TW202330598A - Antigen-binding molecules that specifically bind to baff and il-12/23 and uses thereof - Google Patents

Abstract

The present disclosure relates to antigen binding molecules that specifically bind to BAFF and IL-12/23 and uses thereof.

Description

Antigen-binding molecule specifically binding to BAFF and IL-12/23 and use thereof

This application claims the priority of the Chinese patent application (application number 202110735262.2) filed on June 30, 2021.

The disclosure belongs to the field of biotechnology, and more specifically, the disclosure relates to an antigen-binding molecule that specifically binds BAFF and IL-12/23 and its application.

The statements herein merely provide background information related to the present disclosure and do not necessarily constitute prior art.

BAFF (B cell activating factor) is a 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 trimers. The BAFF on the surface of the cell membrane will be hydrolyzed by protease to form soluble BAFF and enter the blood circulation system. The soluble BAFF has the characteristics of multimerization and can form up to 60-mer . In addition, BAFF can also interact with another protein of the same family, APRIL, to form a heterologous trimer. It is currently known that 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 There are two common receptors, 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. Khamashta, Autoimmun Rev, 2014.13: p.1094-1101). 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 that promote autoimmune responses (Cancro, M.P., D.P.D'Cruz, and M.A. Khamashta, J Clin Invest, 2009.119: p.1066-73).

IL-12 and IL-23 are two cytokines belonging to the same family. Both IL-12 and IL-23 are heterodimeric proteins composed of two subunits (Moschen, A.R., H. Tilg, and T. Raine, Nat Rev Gastroenterol Hepatol, 2019.16: p.185-196; Frieder , J., et al., Clin Pharmacol Ther, 2018.103: p.88-101). IL-12 is composed of two subunits, p35 and p40, while IL-23 is composed of two subunits, p19 and p40. Among them, p40 is a subunit shared by IL-12 and IL-23 (IL-12/23 P40), so antibodies targeting p40 can simultaneously inhibit both IL-12 and IL-23 signaling pathways. The interaction between IL-12 and receptors mainly activates the differentiation of Th1 cells, and at the same time participates in stimulating the secretion of interferon-γ and TNF by various immune cells. The interaction between IL-23 and receptors mainly activates the differentiation of Th17 cells, and stimulates a variety of cells to secrete cytokines such as IL-17, IL-22 and TNF (Lee, E.B., et al., Cutis, 2018.101: p.5-9; Floss, D.M., et al., Cytokine Growth Factor Rev, 2015.26: p.569-578). These cytokines produced by the activation of IL-12/23 signaling pathway further participate in the development of diseases such as systemic lupus erythematosus through their own pathways.

The present disclosure constructs an antigen-binding molecule that specifically binds BAFF, and constructs an antigen-binding molecule that specifically binds BAFF and IL-12 and/or IL-23.

In one aspect, the present disclosure provides an antigen-binding molecule comprising an antigen-binding module 1 that specifically binds BAFF and an antigen-binding module 2 that specifically binds IL-12 and/or IL-23, wherein the antigen-binding module 1 comprises a heavy Chain variable region B-VH and light chain variable region B-VL, the B-VH comprising B-HCDR1, B-HCDR2 and B-HCDR3, the B-VL comprising B-LCDR1, B-LCDR2 and B-LCDR3 , wherein, the B-HCDR1, B-HCDR2 and B-HCDR3 respectively comprise the amino acid sequences of Bv-HCDR1, Bv-HCDR2 and Bv-HCDR3 in SEQ ID NO: 63, and the B-LCDR1, B-LCDR2 and B-LCDR3 respectively comprise the amino acid sequences of Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 in SEQ ID NO: 64, wherein, SEQ ID NO: 63 is:

SEQ ID NO: 64 is:

