu29-19 or Hu29-24.
  • the TACI polypeptide can be any suitable TACI polypeptide.
  • the TACI polypeptide is derived from Chinese patent application 202110348497.6 (invention name: a new type of TACI polypeptide, its fusion protein and its use; application date: 2021/3/31) and will TACI polypeptides described in its patent application (incorporated herein by reference in its entirety) as a priority document, such as the TACI polypeptides shown in Table 9 below:
  • two TACI polypeptides are respectively fused to the N-terminal and C-terminal of the heavy chain of an anti-IL23 antibody (such as Hu29-19 or Hu29-24), and the TACI polypeptide at the C-terminal of the heavy chain of the antibody is fused SS is added to the C-terminus of the polypeptide as a protective amino acid to prevent the TACI polypeptide from being cut by carboxypeptidase, and the structural diagram of the anti-IL23 antibody fusion protein (Hu29-19T and Hu29-24T) constructed and obtained is shown in Figure 1, heavy chain and light chain amino acids The sequence looks like this:
  • the single underline in the sequence is the TACI sequence
  • the double underline is the antibody variable region
  • the italic is the antibody constant region
  • the bold is the linker.
  • control molecule RCT-18 telitacicept, RC18, Rongchang Biotechnology, WHO (SEQ ID: 10932) is shown below.
  • the binding activity of the test molecule to IL-23 p19 and BAFF was detected by the Elisa method (coating the test molecule).
  • the specific method is as follows:
  • the sample to be tested was diluted to 2 ⁇ g/mL with pH 7.4 PBS (B320), added to a 96-well microplate (Corning, 3590) at a volume of 100 ⁇ L/well, and incubated overnight at 4°C. After the liquid was discarded, 300 ⁇ L of 5% skimmed milk (BD, 232100) diluted with PBS was added to each well for blocking, and incubated at 37° C. for 2 hours.
  • TMB chromogenic substrate KPL, 5120-00757
  • 50 ⁇ L 1M H 2 SO 4 50 ⁇ L 1M H 2 SO 4 to each well to stop the reaction, and read it with a microplate reader
  • the absorption value at 450nm was used to fit the binding curve of the antibody to the antigen by software, and the EC 50 value was calculated.
  • the receptor protein was diluted to 2 ⁇ g/mL with pH 7.4 PBS (B320), added to a 96-well microplate (Corning, 3590) at a volume of 100 ⁇ L/well, and incubated overnight at 4°C. After the liquid was discarded, 200 ⁇ L of 1% Casein blocking solution (Thermo, 37528) was added to each well for blocking, and incubated at 37° C. for 2 hours. After the blocking, the blocking solution was discarded, and the plate was washed 3 times with PBST buffer (pH7.4 PBS containing 0.1% tween-20) before use.
  • PBST buffer pH7.4 PBS containing 0.1% tween-20
  • a fixed concentration of biotin-labeled ligand protein was mixed with a serially diluted antibody or fusion protein, pre-incubated at 37°C for 30 minutes, then added to the blocked microtiter plate, and incubated at 37°C for 1.5 hours. After the incubation, the plate was washed 3 times with PBST, 100 ⁇ L of streptavidin-HRP (Invitrogen, 434323, diluted 1:4000) was added to each well, and incubated at 37° C. for 1 hour.
  • streptavidin-HRP Invitrogen, 434323, diluted 1:4000
  • TMB chromogenic substrate KPL, 5120-00757
  • 50 ⁇ L 1M H 2 SO 4 50 ⁇ L 1M H 2 SO 4
  • a microplate reader to Read the absorbance value at 450nm
  • the sources of the ligand proteins used in this test example are as follows: IL-23 (Sino Biological, CT048-H08H), BAFF (Sino biological, 10056-HNCH), APRIL (R&D Systems, 5860-AP-010/CF).
  • the sources of the receptor proteins used are as follows: IL-23R (Sino Biological, 13840-H02H), BAFF-R (Sino biological, 16079-H02H), BCMA (Sino biological, 10620-H02H), TACI (ACROBiosystems, TAI-H5256 ).
  • BaF3 cells (Cobioer, CBP60474) stably expressing IL-23R and IL-12R ⁇ 1 were plated in a 96-well cell plate (Corning, 3903), with a volume of 50 ⁇ L per well and 4000 cells. Mix the serially diluted sample to be tested with a fixed concentration of IL-23 protein (Sino Biological, CT048-H08H), add it to a cell culture plate, and incubate in a 37°C incubator for 72 hours.
  • IL-23 protein Seo Biological, CT048-H08H
  • Hu38-10 0.1161 Hu38-11 0.1683 Hu38-12 0.1838 Hu38-13 0.1866 Hu38-14 0.1522 Hu38-15 0.1493 Hu38-16 0.1458 Hu29-1 0.07111 Hu29-2 0.06227 Hu29-5 0.05531 Hu29-6 0.08897 Hu29-9 0.05608 Hu29-10 0.08601 Hu29-15 0.2158 Hu29-16 0.2251 Hu29-23 0.1184
  • the IL-17 secretion assay was used to detect the inhibition of IL-23-induced T cell differentiation by anti-IL23 antibody and its fusion protein.
  • the experimental method is as follows:
  • the cell suspension was sorted with the Mouse CD4 Cells Kit (Invitrogen, 11415D), and the isolated CD4+ T cells were resuspended with the medium RPMI 1640 Medium (Gibco, 11875119)+10% FBS (Gibco, 10099-141) and counted for later use .
  • Spread the cell suspension in a coated 96-well plate mix a fixed concentration of IL-23 (R&D Systems, 1290-IL-010) with a serially diluted antibody or fusion protein, and pre-incubate for 1 hour before adding to the 96-well plate Incubate for 48 hours in a 37°C cell culture incubator.
  • the B cell proliferation assay was used to detect whether the anti-IL23 antibody fusion protein could inhibit the B cell proliferation induced by BAFF and APRIL.
  • the experimental method is as follows:
  • the cell suspension was sorted with B Cell Isolation Kit (Miltenyi Biotec, 130-090-862), and the isolated B cells were sorted with RPMI 1640 Medium (Gibco, 11875119)+10% FBS (Gibco, 10099-141)+50 ⁇ M 2 -mercaptoethanol (Sigma-Aldrich, M6250) was resuspended and counted, and the cells were plated in a 96-well cell plate (Costar, 3903) for later use.
  • Biosensing chip Protein A (GE, 29127556) to affinity capture a certain amount of the sample to be tested, and then flow through a series of concentration gradient antigens on the surface of the chip, and use Biacore (GE, 8K) to detect the reaction signal in real time to obtain the binding and dissolving off the curve.
  • Biacore GE, 8K
  • the biochip was cleaned and regenerated with 10 mM glycine-hydrochloric acid solution pH 1.5 (GE, BR-1003-54).
  • the experimental data was fitted with a 1:1 model using BIAevaluation version 4.1 software to obtain the affinity value.
  • the relevant antigen proteins used in this test are as follows: human IL-23 (Sino biological, CT048-H08H), human BAFF (Sino biological, 10056-HNCH), human APRIL (R&D Systems, 5860-AP-010/CF), Cynomolgus monkey IL-23 (Acro Biosystems, ILB-CM52W8), cynomolgus monkey BAFF (Kactus, BAF-CM412), cynomolgus monkey APRIL (Kactus, APR-CM410B), mouse IL-23 (R&D Systems, 1887-ML /CF), mouse BAFF (Acro Biosystems, BAF-M521y), mouse APRIL (R&D Systems, 7907-AP/CF).
  • the test results of affinity are shown in Table 19 to Table 25 below:
  • the anti-IL23 antibody and its fusion protein constructed in this disclosure can bind to human IL-23 p19 and cynomolgus monkey IL-23 p19 with high affinity, but not to mouse IL-23 p19;
  • the IL23 antibody fusion protein can bind to human BAFF, cynomolgus monkey BAFF, mouse BAFF, human APRIL, cynomolgus monkey APRIL and mouse APRIL with high affinity.
  • Test Example 7 Animal Model Experiment of Psoriasis Induced by Imiquimod
  • Risankizumab was used as a positive control, and PBS was used as a negative control, and they were administered twice in total. The severity of rash and desquamation of the back skin of the mice was scored every day, and the experiment was ended on the 6th day.
  • Rash severity was scored as follows: no rash, 0 points; reddish, 1 point; red but not dark, 2 points; deep red, 3 points; extremely red, 4 points.
  • the scoring criteria for desquamation are as follows: no desquamation, 0 points; small area of fine dander, 1 point; moderate area of thicker dander, 2 points; large area of rough thickened dander, 3 points; large area of Large pieces of dander, 4 points.
  • Test Example 8 Psoriasis Animal Model Experiment Induced by Human IL-23
  • the in vivo drug efficacy of the anti-IL23 antibody constructed in the present disclosure was evaluated by human IL-23-induced psoriasis animal model.
  • SPF grade female C57BL/6J mice (Weitong Lihua Experimental Animal Technology Co., Ltd.), 6 to 8 weeks old, measured the thickness and weight of the right ear of the mice after anesthesia, and grouped them according to the thickness and weight of the right ear. 6 mice per group. The day of grouping was recorded as day 0. From day 1, 1 ⁇ g of human IL-23 protein (R&D Systems, 1290-IL-500/CF) was intradermally injected into the right ear of the mice every day for a total of 7 days, and the mice in the normal group were injected with PBS.
  • human IL-23 protein R&D Systems, 1290-IL-500/CF
  • the test drug was injected intraperitoneally, and the mouse body weight and the thickness of the right ear were measured and recorded on the 0, 2, 4, 6, and 8 days respectively, and the effect of the test drug was evaluated by the change of the right ear thickness.
  • the mitigation of the model At the end of the experiment on the 8th day, ear pieces with a diameter of 8 mm were collected and weighed.
  • the experimental results are shown in Figures 5, 6, and 7.
  • the experimental results show that the anti-IL23 antibody Hu29-19 constructed in this disclosure has stronger in vivo efficacy than the control molecule Risankizumab in an animal model of psoriasis induced by human IL-23.
  • the ear thickness of the Hu29-19 group was lower than that of the control Risankizumab; the statistical results of the area under the curve showed that at a dose of 3mpk, the effect of Hu29-19 was stronger than that of Risankizumab; the mouse ear weight The statistical results show that the ear weight of the Hu29-19 1mpk group is the same as that of the Risankizumab 3mpk group, and the ear weight of the Hu29-19 3mpk group is lower than that of the Risankizumab group.
  • mice Simultaneously stimulate mice with human IL-23 and human BAFF proteins to induce the production of cytokines such as TNF ⁇ , IL-22 and IgA in mice, and evaluate the in vivo activity of antibodies or fusion proteins by detecting the levels of these cytokines.
  • cytokines such as TNF ⁇ , IL-22 and IgA
  • mice in each group On the fifth day, plasma samples of mice in each group were collected, and the levels of TNF ⁇ , IL-22 and IgA were detected respectively.
  • the source information of the detection kits used in this test case is as follows: Mouse TNF-alpha Quantikine ELISA Kit (R&D Systems, MTA00B), Mouse/Rat IL-22 Quantikine ELISA Kit (R&D Systems, M2200), Mouse IgA ELISA Kit (Abcam, ab157717).
  • RCT-18 and Risankizumab were used as positive controls, and PBS was used as negative controls. 4mpk RCT-18, 8mpk Risankizumab, 8.8mpk Hu29-19T have the same drug molar concentration.
  • the detection results of IgA show that the anti-IL23 antibody fusion protein constructed in the present disclosure inhibits the secretion of IgA significantly stronger than the control molecule RCT-18, while Risankizumab has no inhibitory activity on the secretion of IgA;
  • the detection results of TNF ⁇ show that at the dose of equimolar concentration, the anti-IL23 antibody fusion protein constructed by the present disclosure has stronger activity in inhibiting the secretion of TNF ⁇ than Risankizumab, while RCT-18 does not show significant Inhibitory effect:
  • IL-22 detection results show that at equimolar doses, the anti-IL23 antibody fusion protein constructed in the present disclosure has stronger activity in inhibiting IL-22 secretion than the control risankizumab and RCT-18.
  • Test Example 10 In Vivo Drug Effect in Animal Model of Systemic Lupus Erythematosus
  • MRL/lpr mice will spontaneously develop symptoms of autoimmune diseases as they age, and the symptoms are similar to human systemic lupus erythematosus. This model was used to evaluate the in vivo efficacy of the fusion protein.
  • MRL/lpr mice female, were purchased from Shanghai Slack Experimental Animal Co., Ltd., 5 mice/cage were raised in an SPF-grade environment at a temperature of 20-25°C and a humidity of 40-60%. Mice were acclimatized in the laboratory environment for at least one week before the experiment.
  • mice When the mice reached 9 to 10 weeks of age, the urine protein and dsDNA IgG values were significantly higher than those of normal control mice, and the subcutaneous injection was started (RCT-18 (8mpk), Hu29-19T (17.6mpk), Hu29-19T ( 8.8mpk), Hu29-24T (17.6mpk), negative control (PBS)), wherein RCT-18 8mpk, Hu29-19T 17.6mpk, Hu29-24T 17.6mpk have the same molar concentration.
  • the drug was administered twice a week (once on Monday and once on Thursday), and continued for 8 weeks. Regularly observe the hair and skin condition of the mice, score the severity of the skin damage of the mice, collect urine and plasma, and detect the urine protein content.
  • the kidneys of the mice were taken out, fixed and stained with H&E to observe glomerulus, tubular lesions, and inflammatory cell infiltration in the kidney tissues of the mice in each group, and score the degree of renal lesions.
  • the urine protein was detected by Bradford protein concentration assay, and the detection kit was purchased from Beyond Biotechnology Co., Ltd.
  • the experimental results are shown in Tables 29, 30, 31 and accompanying drawings 11, 12, and 13.
  • the experimental results showed that RCT-18 did not reduce the urinary protein level of mice, and Hu29-19T could significantly reduce the level of urinary protein in both doses.
  • the urinary protein level of Hu29-19T group mice remained the same or lower than that at the beginning of the experiment, and there was no tendency to increase at all.
  • Hu29-24T also significantly reduced urinary protein in mice.
  • the results of the skin damage score of the mice showed that the skin damage score of the mice in the Hu29-19T group had always been the lowest, and Hu29-24T was at an intermediate level, and RCT-18 had no obvious drug effect on this indicator.
  • the renal pathological scoring results of the mice showed that compared with the negative group, the two dose groups of Hu29-19T showed significant statistical differences.

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Abstract

抗IL23抗体融合蛋白及其用途，所述抗IL23抗体融合蛋白包含抗IL23抗体和TAC1多肽，所述抗IL23抗体特异性结合人IL23 p19亚基。所述抗IL23抗体或融合蛋白可用于治疗或改善B细胞障碍或自身免疫性疾病。

Description

抗IL23抗体融合蛋白及用途 技术领域
本披露属于生物技术领域，更具体地，本披露涉及一种抗IL23抗体及其融合蛋白和应用。
背景技术
这里的陈述仅是提供与本披露有关的背景信息，而不必然地构成现有技术。
IL-23(也称IL23)主要由活化的树突状细胞、巨噬细胞及单核细胞等产生，是IL-12异源二聚体细胞因子家族中的一员，由p19(也称IL23 p19亚基)和p40(也称IL23 p40亚基)两个亚基组成，其中p40亚基是与IL-12共同含有的亚基(J Immunol.2018Sep 15；201(6):1605-1613.)。
IL-23通过与其受体相互作用，激活下游信号通路而发挥生物学功能。IL-23受体包括IL-12受体β1和IL-23受体2个亚基(J Immunol.2002 Jun 1；168(11):5699-708.)。IL-23能促进Th17细胞分化，在Th17细胞的增殖与稳定中发挥重要作用，能促进Th17细胞产生IL-17A、IL-17F及IL-22等细胞因子，这些炎症因子作用于角质形成细胞，导致角质形成细胞活化和过度增殖。活化的角质形成细胞通过产生大量细胞因子、趋化因子和抗菌肽等，募集并激活T细胞等免疫细胞，形成免疫应答的级联效应，引起银屑病的症状(Cutis.2018 Mar；101(3S):5-9.)。
系统性红斑狼疮(SLE)是一种免疫反应过度激活引起的自身免疫疾病(Nat Rev Dis Primers.2016 Jun 16；2:16039.)。有多种细胞参与了SLE疾病的发展。源头是中性粒细胞和凋亡的细胞，中性粒细胞释放网状的胞外陷阱，捕获抗菌蛋白，在干扰素的共同作用下刺激pDC分泌大量的I型干扰素。而凋亡的细胞会将核内的高迁移率蛋白和核酸释放出来，作为自身的抗原诱导免疫细胞产生核苷酸抗体，核酸和抗体的免疫复合物与细胞表面的受体作用激活TLR信号，刺激pDC分泌干扰素。干扰素进一步激活mDC产生多种细胞因子，分别激活T细胞和B细胞(Curr Opin Rheumatol.2017 Mar；29(2):178-186.)。其中IL-12和23是两种已知可以激活T细胞分化的细胞因子。T细胞激活后会分泌多种细胞因子，IL-17等细胞因子参与募集炎症细胞并导致组织损伤，IL-21等细胞因子参与B细胞的共刺激。BAFF和APRIL是B细胞的激活因子，它们通过与B细胞表面的受体作用刺激B细胞分化成熟并分泌抗体(Nat Rev Rheumatol.2016 Nov 22；12(12):716-730.)。
TACI是一种膜结合受体，具有包含二个富含半胱氨酸的假性重复片断(cysteine-rich pseudo-repeats)的胞外区、一个跨膜区和与CAML(钙调节剂和亲环蛋白配体)相互作用的胞质区。TACI与B细胞和T细胞的一种亚型相关。TACI受体与肿瘤坏死因子配体家族的BAFF相结合。BAFF是属于TNF家族的一种B细胞 激活因子，BAFF主要在骨髓细胞膜表面表达，以三聚体的形式存在，在细胞膜表面的BAFF会被蛋白酶水解形成可溶性的BAFF进入血液循环系统，可溶性的BAFF具有多聚化的特征，最多可以形成60聚体。另外BAFF也可以跟同家族的另外一个蛋白APRIL作用形成异源的三聚体。目前已知在B细胞表面有三个BAFF的受体，分别为BAFF-R、BCMA和TACI。BAFF与这三个受体作用，参与B细胞的分化成熟、存活和调节。APRIL与BAFF有两个共同的受体，分别为BCMA和TACI，APRIL与这两个受体作用参与B细胞的存活和调节(Samy,E.,et al.,Int Rev Immunol,2017.36:p.3-19；Kamal,A.and M.Khamashta,Autoimmun Rev,2014.13:p.1094-1101)。BAFF对维持B细胞的体内平衡非常重要，BAFF信号通路的过度激活会导致自反应B细胞的存活并产生自身抗体促进自身免疫反应(Cancro,M.P.,D.P.D'Cruz,and M.A.Khamashta,J Clin Invest,2009.119:p.1066-73)。
发明内容
本披露构建了一种抗IL23抗体融合蛋白，其包含抗IL23抗体和TACI多肽，其中所述抗IL23抗体特异性结合人IL23 p19亚基。
根据本披露的抗IL23抗体融合蛋白的一些实施方案，所述抗IL23抗体融合蛋白包含与TACI多肽融合的抗IL23抗体，或由其组成；优选地，所述TACI多肽在所述抗IL23抗体的重链的N段和/或C端与所述抗IL23抗体融合。
在一些实施方案中，所述抗IL23抗体融合蛋白中的所述抗IL23抗体包含重链可变区和轻链可变区，其中，所述重链可变区包含HCDR1、HCDR2和HCDR3，所述轻链可变区包含LCDR1、LCDR2和LCDR3，其中，
(i)所述重链可变区的HCDR1、HCDR2和HCDR3分别包含SEQ ID NO：22、21、20、19、18、17或1中的HCDR1、HCDR2和HCDR3的氨基酸序列，和所述轻链可变区的LCDR1、LCDR2和LCDR3分别包含SEQ ID NO：26、25、24、23或2中的LCDR1、LCDR2和LCDR3的氨基酸序列；或
(ii)所述重链可变区的HCDR1、HCDR2和HCDR3分别包含SEQ ID NO：29、30、31、32或3中的HCDR1、HCDR2和HCDR3的氨基酸序列，和所述轻链可变区的LCDR1、LCDR2和LCDR3分别包含SEQ ID NO：36、35、34、33或4中的LCDR1、LCDR2和LCDR3的氨基酸序列。
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述抗IL23抗体的重链可变区的HCDR1、HCDR2和HCDR3以及轻链可变区的LCDR1、LCDR2和LCDR3是根据选自Kabat、IMGT、Chothia、AbM和Contact的编号规则定义的。在一些实施方案中，所述重链可变区的HCDR1、HCDR2和HCDR3以及轻链可变区的LCDR1、LCDR2和LCDR3是根据Kabat编号规则定义的；在一些实施方案中，所述重链可变区的HCDR1、HCDR2和HCDR3以及轻链可变区的LCDR1、LCDR2和LCDR3是根据IMGT编号规则定义的；在一些实施方案中， 所述重链可变区的HCDR1、HCDR2和HCDR3以及轻链可变区的LCDR1、LCDR2和LCDR3是根据Chothia编号规则定义的；在一些实施方案中，所述重链可变区的HCDR1、HCDR2和HCDR3以及轻链可变区的LCDR1、LCDR2和LCDR3是根据AbM编号规则定义的；在一些实施方案中，所述重链可变区的HCDR1、HCDR2和HCDR3以及轻链可变区的LCDR1、LCDR2和LCDR3是根据Contact编号规则定义的。
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述抗IL23抗体包含重链可变区和轻链可变区，其中：
(i)所述重链可变区的HCDR1包含SEQ ID NO：27、28或5的氨基酸序列，HCDR2包含SEQ ID NO：6的氨基酸序列，和HCDR3包含SEQ ID NO：7的氨基酸序列，和所述轻链可变区的LCDR1包含SEQ ID NO：8的氨基酸序列，LCDR2包含SEQ ID NO：9的氨基酸序列，和LCDR3包含SEQ ID NO：10的氨基酸序列；或
(ii)所述重链可变区的HCDR1包含SEQ ID NO：11的氨基酸序列，HCDR2包含SEQ ID NO：12的氨基酸序列，和HCDR3包含SEQ ID NO：13的氨基酸序列，和所述轻链可变区的LCDR1包含SEQ ID NO：14的氨基酸序列，LCDR2包含SEQ ID NO：37或15的氨基酸序列，和LCDR3包含SEQ ID NO：16的氨基酸序列；
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述抗IL23抗体包含重链可变区和轻链可变区，其中
(i)所述重链可变区的HCDR1包含SEQ ID NO：27的氨基酸序列，HCDR2包含SEQ ID NO：6的氨基酸序列，和HCDR3包含SEQ ID NO：7的氨基酸序列，和所述轻链可变区的LCDR1包含SEQ ID NO：8的氨基酸序列，LCDR2包含SEQ ID NO：9的氨基酸序列，和LCDR3包含SEQ ID NO：10的氨基酸序列；或
(ii)所述重链可变区的HCDR1包含SEQ ID NO：28的氨基酸序列，HCDR2包含SEQ ID NO：6的氨基酸序列，和HCDR3包含SEQ ID NO：7的氨基酸序列，和所述轻链可变区的LCDR1包含SEQ ID NO：8的氨基酸序列，LCDR2包含SEQ ID NO：9的氨基酸序列，和LCDR3包含SEQ ID NO：10的氨基酸序列；或
(iii)所述重链可变区的HCDR1包含SEQ ID NO：11的氨基酸序列，HCDR2包含SEQ ID NO：12的氨基酸序列，和HCDR3包含SEQ ID NO：13的氨基酸序列，和所述轻链可变区的LCDR1包含SEQ ID NO：14的氨基酸序列，LCDR2包含SEQ ID NO：37的氨基酸序列，和LCDR3包含SEQ ID NO：16的氨基酸序列。
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述的抗IL23抗体为鼠源抗体、嵌合抗体或人源化抗体。在一些实施方案中，所述抗体是人源化抗体。
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述抗IL23 抗体包含重链可变区和轻链可变区，其中：
(i)所述重链可变区包含SEQ ID NO：22、21、20、19、18或17中的任一或与其具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列，和所述轻链可变区包含SEQ ID NO：26、25、24或23中的任一或与其具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列；或
(ii)所述重链可变区包含SEQ ID NO：29、30、31或32中的任一或与其具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列，和所述轻链可变区包含SEQ ID NO：36、35、34或33中的任一或与其具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列；或
(iii)所述重链可变区包含SEQ ID NO：1或与其具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列，和所述轻链可变区包含SEQ ID NO：2或与其具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列；或
(iv)所述重链可变区包含SEQ ID NO：3或与其具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列，和所述轻链可变区包含SEQ ID NO：4或与其具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列。
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述抗IL23抗体的重链可变区的框架区上包含选自第1、30、37、44、49、73、89和93位(根据Kabat编号系统编号)中的一个或更多个氨基酸突变，和/或轻链可变区的框架区上包含选自第4、17、36、58、60和68位(根据Kabat编号系统编号)中的一个或更多个氨基酸突变。在所述的抗IL23抗体融合蛋白的一些实施方案中，所述抗IL23抗体的重链可变区的HCDR1包含SEQ ID NO：27、28或5的氨基酸序列，HCDR2包含SEQ ID NO：6的氨基酸序列，和HCDR3包含SEQ ID NO：7的氨基酸序列，和所述轻链可变区的LCDR1包含SEQ ID NO：8的氨基酸序列，LCDR2包含SEQ ID NO：9的氨基酸序列，和LCDR3包含SEQ ID NO：10的氨基酸序列；所述抗体的重链可变区的框架区上包含选自第1E、30T、37V、44G、49G、73N、89R和93V(根据Kabat编号系统编号)中的一个或更多个氨基酸突变，和/或所述轻链可变区的框架区上包含选自4L、17Q、36F、58I、60A和68R(根据Kabat编号系统编号)中的一个或更多个氨基酸突变。
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述抗IL23抗体的重链可变区的框架区上包含选自第1、2、28、44、71和73位(根据Kabat编号系统编号)中的一个或更多个氨基酸突变，和/或轻链可变区的框架区上包含选自第4、36、39、71和100位(根据Kabat编号系统编号)中的一个或更多个氨 基酸突变。在所述的抗IL23抗体融合蛋白的一些实施方案中，所述抗IL23抗体的重链可变区的HCDR1包含SEQ ID NO：11的氨基酸序列，HCDR2包含SEQ ID NO：12的氨基酸序列，和HCDR3包含SEQ ID NO：13的氨基酸序列，和所述轻链可变区的LCDR1包含SEQ ID NO：14的氨基酸序列，LCDR2包含SEQ ID NO：37或15的氨基酸序列，和LCDR3包含SEQ ID NO：16的氨基酸序列；所述抗体的重链可变区的框架区上包含选自第1E、2F、28S、44S、71V和73Q(根据Kabat编号系统编号)中的一个或更多个氨基酸突变，和/或所述轻链可变区的框架区上包含选自4I、36F、39R、71L和100S(根据Kabat编号系统编号)中的一个或更多个氨基酸突变。
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述抗IL23抗体包含重链可变区和轻链可变区，其中，
(i)所述重链可变区包含SEQ ID NO：22、21、20、19、18或17的氨基酸序列，和所述轻链可变区包含SEQ ID NO：26、25、24或23的氨基酸序列；或
(ii)所述重链可变区包含SEQ ID NO：29、30、31或32的氨基酸序列，和所述轻链可变区包含SEQ ID NO：36、35、34或33的氨基酸序列；或
(iii)所述重链可变区包含SEQ ID NO：1的氨基酸序列，和所述轻链可变区包含SEQ ID NO：2的氨基酸序列；或
(iv)所述重链可变区包含SEQ ID NO：3的氨基酸序列，和所述轻链可变区包含SEQ ID NO：4的氨基酸序列。
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述抗IL23抗体包含重链可变区和轻链可变区，其中，
(i)所述重链可变区包含SEQ ID NO：22的氨基酸序列，和所述轻链可变区包含SEQ ID NO：26的氨基酸序列；
(ii)所述重链可变区包含SEQ ID NO：21的氨基酸序列，和所述轻链可变区包含SEQ ID NO：25的氨基酸序列；或
(iii)所述重链可变区包含SEQ ID NO：29的氨基酸序列，和所述轻链可变区包含SEQ ID NO：36的氨基酸序列。
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述的抗IL23抗体进一步包含抗体重链恒定区和轻链恒定区。在一些实施方案中，所述重链恒定区为人IgG重链恒定区；在一些实施方案中，所述重链恒定区选自人IgG1、IgG2、IgG3和IgG4恒定区；在一些实施方案中，所述轻链恒定区选自人抗体κ或λ链恒定区；在一些实施方案中，所述重链恒定区为人IgG1重链恒定区，所述轻链恒定区为人κ轻链恒定区。在一些实施方案中，所述重链恒定区的Fc区具有一个或更多个能够减少Fc区与Fc受体结合的氨基酸取代。在一些实施方案中，所述Fc区具有L234A、L235A突变，和/或S228P突变，和/或YTE突变(M252Y、S254T和T256E)，所述突变编号依据为EU索引。在一些实施方案中，所述重链恒定区 包含SEQ ID NO：38的氨基酸序列，所述轻链恒定区包含SEQ ID NO：39的氨基酸序列。
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述的抗IL23抗体包含重链和轻链，其中，
(i)所述抗IL23抗体的重链包含与SEQ ID NO：42具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列，和所述轻链包含与SEQ ID NO：43具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列；或
(ii)所述抗IL23抗体的重链包含与SEQ ID NO：40具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列，和所述轻链包含与SEQ ID NO：41具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列；或
(iii)所述抗IL23抗体的重链包含与SEQ ID NO：44具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列，和所述轻链包含与SEQ ID NO：45具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列。
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述的抗IL23抗体包含重链和轻链，其中，
(i)所述抗IL23抗体的重链包含SEQ ID NO：42的氨基酸序列，和所述轻链包含SEQ ID NO：43的氨基酸序列；或
(ii)所述抗IL23抗体的重链包含SEQ ID NO：40的氨基酸序列，和所述轻链包含SEQ ID NO：41的氨基酸序列；或
(iii)所述抗IL23抗体的重链包含SEQ ID NO：44的氨基酸序列，和所述轻链包含SEQ ID NO：45的氨基酸序列。
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述TACI多肽具有更好的防止断裂功能。