Wherein, _X1 is selected from S, A, V or G, _X2 is selected from A, I, M or S, _X3 is selected from T or G, _X4 is selected from Q, E, G or K, _X5 is selected from N, G or Q, X ₆ is selected from L or P, X ₇ is selected from H, Q or D, X ₈ is selected from H or D, X ₉ is selected from A or G, X ₁₀ is selected from S or L, X ₁₁ selected from P or S, X ₁₂ selected from Q or H, X ₁₃ selected from D, A or N, X ₁₄ selected from S or I, X ₁₅ selected from K, R or T, X ₁₆ selected from S, W, T or D, X ₁₇ is selected from H, Y or S, X ₁₈ is selected from Y or R, X ₁₉ is selected from G or S, X ₂₀ is selected from A or D, X ₂₁ is selected from E or S, X ₂₂ is selected from S or A, X ₂₃ is selected from V, E, A, N or W, X ₂₄ is selected from G, K, H or R, X ₂₅ is selected from L or V, X ₂₆ is selected from G or C, X ₂₇ is selected from R or G, X ₂₈ is selected from G or C;

Moreover, the B-HCDR1, B-HCDR2, B-HCDR3, B-LCDR1, B-LCDR2 and B-LCDR3 do not contain the following CDR combination: B-HCDR1, B-HCDR2 and B-HCDR3 respectively contain SEQ ID NO: 1 The amino acid sequences of Bv-HCDR1, Bv-HCDR2 and Bv-HCDR3 in, and B-LCDR1, B-LCDR2 and B-LCDR3 respectively comprise Bv-LCDR1, Bv-LCDR2 and Bv-LCDR1 in SEQ ID NO: 2 Amino acid sequence of LCDR3.

In some embodiments, the B-HCDR1, B-HCDR2, B-HCDR3, B-LCDR1, B-LCDR2, B-LCDR3, Bv-HCDR1, Bv-HCDR2, Bv-HCDR3, Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 are defined according to the same numbering convention selected from Kabat, IMGT, Chothia, AbM and Contact.

In some embodiments, the B-HCDR1, B-HCDR2, B-HCDR3, B-LCDR1, B-LCDR2, B-LCDR3, Bv-HCDR1, Bv-HCDR2, Bv-HCDR3, Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 are defined according to the Kabat numbering convention.

In some embodiments, the B-HCDR1, B-HCDR2, B-HCDR3, B-LCDR1, B-LCDR2, B-LCDR3, Bv-HCDR1, Bv-HCDR2, Bv-HCDR3, Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 are defined according to the IMGT numbering rules.

In some embodiments, the B-HCDR1, B-HCDR2, B-HCDR3, B-LCDR1, B-LCDR2, B-LCDR3, Bv-HCDR1, Bv-HCDR2, Bv-HCDR3, Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 are defined according to the Chothia numbering convention.

In some embodiments, the B-HCDR1, B-HCDR2, B-HCDR3, B-LCDR1, B-LCDR2, B-LCDR3, Bv-HCDR1, Bv-HCDR2, Bv-HCDR3, Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 are defined according to the AbM numbering convention.

In some embodiments, the B-HCDR1, B-HCDR2, B-HCDR3, B-LCDR1, B-LCDR2, B-LCDR3, Bv-HCDR1, Bv-HCDR2, Bv-HCDR3, Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 are defined according to the Contact Numbering Rules.