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述TACI多肽为包含SEQ ID NO：58的第48位至第85位氨基酸残基的多肽或其变体；其中，所述变体在选自第49、52、53、57、65、82和83位中的一个或更多个位点上具有氨基酸替换，所述氨基酸替换的位点为相对于序列SEQ ID NO：58的自然顺序编号的氨基酸残基位点。在一些实施方案中，所述TACI多肽的变体具有选自由49T或49R、52S、53E或53Q、57E、65T或65A、82A或82R、和83Y组成的组中的一个或更多个氨基酸替换，所述氨基酸替换的位点为相对于序列SEQ ID NO：58的自然顺序编号的氨基酸残基位点。
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述TACI多肽如SEQ ID NO：58所示或是SEQ ID NO：58的截短片段或其变体；其中所述截 短片段包含SEQ ID NO：58的第48位至第85位氨基酸残基，所述变体在SEQ ID NO：58或其截短片段上具有选自第49、52、53、57、65、82和83位中的一个或更多个氨基酸替换，其中所述氨基酸替换的位点为相对于序列SEQ ID NO：58的自然顺序编号的氨基酸残基位点。
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述TACI多肽的截短片段包含：SEQ ID NO：58的第48位至第86位氨基酸残基；SEQ ID NO：58的第48位至第87位氨基酸残基；或SEQ ID NO：58的第48位至第88位氨基酸残基。
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述TACI多肽的序列如SEQ ID NO：60-63任一项所示。
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述TACI多肽的序列为SEQ ID NO：58的变体或者SEQ ID NO：58的截短片段(例如SEQ ID NO：60、SEQ ID NO：61、SEQ ID NO：62或SEQ ID NO：63序列的截短片段)的变体，所述变体在SEQ ID NO：58或其截短片段序列上具有选自第49、52、53、57、65、82和83位中的任意1个、2个、3个、4个、5个、6个或7个氨基酸的替换，所述氨基酸替换的位点为相对于序列SEQ ID NO：58的自然顺序编号的氨基酸残基位点。
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述TACI多肽在SEQ ID NO：58、SEQ ID NO：60、SEQ ID NO：61、SEQ ID NO：62或SEQ ID NO：63序列的基础上具有选自由49T或49R、52S、53E或53Q、57E、65T或65A、82A或82R、和83Y组成的组中的一个或更多个氨基酸替换(例如1个、2个、3个、4个、5个、6个或7个氨基酸的替换)，其中所述氨基酸替换的位点为相对于序列SEQ ID NO：58的自然顺序编号的氨基酸残基位点。
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述TACI多肽为：在SEQ ID NO：58、SEQ ID NO：60、SEQ ID NO：61、SEQ ID NO：62或SEQ ID NO：63序列的基础上具有选自49T、52S、53E、53Q、57E和82A中的任一个氨基酸替换；在SEQ ID NO：58、SEQ ID NO：60、SEQ ID NO：61、SEQ ID NO：62或SEQ ID NO：63序列的基础上具有49R和65T氨基酸替换；在SEQ ID NO：58、SEQ ID NO：60、SEQ ID NO：61、SEQ ID NO：62或SEQ ID NO：63序列的基础上具有45R和65A氨基酸替换；在SEQ ID NO：58、SEQ ID NO：60、SEQ ID NO：61、SEQ ID NO：62或SEQ ID NO：63序列的基础上具有49R、65T和82R氨基酸替换；在SEQ ID NO：58、SEQ ID NO：60、SEQ ID NO：61、SEQ ID NO：62或SEQ ID NO：63序列的基础上具有53E和57E氨基酸替换；在SEQ ID NO：58、SEQ ID NO：60、SEQ ID NO：61、SEQ ID NO：62或SEQ ID NO：63序列的基础上具有52S、53E和57E氨基酸替换；在SEQ ID NO：58、SEQ ID NO：60、SEQ ID NO：61、SEQ ID NO：62或SEQ ID NO：63序列的基础上具有49T 和82A氨基酸替换；在SEQ ID NO：58、SEQ ID NO：60、SEQ ID NO：61、SEQ ID NO：62或SEQ ID NO：63序列的基础上具有49T和83Y氨基酸替换；在SEQ ID NO：58、SEQ ID NO：60、SEQ ID NO：61、SEQ ID NO：62或SEQ ID NO：63序列的基础上具有49T、82A和83Y氨基酸替换；或者在SEQ ID NO：58、SEQ ID NO：60、SEQ ID NO：61、SEQ ID NO：62或SEQ ID NO：63序列的基础上具有49T、53E、57E和82A氨基酸替换。其中前述氨基酸替换的位点为相对于序列SEQ ID NO：58的自然顺序编号的氨基酸残基位点。
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述TACI多肽的氨基酸序列如SEQ ID NO：51至83中任一所示。
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述TACI多肽的氨基酸序列如SEQ ID NO：60-63和SEQ ID NO：66-83中的任一所示。
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述TACI多肽的氨基酸序列如SEQ ID NO：83所示。
在一些实施方案中，如上任一项所述的抗IL23抗体融合蛋白包括：
第一链：[TACI多肽1]-[连接子1]-[抗IL23抗体的重链]-[连接子2]-[TACI多肽2]，和
第二链：抗IL23抗体的轻链，
其中，所述TACI多肽1和TACI多肽2是相同或不相同的，任选地，所述TACI多肽1和TACI多肽2可以独立地选自本文所述的任一TACI多肽；所述连接子1和连接子2是相同或不相同的；在一些实施方案中，所述连接子为选自(G xS) y连接子，其中，x选自1-5的整数，y选自0-6的整数，在一些实施方案中，所述连接子为选自(G xS) y连接子，其中，x选自1-5的整数，y选自1-6的整数，在一些实施方案中，所述连接子为GGGGSGGGGSGGGGS(如SEQ ID NO：84所示)或GGGS(如SEQ ID NO：85所示)。
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述的抗IL23抗体融合蛋白具有：包含SEQ ID NO：47的氨基酸序列的第一链，和包含SEQ ID NO：43的氨基酸序列的第二链；或所述的抗IL23抗体融合蛋白具有：包含SEQ ID NO：46的氨基酸序列的第一链，和包含SEQ ID NO：41的氨基酸序列的第二链。在一些实施方案中，所述的抗IL23抗体融合蛋白具有2条包含SEQ ID NO：47的氨基酸序列的第一链，和2条包含SEQ ID NO：43的氨基酸序列的第二链；在一些实施方案中，所述的抗IL23抗体融合蛋白具有2条包含SEQ ID NO：46的氨基酸序列的第一链，和2条包含SEQ ID NO：41的氨基酸序列的第二链。
另一方面，本披露提供一种抗IL23抗体，其中所述抗IL23抗体包含重链可变区和轻链可变区，其中，所述重链可变区包含HCDR1、HCDR2和HCDR3，所述轻链可变区包含LCDR1、LCDR2和LCDR3，其中，
(i)所述重链可变区的HCDR1、HCDR2和HCDR3分别包含SEQ ID NO： 22、21、20、19、18、17或1中的HCDR1、HCDR2和HCDR3的氨基酸序列，和所述轻链可变区的LCDR1、LCDR2和LCDR3分别包含SEQ ID NO：26、25、24、23或2中的LCDR1、LCDR2和LCDR3的氨基酸序列；或
(ii)所述重链可变区的HCDR1、HCDR2和HCDR3分别包含SEQ ID NO：29、30、31、32或3中的HCDR1、HCDR2和HCDR3的氨基酸序列，和所述轻链可变区的LCDR1、LCDR2和LCDR3分别包含SEQ ID NO：36、35、34、33或4中的LCDR1、LCDR2和LCDR3的氨基酸序列。
在如上任一项所述的抗IL23抗体的一些实施方案中，其中所述重链可变区的HCDR1、HCDR2和HCDR3以及轻链可变区的LCDR1、LCDR2和LCDR3是根据选自Kabat、IMGT、Chothia、AbM和Contact的编号规则定义的。在一些实施方案中，所述重链可变区的HCDR1、HCDR2和HCDR3以及轻链可变区的LCDR1、LCDR2和LCDR3是根据Kabat编号规则定义的；在一些实施方案中，所述重链可变区的HCDR1、HCDR2和HCDR3以及轻链可变区的LCDR1、LCDR2和LCDR3是根据IMGT编号规则定义的；在一些实施方案中，所述重链可变区的HCDR1、HCDR2和HCDR3以及轻链可变区的LCDR1、LCDR2和LCDR3是根据Chothia编号规则定义的；在一些实施方案中，所述重链可变区的HCDR1、HCDR2和HCDR3以及轻链可变区的LCDR1、LCDR2和LCDR3是根据AbM编号规则定义的；在一些实施方案中，所述重链可变区的HCDR1、HCDR2和HCDR3以及轻链可变区的LCDR1、LCDR2和LCDR3是根据Contact编号规则定义的。
在如上任一项所述的抗IL23抗体的一些实施方案中：
(i)所述重链可变区的HCDR1包含SEQ ID NO：27、28或5的氨基酸序列，HCDR2包含SEQ ID NO：6的氨基酸序列，和HCDR3包含SEQ ID NO：7的氨基酸序列，和所述轻链可变区的LCDR1包含SEQ ID NO：8的氨基酸序列，LCDR2包含SEQ ID NO：9的氨基酸序列，和LCDR3包含SEQ ID NO：10的氨基酸序列；或
(ii)所述重链可变区的HCDR1包含SEQ ID NO：11的氨基酸序列，HCDR2包含SEQ ID NO：12的氨基酸序列，和HCDR3包含SEQ ID NO：13的氨基酸序列，和所述轻链可变区的LCDR1包含SEQ ID NO：14的氨基酸序列，LCDR2包含SEQ ID NO：37或15的氨基酸序列，和LCDR3包含SEQ ID NO：16的氨基酸序列；
在如上任一项所述的抗IL23抗体的一些实施方案中，(i)所述重链可变区的HCDR1包含SEQ ID NO：27的氨基酸序列，HCDR2包含SEQ ID NO：6的氨基酸序列，和HCDR3包含SEQ ID NO：7的氨基酸序列，和所述轻链可变区的LCDR1包含SEQ ID NO：8的氨基酸序列，LCDR2包含SEQ ID NO：9的氨基酸序列，和LCDR3包含SEQ ID NO：10的氨基酸序列；或
(ii)所述重链可变区的HCDR1包含SEQ ID NO：28的氨基酸序列，HCDR2 包含SEQ ID NO：6的氨基酸序列，和HCDR3包含SEQ ID NO：7的氨基酸序列，和所述轻链可变区的LCDR1包含SEQ ID NO：8的氨基酸序列，LCDR2包含SEQ ID NO：9的氨基酸序列，和LCDR3包含SEQ ID NO：10的氨基酸序列；或
(iii)所述重链可变区的HCDR1包含SEQ ID NO：11的氨基酸序列，HCDR2包含SEQ ID NO：12的氨基酸序列，和HCDR3包含SEQ ID NO：13的氨基酸序列，和所述轻链可变区的LCDR1包含SEQ ID NO：14的氨基酸序列，LCDR2包含SEQ ID NO：37的氨基酸序列，和LCDR3包含SEQ ID NO：16的氨基酸序列。
在如上任一项所述的抗IL23抗体的一些实施方案中，所述的抗IL23抗体为鼠源抗体、嵌合抗体或人源化抗体。在一些实施方案中，所述抗体是人源化抗体。
在如上任一项所述的抗IL23抗体的一些实施方案中，抗IL23抗体包含重链可变区和轻链可变区，其中：
(i)所述重链可变区包含SEQ ID NO：22、21、20、19、18或17中的任一或与其具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列，和所述轻链可变区包含SEQ ID NO：26、25、24或23中的任一或与其具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列；或
(ii)所述重链可变区包含SEQ ID NO：29、30、31或32中的任一或与其具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列，和所述轻链可变区包含SEQ ID NO：36、35、34或33中的任一或与其具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列；或
(iii)所述重链可变区包含SEQ ID NO：1或与其具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列，和所述轻链可变区包含SEQ ID NO：2或与其具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列；或
(iv)所述重链可变区包含SEQ ID NO：3或与其具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列，和所述轻链可变区包含SEQ ID NO：4或与其具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列。
在如上任一项所述的抗IL23抗体的一些实施方案中，所述抗IL23抗体的重链可变区的框架区上包含选自第1、30、37、44、49、73、89和93位(根据Kabat编号系统编号)中的一个或更多个氨基酸突变，和/或轻链可变区的框架区上包含选自第4、17、36、58、60和68位(根据Kabat编号系统编号)中的一个或更多个氨基酸突变。在一些实施方案中，所述的抗IL23抗体的重链可变区的HCDR1包含SEQ ID NO：27、28或5的氨基酸序列，HCDR2包含SEQ ID NO：6的氨基酸序列，和HCDR3包含SEQ ID NO：7的氨基酸序列，和所述轻链可变区的LCDR1 包含SEQ ID NO：8的氨基酸序列，LCDR2包含SEQ ID NO：9的氨基酸序列，和LCDR3包含SEQ ID NO：10的氨基酸序列；所述抗体的重链可变区的框架区上包含选自第1E、30T、37V、44G、49G、73N、89R和93V(根据Kabat编号系统编号)中的一个或更多个氨基酸突变，和/或所述轻链可变区的框架区上包含选自4L、17Q、36F、58I、60A和68R(根据Kabat编号系统编号)中的一个或更多个氨基酸突变。
在如上任一项所述的抗IL23抗体的一些实施方案中，所述抗IL23抗体的重链可变区的框架区上包含选自第1、2、28、44、71和73位(根据Kabat编号系统编号)中的一个或更多个氨基酸突变，和/或轻链可变区的框架区上包含选自第4、36、39、71和100位(根据Kabat编号系统编号)中的一个或更多个氨基酸突变。在一些实施方案中，所述的抗IL23抗体的重链可变区的HCDR1包含SEQ ID NO：11的氨基酸序列，HCDR2包含SEQ ID NO：12的氨基酸序列，和HCDR3包含SEQ ID NO：13的氨基酸序列，和所述轻链可变区的LCDR1包含SEQ ID NO：14的氨基酸序列，LCDR2包含SEQ ID NO：37或15的氨基酸序列，和LCDR3包含SEQ ID NO：16的氨基酸序列；所述抗体的重链可变区的框架区上包含选自第1E、2F、28S、44S、71V和73Q(根据Kabat编号系统编号)中的一个或更多个氨基酸突变，和/或所述轻链可变区的框架区上包含选自4I、36F、39R、71L和100S(根据Kabat编号系统编号)中的一个或更多个氨基酸突变。
在一些实施方案中，如上任一项所述的抗IL23抗体包含重链可变区和轻链可变区，其中，
(i)所述重链可变区包含SEQ ID NO：22、21、20、19、18或17的氨基酸序列，和所述轻链可变区包含SEQ ID NO：26、25、24或23的氨基酸序列；或
(ii)所述重链可变区包含SEQ ID NO：29、30、31或32的氨基酸序列，和所述轻链可变区包含SEQ ID NO：36、35、34或33的氨基酸序列；或
(iii)所述重链可变区包含SEQ ID NO：1的氨基酸序列，和所述轻链可变区包含SEQ ID NO：2的氨基酸序列；或
(iv)所述重链可变区包含SEQ ID NO：3的氨基酸序列，和所述轻链可变区包含SEQ ID NO：4的氨基酸序列。
在一些实施方案中，如上任一项所述的抗IL23抗体包含重链可变区和轻链可变区，其中，
(i)所述重链可变区包含SEQ ID NO：22的氨基酸序列，和所述轻链可变区包含SEQ ID NO：26的氨基酸序列；或
(ii)所述重链可变区包含SEQ ID NO：21的氨基酸序列，和所述轻链可变区包含SEQ ID NO：25的氨基酸序列；或
(iii)所述重链可变区包含SEQ ID NO：29的氨基酸序列，和所述轻链可变区包含SEQ ID NO：36的氨基酸序列。
在如上任一项所述的抗IL23抗体的一些实施方案中，所述的抗IL23抗体包含进一步抗体重链恒定区和轻链恒定区。在一些实施方案中，所述重链恒定区为人IgG重链恒定区；在一些实施方案中，所述重链恒定区选自人IgG1、IgG2、IgG3和IgG4恒定区；在一些实施方案中，所述轻链恒定区选自人抗体κ或λ链恒定区；在一些实施方案中，所述重链恒定区为人IgG1重链恒定区，所述轻链恒定区为人κ轻链恒定区。在一些实施方案中，所述重链恒定区的Fc区具有一个或更多个能够减少Fc区与Fc受体结合的氨基酸取代。在一些实施方案中，所述Fc区具有L234A、L235A突变，和/或S228P突变，和/或YTE突变(M252Y、S254T和T256E)，所述突变编号依据为EU索引。在一些实施方案中，所述重链恒定区包含SEQ ID NO：38的氨基酸序列，所述轻链恒定区包含SEQ ID NO：39的氨基酸序列。
在一些实施方案中，如上任一项所述的抗IL23抗体包含重链和轻链，其中，
(i)所述抗IL23抗体的重链包含与SEQ ID NO：42具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列，和轻链包含与SEQ ID NO：43具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列；或
(ii)所述抗IL23抗体的重链包含与SEQ ID NO：40具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列，和轻链包含与SEQ ID NO：41具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列；或
(iii)所述抗IL23抗体的重链包含与SEQ ID NO：44具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列，和轻链包含与SEQ ID NO：45具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列。
在一些实施方案中，如上任一项所述的抗IL23抗体包含重链和轻链，其中
(i)所述抗IL23抗体的重链包含SEQ ID NO：42的氨基酸序列，和轻链包含SEQ ID NO：43的氨基酸序列；或
(ii)所述抗IL23抗体的重链包含SEQ ID NO：40的氨基酸序列，和轻链包含SEQ ID NO：41的氨基酸序列；或
(iii)所述抗IL23抗体的重链包含SEQ ID NO：44的氨基酸序列，和轻链包含SEQ ID NO：45的氨基酸序列。
在一些实施方案中，本披露还提供一种分离的抗IL23抗体或抗IL23抗体融合蛋白，其与前面任一项所述的抗IL23抗体或抗IL23抗体融合蛋白竞争性结合人IL23。
在一些实施方案中，前面任一项所述的抗IL23抗体或抗IL23抗体融合蛋白，其具有一种或更多种以下特性：
A.与人IL-23 p19和食蟹猴IL-23 p19特异性结合，不与鼠IL-23 p19特异性 结合；优选地，以小于6.00E-11M(例如，小于6.00E-11M、小于5.00E-11M、小于4.00E-11M、小于3.00E-11M、小于2.00E-11M、小于1.7E-11M、小于1.6E-11M、小于1.5E-11M或更小)的KD值与人IL-23 p19结合，和/或以小于9.00E-11M(例如，小于9.00E-11M、小于8.00E-11M、小于7.00E-11M、小于6.00E-11M或更小)的KD值与食蟹猴IL-23 p19结合，所述KD值通过表面等离子体共振测定法所测量，例如通过
Figure PCTCN2022123370-appb-000001
表面等离子体共振测定法所测量；在一些实施方案中，所述KD值通过本披露测试6方法检测；
B.具有阻断IL-23/IL-23R结合活性；优选地，阻断人IL-23/IL-23R结合的IC 50值小于0.6nM(例如，小于0.6nM、小于0.5nM、小于0.4nM、小于0.3nM或更小)，所述IC 50值通过Elisa方法检测；在一些实施方案中，所述IC 50值通过本披露测试2方法检测；
C.具有阻断BAFF/BAFF-R结合活性；优选地，阻断BAFF/BAFF-R结合的IC 50值小于7.00nM(例如，小于7.00nM、小于6.00nM、小于5.00nM、小于4.00nM、小于3.00nM、小于2.00nM、小于1.00nM、小于0.9nM或更小)，所述IC 50值通过Elisa方法检测；在一些实施方案中，所述IC 50值通过本披露测试2方法检测；
D.具有阻断BAFF/BCMA结合活性；优选地，阻断BAFF/BCMA结合的IC 50值小于4.50nM(例如，小于4.50nM、小于3.00nM、小于2.00nM、小于1.00nM或更小)，所述IC 50值通过Elisa方法检测；在一些实施方案中，所述IC 50值通过本披露测试2方法检测；
E.具有阻断BAFF/TACI结合活性；优选地，阻断BAFF/TACI结合的IC 50值小于6.00nM(例如，小于6.00nM、小于5.00nM、小于4.00nM、小于3.00nM、小于2.00nM、小于1.00nM或更小)，所述IC 50值通过Elisa方法检测；在一些实施方案中，所述IC 50值通过本披露测试2方法检测；
F.具有阻断APRIL/BCMA结合活性；优选地，阻断APRIL/BCMA结合的IC 50值小于1.00nM(例如，小于1.00nM、小于0.5nM、小于0.4nM、小于0.1nM或更小)，所述IC 50值通过Elisa方法检测；在一些实施方案中，所述IC 50值通过本披露测试2方法检测；
G.具有阻断APRIL/TACI结合活性；优选地，阻断APRIL/TACI结合的IC 50值小于3.00nM(例如，小于3.00nM、小于2.00nM、小于1.50nM或更小)，所述IC 50值通过Elisa方法检测；在一些实施方案中，所述IC 50值通过本披露测试2方法检测；
H.具有抑制IL-17分泌活性；优选地，以小于0.03nM(例如，小于0.03nM、小于0.02nM nM、小于0.01nM或更小)的IC 50值抑制IL-17分泌；在一些实施方案中，所述IC 50值通过本披露测试4方法检测；
I.具有抑制BaF3-IL-23R细胞增殖活性；优选地，以小于0.5nM(例如，小 于0.5nM、小于0.4nM nM、小于0.3nM、小于0.2nM、小于0.1nM或更小)的IC 50值抑制BaF3-IL-23R细胞增殖；所述IC 50值通过PerkinElmer检测；在一些实施方案中，所述IC 50值通过本披露测试3方法检测；
J.具有抑制TNFα、IL-22和IgA细胞因子分泌活性；在一些实施方案中，所述TNFα、IL-22和IgA通过本披露测试9方法检测；或
K.具有抑制B细胞增殖活性；在一些实施方案中，所述TNFα、IL-22和IgA通过本披露测试5方法检测。
本披露还提供一种药物组合物，其包含前面任一项所述的抗IL23抗体融合蛋白或抗IL23抗体，以及一种或多种药学上可接受的载体、稀释剂或赋形剂。
在一些实施方案中，本披露提供一种核酸分子，其编码前面任一项所述的抗IL23抗体融合蛋白或抗IL23抗体。
在一些实施方案中，本披露提供一种表达载体，其包含前述的核酸分子。
在一些实施方案中，本披露提供一种宿主细胞，其含前述的核酸分子。
在一些实施方案中，本披露提供一种宿主细胞，其含前述的表达载体。
在一些实施方案中，本披露提供一种治疗或改善B细胞障碍或自身免疫性疾病的方法，所述方法包括向有需要的受试者施用治疗有效量的前面任一项所述的抗IL23抗体融合蛋白或抗IL23抗体或药物组合物的步骤。
在一些实施方案中，本披露提供前述任一项所述的抗IL23抗体融合蛋白或抗IL23抗体、核酸分子或药物组合物在制备治疗或预防疾病的药物中的用途。
在另一个方面，本披露还提供用作药物的前述任一项所述的抗IL23抗体融合蛋白或抗IL23抗体、核酸分子或组合物。在一些实施方案中，药物用于治疗B细胞障碍或自身免疫性疾病。