In some embodiments, the antigen-binding molecule of any one of the above, the B-HCDR1, B-HCDR2, B-HCDR3, B-LCDR1, B-LCDR2, B-LCDR3 are defined according to the Kabat numbering rules, wherein , B-HCDR1 comprises the amino acid sequence NNAIN (SEQ ID NO: 18), B-HCDR2 comprises the amino acid sequence X ₁ IX ₂ PMFGX ₃ AKYSX ₄ X ₅ FQG (SEQ ID NO: 65), and B-HCDR3 comprises the amino acid sequence The amino acid sequence SRDX ₆ LLFPX ₇ X ₈ X ₉ LX ₁₀ X ₁₁ (SEQ ID NO: 66), B-LCDR1 contains the amino acid sequence X ₁₂ GX ₁₃ X ₁₄ LX ₁₅ X ₁₆ X ₁₇ X ₁₈ AS (SEQ ID NO : 67), B-LCDR2 comprises the amino acid sequence GKNNRPS (SEQ ID NO: 32) and B-LCDR3 comprises the amino acid sequence X ₁₉ SRX ₂₀ X ₂₁ X ₂₂ GX ₂₃ X ₂₄ WX ₂₅ (SEQ ID NO: 68) , wherein, _X1 is selected from S, A, V or G, _X2 is selected from A, I, M or S, _X3 is selected from T or G, _X4 is selected from Q, E, G or K, _X5 is selected from From N, G or Q, X ₆ is selected from L or P, X ₇ is selected from H, Q or D, X ₈ is selected from H or D, X ₉ is selected from A or G, X ₁₀ is selected from S or L, X ₁₁ is selected from P or S, X ₁₂ is selected from Q or H, X ₁₃ is selected from D, A or N, X ₁₄ is selected from S or I, X ₁₅ is selected from K, R or T, X ₁₆ is selected from S, W , T or D, X ₁₇ is selected from H, Y or S, X ₁₈ is selected from Y or R, X ₁₉ is selected from G or S, X ₂₀ is selected from A or D, X ₂₁ is selected from E or S, X ₂₂ is selected from From S or A, X ₂₃ is selected from V, E, A, N or W, X ₂₄ is selected from G, K, H or R, X ₂₅ is selected from L or V; and the B-HCDR1, B-HCDR2, B - HCDR3, B-LCDR1, B-LCDR2 and B-LCDR3 do not contain the following CDR combination: B-HCDR1 contains the amino acid sequence of SEQ ID NO: 18, B-HCDR2 contains the amino acid sequence of SEQ ID NO: 19, B-HCDR3 comprises the amino acid sequence of SEQ ID NO: 20, B-LCDR1 comprises the amino acid sequence of SEQ ID NO: 31, B-LCDR2 comprises the amino acid sequence of SEQ ID NO: 32, and B-LCDR3 comprises Amino acid sequence of SEQ ID NO:33.

In some embodiments, the antigen binding molecule of any one of the above, wherein,

(i) the B-HCDR1, B-HCDR2 and B-HCDR3 respectively comprise the amino acid sequences of Bv-HCDR1, Bv-HCDR2 and Bv-HCDR3 in SEQ ID NO: 47, and the B-LCDR1, B-LCDR2 and B-LCDR3 respectively comprising the amino acid sequences of Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 in SEQ ID NO: 48, or

(ii) the B-HCDR1, B-HCDR2 and B-HCDR3 respectively comprise the amino acid sequences of Bv-HCDR1, Bv-HCDR2 and Bv-HCDR3 in SEQ ID NO: 5, and the B-LCDR1, B-LCDR2 and B-LCDR3 respectively comprising the amino acid sequences of Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 in SEQ ID NO: 12, or

(iii) the B-HCDR1, B-HCDR2 and B-HCDR3 respectively comprise the amino acid sequences of Bv-HCDR1, Bv-HCDR2 and Bv-HCDR3 in SEQ ID NO: 6, and the B-LCDR1, B-LCDR2 and B-LCDR3 respectively comprising the amino acid sequences of Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 in SEQ ID NO: 13, or

(iv) the B-HCDR1, B-HCDR2 and B-HCDR3 respectively comprise the amino acid sequences of Bv-HCDR1, Bv-HCDR2 and Bv-HCDR3 in SEQ ID NO: 7, and the B-LCDR1, B-LCDR2 and B-LCDR3 respectively comprising the amino acid sequences of Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 in SEQ ID NO: 14, or

(v) the B-HCDR1, B-HCDR2 and B-HCDR3 respectively comprise the amino acid sequences of Bv-HCDR1, Bv-HCDR2 and Bv-HCDR3 in SEQ ID NO: 8, and the B-LCDR1, B-LCDR2 and B-LCDR3 respectively comprising the amino acid sequences of Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 in SEQ ID NO: 15, or

(vi) The B-HCDR1, B-HCDR2 and B-HCDR3 respectively comprise the amino acid sequences of Bv-HCDR1, Bv-HCDR2 and Bv-HCDR3 in SEQ ID NO: 9, and the B-LCDR1, B-LCDR2 and B-LCDR3 respectively comprising the amino acid sequences of Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 in SEQ ID NO: 16, or

(vii) the B-HCDR1, B-HCDR2 and B-HCDR3 respectively comprise the amino acid sequences of Bv-HCDR1, Bv-HCDR2 and Bv-HCDR3 in SEQ ID NO: 10, and the B-LCDR1, B-LCDR2 and B-LCDR3 respectively comprising the amino acid sequences of Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 in SEQ ID NO: 17, or

(viii) the B-HCDR1, B-HCDR2 and B-HCDR3 respectively comprise the amino acid sequences of Bv-HCDR1, Bv-HCDR2 and Bv-HCDR3 in SEQ ID NO: 11, and the B-LCDR1, B-LCDR2 and B-LCDR3 respectively comprise the amino acid sequences of Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 in SEQ ID NO:2.