在一些实施方案中，前面任一项所述的疾病是与IL23表达相关的疾病或病症。在一些实施方案中，所述自身免疫性疾病选自：系统性红斑狼疮、重症肌无力、多发性硬化、胰岛素依赖性糖尿病、克罗恩氏病、类风湿关节炎、多关节型青少年类风湿关节炎和银屑病性关节炎；所述B细胞障碍选自：肿瘤、慢性白细胞性白血病、多发性骨髓瘤、非霍奇金淋巴瘤、移植后淋巴组织增生病以及轻链丙球蛋白病。在一些实施方案中，所述疾病为系统性红斑狼疮。
在一些实施方案中，前面任一项所述的治疗，进一步包括向受试者施用另外的治疗药物。
附图说明
图1：Hu29-19T和Hu29-24T结构示意图；
图2：咪喹莫特诱导的银屑病动物的红疹严重程度评分的实验结果；
图3：咪喹莫特诱导的银屑病动物的皮肤脱屑严重程度评分的实验结果；
图4：咪喹莫特诱导的银屑病动物的红疹和皮肤脱屑的综合评分结果；
图5：人IL-23诱导的银屑病动物的右耳厚度实验结果；
图6：人IL-23诱导的银屑病动物的曲线下面积的实验结果(附图中，与阴性对照相比，***P＜0.001)；
图7：人IL-23诱导的银屑病动物的右耳重量的实验结果(附图中，与阴性对照相比，**P＜0.01，***P＜0.001)；
图8：抗IL23抗体融合蛋白抑制IgA的分泌实验结果；
图9：抗IL23抗体融合蛋白抑制TNFα的分泌实验结果；
图10：抗IL23抗体融合蛋白抑制IL-22的分泌实验结果；
图11：系统性红斑狼疮动物药效实验中的肾脏病理学评分结果；
图12：系统性红斑狼疮动物药效实验中的皮肤损伤评分结果；
图13：系统性红斑狼疮动物药效实验中的尿蛋白检测结果。
具体实施方式
术语
为了更容易理解本披露，以下对某些技术和科学术语进行了描述。除非在本文中另有明确定义，本文使用的全部技术和科学术语具有与本领域的普通技术人员通常所理解的相同含义。
说明书和权利要求书中所用的单数形式“一个”、“一种”和“所述”包括复数指代，除非上下文清楚表明并非如此。
除非上下文另外清楚要求，否则在专利说明书和权利要求书中，应将词语“包含”、“具有”、“包括”等理解为“包括但不仅限于”的意义，而不是排他性或穷举性意义。
术语“细胞因子”是由一个细胞群体释放的、作为细胞间介质作用于其它细胞的蛋白质的一般术语。这样的细胞因子的例子包括淋巴因子、单核因子、趋化因子和传统的多肽激素。示例性的细胞因子包括：IL-2、IFN-γ、IL-6、TNFα、IL-17和IL-5。
本披露所述的TACI是一种膜结合受体，野生型人TACI包含两个富含半胱氨酸的假性重复片断(cysteine-rich pseudo-repeats)的胞外区、一个跨膜区和与CAML(钙调节剂和亲环蛋白配体)相互作用的胞质区。野生型人TACI胞外区(第1-165位)参见本披露的SEQ ID NO：51。本披露中的“TACI胞外结构域”与“TACI胞外区”可相互替换。
IL-23(也称IL23)主要由活化的树突状细胞、巨噬细胞及单核细胞等产生，是IL-12异源二聚体细胞因子家族中的一员，由p19(也称IL23 p19亚基)和p40(也称IL23 p40亚基)两个亚基组成，其中p40亚基是与IL-12共同含有的亚基(J Immunol.2018 Sep 15；201(6):1605-1613.)。
术语“和/或”，意指包含“和”与“或”两种含义。例如短语“A、B和/或C”旨在涵 盖以下方面中的每一个：A、B和C；A、B或C；A或C；A或B；B或C；A和C；A和B；B和C；A(单独)；B(单独)；和C(单独)。
本披露所用氨基酸三字母代码和单字母代码如J.biol.chem，243，p3558(1968)中所述。
术语“氨基酸”是指天然存在的和合成的氨基酸，以及以与天然存在的氨基酸类似的方式起作用的氨基酸类似物和氨基酸模拟物。天然存在的氨基酸是由遗传密码编码的那些氨基酸，以及后来修饰的那些氨基酸，例如羟脯氨酸、γ-羧基谷氨酸和O-磷酸丝氨酸。氨基酸类似物是指与天然存在的氨基酸具有相同基本化学结构(即与氢、羧基、氨基和R基团结合的α碳)的化合物，例如高丝氨酸、正亮氨酸、甲硫氨酸亚砜、甲硫氨酸甲基锍。此类类似物具有修饰的R基团(例如，正亮氨酸)或修饰的肽骨架，但保留与天然存在的氨基酸相同的基本化学结构。氨基酸模拟物是指具有与氨基酸的一般化学结构不同的结构，但是以与天然存在的氨基酸类似的方式起作用的化学化合物。
术语“氨基酸突变”包括氨基酸取代(也称氨基酸替换)、缺失、插入和修饰。可以进行取代、缺失、插入和修饰的任意组合来实现最终构建体，只要最终构建体拥有期望的特性，例如降低或对Fc受体的结合。氨基酸序列缺失和插入包括在多肽链的氨基端和/或羧基端的缺失和插入。具体的氨基酸突变可以是氨基酸取代。在一个实施方式中，氨基酸突变是非保守性的氨基酸取代，即将一个氨基酸用具有不同结构和/或化学特性的另一种氨基酸替换。氨基酸取代包括由非天然存在的氨基酸或由20种天然氨基酸的衍生物(例如4-羟脯氨酸、3-甲基组氨酸、鸟氨酸、高丝氨酸、5-羟赖氨酸)替换。可以使用本领域中公知的遗传或化学方法生成氨基酸突变。遗传方法可以包括定点诱变、PCR，基因合成等。预计基因工程以外的改变氨基酸侧链基团的方法，如化学修饰也可能是可用的。本文中可使用各种名称来指示同一氨基酸突变。本文中，可采用位置+氨基酸残基的方式表示特定位点的氨基酸残基，例如366W，表示在366位点上的氨基酸残基为W。T366W则表示第366位点上的氨基酸残基由原来的T突变为了W。
术语“抗体”以最广义使用，并且涵盖各种抗体结构，包括但不限于单克隆抗体，多克隆抗体；单特异性抗体，多特异性抗体(例如双特异性抗体)；全长抗体和抗体片段(或抗原结合片段，或抗原结合部分)，只要它们展现出期望的抗原结合活性。“天然抗体”指天然存在的免疫球蛋白分子。例如，天然IgG抗体是约150,000道尔顿的异四聚糖蛋白，由二硫键结合的两条相同轻链和两条相同重链构成。从N至C端，每条重链具有一个可变区(VH)，又称作可变重域、重链可变区，接着是重链恒定区，天然IgG重链恒定区通常含三个恒定域(CH1、CH2和CH3)。类似地，从N至C端，每条轻链具有一个可变区(VL)，又称作可变轻域，或轻链可变域，接着是一个恒定轻域(轻链恒定区、CL)。术语“全长抗体”、“完整抗体”和“全抗体”在本文可互换使用，指具有与天然抗体结 构基本类似的结构或具有如本文所限定的Fc区的重链的抗体。天然完整抗体轻链包括轻链可变区VL及恒定区CL，VL处于轻链的氨基末端，轻链恒定区包括κ链及λ链；重链包括可变区VH及恒定区(CH1、CH2及CH3)，VH处于重链的氨基末端，恒定区处于羧基末端，其中CH3最接近多肽的羧基末端，重链可属于任何同种型，包括IgG(包括IgG1、IgG2、IgG3及IgG4亚型)、IgA(包括IgA1及IgA2亚型)、IgM及IgE。
术语抗体“可变区”或“可变域”指抗体重链或轻链中涉及抗体结合抗原的域。本文中，抗体重链可变区(VH)和轻链可变区(VL)各包含四个保守的框架区(FR)和三个互补决定区(CDR)。其中，术语“互补决定区”或“CDR”指可变结构域内主要促成与抗原结合的区域；“框架”或“FR”是指除CDR残基之外的可变结构域残基。VH包含3个CDR区：HCDR1、HCDR2和HCDR3；VL包含3个CDR区：LCDR1、LCDR2和LCDR3。每个VH和VL由从氨基末端(也称N末端)排到羧基末端(也称C末端)按以下顺序排列的三个CDR和四个FR构成：FR1、CDR1、FR2、CDR2、FR3、CDR3、FR4。
可以通过各种公知方案来确定CDR的氨基酸序列边界，例如：“Kabat”编号规则(参见Kabat等(1991)，“Sequences of Proteins of Immunological Interest”，第5版，Public Health Service，National Institutes of Health，Bethesda，MD)、“Chothia”编号规则、“ABM”编号规则、“contact”编号规则(参见Martin,ACR.Protein Sequence and Structure Analysis of Antibody Variable Domains[J].2001)和ImMunoGenTics(IMGT)编号规则(Lefranc，M.P.等，Dev.Comp.Immunol.，27，55-77(2003)；Front Immunol.2018 Oct 16；9:2278)等；各种编号系统之间的对应关系是本领域技术人员熟知的，示例性的，如下表1中所示。
表1.CDR编号系统之间的关系
CDR IMGT Kabat AbM Chothia Contact
HCDR1 27-38 31-35 26-35 26-32 30-35
HCDR2 56-65 50-65 50-58 52-56 47-58
HCDR3 105-117 95-102 95-102 95-102 93-101
LCDR1 27-38 24-34 24-34 24-34 30-36
LCDR2 56-65 50-56 50-56 50-56 46-55
LCDR3 105-117 89-97 89-97 89-97 89-96
除非另有说明，本披露实施例中的可变区和CDR均适用“Kabat”编号规则。
术语“抗体片段”指不同于完整抗体的分子，其包含完整抗体的部分，所述部分与完整抗体所结合的抗原相结合。抗体片段的实例包括但不限于Fv、Fab、Fab’、Fab’-SH、F(ab′) 2、单域抗体、单链Fab(scFab)、双抗体、线性抗体、单链抗体分子(例如scFv)；以及由抗体片段形成的多特异性抗体。
术语“Fc区”或“片段可结晶区”用于定义抗体重链的C末端区域，包括天然Fc 区和改造的Fc区。在一些实施方式中，Fc区包含了相同或不同的两个亚基。在一些实施方式中，人IgG重链的Fc区定义为从Cys226位置处的氨基酸残基或从Pro230延伸至其羧基末端。用于本文所述抗体的合适Fc区包括人IgG1、IgG2(IgG2A、IgG2B)、IgG3和IgG4的Fc区。在一些实施方式中，Fc区的边界还可以变化，例如缺失Fc区的C末端赖氨酸(根据EU编号系统的残基447)或缺失Fc区的C末端甘氨酸和赖氨酸(根据EU编号系统的残基446和447)。除非另有说明，Fc区的编号规则为EU编号系统，又称作EU索引。
术语“嵌合”抗体指抗体中的重和/或轻链的一部分自特定的来源或物种衍生，而重和/或轻链的剩余部分自另外的不同来源或物种衍生的抗体。
术语“人源化”抗体是保留非人抗体的反应性同时在人中具有较低免疫原性的抗体。例如，可以通过保留非人CDR区并用其人对应物(即，恒定区以及可变区的框架区部分)替换抗体的其余部分来实现。
术语“人抗体”、“人源抗体”、“全人抗体”、“完全人抗体”可以互换使用，意指可变区及恒定区是人序列的抗体。该术语涵盖源自人基因但具有，例如，降低可能的免疫原性、增加亲和力、消除可能会引起不期望的折叠的半胱氨酸或糖基化位点等序列已发生改变的抗体。该术语涵盖这些在非人细胞(其可能会赋予不具人细胞特征的糖基化)中重组产生的抗体。该术语亦涵盖已在含有一些或所有人免疫球蛋白重链及轻链基因座的转基因小鼠中饲养的抗体。人抗体的含义明确排除包含非人抗原结合残基的人源化抗体。
术语“亲和力”是指分子(例如，抗体)的单个结合部位与其结合配体(例如，抗原)之间非共价相互作用的总体的强度。除非另外指明，如本文所用，结合“亲和力”是指内部结合亲和力，其反映出结合对(例如，抗体与抗原)的成员之间1:1相互作用。分子X对其配体Y的亲和力通常可以由解离常数(KD)表示。亲和力可以通过本领域已知的常规方法(包括本文所述的那些方法)测量。
如本文所使用的，术语“kassoc”或“ka”指特定抗体-抗原相互作用的缔合速率，术语“kdis”或“kd”指特定抗体-抗原相互作用的解离速率。术语“KD”指解离常数，其获得自kd与ka的比率(即kd/ka)并且表示为摩尔浓度(M)。可以使用本领域公知的方法测定抗体的KD值。例如，使用生物传感系统例如系统测量表面等离子体共振(例如Biacore)，或通过溶液平衡滴定法(SET)测量溶液中的亲和力。
术语“效应子功能”指那些可归于抗体Fc区(天然序列Fc区或氨基酸序列突变的Fc区)且随抗体同种型而变化的生物学活性。抗体效应子功能的例子包括但不限于：C1q结合和补体依赖性细胞毒性、Fc受体结合、抗体依赖性细胞介导的细胞毒性(ADCC)、吞噬作用、细胞表面受体(例如B细胞受体)下调；和B细胞活化。
术语“单克隆抗体”指基本上均质的抗体的群，即在该群中包含的抗体分子 的氨基酸序列是相同的，除了可能少量存在的天然突变以外。相比之下，多克隆抗体制剂通常包含在其可变结构域具有不同氨基酸序列的多种不同抗体，其通常特异性针对不同表位。“单克隆”表示从基本上均质的抗体群体获得的抗体的特征，并且不应解释为要求通过任何特定方法来生产抗体。在一些实施方式中，本披露提供的抗体是单克隆抗体。
术语“抗原”是指能够由诸如抗原结合蛋白(包括例如抗体)的选择性结合剂结合，且另外能够用于动物中以产生能够结合该抗原的抗体的分子或分子部分。抗原可具有一个或多个能够与不同的抗原结合蛋白(例如抗体)相互作用的表位。
术语“表位”指能够与抗体或其抗原结合片段特异性结合的抗原上的区域(area或region)。表位可以由连续氨基酸串(线性表位)形成或包含非连续氨基酸(构象表位)，例如因抗原的折叠(即通过蛋白质性质的抗原的三级折叠)而变成空间接近。构象表位和线性表位的差别在于：在变性溶剂的存在下，抗体对构象表位的结合丧失。表位包含处于独特空间构象的至少3，至少4，至少5，至少6，至少7，或8-10个氨基酸。筛选结合特定表位的抗体(即那些结合相同表位的)可以使用本领域例行方法来进行，例如但不限于丙氨酸扫描，肽印迹，肽切割分析，表位切除，表位提取，抗原的化学修饰(见Prot.Sci.9(2000)487-496)，和交叉阻断。
术语“能够特异性结合”、“特异性结合”或“结合”是指相比其他抗原或表位，抗体能够以更高的亲和力结合至某个抗原或该抗原内的表位。通常地，抗体以约1×10 -7M或更小(例如约1×10 -8M、1×10 -9M、1×10 -10M、1×10 -11M或更小)的平衡解离常数(KD)结合抗原或抗原内的表位。在一些实施方式中，抗体与抗原结合的KD为该抗体结合至非特异性抗原(例如BSA、酪蛋白)的KD的10％或更低(例如1％)。可使用已知的方法来测量KD，例如通过
Figure PCTCN2022123370-appb-000002
表面等离子体共振测定法所测量的。然而，特异性结合至抗原或抗原内的表位的抗体可能对其它相关的抗原具有交叉反应性，例如，对来自其它物种(同源)(诸如人或猴，例如食蟹猕猴(Macaca fascicularis)(cynomolgus，cyno)、黑猩猩(Pan troglodytes)(chimpanzee，chimp))或狨猴(Callithrix jacchus)(commonmarmoset，marmoset)的相应抗原具有交叉反应性。
术语“抗IL23抗体”和“结合IL23的抗体”是指能够以足够的亲和力结合IL23的抗体。在一个实施例中，与无关的、非IL23蛋白的抗体的结合程度小于该抗体与IL23和IL12结合的约10％，所述结合通过
Figure PCTCN2022123370-appb-000003
表面等离子体共振测定法测量。在某些实施例中，与IL23蛋白结合的抗体具有以下解离常数(KD)<约1μM、<约100nM、<约10nM、<约1nM、<约0.1nM、<约0.01nM或<约0.001nM。在某些实施例中，抗IL23抗体结合人或食蟹猴IL23 p19亚基的抗原表位。
术语“连接子”、“Linker”或“接头”指连接两个多肽片段的连接单元，通常具有一定的柔性，接头的使用不会使蛋白质结构域原有的功能丧失。在本文中，同一 结构中出现的连接子可以是相同或不同的。连接子可以是肽连接子，其包含一个或多个氨基酸，典型的约1-30个、2-24个或3-15个氨基酸。应用于本文的连接子可以是相同或不同的。
术语“抗体依赖性细胞的细胞毒性”、“抗体依赖性细胞介导的细胞毒性”或“ADCC”是诱导细胞死亡的机制，该机制依赖于抗体包被靶细胞与具有裂解活性的效应细胞(诸如自然杀伤细胞(NK)、单核细胞、巨噬细胞和中性粒细胞)经由效应细胞上表达的Fcγ受体(FcγR)发生的相互作用。例如，NK细胞表达FcγRIIIa，而单核细胞表达FcγRI、FcγRII和FcγRIIIa。本文提供的抗体的ADCC活性可使用体外测定，使用表达抗原的细胞作为靶细胞和NK细胞作为效应细胞进行评定。根据从裂解的细胞中释放的标记物(例如放射性底物、荧光染料或天然胞内蛋白)来检测细胞裂解。
术语“抗体依赖性细胞吞噬作用”(“ADCP”)是指通过吞噬细胞(诸如巨噬细胞或树突状细胞)的内化作用消除抗体包被的靶细胞的机制。
术语“补体依赖性细胞毒性”或“CDC”是指诱导细胞死亡的机制，其中靶结合抗体的Fc效应域结合并激活补体成分C1q，C1q继而激活补体级联，从而导致靶细胞死亡。补体的激活也可导致补体成分沉积在靶细胞表面上，这些补体成分通过结合白细胞上的补体受体(例如，CR3)来促进CDC。
术语“核酸”在本文中可与术语“多核苷酸”互换使用，并且是指呈单链或双链形式的脱氧核糖核苷酸或核糖核苷酸及其聚合物。所述术语涵盖含有已知核苷酸类似物或修饰的骨架残基或连接的核酸，所述核酸是合成的、天然存在的和非天然存在的，具有与参考核酸相似的结合特性，并且以类似于参考核苷酸的方式代谢。此类类似物的实例包括但不限于硫代磷酸酯、氨基磷酸酯、甲基膦酸酯、手性-甲基膦酸酯、2-O-甲基核糖核苷酸、肽-核酸(PNA)。“分离的”核酸指已经与其天然环境的组分分开的核酸分子。分离的核酸包括在下述细胞中含有的核酸分子，所述细胞通常含有该核酸分子，但该核酸分子存在于染色体外或存在于不同于其天然染色体位置的染色体位置处。编码多肽或融合蛋白的分离的核酸指编码多肽或融合蛋白的一个或更多个核酸分子，包括在单一载体或分开的载体中的这样的一个或更多个核酸分子，和存在于宿主细胞中一个或更多个位置的这样的一个或更多个核酸分子。除非另有说明，否则特定的核酸序列还隐含地涵盖其保守修饰的变体(例如，简并密码子取代)和互补序列以及明确指明的序列。具体地，如下详述，简并密码子取代可以通过产生如下序列而获得，在这些序列中，一个或多个所选的(或全部)密码子的第三位被混合碱基和/或脱氧肌苷残基取代。
术语“多肽”和“蛋白质”在本文中可互换使用，指氨基酸残基的聚合物。该术语适用于氨基酸聚合物，其中一个或多个氨基酸残基是相应天然存在的氨基酸的人工化学模拟物，以及适用于天然存在的氨基酸聚合物和非天然存在的氨基酸聚合物。除非另外说明，否则特定的多肽序列还隐含地涵盖其保守修饰的变体。
术语序列“同一性”指，当对两条序列进行最佳比对时，必要时引入间隙，以获取最大序列同一性百分比，且不将任何保守性取代视为序列同一性的一部分，两条序列的氨基酸/核酸在等价位置相同的程度(百分比)。为测定序列同一性百分比，比对可以通过本领域技术已知的技术来实现，例如使用公开可得到的计算机软件，诸如BLAST、BLAST-2、ALIGN、ALIGN-2或Megalign(DNASTAR)软件。本领域技术人员可确定适用于测量比对的参数，包括在所比较的序列全长上达成最大比对所需的任何算法。
本披露中，例如“相对于XX序列的位点”，意指待测序列与XX序列进行最佳比对，获得最高百分比同一性，此时待测序列与XX序列对应位置的位点为两条序列的相对位点。例如TACI的胞外区序列SEQ ID NO：51和SEQ ID NO：60上相对于SEQ ID NO：58的自然顺序氨基酸残基位点如表2所示：
表2.SEQ ID NO：58位点与SEQ ID NO：51或/SEQ ID NO：60相应位点
Figure PCTCN2022123370-appb-000004
备注：例如SEQ ID NO：60上第2位(自然顺序)残基位点与序列SEQ ID NO：58的第49位(自然顺序)残基位点为相应位点。
术语“融合”或“连接”是指部件(例如TACI多肽和抗体重链)直接地或经由一个或多个连接子通过共价键连接。当连接子是肽连接子时，所述共价键是肽键。
术语“抗IL23抗体融合蛋白”是指抗IL23抗体与活性蛋白融合的蛋白质。例如，TACI多肽融合到抗IL23抗体的重链末端而形成蛋白。
术语“载体”意指能够转运与其连接的另一多核苷酸的多核苷酸分子。一种类型的载体是“质粒”，其是指环状双链DNA环，其中可以连接附加的DNA区段。另 一种类型的载体是病毒载体，例如腺相关病毒载体(AAV或AAV2)，其中另外的DNA区段可以连接到病毒基因组中。某些载体能够在引入它们的宿主细胞中自主复制(例如，具有细菌复制起点的细菌载体和附加型哺乳动物载体)。其他载体(例如，非附加型哺乳动物载体)可以在引入宿主细胞中后整合到宿主细胞的基因组中，从而与宿主基因组一起复制。术语“表达载体”或“表达构建体”是指可对宿主细胞进行转化，且含有指导和/或控制(连同宿主细胞一起)与其可操作地连接的一个或多个异源编码区的表达的核酸序列的载体。表达构建体可以包括但不限于影响或控制转录、翻译且在存在内含子时影响与其可操作地连接的编码区的RNA剪接的序列。
术语“宿主细胞”、“宿主细胞系”和“宿主细胞培养物”可互换使用，并且指已经导入外源核酸的细胞，包括此类细胞的后代。宿主细胞包括“转化体”和“经转化的细胞”，其包括原代的经转化的细胞及自其衍生的后代，而不考虑传代的次数。后代在核酸内容物上可以与亲本细胞不完全相同，而是可以含有突变。本文中包括具有与在初始转化细胞中筛选或选择的相同功能或生物学活性的突变体后代。宿主细胞包括原核和真核宿主细胞，其中真核宿主细胞包括但不限于哺乳动物细胞、昆虫细胞系植物细胞和真菌细胞。哺乳动物宿主细胞包括人、小鼠、大鼠、犬、猴、猪、山羊、牛、马和仓鼠细胞，包括但不限于中国仓鼠卵巢(CHO)细胞、NSO、SP2细胞、HeLa细胞、幼仓鼠肾(BHK)细胞、猴肾细胞(COS)、人肝细胞癌细胞(例如，Hep G2)、A549细胞、3T3细胞和HEK-293细胞。真菌细胞包括酵母和丝状真菌细胞，包括例如巴氏毕赤酵母(Pichiapastoris)、芬兰毕赤酵母(Pichia finlandica)、海藻毕赤酵母(Pichia trehalophila)、科克拉马毕赤酵母(Pichia koclamae)、膜状毕赤酵母(Pichia membranaefaciens)、小毕赤酵母(Pichia minuta)(Ogataea minuta、Pichia lindneri)、仙人掌毕赤酵母(Pichiaopuntiae)、耐热毕赤酵母(Pichia thermotolerans)、柳毕赤酵母(Pichia salictaria)、Pichia guercuum、皮杰普毕赤酵母(Pichia pijperi)、具柄毕赤酵母(Pichia stiptis)、甲醇毕赤酵母(Pichia methanolica)、毕赤酵母属、酿酒酵母(Saccharomycescerevisiae)、酿酒酵母属、多形汉逊酵母(Hansenula polymorpha)、克鲁维酵母属、乳酸克鲁维酵母(Kluyveromyces lactis)、白色念珠菌(Candida albicans)、构巢曲霉(Aspergillus nidulans)、黑曲霉(Aspergillus niger)、米曲霉(Aspergillus oryzae)、里氏木霉(Trichoderma reesei)、勒克氏菌(Chrysosporium lucknowense)、镰刀菌属(Fusarium sp.)、禾谷镰刀菌(Fusarium gramineum)、菜镰刀菌(Fusarium venenatum)、小立碗藓(Physcomitrella patens)和粗糙脉孢菌(Neurospora crassa)。毕赤酵母属、任何酿酒酵母属、多形汉逊酵母(Hansenula polymorpha)、任何克鲁维酵母属、白色念珠菌(Candida albicans)、任何曲霉属、里氏木霉(Trichoderma reesei)、勒克霉菌(Chrysosporium lucknowense)、任何镰刀菌属、解脂耶氏酵母(Yarrowia lipolytica)和粗糙脉孢菌(Neurospora crassa)。
如在本申请中所使用的，表述“细胞”、“细胞系”和“细胞培养物”可以互换使用，并且所有这样的名称均包括子代。因而，词语“转化体”和“转化的细胞”包括原代受试者细胞和来源于其的培养物，而与传代的次数无关。还应理解的是，由于有意或无意的突变，使得并非所有子代均具有完全相同的DNA内容物。包括与筛选出其的原始转化细胞具有相同功能或生物活性的突变子代。
“任选”或“任选地”意味着随后所描述地事件或环境可以但不必发生，该说明包括该事件或环境发生或不发生的场合。
术语“药物组合物”表示含有一种或多种本文所述的抗IL23抗体融合蛋白与其他化学组分的混合物，所述其他组分例如生理学/可药用的载体和赋形剂。
术语“药学上可接受的载体”指药学配制剂中与活性成分不同的,且对受试者无毒的成分。药学可接受载剂包括但不限于缓冲剂、赋形剂、稳定剂或防腐剂。
术语“受试者”或“个体”包括人类和非人类动物。非人动物包括所有脊椎动物(例如哺乳动物和非哺乳动物)例如非人灵长类(例如，食蟹猴)、绵羊、狗、牛、鸡、两栖动物和爬行动物。除非指出时，否则所述术语“患者”或“受试者”在本文中可互换地使用。如本文所使用的，术语“食蟹猴(cyno)”或“食蟹猴(cynomolgus)”是指食蟹猴(Macaca fascicularis)。在某些实施方案中，个体或受试者是人。
“施用”或“给予”，当其应用于动物、人、实验受试者、细胞、组织、器官或生物流体时，是指外源性药物、治疗剂、诊断剂或组合物与动物、人、受试者、细胞、组织、器官或生物流体的接触。
术语“样本”是指从受试者分离的类似流体、细胞、或组织的采集物，以及存在于受试者体内的流体、细胞或组织。示例性样本为生物流体，诸如血液、血清和浆膜液、血浆、淋巴液、尿液、唾液、囊液、泪液、排泄物、痰、分泌组织和器官的粘膜分泌物、阴道分泌物、腹水、胸膜、心包、腹膜、腹腔和其它体腔的流体、由支气管灌洗液收集的流体、滑液、与受试者或生物来源接触的液体溶液，例如细胞和器官培养基(包括细胞或器官条件培养基)、灌洗液等，组织活检样本、细针穿刺、手术切除的组织、器官培养物或细胞培养物。
“治疗(treatment或treat)”和“处理”(及其语法变型)指试图改变所治疗个体的天然过程的临床干预，并且可以为了预防或者在临床病理学的过程期间实施。治疗的期望效果包括但不限于预防疾病的发生或再发生，减轻症状，减轻/减少疾病的任何直接或间接病理后果，预防转移，降低疾病进展速率，改善或减轻疾病状态，和消退或改善的预后。在一些实施方案中，使用本披露的抗体来延迟疾病的形成或减缓疾病的进展。
“有效量”一般是足以降低症状的严重程度及/或频率、消除这些症状及/或潜在病因、预防症状及/或其潜在病因出现及/或改良或改善由疾病状态引起或与其相关的损伤(例如肺病)的量。在一些实施例中，有效量是治疗有效量或预防有效量。“治疗有效量”是足以治疗疾病状态或症状、尤其与该疾病状态相关的状态或症状， 或者以其他方式预防、阻碍、延迟或逆转该疾病状态或以任何方式与该疾病相关的任何其他不理想症状的进展的量。“预防有效量”是当给予受试者时将具有预定预防效应，例如预防或延迟该疾病状态的发作(或复发)，或者降低该疾病状态或相关症状的发作(或复发)可能性的量。完全治疗或预防效未必在给予一个剂量之后便发生，可能在给予一系列剂量之后发生。因而，治疗或预防有效量可以一次或多次给予的方式给予。“治疗有效量”和“预防有效量”可取决于多种因素变化：诸如个体的疾病状态、年龄、性别和体重，以及治疗剂或治疗剂组合在个体中引发期望的应答的能力。有效治疗剂或治疗剂组合的示例性指标包括例如患者改善的健康状况。
本披露的抗IL23抗体
在一个方面中，本披露设计了一种新的抗IL23抗体及其融合蛋白。其具有以下一种或多种特性：
A.与人IL-23 p19和食蟹猴IL-23 p19特异性结合，不与鼠IL-23 p19特异性结合；优选地，以小于6.00E-11M(例如，小于6.00E-11M、小于5.00E-11M、小于4.00E-11M、小于3.00E-11M、小于2.00E-11M、小于1.7E-11M、小于1.6E-11M、小于1.5E-11M或更小)的KD值与人IL-23 p19结合，和/或以小于9.00E-11M(例如，小于9.00E-11M、小于8.00E-11M、小于7.00E-11M、小于6.00E-11M或更小)的KD值与食蟹猴IL-23 p19结合，所述KD值通过表面等离子体共振测定法所测量，例如通过
Figure PCTCN2022123370-appb-000005
表面等离子体共振测定法所测量；在一些实施方案中，所述检测
B.具有阻断IL-23/IL-23R结合活性；优选地，阻断人IL-23/IL-23R结合的IC 50值小于0.6nM(例如，小于0.6nM、小于0.5nM、小于0.4nM、小于0.3nM或更小)，所述IC 50值通过Elisa方法检测；
C.具有抑制IL-17分泌活性；优选地，以小于0.03nM(例如，小于0.03nM、小于0.02nM nM、小于0.01nM或更小)的IC 50值抑制IL-17分泌；所述IC 50值通过Elisa检测；
D.具有抑制BaF3-IL-23R细胞增殖活性；优选地，以小于0.5nM(例如，小于0.5nM、小于0.4nM nM、小于0.3nM、小于0.2nM、小于0.1nM或更小)的IC 50值抑制BaF3-IL-23R细胞增殖；所述IC 50值通过PerkinElmer检测；
E.具有抑制TNFα和/或IL-22分泌活性；或
F.在咪喹莫特诱导的银屑病动物模型、人IL-23诱导的银屑病动物模型和/或系统性红斑狼疮动物模型中具有强的体内药效。
示例性的抗IL23抗体
一方面，本披露提供一种抗IL23抗体，其中所述抗IL23抗体包含重链可变区和轻链可变区，其中，所述重链可变区包含HCDR1、HCDR2和HCDR3，所述轻链可变区包含LCDR1、LCDR2和LCDR3，其中，
在一些实施方案中，如上任一项所述的抗IL23抗体的所述重链可变区的HCDR1、HCDR2和HCDR3以及轻链可变区的LCDR1、LCDR2和LCDR3是根据选自Kabat编号规则定义的，其中：
(i)所述重链可变区的HCDR1如SEQ ID NO：27、28或5所示，HCDR2如SEQ ID NO：6所示，和HCDR3如SEQ ID NO：7所示，和所述轻链可变区的LCDR1如SEQ ID NO：8所示，LCDR2如SEQ ID NO：9所示，和LCDR3如SEQ ID NO：10所示；或
(ii)所述重链可变区的HCDR1如SEQ ID NO：11所示，HCDR2如SEQ ID NO：12所示，和HCDR3如SEQ ID NO：13所示，和所述轻链可变区的LCDR1如SEQ ID NO：14所示，LCDR2如SEQ ID NO：37或15所示，和LCDR3如SEQ ID NO：16所示。
在一些实施方案中，如上任一项所述的抗IL23抗体包含重链可变区和轻链可变区，其中
(i)所述重链可变区的HCDR1如SEQ ID NO：27所示，HCDR2如SEQ ID NO：6所示，和HCDR3如SEQ ID NO：7所示，和所述轻链可变区的LCDR1如SEQ ID NO：8所示，LCDR2如SEQ ID NO：9所示，和LCDR3如SEQ ID NO：10所示；或
(ii)所述重链可变区的HCDR1如SEQ ID NO：28所示，HCDR2如SEQ ID NO：6所示，和HCDR3如SEQ ID NO：7所示，和所述轻链可变区的LCDR1如SEQ ID NO：8所示，LCDR2如SEQ ID NO：9所示，和LCDR3如SEQ ID NO：10所示；或
(iii)所述重链可变区的HCDR1如SEQ ID NO：11所示，HCDR2如SEQ ID NO：12所示，和HCDR3如SEQ ID NO：13所示，和所述轻链可变区的LCDR1如SEQ ID NO：14所示，LCDR2如SEQ ID NO：37所示，和LCDR3如SEQ ID NO：16所示。
在一些实施方案中，如前任一项所述的抗IL23抗体是鼠源、嵌合或人源化的。在一些实施方案中，所述抗IL23抗体是人源化的。在一些实施方案中，所述抗IL23抗体的重链可变区具有来源于IGHV2-26*01的FR1、FR2、FR3和来源于IGHJ6*01的FR4，并且其是未被取代的或具有选自1E、30T、37V、44G、49G、73N、89R和93V组成的组中的一个或多个氨基酸取代；和/或所述轻链可变区具有来源于IGKV4-1*01的FR1、FR2、FR3和来源于IGKJ4*01的FR4，并且其是未被取代的或具有选自4L、17Q、36F、58I、60A和68R组成的组中的一个或多个氨基酸取代。在一些实施方案中，其中抗IL23抗体，所述重链可变区的HCDR1如SEQ ID NO：27、28或5所示，HCDR2如SEQ ID NO：6所示，和HCDR3如SEQ ID NO：7所示，和所述轻链可变区的LCDR1如SEQ ID NO：8所示，LCDR2如SEQ ID NO：9所示，和LCDR3如SEQ ID NO：10所示。上述可变区和CDR是根据Kabat编 号规则定义的。
在一些实施方案中，如前任一项所述的抗IL23抗体是人源化抗体，在一些实施方案中，所述抗IL23抗体的重链可变区具有来源于IGHV1-3*01的FR1、FR2、FR3和来源于IGHJ1*01的FR4，并且其是未被取代的或具有选自1E、2F、28S、44S、71V和73Q组成的组中的一个或多个氨基酸取代；和/或所述轻链可变区具有来源于IGKV2-40*01的FR1、FR2、FR3和来源于IGKJ2*01的FR4，并且其是未被取代的或具有选自4I、36F、39R、71L和100S组成的组中的一个或多个氨基酸取代。在一些实施方案中，所述抗IL23抗体的重链可变区的HCDR1如SEQ ID NO：11所示，HCDR2如SEQ ID NO：12所示，和HCDR3如SEQ ID NO：13所示，和所述轻链可变区的LCDR1如SEQ ID NO：14所示，LCDR2如SEQ ID NO：37或15所示，和LCDR3如SEQ ID NO：16所示。上述可变区和CDR是根据Kabat编号规则定义的。
在一些实施方案中，如上任一项所述的抗IL23抗体包含重链可变区和轻链可变区，其中，
(i)所述重链可变区如SEQ ID NO：22、21、20、19、18或17所示，和所述轻链可变区如SEQ ID NO：26、25、24或23所示；或
(ii)所述重链可变区如SEQ ID NO：29、30、31或32所示，和所述轻链可变区如SEQ ID NO：36、35、34或33所示；或
(iii)所述重链可变区如SEQ ID NO：1所示，和所述轻链可变区如SEQ ID NO：2所示；或
(iv)所述重链可变区如SEQ ID NO：3所示，和所述轻链可变区如SEQ ID NO：4所示。
在一些实施方案中，如上任一项所述的抗IL23抗体包含重链可变区和轻链可变区，其中，
(i)所述重链可变区如SEQ ID NO：22所示，和所述轻链可变区如SEQ ID NO：26所示；
(ii)所述重链可变区如SEQ ID NO：21所示，和所述轻链可变区如SEQ ID NO：25所示；或
(iii)所述重链可变区如SEQ ID NO：29所示，和所述轻链可变区如SEQ ID NO：36所示。
在一些实施方案中，如上任一项所述的抗IL23抗体进一步包含抗体重链恒定区和轻链恒定区。在一些实施方案中，所述重链恒定区为人IgG重链恒定区；在一些实施方案中，所述重链恒定区选自人IgG1、IgG2、IgG3和IgG4恒定区；在一些实施方案中，所述轻链恒定区选自人抗体κ或λ链恒定区；在一些实施方案中，所述重链恒定区为人IgG1重链恒定区，所述轻链恒定区为人κ轻链恒定区。在一些实施方案中，所述重链恒定区的Fc区具有一个或更多个能够减少Fc区与 Fc受体结合的氨基酸取代。在一些实施方案中，所述Fc区具有L234A、L235A突变，和/或S228P突变，和/或YTE突变(M252Y、S254T和T256E)，所述突变编号依据为EU索引。在一些实施方案中，所述Fc区具有L234A、L235A突变。在一些实施方案中，所述重链恒定区包含SEQ ID NO：38的氨基酸序列，所述轻链恒定区包含SEQ ID NO：39的氨基酸序列。
在一些实施方案中，如上任一项所述的抗IL23抗体包含重链和轻链，其中，
(i)所述抗IL23抗体的重链包含与SEQ ID NO：42具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列，和轻链包含与SEQ ID NO：43具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列；或
(ii)所述抗IL23抗体的重链包含与SEQ ID NO：40具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列，和轻链包含与SEQ ID NO：41具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列；或
(iii)所述抗IL23抗体的重链包含与SEQ ID NO：44具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列，和轻链包含与SEQ ID NO：45具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列。
在一些实施方案中，如上任一项所述的抗IL23抗体包含重链和轻链，其中
(i)所述抗IL23抗体的重链如SEQ ID NO：42所示，和轻链如SEQ ID NO：43所示；或
(ii)所述抗IL23抗体的重链如SEQ ID NO：40所示，和轻链如SEQ ID NO：41所示；或
(iii)所述抗IL23抗体的重链如SEQ ID NO：44所示，和轻链如SEQ ID NO：45所示。
本披露的抗IL23抗体融合蛋白
本披露构建了一种抗IL23抗体融合蛋白，其包含抗IL23抗体和TACI多肽，其中所述抗IL23抗体特异性结合人IL23，优选特异性结合人IL23 p19亚基。在一些实施方案中，前面任一项所述的抗IL23抗体或抗IL23抗体融合蛋白，其具有一种或更多种以下特性：
A.与人IL-23 p19和食蟹猴IL-23 p19特异性结合，不与鼠IL-23 p19特异性结合；优选地，以小于6.00E-11M(例如，小于6.00E-11M、小于5.00E-11M、小于4.00E-11M、小于3.00E-11M、小于2.00E-11M、小于1.7E-11M、小于1.6E-11M、小于1.5E-11M或更小)的KD值与人IL-23 p19结合，和/或以小于9.00E-11M(例如，小于9.00E-11M、小于8.00E-11M、小于7.00E-11M、小于6.00E-11M或更小) 的KD值与食蟹猴IL-23 p19结合，所述KD值通过表面等离子体共振测定法所测量，例如通过
Figure PCTCN2022123370-appb-000006
表面等离子体共振测定法所测量；
B.具有阻断IL-23/IL-23R结合活性；优选地，阻断人IL-23/IL-23R结合的IC 50值小于0.6nM(例如，小于0.6nM、小于0.5nM、小于0.4nM、小于0.3nM或更小)，所述IC 50值通过Elisa方法检测；
C.具有阻断BAFF/BAFF-R结合活性；优选地，阻断BAFF/BAFF-R结合的IC 50值小于7.00nM(例如，小于7.00nM、小于6.00nM、小于5.00nM、小于4.00nM、小于3.00nM、小于2.00nM、小于1.00nM、小于0.9nM或更小)，所述IC 50值通过Elisa方法检测；
D.具有阻断BAFF/BCMA结合活性；优选地，阻断BAFF/BCMA结合的IC 50值小于4.50nM(例如，小于4.50nM、小于3.00nM、小于2.00nM、小于1.00nM或更小)，所述IC 50值通过Elisa方法检测；
E.具有阻断BAFF/TACI结合活性；优选地，阻断BAFF/TACI结合的IC 50值小于6.00nM(例如，小于6.00nM、小于5.00nM、小于4.00nM、小于3.00nM、小于2.00nM、小于1.00nM或更小)，所述IC 50值通过Elisa方法检测；
F.具有阻断APRIL/BCMA结合活性；优选地，阻断APRIL/BCMA结合的IC 50值小于1.00nM(例如，小于1.00nM、小于0.5nM、小于0.4nM、小于0.1nM或更小)，所述IC 50值通过Elisa方法检测；
G.具有阻断APRIL/TACI结合活性；优选地，阻断APRIL/TACI结合的IC 50值小于3.00nM(例如，小于3.00nM、小于2.00nM、小于1.50nM或更小)，所述IC 50值通过Elisa方法检测；
H.具有抑制IL-17分泌活性；优选地，以小于0.03nM(例如，小于0.03nM、小于0.02nM nM、小于0.01nM或更小)的IC 50值抑制IL-17分泌；所述IC 50值通过Elisa检测；
I.具有抑制BaF3-IL-23R细胞增殖活性；优选地，以小于0.5nM(例如，小于0.5nM、小于0.4nM nM、小于0.3nM、小于0.2nM、小于0.1nM或更小)的IC 50值抑制BaF3-IL-23R细胞增殖；所述IC 50值通过PerkinElmer检测；
J.具有抑制TNFα、IL-22和IgA细胞因子分泌活性；或
K.具有抑制B细胞增殖活性。
示例性的抗IL23抗体融合蛋白
在所述抗IL23抗体融合蛋白的一些实施方案中，其中所述抗IL23抗体包含重链可变区和轻链可变区，其中，所述重链可变区包含HCDR1、HCDR2和HCDR3，所述轻链可变区包含LCDR1、LCDR2和LCDR3，其中，所述重链可变区的HCDR1、HCDR2和HCDR3以及轻链可变区的LCDR1、LCDR2和LCDR3是根据选自Kabat编号规则定义的，其中：
(i)所述重链可变区的HCDR1如SEQ ID NO：27、28或5所示，HCDR2 如SEQ ID NO：6所示，和HCDR3如SEQ ID NO：7所示，和所述轻链可变区的LCDR1如SEQ ID NO：8所示，LCDR2如SEQ ID NO：9所示，和LCDR3如SEQ ID NO：10所示；或
(ii)所述重链可变区的HCDR1如SEQ ID NO：11所示，HCDR2如SEQ ID NO：12所示，和HCDR3如SEQ ID NO：13所示，和所述轻链可变区的LCDR1如SEQ ID NO：14所示，LCDR2如SEQ ID NO：37或15所示，和LCDR3如SEQ ID NO：16所示。
在一些实施方案中，其中
(i)所述重链可变区的HCDR1如SEQ ID NO：27所示，HCDR2如SEQ ID NO：6所示，和HCDR3如SEQ ID NO：7所示，和所述轻链可变区的LCDR1如SEQ ID NO：8所示，LCDR2如SEQ ID NO：9所示，和LCDR3如SEQ ID NO：10所示；或
(ii)所述重链可变区的HCDR1如SEQ ID NO：28所示，HCDR2如SEQ ID NO：6所示，和HCDR3如SEQ ID NO：7所示，和所述轻链可变区的LCDR1如SEQ ID NO：8所示，LCDR2如SEQ ID NO：9所示，和LCDR3如SEQ ID NO：10所示；或
(iii)所述重链可变区的HCDR1如SEQ ID NO：11所示，HCDR2如SEQ ID NO：12所示，和HCDR3如SEQ ID NO：13所示，和所述轻链可变区的LCDR1如SEQ ID NO：14所示，LCDR2如SEQ ID NO：37所示，和LCDR3如SEQ ID NO：16所示。
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述的抗IL23抗体为鼠源抗体、嵌合抗体或人源化抗体。在一些实施方案中，所述抗IL23抗体是人源化抗体。在一些实施方案中，所述抗IL23抗体的重链可变区具有来源于IGHV2-26*01的FR1、FR2、FR3和来源于IGHJ6*01的FR4，并且其是未被取代的或具有选自1E、30T、37V、44G、49G、73N、89R和93V组成的组中的一个或多个氨基酸取代；和/或所述轻链可变区具有来源于IGKV4-1*01的FR1、FR2、FR3和来源于IGKJ4*01的FR4，并且其是未被取代的或具有选自4L、17Q、36F、58I、60A和68R组成的组中的一个或多个氨基酸取代。在一些实施方案中，所述抗IL23抗体的重链可变区的HCDR1如SEQ ID NO：27、28或5所示，HCDR2如SEQ ID NO：6所示，和HCDR3如SEQ ID NO：7所示，和所述轻链可变区的LCDR1如SEQ ID NO：8所示，LCDR2如SEQ ID NO：9所示，和LCDR3如SEQ ID NO：10所示。上述可变区和CDR是根据Kabat编号规则定义的。
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述的抗IL23抗体为人源化抗体，所述抗IL23抗体的重链可变区具有来源于IGHV1-3*01的FR1、FR2、FR3和来源于IGHJ1*01的FR4，并且其是未被取代的或具有选自1E、2F、28S、44S、71V和73Q组成的组中的一个或多个氨基酸取代；和/或所述轻链可变 区具有来源于IGKV2-40*01的FR1、FR2、FR3和来源于IGKJ2*01的FR4，并且其是未被取代的或具有选自4I、36F、39R、71L和100S组成的组中的一个或多个氨基酸取代。在一些实施方案中，所述抗IL23抗体的重链可变区的HCDR1如SEQ ID NO：11所示，HCDR2如SEQ ID NO：12所示，和HCDR3如SEQ ID NO：13所示，和所述轻链可变区的LCDR1如SEQ ID NO：14所示，LCDR2如SEQ ID NO：37或15所示，和LCDR3如SEQ ID NO：16所示。上述可变区和CDR是根据Kabat编号规则定义的。
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述抗IL23抗体包含重链可变区和轻链可变区，其中，
(i)所述重链可变区如SEQ ID NO：22、21、20、19、18或17所示，和所述轻链可变区如SEQ ID NO：26、25、24或23所示；或
(ii)所述重链可变区如SEQ ID NO：29、30、31或32所示，和所述轻链可变区如SEQ ID NO：36、35、34或33所示；或
(iii)所述重链可变区如SEQ ID NO：1所示，和所述轻链可变区如SEQ ID NO：2所示；或
(iv)所述重链可变区如SEQ ID NO：3所示，和所述轻链可变区如SEQ ID NO：4所示。
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述抗IL23抗体包含重链可变区和轻链可变区，其中，
(i)所述重链可变区如SEQ ID NO：22所示，和所述轻链可变区如SEQ ID NO：26所示；或
(ii)所述重链可变区如SEQ ID NO：21所示，和所述轻链可变区如SEQ ID NO：25所示；或
(iii)所述重链可变区如SEQ ID NO：29所示，和所述轻链可变区如SEQ ID NO：36所示。
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述的抗IL23抗体进一步包含抗体重链恒定区和轻链恒定区。在一些实施方案中，所述重链恒定区为人IgG重链恒定区；在一些实施方案中，所述重链恒定区选自人IgG1、IgG2、IgG3和IgG4恒定区；在一些实施方案中，所述轻链恒定区选自人抗体κ或λ链恒定区；在一些实施方案中，所述重链恒定区为人IgG1重链恒定区，所述轻链恒定区为人κ轻链恒定区。在一些实施方案中，所述重链恒定区的Fc区具有一个或更多个能够减少Fc区与Fc受体结合的氨基酸取代。在一些实施方案中，所述Fc区具有L234A、L235A突变，和/或S228P突变，和/或YTE突变(M252Y、S254T和T256E)，所述突变编号依据为EU索引。在一些实施方案中，所述重链恒定区包含SEQ ID NO：38的氨基酸序列，所述轻链恒定区包含SEQ ID NO：39的氨基酸序列。
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述抗IL23抗体包含重链和轻链，其中，
(i)所述抗IL23抗体的重链包含与SEQ ID NO：42具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列，和轻链包含与SEQ ID NO：43具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列；或
(ii)所述抗IL23抗体的重链包含与SEQ ID NO：40具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列，和轻链包含与SEQ ID NO：41具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列；或
(iii)所述抗IL23抗体的重链包含与SEQ ID NO：44具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列，和轻链包含与SEQ ID NO：45具有至少90％(例如至少90％、95％、96％、97％、98％或99％)序列同一性的氨基酸序列。
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述抗IL23抗体包含重链和轻链，其中
(i)所述抗IL23抗体的重链如SEQ ID NO：42所示，和轻链如SEQ ID NO：43所示；或
(ii)所述抗IL23抗体的重链如SEQ ID NO：40所示，和轻链如SEQ ID NO：41所示；或
(iii)所述抗IL23抗体的重链如SEQ ID NO：44所示，和轻链如SEQ ID NO：45所示。
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述TACI多肽相比野生型的TACI多肽(序列如SEQ ID NO：51)不容易发生断裂。
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述TACI多肽如SEQ ID NO：58所示或是SEQ ID NO：58的截短片段或其变体；其中所述截短片段包含SEQ ID NO：58的第48位至第85位氨基酸残基，所述变体在SEQ ID NO：58或其截短片段的基础上具有选自第49、52、53、57、65、82和83位中的一个或更多个氨基酸替换，其中所述氨基酸替换的位点为相对于序列SEQ ID NO：58的自然顺序编号的氨基酸残基位点。
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述TACI多肽的截短片段包含：SEQ ID NO：58的第48位至第86位氨基酸残基；SEQ ID NO：58的第48位至第87位氨基酸残基；或SEQ ID NO：58的第48位至第88位氨基酸残基。
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述TACI多肽的序列如SEQ ID NO：60-63任一项所示。
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述TACI多肽的序列为SEQ ID NO：58变体或者SEQ ID NO：58的截短片段(例如SEQ ID NO：60、SEQ ID NO：61、SEQ ID NO：62或SEQ ID NO：63)的变体，所述变体在SEQ ID NO：58或其截短片段序列的基础上具有选自第49、52、53、57、65、82和83位中的任意1个、2个、3个、4个、5个、6个或7个氨基酸的替换，所述氨基酸替换的位点为相对于序列SEQ ID NO：58的自然顺序编号的氨基酸残基位点。