In some embodiments, the antigen binding molecule of any one of the above, the B-HCDR1, B-HCDR2, B-HCDR3, B-LCDR1, B-LCDR2, B-LCDR3, Bv- HCDR1, Bv-HCDR2, Bv-HCDR3, Bv-LCDR1, Bv-LCDR2, and Bv-LCDR3 are defined according to the Kabat numbering rule, where,

(i) The B-HCDR1 comprises the amino acid sequence of SEQ ID NO: 18, the B-HCDR2 comprises the amino acid sequence of SEQ ID NO: 28, and the B-HCDR3 comprises the amino acid sequence of SEQ ID NO: 29 , and the B-LCDR1 comprises the amino acid sequence of SEQ ID NO: 43, the B-LCDR2 comprises the amino acid sequence of SEQ ID NO: 32, and the B-LCDR3 comprises the amino acid sequence of SEQ ID NO: 44 ;or

(ii) The B-HCDR1 comprises the amino acid sequence of SEQ ID NO: 18, the B-HCDR2 comprises the amino acid sequence of SEQ ID NO: 21, and the B-HCDR3 comprises the amino acid sequence of SEQ ID NO: 20 , and the B-LCDR1 comprises the amino acid sequence of SEQ ID NO: 34, the B-LCDR2 comprises the amino acid sequence of SEQ ID NO: 32, and the B-LCDR3 comprises the amino acid sequence of SEQ ID NO: 35 ;or

(iii) The B-HCDR1 comprises the amino acid sequence of SEQ ID NO: 18, the B-HCDR2 comprises the amino acid sequence of SEQ ID NO: 22, and the B-HCDR3 comprises the amino acid sequence of SEQ ID NO: 23 , and the B-LCDR1 comprises the amino acid sequence of SEQ ID NO: 31, the B-LCDR2 comprises the amino acid sequence of SEQ ID NO: 32, and the B-LCDR3 comprises the amino acid sequence of SEQ ID NO: 36 ;or

(iv) The B-HCDR1 comprises the amino acid sequence of SEQ ID NO: 18, the B-HCDR2 comprises the amino acid sequence of SEQ ID NO: 24, and the B-HCDR3 comprises the amino acid sequence of SEQ ID NO: 20 , and the B-LCDR1 comprises the amino acid sequence of SEQ ID NO: 37, the B-LCDR2 comprises the amino acid sequence of SEQ ID NO: 32, and the B-LCDR3 comprises the amino acid sequence of SEQ ID NO: 38 ;or

(v) The B-HCDR1 comprises the amino acid sequence of SEQ ID NO: 18, the B-HCDR2 comprises the amino acid sequence of SEQ ID NO: 25, and the B-HCDR3 comprises the amino acid sequence of SEQ ID NO: 20 , and the B-LCDR1 comprises the amino acid sequence of SEQ ID NO: 39, the B-LCDR2 comprises the amino acid sequence of SEQ ID NO: 32, and the B-LCDR3 comprises the amino acid sequence of SEQ ID NO: 40 ;or

(vi) The B-HCDR1 comprises the amino acid sequence of SEQ ID NO: 18, the B-HCDR2 comprises the amino acid sequence of SEQ ID NO: 26, and the B-HCDR3 comprises the amino acid sequence of SEQ ID NO: 27 , and the B-LCDR1 comprises the amino acid sequence of SEQ ID NO: 41, the B-LCDR2 comprises the amino acid sequence of SEQ ID NO: 32, and the B-LCDR3 comprises the amino acid sequence of SEQ ID NO: 42 ;or

(vii) The B-HCDR1 comprises the amino acid sequence of SEQ ID NO: 18, the B-HCDR2 comprises the amino acid sequence of SEQ ID NO: 30, and the B-HCDR3 comprises the amino acid sequence of SEQ ID NO: 20 , and the B-LCDR1 comprises the amino acid sequence of SEQ ID NO: 31, the B-LCDR2 comprises the amino acid sequence of SEQ ID NO: 32, and the B-LCDR3 comprises the amino acid sequence of SEQ ID NO: 33 .

In some embodiments, the antigen-binding molecule of any one of the above, wherein the B-VH comprises at least 90% (e.g., at least 90%, 95%, 96%, 97%, 98%) of SEQ ID NO: 63 % or 99%) sequence identity of an amino acid sequence comprising at least 90% (e.g., at least 90%, 95%, 96%, 97%, 98% or 99%) to SEQ ID NO: 64 The amino acid sequence of sequence identity, wherein SEQ ID NO: 63 is:

SEQ ID NO: 64 is:

，其中，X₁選自S、A、V或G，X₂選自A、I、M或S，X₃選自T或G，X₄選自Q、E、G或K，X₅選自N、G或Q，X₆選自L或P，X₇選自H、Q或D，X₈選自H或D，X₉選自A或G，X₁₀選自S或L，X₁₁選自P或S，X₁₂選自Q或H，X₁₃選自D、A或N，X₁₄選自S或I，X₁₅選自K、R或T，X₁₆選自S、W、T或D，X₁₇選自H、Y或S，X₁₈選自Y或R，X₁₉選自G或S，X₂₀選自A或D，X₂₁選自E或S，X₂₂選自S或A，X₂₃選自V、E、A、N或W，X₂₄選自G、K、H或R，X₂₅選自L或V，X₂₆選自G或C，X₂₇選自R或G，X₂₈選自G或C；並且，該B-VH和B-VL不包含如下可變區組合：B-VH包含SEQ ID NO：1的胺基酸序列，且B-VL包含SEQ ID NO：2的胺基酸序列。

, wherein, _X1 is selected from S, A, V or G, _X2 is selected from A, I, M or S, _X3 is selected from T or G, _X4 is selected from Q, E, G or K, _X5 is selected from From N, G or Q, X ₆ is selected from L or P, X ₇ is selected from H, Q or D, X ₈ is selected from H or D, X ₉ is selected from A or G, X ₁₀ is selected from S or L, X ₁₁ is selected from P or S, X ₁₂ is selected from Q or H, X ₁₃ is selected from D, A or N, X ₁₄ is selected from S or I, X ₁₅ is selected from K, R or T, X ₁₆ is selected from S, W , T or D, X ₁₇ is selected from H, Y or S, X ₁₈ is selected from Y or R, X ₁₉ is selected from G or S, X ₂₀ is selected from A or D, X ₂₁ is selected from E or S, X ₂₂ is selected from From S or A, X ₂₃ is selected from V, E, A, N or W, X ₂₄ is selected from G, K, H or R, X ₂₅ is selected from L or V, X ₂₆ is selected from G or C, X ₂₇ is selected from From R or G, X ₂₈ is selected from G or C; and, the B-VH and B-VL do not comprise the following variable region combinations: B-VH comprises the amino acid sequence of SEQ ID NO: 1, and B-VL Comprising the amino acid sequence of SEQ ID NO:2.

In some embodiments, the antigen binding molecule of any one of the above, wherein,

(i) the B-VH 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: 47, and the B-VL 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: 48, or

(ii) the B-VH 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: 5, and the B-VL 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: 12, or

(iii) the B-VH 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: 6, and the B-VL 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: 13, or

(iv) the B-VH 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: 7, and the B-VL 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: 14, or

(v) the B-VH 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: 8, and the B-VL 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: 15, or

(vi) the B-VH 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: 9, and the B-VL 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: 16, or

(vii) the B-VH 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: 10, and the B- VL 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: 17, or

(viii) the B-VH 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: 11, and the B-VL 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: 2;