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述TACI多肽在SEQ ID NO：58、SEQ ID NO：60、SEQ ID NO：61、SEQ ID NO：62或SEQ ID NO：63序列的基础上具有选自由49T或49R、52S、53E或53Q、57E、65T或65A、82A或82R、和83Y组成的组中的一个或更多个氨基酸替换(例如1个、2个、3个、4个、5个、6个或7个氨基酸的替换)，其中所述氨基酸替换的位点为相对于序列SEQ ID NO：58的自然顺序编号的氨基酸残基位点。
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，其中所述TACI多肽在SEQ ID NO：58、SEQ ID NO：60、SEQ ID NO：61、SEQ ID NO：62或SEQ ID NO：63序列的基础上具有选自49T、52S、53E、53Q、57E和82A中的任一个氨基酸替换；在SEQ ID NO：58、SEQ ID NO：60、SEQ ID NO：61、SEQ ID NO：62或SEQ ID NO：63序列的基础上具有49R和65T氨基酸替换；在SEQ ID NO：58、SEQ ID NO：60、SEQ ID NO：61、SEQ ID NO：62或SEQ ID NO：63序列的基础上具有45R和65A氨基酸替换；在SEQ ID NO：58、SEQ ID NO：60、SEQ ID NO：61、SEQ ID NO：62或SEQ ID NO：63序列的基础上具有49R、65T和82R氨基酸替换；在SEQ ID NO：58、SEQ ID NO：60、SEQ ID NO：61、SEQ ID NO：62或SEQ ID NO：63序列的基础上具有53E和57E氨基酸替换；在SEQ ID NO：58、SEQ ID NO：60、SEQ ID NO：61、SEQ ID NO：62或SEQ ID NO：63序列的基础上具有52S、53E和57E氨基酸替换；在SEQ ID NO：58、SEQ ID NO：60、SEQ ID NO：61、SEQ ID NO：62或SEQ ID NO：63序列的基础上具有49T和82A氨基酸替换；在SEQ ID NO：58、SEQ ID NO：60、SEQ ID NO：61、SEQ ID NO：62或SEQ ID NO：63序列的基础上具有49T和83Y氨基酸替换；在SEQ ID NO：58、SEQ ID NO：60、SEQ ID NO：61、SEQ ID NO：62或SEQ ID NO：63序列的基础上具有49T、82A和83Y氨基酸替换；或者在SEQ ID NO：58、SEQ ID NO：60、SEQ ID NO：61、SEQ ID NO：62或SEQ ID NO：63序列的基础上具有49T、53E、57E和82A氨基酸替换。其中前述氨基酸替换的位点为相对于序列SEQ ID NO：58的自然顺序编号的氨基酸残基位点。
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述TACI多肽的序列如SEQ ID NO：60-63和SEQ ID NO：66-83中的任一所示。在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述TACI多肽序列如SEQ ID  NO：83所示。
在一些实施方案中，如上任一项所述的抗IL23抗体融合蛋白包括：
第一链，其从N端到C端依次为：[TACI多肽1]-[连接子1]-[抗IL23抗体重链]-[连接子2]-[TACI多肽2]，和
第二链，其为抗IL23抗体轻链，
其中，所述TACI多肽1和TACI多肽2可以相同或不相同，并且所述TACI多肽1和TACI多肽2可以独立地选自如上所述的任一TACI多肽；所述连接子1和连接子2可以相同或不相同；在一些实施方案中，所述TACI多肽1和TACI多肽2相同；在一些实施方案中，所述连接子为选自(G xS) y连接子，其中，X选自1-5的整数，Y选自0-6的整数，在一些实施方案中，所述连接子为GGGGSGGGGSGGGGS(如SEQ ID NO：84所示)或GGGS(如SEQ ID NO：85所示)。
在如上任一项所述的抗IL23抗体融合蛋白的一些实施方案中，所述的抗IL23抗体融合蛋白的第一链如SEQ ID NO：47所示，且第二链如SEQ ID NO：43所示；或第一链如SEQ ID NO：46所示，且第二链如SEQ ID NO：41所示。在一些实施方案中，如上任一项所述的抗IL23抗体融合蛋白具有2条如SEQ ID NO：47的第一链，和2条如SEQ ID NO：43的第二链；在一些实施方案中，所述的抗IL23抗体融合蛋白具有2条如SEQ ID NO：46的第一链，和2条如SEQ ID NO：41的第二链。
抗IL23抗体或其融合蛋白的变体
在某些实施方案中，涵盖本文中提供的抗IL23抗体或其融合蛋白的氨基酸序列变体。例如，可以期望改善抗体的结合亲和力和/或其它生物学特性。可以通过将合适的修饰引入编码抗体的核苷酸序列中，或者通过肽合成来制备抗体的氨基酸序列变体。此类修饰包括例如对抗IL23抗体或其融合蛋白的氨基酸序列内的残基的删除、和/或插入、和/或取代。可以进行删除、插入、和取代的任何组合以得到最终的构建体，只要最终的构建体拥有期望的特征，例如抗原结合特性。
取代、插入、和删除变体
在某些实施方案中，提供了具有一处或多处氨基酸取代的抗体变体。取代诱变感兴趣的位点包括CDR和FR。保守取代在表3中在“优选的取代”的标题下显示。更实质的变化在表3中在“示例性取代”的标题下提供，并且如下文参照氨基酸侧链类别进一步描述的。可以将氨基酸取代引入感兴趣的抗体中，并且对产物筛选期望的活性，例如保留/改善的抗原结合，降低的免疫原性，或改善的ADCC或CDC。
表3.氨基酸的取代
原始残基 示例性取代 优选的取代
Ala(A) Val；Leu；Ile Val
Arg(R) Lys；Gln；Asn Lys
Asn(N) Gln；His；Asp,Lys；Arg Gln
Asp(D) Glu；Asn Glu
Cys(C) Ser；Ala Ser
Gln(Q) Asn；Glu Asn
Glu(E) Asp；Gln Asp
Gly(G) Ala Ala
His(H) Asn；Gln；Lys；Arg Arg
Ile(I) Leu；Val；Met；Ala；Phe；正亮氨酸 Leu
Leu(L) 正亮氨酸；Ile；Val；Met；Ala；Phe Ile
Lys(K) Arg；Gln；Asn Arg
Met(M) Leu；Phe；Ile Leu
Phe(F) Trp；Leu；Val；Ile；Ala；Tyr Tyr
Pro(P) Ala Ala
Ser(S) Thr Thr
Thr(T) Ser Ser
Trp(W) Tyr；Phe Tyr
Tyr(Y) Trp；Phe；Thr；Ser Phe
Val(V) Ile；Leu；Met；Phe；Ala；正亮氨酸 Leu
依照常见的侧链特性，氨基酸可以如下分组:
(1)疏水性的：正亮氨酸，Met，Ala，Val，Leu，Ile；
(2)中性，亲水性的：Cys，Ser，Thr，Asn，Gln；
(3)酸性的：Asp，Glu；
(4)碱性的：His，Lys，Arg；
(5)影响链取向的残基：Gly，Pro；
(6)芳香族的：Trp，Tyr，Phe。
非保守取代会需要用这些类别之一的成员替换另一个类别的成员。
一类取代变体涉及取代亲本抗体(例如人源化或人抗体)的一个或多个CDR残基。一般地，经选择用于进一步研究的所得变体相对于亲本抗体会具有某些生物学特性(例如升高的亲和力，降低的免疫原性)的改变(例如改善)，和/或会基本上保留亲本抗体的某些生物学特性。一种例示性的取代变体是亲和力成熟的抗体，可以例如使用基于噬菌体展示的亲和力成熟技术(如本文所述的那些技术)，便利地产生所述抗体。简言之，将一个或多个CDR残基突变，并将变体抗体在噬菌体上展示，并对其筛选特定的生物学活性(例如结合亲和力)。可以对CDR做出改变(例如取代)，例如以改善抗体亲和力。可以对CDR“热点”，即在体细胞成熟过程期间以高频率经历突变的密码子所编码的残基，和/或接触抗原的残基做出此类改变，同时对所得的变体VH或VL测试结合亲和力。在亲和力成熟的一些实施方案中，通过多种方法(例如易错PCR、链改组、或寡核苷酸指导的诱变)的任一种，将多样性引入所选择用于成熟的可变基因中。然后，创建次级文库。 然后，筛选文库以鉴定具有期望的亲和力的任何抗体变体。另一种引入多样性的方法涉及CDR定向的方法，其中将几个CDR残基(例如一次4-6个残基)随机化。可以例如使用丙氨酸扫描诱变或建模来特异性鉴定涉及抗原结合的CDR残基。特别地，经常靶向HCDR3和LCDR3。
在某些实施方案中，取代、插入或缺失可以在一个或多个CDR内发生，只要此类变化不实质性降低抗体结合抗原的能力。例如，可以对CDR做出保守变化(例如保守取代，如本文中提供的)，其不实质性降低结合亲和力。此类变化可以例如在CDR中的抗原接触残基外部。在上文提供的变体VH和VL序列的某些实施方案中，每个CDR是未改变的，或者含有不超过1、2或3处氨基酸取代。
一种可用于鉴定抗体中可以作为诱变靶位的残基或区域的方法称作“丙氨酸扫描诱变”。在这种方法中，鉴定一个残基或靶残基组(例如带电荷的残基，诸如Arg、Asp、His、Lys和Glu)，并且用中性或带负电荷的氨基酸(例如，Ala或聚丙氨酸)替换以确定该抗体与抗原的相互作用是否受影响。可以在对初始取代显示功能敏感性的氨基酸位置引入进一步的取代。此外，可通过研究抗原-抗体复合物的晶体结构来鉴定抗体与抗原间的接触点。这些接触残基及邻近残基可以作为取代候选物被打靶或消除。可以筛选变体以确定它们是否含有期望的特性。
氨基酸序列插入包括长度范围为1个残基至含有100或更多个残基的多肽的氨基和/或羧基端融合，和单个或多个氨基酸残基的序列内插入。末端插入的例子包括具有N端甲硫氨酰基残基的抗体。抗体分子的其它插入变体包括抗体的N或C端与酶或延长抗体的血清半衰期的多肽的融合物。
Fc区的修饰
在一个方面，本披露的抗IL23抗体或抗IL23抗体融合蛋白的Fc区包含一个或多个氨基酸取代，所述一个或多个氨基酸取代减少其与Fc受体的结合，例如其与Fcγ受体的结合，并且降低或消除效应子功能。天然IgG Fc区，具体地是IgG 1Fc区或IgG 4Fc区，可能导致本披露的融合蛋白靶向表达Fc受体的细胞，而不是表达抗原的细胞。在一些实施方案中，本披露改造的Fc区表现出降低的对Fc受体的结合亲和力和/或降低的效应子功能。在一些实施方案中，改造的Fc区与天然Fc区相比，对Fc受体的结合亲和力下降50％、80％、90％或95％以上。在一些实施方案中，所述的Fc受体是Fcγ受体。在一些实施方案中，所述Fc受体是人Fcγ受体，例如FcγRI、FcγRIIa、FcγRIIB、FcγRIIIa。在一些实施方案中，改造的Fc区与天然Fc区相比，对补体，如C1q的结合亲和力也降低。在一些实施方案中，改造的Fc区与天然Fc区相比，对新生儿Fc受体(FcRn)的结合亲和力不降低。在一些实施例中，改造的Fc区具有降低的效应子功能，所述降低的效应子功能可以包括但不限于以下中的一个或多个：降低的补体依赖性细胞毒性(CDC)、降低的抗体依赖性细胞介导的细胞毒性(ADCC)、降低的抗体依赖性细胞吞噬(ADCP)、减少的细胞因子分泌、减少的免疫复合物介导的抗原呈递细胞的抗原摄取、减少 的与NK细胞的结合、减少的与巨噬细胞的结合、减少的与单核细胞的结合、减少的与多形核细胞的结合、减少的直接信号传导诱导性细胞凋亡、降低的树突细胞成熟或减少的T细胞引发。对于IgG 1Fc区，在238、265、269、270、297、327和329等位置的氨基酸残基取代可降低的效应子功能。在一些实施方案中，所述Fc区是人IgG 1Fc区，并且在234和235位置的氨基酸残基为A，编号依据为EU索引。对于IgG 4Fc区，在228等位置的氨基酸残基取代可降低的效应子功能。
抗IL23抗体或抗IL23抗体融合蛋白可包含与Fc区的两个亚基融合的不同结合域，因此可能导致不期望的同源二聚化。为了提高产率和纯度，可以在本披露的融合蛋白的Fc区中引入促进异源二聚化的修饰将是有利的。在一些实施方式中，本披露的Fc区包含根据杵臼(knob-into-hole，KIH)技术的改造，该方法涉及在第一亚基的界面处引入凸起结构(knob)以及在第二亚基的界面处引入孔结构(hole)。使得所述凸起结构可以定位在孔结构中，促进异源二聚体的形成并抑制同源二聚体的产生。凸起结构是通过用较大侧链(例如酪氨酸或色氨酸)取代来自第一亚基的界面的小氨基酸侧链而构建的。而孔结构是通过用较小的氨基酸侧链(例如丙氨酸或苏氨酸)取代大氨基酸侧链而在第二亚基的界面中创建的。凸起结构和孔结构通过改变编码多肽的核酸来制备，可选的氨基酸取代如下表4所示：
表4.KIH突变组合
Figure PCTCN2022123370-appb-000007
除了杵臼技术外，用于修饰多特异性抗体的重链的CH3结构域以实现异源二聚化的其他技术也是本领域中已知的，例如WO96/27011、WO98/050431、EP1870459、WO2007/110205、WO 007/147901、WO2009/089004、WO2010/129304、WO2011/90754、WO2011/143545、WO2012/058768、WO2013/157954和WO013/096291。
Fc区的C末端可以是以氨基酸残基PGK结束的完整C末端；也可以是缩短的C末端，例如在所述缩短的C末端中已经去除了一个或两个C末端氨基酸残基。在一个优选的方面中，重链的C末端是以PG结束的缩短的C末端。因此，在一些实施方式中，完整抗体的组合物可以包括去除了所有K447残基和/或G446+K447残基的抗体群体。在一些实施方式中，完整抗体的组合物可以包括没有去除K447 残基和/或G446+K447残基的抗体群体。在一些实施方式中，完整抗体的组合物具有带有和不带有K447残基和/或G446+K447残基的抗体混合物的抗体群体。
重组方法
抗IL23抗体或抗IL23抗体融合蛋白可以使用重组方法来产生。对于这些方法，提供编码多肽或融合蛋白的一个或更多个分离的核酸。
在一个实施方案中，本披露提供了编码如前所述的抗IL23抗体或抗IL23抗体融合蛋白的分离的核酸。此类核酸可以给自独立的编码前述的任一多肽链。在另一方面中，本披露提供了包含此类核酸的一种或多种载体(例如表达载体)。在另一方面中，本披露提供了包含此类核酸的宿主细胞。在一个实施方案中，提供制备多肽或融合蛋白的方法，其中所述方法包括，在适合表达的条件下，培养包含编码所述多肽或融合蛋白的核酸的宿主细胞，如上文所提供的，和任选地从宿主细胞(或宿主细胞培养基)回收所述抗IL23抗体或抗IL23抗体融合蛋白。
为了重组产生抗IL23抗体或抗IL23抗体融合蛋白，将编码蛋白的核酸分离并插入一个或更多个载体中，用于在宿主细胞中进一步克隆和/或表达。此类核酸可以使用常规程序容易地分离和测序，或者通过重组方法产生或通过化学合成获得。
用于克隆或表达编码抗IL23抗体或抗IL23抗体融合蛋白的载体的适当宿主细胞包括本文描述的原核或真核细胞。例如，可在细菌中产生，特别是当不需要糖基化和Fc效应子功能时。在表达后，可以在可溶级分中从细菌细胞糊状物分离，并且可进一步纯化。
除了原核生物以外，真核微生物诸如丝状真菌或酵母也是用于编码融合蛋白的载体的合适的克隆或表达宿主，包括真菌和酵母菌株。适于表达融合蛋白的合适的宿主细胞也可源自多细胞生物体(无脊椎动物和脊椎动物)；无脊椎动物细胞的例子包括植物和昆虫细胞。已经鉴定了许多杆状病毒株，其可与昆虫细胞联合使用，特别是用于草地贪夜蛾(Spodoptera frugiperda)细胞的转染；还可利用植物细胞培养物作为宿主，例如US5959177、US 6040498、US6420548、US 7125978和US6417429；也可将脊椎动物细胞用作宿主，例如适应于在悬浮液中生长的哺乳动物细胞系。适宜的哺乳动物宿主细胞系的其它例子是经SV40转化的猴肾CVl系(COS-7)；人胚肾系(293或293T细胞)；幼仓鼠肾细胞(BHK)；小鼠塞托利(sertoli)细胞(TM4细胞)；猴肾细胞(CV1)；非洲绿猴肾细胞(VERO-76)；人宫颈癌细胞(HELA)；犬肾细胞(MDCK)；水牛鼠(buffalo rat)肝细胞(BRL3A)；人肺细胞(W138)；人肝细胞(Hep G2)；小鼠乳房肿瘤(MMT 060562)；TRI细胞；MRC 5细胞；和FS4细胞。其它适宜的哺乳动物宿主细胞系包括中国仓鼠卵巢(CHO)细胞，包括DHFR-CHO细胞；以及骨髓瘤细胞系，如Y0、NS0和Sp2/0。关于适合产生抗体的某些哺乳动物宿主细胞系的综述参见例如Yazaki，P.和Wu，A.M.，Methods in Molecular Biology，Vol.248，Lo，B.K.C.(编)，Humana Press，Totowa，NJ(2004)，第255-268页。
测定
本文提供的抗IL23抗体或抗IL23抗体融合蛋白可以通过本领域已知的多种测定法对其物理/化学特征和/或生物学活性进行鉴定、筛选或表征。在一个方面中，例如通过已知方法如ELISA、蛋白印迹法等，测试本披露的抗IL23抗体或抗IL23抗体融合蛋白活性。
治疗方法与施用途径
本文提供的任何抗IL23抗体或抗IL23抗体融合蛋白可用于治疗方法。在又一个方面，本披露提供抗IL23抗体或抗IL23抗体融合蛋白在药物的制造或制备中的用途。在一些实施方案中，所述B细胞障碍或自身免疫性疾病是与IL23相关的疾病或病症。在一些实施方案中，所述自身免疫性疾病选自：系统性红斑狼疮、重症肌无力、多发性硬化、胰岛素依赖性糖尿病、克罗恩氏病、类风湿关节炎、多关节型青少年类风湿关节炎和银屑病性关节炎；所述B细胞障碍选自：肿瘤、慢性白细胞性白血病、多发性骨髓瘤、非霍奇金淋巴瘤、移植后淋巴组织增生病以及轻链丙球蛋白病。在一些实施方案中，所述自身免疫性疾病为系统性红斑狼疮。在一个此类实施方案中，所述用途进一步包括向受试者施用有效量的至少一种另外的治疗剂(例如一种、两种、三种、四种、五种或六种另外的治疗剂)。根据任意以上实施方案的“受试者”可以是人。
在又一个的方面，提供包含所述抗IL23抗体或抗IL23抗体融合蛋白的药物组合物，例如，其用于以上任何制药用途或治疗方法。在一个实施方案中，药物组合物包含本文提供的任何多肽或融合蛋白和药学上可接受的载体。在另一个实施方案中，药物组合物还包含至少一种另外的治疗剂。
本披露的抗IL23抗体或抗IL23抗体融合蛋白可单独使用或与其他试剂联合用于治疗。例如，本披露的抗IL23抗体或抗IL23抗体融合蛋白可与至少一种另外的治疗剂共同施用。
本披露的抗IL23抗体或抗IL23抗体融合蛋白(和任何另外的治疗剂)可通过任何合适的手段施用，包括肠胃外、肺内和鼻内，并且如果需要局部治疗，则病灶内施用。肠胃外输注包括肌肉内、静脉内、动脉内、腹膜内或皮下施用。给药可以通过任何适当的途径，例如，通过注射，诸如静脉内或皮下注射，这部分取决于施用是短期的还是长期的。本文考虑多种给药时间方案，包括但不限于，单次或在多个时间点多次施用，推注施用和脉冲输注。
本披露的抗IL23抗体或抗IL23抗体融合蛋白将以符合良好医疗实践的方式配制、给药和施用。在此背景下考虑的因素包括所治疗的具体病症、所治疗的具体哺乳动物、个体患者的临床状况、病症的起因、试剂的递送部位、施用方法、施用时间安排以及医学从业者已知的其他因素。多肽或融合蛋白可以与或不与目前用于预防或治疗所述病症的一种或更多种试剂一起配制。此类其它试剂的有效量 取决于药物组合物中存在的量、病症或治疗的类型以及其它因素。这些通常以与本文所述相同的剂量和施用路径使用，或以本文所述剂量的约1至99％使用，或以其它剂量使用，并通过经验/临床确定为合适的任何途径使用。
为了预防或治疗疾病，本披露的抗IL23抗体或抗IL23抗体融合蛋白(当单独使用或与一种或更多种其他另外的治疗剂组合使用时)的适当的剂量将取决于待治疗的疾病的类型，治疗分子的类型，疾病的严重性和病程，是为预防还是治疗目的施用，之前的治疗，患者的临床病史和对治疗分子的响应，和主治医师的判断。治疗分子恰当地以一次或经过一系列治疗施用于患者。
制品
在本披露的另一方面中，提供一种制品，所述制品包含可用于治疗、预防和/或诊断上述病症的材料。该制品包含容器和在容器上或与容器联合的标签或包装插页(package insert)。合适的容器包括，例如，瓶子、管形瓶、注射器、IV溶液袋等。容器可以自各种材料诸如玻璃或塑料形成。容器装有单独或与另一种组合物组合有效治疗，预防和/或诊断疾患的组合物，并且可具有无菌的存取口(例如，容器可以是具有由皮下注射针可刺穿的塞子的静脉内溶液袋或管形瓶)。组合物中的至少一种活性试剂是本披露的抗IL23抗体或抗IL23抗体融合蛋白。标签或包装插页指示使用该组合物是来治疗选择的病况。此外，制品可以包含：(a)其中装有组合物的第一容器，其中所述组合物包含本披露的抗IL23抗体或抗IL23抗体融合蛋白；和(b)其中装有组合物的第二容器，其中所述组合物包含另外的细胞毒性剂或其他方面的治疗剂。本披露的该实施方案中的制品可进一步包含包装插页，所述包装插页指示所述组合物可以用于治疗特定病况。备选地，或另外地，制品可进一步包含第二(或第三)容器，所述第二(或第三)容器包含药学上可接受的缓冲液。从商业和用户立场，它可进一步包括所需的其他材料，包括其他缓冲剂、稀释剂、滤器、针头和注射器。
实施例与测试例
以下结合实施例和测试例进一步描述本披露，但这些实施例和测试例并非限制着本披露的范围。本披露实施例和测试例中未注明具体条件的实验方法，通常按照常规条件，如冷泉港的抗体技术实验手册，分子克隆手册；或按照原料或商品制造厂商所建议的条件。未注明具体来源的试剂，为市场购买的常规试剂。
实施例1：小鼠抗人IL-23 p19单克隆抗体的制备
使用人IL-23蛋白(Sino Biological，CT048-H08H)免疫SJL小鼠。免疫3次后取血测定血清中抗体的效价，选择血清中抗体滴度高并且滴度趋于平台的小鼠进行脾细胞融合，将融合好的杂交瘤细胞铺在96孔细胞培养板中，置于37℃，5％CO 2培养箱中进行培养。取细胞培养上清液通过酶联免疫吸附分析法(ELISA)进 行检测。将筛选出的阳性克隆进行扩增冻存保种和二到三次亚克隆直至获得单细胞克隆。选择出的杂交瘤克隆用无血清细胞培养法进一步制备和纯化抗体。得到的杂交瘤抗体用ELISA检测抗体与人IL-23蛋白的结合和对受体的阻断情况(方法见本披露测试例1和测试例2)，挑选出结合活性和阻断活性好的杂交瘤细胞株。
选择单克隆杂交瘤细胞株mAb29和mAb38克隆单克隆抗体的序列。过程如下：收集对数生长期杂交瘤细胞，用Trizol(Invitrogen,Cat#15596-018)提取RNA，反转录为cDNA。用cDNA为模板进行PCR扩增后送测序公司测序，得到的DNA序列对应的抗体的可变区氨基酸序列如下：
mAb29重链可变区的氨基酸序列：
Figure PCTCN2022123370-appb-000008
mAb29轻链可变区的氨基酸序列：
Figure PCTCN2022123370-appb-000009
mAb38重链可变区的氨基酸序列：
Figure PCTCN2022123370-appb-000010
mAb38轻链可变区的氨基酸序列：
Figure PCTCN2022123370-appb-000011
表5.抗体CDR序列表
Figure PCTCN2022123370-appb-000012
Figure PCTCN2022123370-appb-000013
备注：表中CDR是根据Kabat编号系统确认的。
将上述mAb29和mAb38候选分子可变区序列分别通过PCR扩增VH/VK序列，再与表达载体pHr(带信号肽及hIgG1/hkappa恒定区基因(CH1-Fc/CL)片段)进行同源重组，示范性地，人重链IgG1恒定区序列如SEQ ID NO：38所示，人轻链κ恒定区序列如SEQ ID NO：39所示，构建重组嵌合抗体全长表达质粒VH-CH1-Fc-pHr/VL-CL-pHr，进而获得其嵌合抗体Ch29和Ch38。
实施例2：鼠源抗人IL-23单克隆抗体的人源化
通过比对IMGT人类抗体重轻链可变区种系基因数据库，分别挑选同源性高的重轻链可变区种系基因作为模板，将鼠源抗体的CDR分别嫁接(Grafted)到相应的人源模板中，形成次序为FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4的可变区序列，然后将可变区序列与人恒定区序列融合，获得人源化抗体。以下示范性地描述mAb29和mAb38鼠源抗体的人源化，示例中抗体的CDR氨基酸残基由Kabat编号系统确定并注释。
1、鼠源抗体mAb29的人源化
挑选同源性高的重轻链可变区种系基因作为模板。鼠源抗体mAb29的人源化轻链模板选IGKV4-1*01和IGKJ4*01，即选择人种系轻链IGKV4-1*01的FR1，FR2，FR3，和IGKJ4*01的JK4区(作为FR4)作为人源化抗体轻链框架区；人源化重链模板选IGHV2-26*01和IGHJ6*01，即选择人种系重链IGHV2-26*01的FR1，FR2，FR3，和IGHJ6*01的JH6区(作为FR4)作为人源化抗体轻链框架区。首先，将鼠源抗体mAb29的CDR分别移植到选择的相应人源模板中，替换人源模板的CDR区；然后，对人源化抗体的轻链可变区的第4、17、36、58、60和/或68位(根据Kabat编号系统编号)氨基酸残基进行突变，重链可变区的的第1、30、37、44、49、73、89和/或93(根据Kabat编号系统编号)氨基酸残基进行突变，此外，还对重链可变区的HCDR1：SYAIS(SEQ ID NO：5)的第1位氨基酸残基突变为N或Q，获得新的HCDR1：NYAIS(SEQ ID NO：27)或QYAIS(SEQ ID NO：28)，mAb29的人源化抗体可变区序列如下：
>hAb29VH1的氨基酸序列(Graft+Q1E、A49G、A93V)
Figure PCTCN2022123370-appb-000014
>hAb29VH2的氨基酸序列(Graft+Q1E、S30T、A49G、T73N、A93V)
Figure PCTCN2022123370-appb-000015
Figure PCTCN2022123370-appb-000016
>hAb29VH3的氨基酸序列(Graft+Q1E、S30T、I37V、A49G、T73N、T89R、A93V)
Figure PCTCN2022123370-appb-000017
>hAb29VH4的氨基酸序列(Graft+S30T、I37V、A44G、A49G、T73N、T89R、A93V)
Figure PCTCN2022123370-appb-000018
>hAb29VH5的氨基酸序列(Graft+S30T、I37V、A44G、A49G、T73N、T89R、A93V+S31N)：
Figure PCTCN2022123370-appb-000019
>hAb29VH6的氨基酸序列(Graft+S30T、I37V、A44G、A49G、T73N、T89R、A93V+S31Q)：
Figure PCTCN2022123370-appb-000020
>hAb29VL1的氨基酸序列(Graft+Y36F、G68R)
Figure PCTCN2022123370-appb-000021
>hAb29VL2的氨基酸序列(Graft+M4L、Y36F、V58I、G68R)
Figure PCTCN2022123370-appb-000022
>hAb29VL3的氨基酸序列(Graft+M4L、Y36F、V58I、D60A、G68R)
Figure PCTCN2022123370-appb-000023
>hAb29VL4的氨基酸序列(Graft+M4L、E17Q、Y36F、V58I、D60A、G68R)
Figure PCTCN2022123370-appb-000024
注：序列顺序为FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4，序列中下划线部分为CDR区(CDR氨基酸残基由Kabat编号系统确定)，其余部分为FR区。
2、鼠源抗体mAb38的人源化
鼠源抗体mAb38的人源化轻链模板选IGKV2-40*01和IGKJ2*01，即选择人种系轻链IGKV2-40*01的FR1，FR2，FR3，和IGKJ2*01的JK2区(作为FR4)作为人源化抗体轻链框架区；人源化重链模板选IGHV1-3*01和IGHJ1*01，即选择人种系重链IGHV1-3*01的FR1，FR2，FR3，和IGHJ1*01的JH1区(作为FR4)作为人源化抗体重链框架区。首先，将鼠源抗体mAb38的CDR分别移植到选择的相应的人源模板中，替换人源模板的CDR区，然后，对人源化抗体的轻链可变区的第4、36、39、71和/或100位(根据Kabat编号系统编号)氨基酸残基进行突变，重链可变区的的第1、2、28、44、71和/或73(根据Kabat编号系统编号)氨基酸残基进行突变，此外，还对轻链可变区的LCDR2：LMSTRAS(SEQ ID NO：15)的第3位氨基酸残基突变为Q，获得新的LCDR2：LMQTRAS(SEQ ID NO：37)，mAb38的人源化抗体可变区序列如下：
>hAb38VH1的氨基酸序列(Graft+Q1E、V2F、R71V)
Figure PCTCN2022123370-appb-000025
>hAb38VH2的氨基酸序列(Graft+Q1E、T28S、R71V)
Figure PCTCN2022123370-appb-000026
>hAb38VH3的氨基酸序列(Graft+Q1E、V2F、T28S、R71V、T73Q)
Figure PCTCN2022123370-appb-000027
>hAb38VH4的氨基酸序列(Graft+Q1E、V2F、T28S、R44S、R71V、T73Q)
Figure PCTCN2022123370-appb-000028
>hAb38VL1的氨基酸序列(Graft+Q100S)
Figure PCTCN2022123370-appb-000029
>hAb38VL2的氨基酸序列(Graft+Y36F、F71L、Q100S)
Figure PCTCN2022123370-appb-000030
>hAb38VL3的氨基酸序列(Graft+M4I、Y36F、K39R、F71L、Q100S)
Figure PCTCN2022123370-appb-000031
>hAb38VL4的氨基酸序列(Graft+Y36F、F71L、Q100S+S52Q)
Figure PCTCN2022123370-appb-000032
注：序列顺序为FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4，序列中下划线部分为CDR区(CDR氨基酸残基由Kabat编号系统确定)，其余部分为FR区。
3、人源化抗体的制备
分别构建抗体轻链和重链的表达载体，将人源化的抗体轻/重链分别交叉配对组合，转染293E细胞后收集培养上清纯化即得到人源化的全长抗体。人源化抗体重链恒定区可选自IgG1、IgG2、IgG3、IgG4恒定区，示例性的，使用人重链IgG1恒定区，人源化抗体轻链恒定区可选自选自人源κ、λ链恒定区，示例性的抗体的恒定区序列如下：
人IgG1重链恒定区(含L234A，L235A突变)的氨基酸序列
Figure PCTCN2022123370-appb-000033
Figure PCTCN2022123370-appb-000034
人轻链κ恒定区的氨基酸序列：
Figure PCTCN2022123370-appb-000035
示例性的，将前述mAb29的人源化抗体重链可变区与人重链IgG1恒定区(序列如SEQ ID NO：38所示)融合形成抗体全长重链，将人源化抗体轻链可变区与人轻链κ恒定区(序列如SEQ ID NO：39所示)融合形成抗体全长轻链，得到如下表6所示的mAb29人源化抗体：
表6.mAb29人源化抗体
Figure PCTCN2022123370-appb-000036
另外，将前述来自mAb38的人源化抗体重链可变区与人重链IgG1恒定区(序列如SEQ ID NO：38所示)融合形成抗体全长重链，将人源化抗体轻链可变区与人轻链κ恒定区(序列如SEQ ID NO：39所示)融合形成抗体全长轻链，得到如下表7所示的mAb38人源化抗体：
表7.mAb38人源化抗体
Figure PCTCN2022123370-appb-000037
示例性的，人源化抗体轻/重链全长序列如下表8所示：
表8.人源化抗体的轻/重链序列
Figure PCTCN2022123370-appb-000038
Figure PCTCN2022123370-appb-000039
实施例3：抗IL23抗体融合蛋白的构建
将前述抗IL23抗体与TACI多肽融合，构建抗IL23抗体融合蛋白。示例性地，抗IL23抗体为Hu29-19或Hu29-24。TACI多肽可以是任意适宜的TACI多肽，示例性地，TACI多肽为来源于中国专利申请202110348497.6(发明名称：一种新型TACI多肽，其融合蛋白及用途；申请日：2021/3/31)及将其作为优先权文件的专利申请(通过援引完整收入本文)中所述的TACI多肽，例如下表9所示的TACI 多肽：
表9.TACI多肽的氨基酸序列
Figure PCTCN2022123370-appb-000040
Figure PCTCN2022123370-appb-000041
示例性的，将2条TACI多肽(例如TACI-9-15c)分别融合到抗IL23抗体(例如Hu29-19或Hu29-24)的重链的N端和C端，融合抗体重链C端的TACI多肽的C端添加SS作为保护氨基酸，防止TACI多肽被羧肽酶切除，构建获得抗IL23抗体融合蛋白(Hu29-19T和Hu29-24T)的结构示意图见附图1，重链和轻链氨基酸序列如下所示：
>Hu29-19T重链的氨基酸序列(SEQ ID NO：46)
Figure PCTCN2022123370-appb-000042
>Hu29-19T轻链(同Hu29-19轻链，SEQ ID NO：41)
>Hu29-24T重链的氨基酸序列(SEQ ID NO：47)
Figure PCTCN2022123370-appb-000043
>Hu29-24T轻链(同Hu29-24轻链，SEQ ID NO：43)
注：序列中单下划线为TACI序列，双下划线为抗体可变区，斜体为抗体恒定区，粗体为连接子。
本披露中对照分子Risankizumab(参见WHO Drug Information Vol.30,No.1, 2016)的序列如下所示：
>Risankizumab重链的氨基酸序列(SEQ ID NO：48)
Figure PCTCN2022123370-appb-000044
>Risankizumab轻链的氨基酸序列(SEQ ID NO：49)
Figure PCTCN2022123370-appb-000045
对照分子RCT-18(telitacicept，RC18，荣昌生物，WHO(序列编号10932))的序列如下所示。
>RCT-18的氨基酸序列(SEQ ID NO：50)
Figure PCTCN2022123370-appb-000046
注：序列中下划线为TACI片段，其余为Fc。
测试例
测试例1：Elisa结合实验
通过Elisa的方法(包被待测分子)检测待测分子对IL-23 p19和BAFF的结合活性。具体方法如下：
用pH 7.4的PBS(源培生物，B320)缓冲液将待测样品稀释至2μg/mL，以100μL/孔的体积加入96孔酶标板(Corning，3590)中，4℃过夜孵育。弃去液体后，每孔加入300μL用PBS稀释的5％脱脂牛奶(BD，232100)进行封闭，37℃孵育2小时。封闭结束后，弃去封闭液，并用PBST缓冲液(pH 7.4 PBS含0.1％tween-20)洗板3次后，每孔加入100μL梯度稀释的IL-23(Sino Biological，CT048-H08H)或BAFF(ACROBiosystems，BAF-H52D4)溶液，于37℃孵育1小时。孵育结束后用PBST洗板3次。检测与IL-23和BAFF的结合时，每孔加入 100μL Anti-His-HRP(Sino biological，105327-MM02T-H，1:2000稀释)，37℃孵育1小时。用PBST洗板3次后，每孔加入100μL TMB显色底物(KPL，5120-0077)，室温孵育10～15分钟，每孔加入50μL 1M H 2SO 4终止反应，用酶标仪读取在450nm处的吸收值，用软件拟合出抗体与抗原的结合曲线，计算出EC 50值。
抗IL23抗体及其融合蛋白对人IL-23的结合实验结果见下表10：
表10：对IL-23蛋白的结合实验结果
Figure PCTCN2022123370-appb-000047
实验结果显示，本披露的抗IL23抗体及其融合蛋白能与人IL-23蛋白特异性结合，抗IL23抗体及其融合蛋白还能与人BAFF蛋白特异性结合，结合活性比对照RCT-18强。
测试例2：配体和受体的阻断实验
通过Elisa方法检测抗IL23抗体及其融合蛋白对IL-23/IL-23R、BAFF/BAFF-R、BAFF/BCMA、BAFF/TACI、APRIL/BCMA、APRIL/TACI等的受体与配体的阻断活性。具体方法如下：
用pH 7.4的PBS(源培生物，B320)缓冲液将受体蛋白稀释至2μg/mL，以100μL/孔的体积加入96孔酶标板(Corning，3590)中，4℃过夜孵育。弃去液体后，每孔加入200μL 1％Casein封闭液(Thermo，37528)进行封闭，37℃孵育2小时。封闭结束后，弃去封闭液，并用PBST缓冲液(pH7.4 PBS含0.1％tween-20)洗板3次后备用。将固定浓度的生物素(Biotin)标记的配体蛋白与梯度稀释的抗体或融合蛋白混合后37℃预孵育30分钟后加入封闭好的酶标板中，37℃孵育1.5小时。孵育结束后用PBST洗板3次，每孔加入100μL streptavidin-HRP(Invitrogen，434323，1:4000稀释)，37℃孵育1小时。去上清，用PBST洗板3次后每孔加入100μL TMB显色底物(KPL，5120-0077)，室温孵育10～15分钟，每孔加入50μL1M H 2SO 4终止反应，用酶标仪读取在450nm处的吸收值，用软件拟合出抑制配体和受体结合的曲线，计算出IC 50值。本测试例中所用配体蛋白的来源信息如下：IL-23(Sino Biological，CT048-H08H)、BAFF(Sino biological，10056-HNCH)，APRIL(R&D Systems，5860-AP-010/CF)。所用受体蛋白的来源信息如下：IL-23R(Sino Biological，13840-H02H)，BAFF-R(Sino biological，16079-H02H)，BCMA (Sino biological，10620-H02H)，TACI(ACROBiosystems，TAI-H5256)。
抗IL23抗体及其融合蛋白对IL-23/IL-23R的阻断实验结果见下表11：
表11.抗IL23抗体及其融合蛋白对IL-23/IL-23R的阻断实验结果
样品 IL-23/IL-23R阻断活性IC 50(nM)
Ch29 0.3858
Ch38 0.3613
Hu29-19 0.4783
Hu29-24 0.4877
Hu38-4 0.4274
Hu29-19T 0.4968
Hu29-24T 0.3137
Risankizumab 0.6104
抗IL23抗体融合蛋白阻断BAFF蛋白与其受体结合的实验结果见下表12：
表12.抗IL23抗体融合蛋白对BAFF蛋白的阻断实验结果
Figure PCTCN2022123370-appb-000048
抗IL23抗体融合蛋白阻断APRIL蛋白与其受体结合的实验结果见下表13：
表13.抗IL23抗体融合蛋白对APRIL蛋白的阻断实验结果
Figure PCTCN2022123370-appb-000049
其它抗IL23抗体对IL-23/IL-23R的阻断实验结果见下表14：
表14.抗IL23抗体对IL-23/IL-23R的阻断实验结果
样品 IL-23/IL-23R阻断活性IC 50(nM)
Hu29-1 0.4743
Hu29-2 0.3972
Hu29-5 0.3881
Hu29-6 0.4367
Hu29-9 0.3966
Hu29-10 0.4242
Hu29-15 0.4104
Hu38-2 0.465
Hu38-5 0.2279
Hu38-9 0.4963
Hu38-13 0.455
实验结果显示，本披露构建的抗IL23抗体或抗IL23抗体融合蛋白能有效阻断IL-23/IL-23R结合，抗IL23抗体融合蛋白阻断BAFF与其受体或APRIL与其受体的活性显著强于对照RCT-18。
测试例3：BaF3-IL-23R细胞增殖实验
通过BaF3-IL-23R细胞增殖实验检测抗体和融合蛋白的体外活性。实验方法如下：
将稳定表达IL-23R和IL-12Rβ1的BaF3细胞(Cobioer，CBP60474)铺在96孔细胞板(Corning，3903)中，每孔50μL体积，4000个细胞。将梯度稀释的待测样品与固定浓度的IL-23蛋白(Sino Biological，CT048-H08H)混合，加入细胞培养板中，37℃培养箱中培养72小时。然后取出细胞培养板，每孔加入50μL Celltiter Glo检测液(Promega，G755B)，置于摇床中10min，取出置于室温静置10分钟，用酶标仪(PerkinElmer，Victor3)检测生物发光信号。数据用软件拟合出抑制曲线，计算IC 50值。
抗IL23抗体及其融合蛋白抑制BaF3-IL-23R细胞增殖的实验结果见下表15：
表15.抗IL23抗体及其融合蛋白抑制BaF3-IL-23R细胞增殖实验结果
样品名称 抑制BaF3-IL-23R细胞增殖的活性IC 50(nM)
Ch29 0.1753
Ch38 0.2221
Hu29-19 0.1938
Hu29-24 0.1784
Hu29-19T 0.1385
Hu29-24T 0.2316
其它抗IL23抗体抑制BaF3-IL-23R细胞增殖实验结果见下表16：
表16.抗IL23抗体抑制BaF3-IL-23R细胞增殖实验结果
样品 抑制BaF3-IL-23R细胞增殖的活性IC 50(nM)
Hu38-1 0.1675
Hu38-2 0.1384
Hu38-3 0.1736
Hu38-4 0.1219
Hu38-5 0.1477
Hu38-6 0.1358
Hu38-7 0.1794
Hu38-8 0.1743
Hu38-9 0.1976
Hu38-10 0.1161
Hu38-11 0.1683
Hu38-12 0.1838
Hu38-13 0.1866
Hu38-14 0.1522
Hu38-15 0.1493
Hu38-16 0.1458
Hu29-1 0.07111
Hu29-2 0.06227
Hu29-5 0.05531
Hu29-6 0.08897
Hu29-9 0.05608
Hu29-10 0.08601
Hu29-15 0.2158
Hu29-16 0.2251
Hu29-23 0.1184
实验结果显示，本披露的抗IL23抗体及其融合蛋白均具有强的抑制BaF3-IL-23R细胞增殖活性。
测试例4：IL-17分泌实验
通过IL-17分泌实验检测抗IL23抗体及其融合蛋白抑制IL-23诱导的T细胞分化情况。实验方法如下：
在96孔板(Corning，3599)中每孔加入100μL 2μg/mL anti-mouse CD3抗体(BioLegend，100238)和2μg/mL anti-mouse CD28抗体(BioLegend，102116)，37℃孵育1小时，用PBS洗涤2次后备用。取小鼠脾脏进行研磨，4℃离心5分钟收集下层细胞，用洗涤溶液(PBS+2％FBS+2mM EDTA)清洗一次并离心，去上清后加入RBC Lysis Buffer(Invitrogen，00-4333-57)，室温静置5分钟至红细胞完全裂解。再次离心并重悬细胞进行计数。细胞悬液用Mouse CD4 Cells Kit(Invitrogen，11415D)进行分选，将分离的CD4+T细胞用培养基RPMI 1640Medium(Gibco，11875119)+10％FBS(Gibco，10099-141)重悬并计数备用。将细胞悬液铺在包被好的96孔板中，将固定浓度的IL-23(R&D Systems，1290-IL-010)与梯度稀释的抗体或融合蛋白混合预孵育1小时后加入96孔板中，37℃细胞培养箱中培养48小时。取出96孔板，1000rpm离心3分钟，收集上清液，用Mouse IL-17 DuoSet ELISA Kit(R&D Systems，DY421)检测上清中IL-17的含量。实验结果见下表17：
表17.抑制IL-17分泌实验结果
样品名称 抑制IL-17分泌的IC 50(nM)
Hu29-15 0.01954
Hu29-19 0.01775
Hu29-24 0.02890
Hu29-19T 0.01648
Hu29-24T 0.01278
实验结果显示，本披露的抗IL23抗体及其融合蛋白均具有强的抑制IL-17分泌的活性。
测试例5：B细胞增殖实验
通过B细胞增殖实验检测抗IL23抗体融合蛋白是否能抑制BAFF和APRIL诱导的B细胞增殖。实验方法如下：
取小鼠脾脏进行研磨，4℃离心5分钟收集下层细胞，用洗涤溶液(PBS+2％FBS+2mM EDTA)清洗一次并离心，去上清后加入RBC Lysis Buffer(Invitrogen，00-4333-57)，室温静置5分钟至红细胞完全裂解。再次离心并重悬细胞进行计数。细胞悬液用B Cell Isolation Kit(Miltenyi Biotec，130-090-862)进行分选，将分离的B细胞用RPMI 1640 Medium(Gibco，11875119)+10％FBS(Gibco，10099-141)+50μM 2-mercaptoethanol(Sigma-Aldrich，M6250)重悬并计数，细胞铺在96孔细胞板(Costar，3903)中备用。将BAFF(R&D Systems，7537-BF)或APRIL(R&D Systems，5860-AP)蛋白稀释至固定浓度，并加入梯度稀释的抗体或融合蛋白混匀，37℃预孵育30分钟后加入96孔细胞板中，37℃细胞培养箱中培养48小时。取出细胞培养板，每孔加入50μL Celltiter Glo检测液(Promega，G7573)，室温孵育10分钟，用酶标仪检测生物发光信号，将检测结果用软件拟合出抑制曲线，计算出IC50值。实验结果见下表18：
表18.抑制B细胞增殖的实验结果
Figure PCTCN2022123370-appb-000050
实验结果显示，本披露的抗IL23抗体融合蛋白对BAFF和APRIL的抑制活性显著的强于对照RCT-18。
测试例6：亲和力测试
用生物传感芯片Protein A(GE，29127556)亲和捕获一定量的待测样品，然后于芯片表面流经一系列浓度梯度的抗原，利用Biacore(GE，8K)实时检测反应信号从而获得结合和解离曲线。在每个循环解离完成后，用10mM甘氨酸-盐酸溶液pH 1.5(GE，BR-1003-54)将生物芯片洗净再生。实验数据用BIAevaluation version 4.1软件以1:1模型进行拟合，从而得出亲和力数值。本测试中用到的相关抗原蛋白如下：人IL-23(Sino biological，CT048-H08H)，人BAFF(Sino biological，10056-HNCH)，人APRIL(R&D Systems，5860-AP-010/CF)，食蟹猴IL-23(Acro  Biosystems，ILB-CM52W8)，食蟹猴BAFF(Kactus，BAF-CM412)，食蟹猴APRIL(Kactus，APR-CM410B)，鼠IL-23(R&D Systems，1887-ML/CF)，鼠BAFF(Acro Biosystems，BAF-M521y)，鼠APRIL(R&D Systems，7907-AP/CF)。亲和力的测试结果见下表19至表25：
表19.抗IL23抗体及其融合蛋白结合人IL-23 p19的亲和力实验结果
Figure PCTCN2022123370-appb-000051
表20.抗IL23抗体及其融合蛋白结合食蟹猴或鼠IL-23 p19的亲和力实验结果
Figure PCTCN2022123370-appb-000052
表21.抗IL23抗体融合蛋白结合人BAFF的亲和力实验结果
Figure PCTCN2022123370-appb-000053
表22.抗IL23抗体融合蛋白结合食蟹猴或鼠BAFF的亲和力实验结果
Figure PCTCN2022123370-appb-000054
表23.抗IL23抗体融合蛋白结合人APRIL的亲和力实验结果
Figure PCTCN2022123370-appb-000055
表24.抗IL23抗体融合蛋白结合食蟹猴或鼠APRIL的亲和力实验结果
Figure PCTCN2022123370-appb-000056
表25.抗IL23抗体结合人IL-23 p19的亲和力实验结果
Figure PCTCN2022123370-appb-000057
实验结果表明，本披露构建的抗IL23抗体及其融合蛋白能以高亲和力与人 IL-23 p19和食蟹猴IL-23 p19结合，但不与鼠IL-23 p19结合；另外，本披露构建抗IL23抗体融合蛋白能以高亲和力与人BAFF、食蟹猴BAFF、鼠BAFF、人APRIL、食蟹猴APRIL和鼠APRIL结合。
测试例7：咪喹莫特诱导的银屑病动物模型实验
通过咪喹莫特乳膏(IMQ)诱导的银屑病动物模型实验来评估抗IL23抗体的体内药效。
SPF级雌性C57BL/6hIL-23A/hIL12B转基因小鼠(百奥赛图江苏基因生物技术有限公司)，8～9周龄，随机分组，每组5只小鼠。实验开始前对小鼠的背部进行脱毛处理。从第0天到第5天，假手术组在小鼠的后背皮肤均匀涂抹80mg凡士林(联合利华)；动物模型组在小鼠的后背皮肤均匀涂抹80mg咪喹莫特乳膏(贝美森)，连续涂抹6天。分别在第0天和第3天，在涂抹咪喹莫特乳膏前1小时腹腔注射待测样品(Risankizumab、Hu29-19、Hu29-24和Hu38-4的给药剂量都为20mpk)。Risankizumab作为阳性对照，PBS作为阴性对照，一共给药2次。每天对小鼠背部皮肤的红疹和脱屑严重程度进行评分，在第6天结束实验。
红疹严重程度的评分标准如下：无红疹，0分；微红，1分；红但不暗，2分；深红，3分；极红，4分。脱屑的评分标准如下：无脱屑，0分；小面积细微皮屑，1分；适中面积较厚的皮屑，2分；较大面积的粗糙增厚皮屑，3分；大面积的大块皮屑，4分。
实验结果见附图2至图4，实验结果表明，相比阳性对照Risankizumab，本披露构建的抗IL23抗体的体内药效更强，红疹严重程度和皮肤脱屑严重程度的评分均低于Risankizumab。
测试例8：人IL-23诱导的银屑病动物模型实验