In some embodiments, the antigen-binding molecule of any one of the above, wherein the B-VH comprises the amino acid sequence of SEQ ID NO: 47, and the B-VL comprises the amino acid sequence of SEQ ID NO: 48 sequence. In some embodiments, the B-VH comprises the amino acid sequence of SEQ ID NO: 5, and the B-VL comprises the amino acid sequence of SEQ ID NO: 12; or the B-VH comprises the amino acid sequence of SEQ ID NO: 6 and the B-VL comprises the amino acid sequence of SEQ ID NO: 13; or the B-VH comprises the amino acid sequence of SEQ ID NO: 7, and the B-VL comprises the amino acid sequence of SEQ ID NO: 14; or the B-VH comprises the amino acid sequence of SEQ ID NO: 8, and the B-VL comprises the amino acid sequence of SEQ ID NO: 15; or the B-VH comprises the amino acid sequence of SEQ ID NO : the amino acid sequence of 9, and the B-VL comprises the amino acid sequence of SEQ ID NO: 16; or the B-VH comprises the amino acid sequence of SEQ ID NO: 10, and the B-VL comprises the amino acid sequence of SEQ ID The amino acid sequence of NO:17; or the B-VH comprises the amino acid sequence of SEQ ID NO:11, and the B-VL comprises the amino acid sequence of SEQ ID NO:2.

In some embodiments, the antigen-binding molecule according to any one of the above, wherein the antigen-binding moiety 1 that specifically binds to BAFF is scFv. In some embodiments, the scFv molecule has the following structure from N-terminus to C-terminus: VL-linker-VH or VH-linker-VL. In some embodiments, the linker is a peptide linker having the structure "L ₁ -(GGGGS)nL ₂ ", wherein L ₁ is a bond, A, GS, GGS or GGGS, and n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, L2 is a bond, G, GG, GGG or GGGG, and the peptide linker is not a bond. In some embodiments, the peptide linker is 3-15 amino acid residues in length. In some embodiments, the linker structure is: (G _x S) _y , wherein x is selected from an integer of 1-5 (eg, 1, 2, 3, 4 or 5), and y is selected from an integer of 1-6 ( For example, 1, 2, 3, 4, 5 or 6). In some embodiments, the linker is GGGGSGGGGSGGGGS (SEQ ID NO: 70).

In some embodiments, the antigen-binding molecule of any one of the above, wherein the antigen-binding moiety 2 comprises a heavy chain variable region P-VH and a light chain variable region P-VL, the P-VH comprising P-HCDR1 , P-HCDR2 and P-HCDR3, the P-VL comprises P-LCDR1, P-LCDR2 and P-LCDR3; the P-HCDR1, P-HCDR2 and P-HCDR3 respectively comprise Pv-HCDR1 in SEQ ID NO:51 , the amino acid sequences of Pv-HCDR2 and Pv-HCDR3, and the P-LCDR1, P-LCDR2 and P-LCDR3 respectively comprise the amine groups of Pv-LCDR1, Pv-LCDR2 and Pv-LCDR3 in SEQ ID NO:52 acid sequence; the P-HCDR1, P-HCDR2, P-HCDR3, P-LCDR1, P-LCDR2, P-LCDR3, Pv-HCDR1, Pv-HCDR2, Pv-HCDR3, Pv-LCDR1, Pv-LCDR2 and Pv- LCDR3 is defined according to the same numbering convention selected from Kabat, IMGT, Chothia, AbM and Contact.

In some embodiments, the P-HCDR1, P-HCDR2, P-HCDR3, P-LCDR1, P-LCDR2, P-LCDR3, Pv-HCDR1, Pv-HCDR2, Pv-HCDR3, Pv-LCDR1, Pv-LCDR2 and Pv-LCDR3 are defined according to the Kabat numbering convention.

In some embodiments, the P-HCDR1, P-HCDR2, P-HCDR3, P-LCDR1, P-LCDR2, P-LCDR3, Pv-HCDR1, Pv-HCDR2, Pv-HCDR3, Pv-LCDR1, Pv-LCDR2 and Pv-LCDR3 are defined according to the IMGT numbering rules.

In some embodiments, the P-HCDR1, P-HCDR2, P-HCDR3, P-LCDR1, P-LCDR2, P-LCDR3, Pv-HCDR1, Pv-HCDR2, Pv-HCDR3, Pv-LCDR1, Pv-LCDR2 and Pv-LCDR3 are defined according to the Chothia numbering convention.

In some embodiments, the P-HCDR1, P-HCDR2, P-HCDR3, P-LCDR1, P-LCDR2, P-LCDR3, Pv-HCDR1, Pv-HCDR2, Pv-HCDR3, Pv-LCDR1, Pv-LCDR2 and Pv-LCDR3 are defined according to the AbM numbering convention.