通过人IL-23诱导的银屑病动物模型评价本披露构建的抗IL23抗体的体内药效。SPF级雌性C57BL/6J小鼠(维通利华实验动物技术有限公司)，6～8周龄，将小鼠麻醉后测量小鼠右耳厚度和体重，根据右耳厚度和体重数据进行分组，每组6只小鼠。分组当天记为第0天。从第1天开始，每天给小鼠右耳皮内注射1μg的人IL-23蛋白(R&D Systems，1290-IL-500/CF)，共注射7天，正常组小鼠注射PBS。在第0天和第3天腹腔注射受试药物，分别在第0，2，4，6，8天测量和记录小鼠体重和右耳厚度，通过右耳厚度的变化来评价受试药物对该模型的缓解作用。第8天结束实验，收集直径为8mm的耳片并称重。
实验结果见附图5、6、7，实验结果表明，本披露构建的抗IL23抗体Hu29-19在人IL-23诱导的银屑病动物模型中的体内药效强于对照分子Risankizumab，其中：在剂量都为3mpk的情况下，Hu29-19组的耳朵厚度低于对照Risankizumab；曲线下面积的统计结果表明，在3mpk的剂量下，Hu29-19的作用效果要强于 Risankizumab；小鼠耳朵重量的统计结果表明：Hu29-19 1mpk组的耳朵重量与Risankizumab 3mpk组相同，Hu29-19 3mpk组的耳朵重量要低于Risankizumab。
测试例9：体内抑制细胞因子分泌活性评价
用人IL-23和人BAFF两种蛋白同时刺激小鼠，诱导小鼠体内产生TNFα、IL-22和IgA等细胞因子，通过检测这些细胞因子的水平来评价抗体或融合蛋白的体内活性。
SPF级雌性C57BL/6小鼠(浙江维通利华实验动物技术有限公司)，8周龄，将小鼠进行随机分组，每组5只，通过腹腔混合注射人IL-23(Sino biological，CT048-H08H)(2μg/小鼠)和人BAFF(Sino biological，10056-HNCH)(1mg/kg)两种蛋白，每天注射一次，持续四天。分别在第一天和第三天注射蛋白前一小时腹腔注射待测样品(RCT-18(4mpk)，Risankizumab(8mpk)，Hu29-19T(8.8mpk)，Hu29-24T(4.4mpk))。第五天收集各组小鼠血浆样本，分别检测TNFα、IL-22和IgA的水平。本测试例中所用的检测试剂盒来源信息如下：Mouse TNF-alpha Quantikine ELISA Kit(R&D Systems，MTA00B)，Mouse/Rat IL-22 Quantikine ELISA Kit(R&D Systems，M2200)，Mouse IgA ELISA Kit(Abcam，ab157717)。RCT-18和Risankizumab作为阳性对照，PBS作为阴性对照。4mpk RCT-18，8mpk Risankizumab，8.8mpk Hu29-19T的药物摩尔浓度相同。
实验结果见表26、27、28和附图8、9、10，其中：IgA的检测结果表明，本披露构建的抗IL23抗体融合蛋白抑制IgA的分泌活性显著强于对照分子RCT-18，而Risankizumab对IgA的分泌没有抑制活性；TNFα的检测结果表明，在等摩尔浓度的剂量下，本披露构建的抗IL23抗体融合蛋白抑制TNFα分泌的活性强于Risankizumab，而RCT-18没有表现出显著的抑制效果；IL-22的检测结果表明，在等摩尔浓度的剂量下，本披露构建的抗IL23抗体融合蛋白抑制IL-22分泌的活性强于对照Risankizumab和RCT-18。
表26.小鼠血浆IgA检测结果(μg/mL)
Figure PCTCN2022123370-appb-000058
表27.小鼠血浆TNFα检测结果(pg/mL)
Figure PCTCN2022123370-appb-000059
表28.小鼠血浆IL-22检测结果(pg/mL)
Figure PCTCN2022123370-appb-000060
测试例10：系统性红斑狼疮动物模型中的体内药效
MRL/lpr小鼠随着周龄的增长，会自发的产生自身免疫疾病的症状，其症状与人类系统性红斑狼疮相似。通过该模型来评价融合蛋白的体内药效。
MRL/lpr小鼠，雌性，购自上海斯莱克实验动物有限责任公司，5只/笼饲养于SPF级环境，温度20-25℃；湿度40-60％。实验前小鼠在实验室环境至少适应一周。待小鼠周龄达到9～10周，尿蛋白和dsDNA IgG值显著高于正常对照小鼠时开始皮下注射给药(RCT-18(8mpk)，Hu29-19T(17.6mpk)，Hu29-19T(8.8mpk)，Hu29-24T(17.6mpk)，阴性对照(PBS))，其中RCT-18 8mpk，Hu29-19T 17.6mpk，Hu29-24T 17.6mpk的摩尔浓度相同。每周给药2次(周一和周四分别给药1次)，持续给药8周。定期观察小鼠毛发和皮肤状态，对小鼠的皮肤损伤严重程度进行评分，收集尿液和血浆，检测尿蛋白含量。实验结束后取小鼠的肾脏，固定后进行H&E染色，观察各组小鼠肾脏组织肾小球、肾小管病变、炎性细胞浸润，并对肾脏病变程度进行评分。本测试例中尿蛋白检测采用Bradford蛋白浓度测定，检测试剂盒购自碧云天生物技术有限公司。
实验结果见表29、30、31和附图11、12、13所示，实验结果表明，RCT-18没有降低小鼠的尿蛋白水平，Hu29-19T在两个剂量下都能显著的降低小鼠的尿蛋白水平，并且整个实验过程中Hu29-19T组小鼠的尿蛋白水平一直保持和实验开始时相同或更低，完全没有升高的趋势。Hu29-24T也能显著降低小鼠的尿蛋白。小鼠皮肤损伤的评分结果表明，Hu29-19T组小鼠的皮肤损伤评分一直都是最低的，Hu29-24T处于中间水平，RCT-18在这个指标上没有显示出明显的药效。小鼠的肾脏病理学评分结果表明，跟阴性组相比，Hu29-19T两个剂量组显示出了显著的统计学差异。
表29.小鼠肾脏病理评分
Figure PCTCN2022123370-appb-000061
表30.小鼠皮肤损伤评分平均值
Figure PCTCN2022123370-appb-000062
表31.小鼠尿蛋白检测结果(mg/dL)
Figure PCTCN2022123370-appb-000063
测试例11.大鼠体内药代动力学实验
用SD大鼠进行体内药代动力学测试。雄性SD大鼠(浙江维通利华实验动物技术有限公司)随机分组，每组4只，静脉注射给药，给药组于给药前及给药后5分钟、8小时、24小时、48小时、84小时、9天、10天、14天、21天、28天采集全血0.2mL，不加抗凝，取血后在4℃放置30分钟，1000g离心15分钟，取上层血清置于EP管中，于-80℃保存。用ELISA法检测血清中的血药浓度，用Winnolin软件计算受试药物的药代动力学参数和体内半衰期。
大鼠体内药代动力学实验结果见下表32，实验结果表明，本披露的抗体和融合蛋白具备较好的稳定性，体内半衰期超过8天。
表32.大鼠体内药代动力学实验结果
Figure PCTCN2022123370-appb-000064
虽然为了清楚的理解，已经借助于附图和实例详细描述了上述发明，但是描述和实例不应当解释为限制本披露的范围。本文中引用的所有专利和科学文献的公开内容通过引用完整地清楚结合。

Claims (19)

  1. 一种抗IL23抗体融合蛋白，其包含抗IL23抗体和TACI多肽，其中所述抗IL23抗体特异性结合人IL23 p19亚基；
    优选地，其中所述抗IL23抗体包含重链可变区和轻链可变区，其中，所述重链可变区包含HCDR1、HCDR2和HCDR3，所述轻链可变区包含LCDR1、LCDR2和LCDR3，其中，
    i)所述重链可变区的HCDR1、HCDR2和HCDR3分别包含SEQ ID NO：21、22、20、19、18、17或1中的HCDR1、HCDR2和HCDR3的氨基酸序列，和所述轻链可变区的LCDR1、LCDR2和LCDR3分别包含SEQ ID NO：25、26、24、23或2中的LCDR1、LCDR2和LCDR3的氨基酸序列；或
    ii)所述重链可变区的HCDR1、HCDR2和HCDR3分别包含SEQ ID NO：29、30、31、32或3中的HCDR1、HCDR2和HCDR3的氨基酸序列，和所述轻链可变区的LCDR1、LCDR2和LCDR3分别包含SEQ ID NO：36、35、34、33或4中的LCDR1、LCDR2和LCDR3的氨基酸序列。
  2. 根据权利要求1所述的抗IL23抗体融合蛋白，其中：
    i)所述重链可变区的HCDR1包含SEQ ID NO：27、28或5的氨基酸序列，HCDR2包含SEQ ID NO：6的氨基酸序列，和HCDR3包含SEQ ID NO：7的氨基酸序列，和所述轻链可变区的LCDR1包含SEQ ID NO：8的氨基酸序列，LCDR2包含SEQ ID NO：9的氨基酸序列，和LCDR3包含SEQ ID NO：10的氨基酸序列；或
    ii)所述重链可变区的HCDR1包含SEQ ID NO：11的氨基酸序列，HCDR2包含SEQ ID NO：12的氨基酸序列，和HCDR3包含SEQ ID NO：13的氨基酸序列，和所述轻链可变区的LCDR1包含SEQ ID NO：14的氨基酸序列，LCDR2包含SEQ ID NO：37或15的氨基酸序列，和LCDR3包含SEQ ID NO：16的氨基酸序列；
    优选地，
    i)所述重链可变区的HCDR1包含SEQ ID NO：27的氨基酸序列，HCDR2包含SEQ ID NO：6的氨基酸序列，和HCDR3包含SEQ ID NO：7的氨基酸序列，和所述轻链可变区的LCDR1包含SEQ ID NO：8的氨基酸序列，LCDR2包含SEQ ID NO：9的氨基酸序列，和LCDR3包含SEQ ID NO：10的氨基酸序列；或
    ii)所述重链可变区的HCDR1包含SEQ ID NO：28的氨基酸序列，HCDR2包含SEQ ID NO：6的氨基酸序列，和HCDR3包含SEQ ID NO：7的氨基酸序列，和所述轻链可变区的LCDR1包含SEQ ID NO：8的氨基酸序列，LCDR2包含SEQ ID NO：9的氨基酸序列，和LCDR3包含SEQ ID NO：10的氨基酸序列；或
    iii)所述重链可变区的HCDR1包含SEQ ID NO：11的氨基酸序列，HCDR2包含SEQ ID NO：12的氨基酸序列，和HCDR3包含SEQ ID NO：13的氨基酸序列，和所述轻链可变区的LCDR1包含SEQ ID NO：14的氨基酸序列，LCDR2包含SEQ ID NO：37的氨基酸序列，和LCDR3包含SEQ ID NO：16的氨基酸序列。
  3. 根据权利要求1或2所述的抗IL23抗体融合蛋白，其中：
    i)所述重链可变区包含SEQ ID NO：21、22、20、19、18或17中的任一或与其具有至少90％序列同一性的氨基酸序列，和所述轻链可变区包含SEQ ID NO：25、26、24或23中的任一或与其具有至少90％序列同一性的氨基酸序列；或
    ii)所述重链可变区包含SEQ ID NO：29、30、31或32中的任一或与其具有至少90％序列同一性的氨基酸序列，和所述轻链可变区包含SEQ ID NO：36、35、34或33中的任一或与其具有至少90％序列同一性的氨基酸序列；或
    iii)所述重链可变区包含SEQ ID NO：1或与其具有至少90％序列同一性的氨基酸序列，和所述轻链可变区包含SEQ ID NO：2或与其具有至少90％序列同一性的氨基酸序列；或
    iv)所述重链可变区包含SEQ ID NO：3或与其具有至少90％序列同一性的氨基酸序列，和所述轻链可变区包含SEQ ID NO：4或与其具有至少90％序列同一性的氨基酸序列；
    优选地，i)所述重链可变区包含SEQ ID NO：21的氨基酸序列，和所述轻链可变区包含SEQ ID NO：25的氨基酸序列；或
    ii)所述重链可变区包含SEQ ID NO：22的氨基酸序列，和所述轻链可变区包含SEQ ID NO：26的氨基酸序列；或
    iii)所述重链可变区包含SEQ ID NO：29的氨基酸序列，和所述轻链可变区包含SEQ ID NO：36的氨基酸序列。
  4. 根据权利要求1至3中任一项所述的抗IL23抗体融合蛋白，其中所述的抗IL23抗体进一步包含抗体重链恒定区和轻链恒定区；
    优选地，所述重链恒定区选自人IgG1、IgG2、IgG3和IgG4恒定区，所述轻链恒定区选自人抗体κ或λ链恒定区；
    更优选地，所述重链恒定区包含SEQ ID NO：38的氨基酸序列，所述轻链恒定区包含SEQ ID NO：39的氨基酸序列。
  5. 根据权利要求1至4中任一项所述的抗IL23抗体融合蛋白，所述的抗IL23抗体包含重链和轻链，其中
    i)所述抗IL23抗体的重链包含SEQ ID NO：40或与其具有至少90％序列同一性的氨基酸序列，和所述抗IL23抗体的轻链包含SEQ ID NO：41或与其具有至 少90％序列同一性的氨基酸序列；或
    ii)所述抗IL23抗体的重链包含SEQ ID NO：42或与其具有至少90％序列同一性的氨基酸序列，和所述抗IL23抗体的轻链包含SEQ ID NO：43或与其具有至少90％序列同一性的氨基酸序列；或
    iii)所述抗IL23抗体的重链包含SEQ ID NO：44或与其具有至少90％序列同一性的氨基酸序列，和所述抗IL23抗体的轻链包含SEQ ID NO：45或与其具有至少90％序列同一性的氨基酸序列；
    优选地，
    i)所述抗IL23抗体的重链包含SEQ ID NO：40的氨基酸序列，和所述抗IL23抗体的轻链包含SEQ ID NO：41的氨基酸序列；或
    ii)所述抗IL23抗体的重链包含SEQ ID NO：42的氨基酸序列，和所述抗IL23抗体的轻链包含SEQ ID NO：43的氨基酸序列；或
    iii)所述抗IL23抗体的重链包含SEQ ID NO：44的氨基酸序列，和所述抗IL23抗体的轻链包含SEQ ID NO：45的氨基酸序列。
  6. 根据权利要求1至5中任一项所述的抗IL23抗体融合蛋白，其中所述TACI多肽为包含SEQ ID NO：58的第48位至第85位氨基酸残基的多肽或其变体；其中，所述变体在选自第49、52、53、57、65、82和83位中的一个或更多个位点上具有氨基酸替换，所述氨基酸替换的位点为相对于序列SEQ ID NO：58的自然顺序编号的氨基酸残基位点；
    优选地，所述变体具有选自由49T或49R、52S、53E或53Q、57E、65T或65A、82A或82R、和83Y组成的组中的一个或更多个氨基酸替换，所述氨基酸替换的位点为相对于序列SEQ ID NO：58的自然顺序编号的氨基酸残基位点。
  7. 根据权利要求1至6中任一项所述的抗IL23抗体融合蛋白，其中所述TACI多肽的氨基酸序列如SEQ ID NO：51至83中任一所示；
    优选地，所述TACI多肽的氨基酸序列如SEQ ID NO：83所示。
  8. 根据权利要求1至7中任一项所述的抗IL23抗体融合蛋白，其包括：
    第一链：[TACI多肽1]-[连接子1]-[抗IL23抗体的重链]-[连接子2]-[TACI多肽2]，和
    第二链：抗IL23抗体的轻链，
    其中，所述TACI多肽1和TACI多肽2是相同或不相同的，其独立地选自权利要求6和7中所定义的任一TACI多肽；所述连接子1和连接子2是相同或不相同的；
    优选地，所述连接子为选自(G xS) y连接子，其中，x选自1-5的整数，y选 自0-6的整数，更优选地，所述连接子为如SEQ ID NO：84或85所示连接子。
  9. 根据权利要求1至8中任一项所述的抗IL23抗体融合蛋白，其中，
    所述的抗IL23抗体融合蛋白具有：包含SEQ ID NO：46的氨基酸序列的第一链，和包含SEQ ID NO：41的氨基酸序列的第二链；或
    所述的抗IL23抗体融合蛋白具有：包含SEQ ID NO：47的氨基酸序列的第一链，和包含SEQ ID NO：43的氨基酸序列的第二链。
  10. 一种抗IL23抗体，其包含重链可变区和轻链可变区，所述重链可变区包含HCDR1、HCDR2和HCDR3，所述轻链可变区包含LCDR1、LCDR2和LCDR3，其中，
    i)所述重链可变区的HCDR1、HCDR2和HCDR3分别包含SEQ ID NO：21、22、20、19、18、17或1中的HCDR1、HCDR2和HCDR3的氨基酸序列，和所述轻链可变区的LCDR1、LCDR2和LCDR3分别包含SEQ ID NO：25、26、24、23或2中的LCDR1、LCDR2和LCDR3的氨基酸序列；或
    ii)所述重链可变区的HCDR1、HCDR2和HCDR3分别包含SEQ ID NO：29、30、31、32或3中的HCDR1、HCDR2和HCDR3的氨基酸序列，和所述轻链可变区的LCDR1、LCDR2和LCDR3分别包含SEQ ID NO：36、35、34、33或4中的LCDR1、LCDR2和LCDR3的氨基酸序列。
  11. 根据权利要求10所述的抗IL23抗体，其中：
    i)所述重链可变区的HCDR1包含SEQ ID NO：27、28或5的氨基酸序列，HCDR2包含SEQ ID NO：6的氨基酸序列，和HCDR3包含SEQ ID NO：7的氨基酸序列，和所述轻链可变区的LCDR1包含SEQ ID NO：8的氨基酸序列，LCDR2包含SEQ ID NO：9的氨基酸序列，和LCDR3包含SEQ ID NO：10的氨基酸序列；或
    ii)所述重链可变区的HCDR1包含SEQ ID NO：11的氨基酸序列，HCDR2包含SEQ ID NO：12的氨基酸序列，和HCDR3包含SEQ ID NO：13的氨基酸序列，和所述轻链可变区的LCDR1包含SEQ ID NO：14的氨基酸序列，LCDR2包含SEQ ID NO：37或15的氨基酸序列，和LCDR3包含SEQ ID NO：16的氨基酸序列；
    优选地，
    i)所述重链可变区的HCDR1包含SEQ ID NO：27的氨基酸序列，HCDR2包含SEQ ID NO：6的氨基酸序列，和HCDR3包含SEQ ID NO：7的氨基酸序列，和所述轻链可变区的LCDR1包含SEQ ID NO：8的氨基酸序列，LCDR2包含SEQ ID NO：9的氨基酸序列，和LCDR3包含SEQ ID NO：10的氨基酸序列；或
    ii)所述重链可变区的HCDR1包含SEQ ID NO：28的氨基酸序列，HCDR2包含SEQ ID NO：6的氨基酸序列，和HCDR3包含SEQ ID NO：7的氨基酸序列，和所述轻链可变区的LCDR1包含SEQ ID NO：8的氨基酸序列，LCDR2包含SEQ ID NO：9的氨基酸序列，和LCDR3包含SEQ ID NO：10的氨基酸序列；或
    iii)所述重链可变区的HCDR1包含SEQ ID NO：11的氨基酸序列，HCDR2包含SEQ ID NO：12的氨基酸序列，和HCDR3包含SEQ ID NO：13的氨基酸序列，和所述轻链可变区的LCDR1包含SEQ ID NO：14的氨基酸序列，LCDR2包含SEQ ID NO：37的氨基酸序列，和LCDR3包含SEQ ID NO：16的氨基酸序列。
  12. 根据权利要求10或11所述的抗IL23抗体，其中：
    i)所述重链可变区包含SEQ ID NO：21、22、20、19、18或17中的任一或与其具有至少90％序列同一性的氨基酸序列，和所述轻链可变区包含SEQ ID NO：25、26、24或23中的任一或与其具有至少90％序列同一性的氨基酸序列；或
    ii)所述重链可变区包含SEQ ID NO：29、30、31或32中的任一或与其具有至少90％序列同一性的氨基酸序列，和所述轻链可变区包含SEQ ID NO：36、35、34或33中的任一或与其具有至少90％序列同一性的氨基酸序列；或
    iii)所述重链可变区包含SEQ ID NO：1或与其具有至少90％序列同一性的氨基酸序列，和所述轻链可变区包含SEQ ID NO：2或与其具有至少90％序列同一性的氨基酸序列；或
    iv)所述重链可变区包含SEQ ID NO：3或与其具有至少90％序列同一性的氨基酸序列，和所述轻链可变区包含SEQ ID NO：4或与其具有至少90％序列同一性的氨基酸序列；
    优选地，i)所述重链可变区包含SEQ ID NO：21的氨基酸序列，和所述轻链可变区包含SEQ ID NO：25的氨基酸序列；或
    ii)所述重链可变区包含SEQ ID NO：22的氨基酸序列，和所述轻链可变区包含SEQ ID NO：26的氨基酸序列；或
    iii)所述重链可变区包含SEQ ID NO：29的氨基酸序列，和所述轻链可变区包含SEQ ID NO：36的氨基酸序列。
  13. 根据权利要求10至12中任一项所述的抗IL23抗体，其中所述的抗IL23抗体进一步包含抗体重链恒定区和轻链恒定区；
    优选地，所述重链恒定区选自人IgG1、IgG2、IgG3和IgG4恒定区，所述轻链恒定区选自人抗体κ或λ链恒定区；
    更优选地，所述重链恒定区包含SEQ ID NO：38的氨基酸序列，所述轻链恒定区包含SEQ ID NO：39的氨基酸序列。
  14. 根据权利要求10至13中任一项所述的抗IL23抗体，所述的抗IL23抗体包含重链和轻链，其中，
    i)所述抗IL23抗体的重链包含SEQ ID NO：40或与其具有至少90％序列同一性的氨基酸序列，和所述抗IL23抗体的轻链包含SEQ ID NO：41或与其具有至少90％序列同一性的氨基酸序列；或
    ii)所述抗IL23抗体的重链包含SEQ ID NO：42或与其具有至少90％序列同一性的氨基酸序列，和所述抗IL23抗体的轻链包含SEQ ID NO：43或与其具有至少90％序列同一性的氨基酸序列；或
    iii)所述抗IL23抗体的重链包含SEQ ID NO：44或与其具有至少90％序列同一性的氨基酸序列，和所述抗IL23抗体的轻链包含SEQ ID NO：45或与其具有至少90％序列同一性的氨基酸序列；
    优选地，
    i)所述抗IL23抗体的重链包含SEQ ID NO：40的氨基酸序列，和所述抗IL23抗体的轻链包含SEQ ID NO：41的氨基酸序列；或
    ii)所述抗IL23抗体的重链包含SEQ ID NO：42的氨基酸序列，和所述抗IL23抗体的轻链包含SEQ ID NO：43的氨基酸序列；或
    iii)所述抗IL23抗体的重链包含SEQ ID NO：44的氨基酸序列，和所述抗IL23抗体的轻链包含SEQ ID NO：45的氨基酸序列。
  15. 根据权利要求1至9中任一项所述的抗IL23抗体融合蛋白或权利要求10至14中任一项所述的抗IL23抗体，其具有一种或更多种以下特性：
    A.与人IL-23 p19和食蟹猴IL-23 p19特异性结合，不与鼠IL-23 p19特异性结合；优选地，以小于6.00E-11M的KD值与人IL-23 p19结合，和/或以小于9.00E-11M的KD值与食蟹猴IL-23 p19结合，所述KD值通过
    Figure PCTCN2022123370-appb-100001
    表面等离子体共振测定法所测量；
    B.具有阻断IL-23/IL-23R结合活性；优选地，阻断人IL-23/IL-23R结合的IC 50值小于0.60nM，所述IC 50值通过Elisa方法检测；
    C.具有阻断BAFF/BAFF-R结合活性；优选地，阻断BAFF/BAFF-R结合的IC 50值小于7.00nM，所述IC 50值通过Elisa方法检测；
    D.具有阻断BAFF/BCMA结合活性；优选地，阻断BAFF/BCMA结合的IC 50值小于4.50nM，所述IC 50值通过Elisa方法检测；
    E.具有阻断BAFF/TACI结合活性；优选地，阻断BAFF/TACI结合的IC 50值小于6.00nM，所述IC 50值通过Elisa方法检测；
    F.具有阻断APRIL/BCMA结合活性；优选地，阻断APRIL/BCMA结合的IC 50值小于1.00nM，所述IC 50值通过Elisa方法检测；
    G.具有阻断APRIL/TACI结合活性；优选地，阻断APRIL/TACI结合的IC 50 值小于3.00nM，所述IC 50值通过Elisa方法检测；
    H.具有抑制IL-17分泌活性；优选地，以小于0.03nM的IC 50值抑制IL-17分泌；所述IC 50值通过Elisa检测；
    I.具有抑制BaF3-IL-23R细胞增殖活性；优选地，以小于0.5nM的IC 50值抑制BaF3-IL-23R细胞增殖；所述IC 50值通过PerkinElmer检测；
    J.具有抑制TNFα、IL-22和IgA细胞因子分泌活性；或
    K.具有抑制B细胞增殖活性。
  16. 一种药物组合物，其包含：权利要求1至9和15中任一项所述的抗IL23抗体融合蛋白或权利要求10至15中任一项所述的抗IL23抗体，以及
    一种或多种药学上可接受的载体、稀释剂或赋形剂。
  17. 一种核酸分子，其编码权利要求1至9和15中任一项所述的抗IL23抗体融合蛋白或权利要求10至15中任一项所述的抗IL23抗体。
  18. 一种宿主细胞，其含有权利要求17所述的核酸分子。
  19. 一种治疗或改善B细胞障碍或自身免疫性疾病的方法，所述方法包括向有需要的受试者施用如权利要求1至9和15中任一项所述的抗IL23抗体融合蛋白或权利要求10至15中任一项所述的抗IL23抗体或权利要求16所述药物组合物的步骤；
    优选地，所述B细胞障碍或自身免疫性疾病是与IL23表达相关的疾病或病症；
    更优选地，所述自身免疫性疾病选自：系统性红斑狼疮、重症肌无力、多发性硬化、胰岛素依赖性糖尿病、克罗恩氏病、类风湿关节炎、多关节型青少年类风湿关节炎和银屑病性关节炎；所述B细胞障碍选自：肿瘤、慢性白细胞性白血病、多发性骨髓瘤、非霍奇金淋巴瘤、移植后淋巴组织增生病和轻链丙球蛋白病；
    最优选地，所述自身免疫性疾病为系统性红斑狼疮。
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