In some embodiments, the P-HCDR1, P-HCDR2, P-HCDR3, P-LCDR1, P-LCDR2, P-LCDR3, Pv-HCDR1, Pv-HCDR2, Pv-HCDR3, Pv-LCDR1, Pv-LCDR2 and Pv-LCDR3 are defined according to the Contact Numbering Rules.

In some embodiments, the antigen-binding molecule of any one of the above, the P-HCDR1, P-HCDR2, P-HCDR3, P-LCDR1, P-LCDR2, P-LCDR3 are defined according to the Kabat numbering rules, the P-HCDR1 comprises the amino acid sequence of SEQ ID NO: 53, the P-HCDR2 comprises the amino acid sequence of SEQ ID NO: 54, the P-HCDR3 comprises the amino acid sequence of SEQ ID NO: 55, and the P- LCDR1 comprises the amino acid sequence of SEQ ID NO:56, the P-LCDR2 comprises the amino acid sequence of SEQ ID NO:57, and the P-LCDR3 comprises the amino acid sequence of SEQ ID NO:58.

In some embodiments, the antigen binding molecule of any one of the above, the heavy chain variable region P-VH comprises at least 90% (eg, at least 90%, 95%, 96%, 97%) of SEQ ID NO: 51 %, 98% or 99%) sequence identity of the amino acid sequence, the light chain variable region The P-VL 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:52.

In some embodiments, the antigen-binding molecule according to any one of the above, the heavy chain variable region P-VH comprises the amino acid sequence of SEQ ID NO: 51, and the light chain variable region P-VL comprises SEQ ID NO: Amino acid sequence of 52.

In some embodiments, the antigen-binding molecule of any one of the above, which comprises a heavy chain constant region CH and a light chain constant region CL. In some embodiments, the heavy chain constant region CH is a human IgG heavy chain constant region and the light chain constant region CL is a human lambda or kappa light chain constant region. In some embodiments, the heavy chain constant region CH is a human IgG1 heavy chain constant region and the light chain constant region CL is a human lambda light chain constant region. In some embodiments, the heavy chain constant region CH is a human IgG1 heavy chain constant region, and the light chain constant region CL is a human kappa light chain constant region. In some embodiments, the heavy chain constant region CH comprises the amino acid sequence of SEQ ID NO:59, and the light chain constant region CL comprises the amino acid sequence of SEQ ID NO:60.

In some embodiments, the antigen-binding molecule of any one of the above, which comprises a heavy chain constant region CH and a light chain constant region CL, wherein the heavy chain constant region CH comprises an Fc region. In some embodiments, the Fc region comprises a first subunit Fc1 and a second subunit Fc2 capable of associating with each other. In some embodiments, the Fc region is an IgG Fc region. In some embodiments, the Fc region is an IgG1 Fc region. In some embodiments, the Fc region has one or more amino acid substitutions that reduce homodimerization; and/or the Fc region has one or more amines that reduce binding of the Fc region to an Fc receptor amino acid substitution. In some embodiments, the Fc region has YTE mutations (M252Y, S254T, and T256E), L234A, L235A mutations, and/or S228P mutations, the mutations numbered according to the EU index. In some embodiments, the Fc region comprises a The first subunit is associated with a second subunit, the first subunit and/or the second subunit having one or more amino acid substitutions that reduce homodimerization. In some embodiments, the first subunit has a raised structure according to the pestle and socket technique, the second subunit has a pore structure according to the pestle and socket technique, or the first subunit has a pore structure according to the pestle and socket technique, the second The subunits have a raised structure according to the pestle and socket technique. In some embodiments, the first subunit has one or more amino acid substitutions at a position selected from 354, 356, 358, and 366, and the second subunit has a position selected from 349, 356, 358, 366, One or more amino acid substitutions at positions 368 and 407. In some embodiments, the second subunit has one or more amino acid substitutions at positions selected from 354, 356, 358, and 366, and the first subunit has substitutions at positions selected from 349, 356, 358, 366, One or more amino acid substitutions at positions 368 and 407. In some embodiments, the first subunit has one or more amino acid substitutions selected from 354C, 356E, 358M, and 366W, and the second subunit has an amino acid substitution selected from 349C, 356E, 358M, 366S, 368A, and 407V. One or more amino acid substitutions. In some embodiments, the second subunit has one or more amino acid substitutions selected from 354C, 356E, 358M, and 366W, and the first subunit has one or more amino acid substitutions selected from 349C, 356E, 358M, 366S, 368A, and 407V. One or more amino acid substitutions. In some embodiments, the first subunit includes amino acid substitutions of 354C, 356E, 358M, and 366W and the second subunit includes amino acid substitutions of 349C, 356E, 358M, 366S, 368A, and 407V. In some embodiments, the second subunit includes amino acid substitutions of 354C, 356E, 358M, and 366W and the first subunit includes amino acid substitutions of 349C, 356E, 358M, 366S, 368A, and 407V.

In some embodiments, in the antigen-binding molecule according to any one of the above, the antigen-binding module 1 is scFv, and the antigen-binding module 2 comprises a heavy chain variable region P-VH, a light chain variable region P-VL, Full-length antibody to the heavy chain constant region CH and light chain constant region CL, the antigen binds Module 1 is fused directly or via a linker to the N-terminus of the variable region of antigen-binding module 2 or the C-terminus of the constant region of antigen-binding module 2; in some embodiments, the N-terminus of the scFv is directly or via a linker Fused to the C-terminus of the heavy chain constant region CH of antigen binding module 2; end.

In some embodiments, the antigen-binding molecule according to any one of the above, the antigen-binding molecule comprises a first chain having a structure represented by formula (a) and a second chain having a structure represented by formula (b); or comprising A first chain having a structure shown in formula (c) and a second chain having a structure shown in formula (b), wherein,

Formula (a): [P-VH]-[CH]-[Linker 1]-[B-VH]-[Linker 2]-[B-VL],

Formula (b): [P-VL]-[CL],

Formula (c): [P-VH]-[CH]-[Linker 1]-[B-VL]-[Linker 2]-[B-VH].

In some embodiments, in formula (a) and formula (c), linker 1 and linker 2 are the same or different peptide linkers. In some embodiments, the peptide linker independently has the structure L ₁ -(GGGGS)nL ₂ , wherein L ₁ is a bond, A, GS, GGS or GGGS, and n is 0, 1, 2, 3, 4 , 5, 6, 7, 8, 9 or 10, L2 is a bond, G, GG, GGG or GGGG, and the peptide linker is not a bond. In some embodiments, the peptide linker is 3-15 amino acid residues in length. In some embodiments, the linker 1 and linker 2 are each independently selected from a linker of formula (d), wherein formula (d): (G _x S) _y , wherein x is selected from an integer of 1-5 (eg 1, 2, 3, 4 or 5), y is selected from an integer of 1-6 (eg, 1, 2, 3, 4, 5 or 6). In some embodiments, the linker 1 is selected from: GGGS (SEQ ID NO: 69), and the linker 2 is selected from: GGGGSGGGGSGGGGS (SEQ ID NO: 70). In some embodiments, the antigen-binding molecule, wherein the first chain comprises at least 90% (eg, at least 90%, 95%, 96%, 97%, 98%, or 99%) of SEQ ID NO: 61 ) sequence identity, and the second strand comprises at least 90% (e.g., at least 90%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 50 or the first strand comprises amino acids having at least 90% (e.g., at least 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO: 62 sequence, and the second strand 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:50. In some embodiments, the antigen binding molecule has a first strand comprising the amino acid sequence of SEQ ID NO: 61 and a second strand comprising the amino acid sequence of SEQ ID NO: 50, or comprises SEQ ID NO: The first strand of the amino acid sequence of 62 and the second strand of the amino acid sequence comprising SEQ ID NO:50.

In some embodiments, the antigen binding molecule has:

(i) two first strands comprising the amino acid sequence of SEQ ID NO: 61 and two second strands comprising the amino acid sequence of SEQ ID NO: 50, or

(ii) Two first strands comprising the amino acid sequence of SEQ ID NO:62 and two second strands comprising the amino acid sequence of SEQ ID NO:50.

In another aspect, the present disclosure provides an antigen-binding molecule that specifically binds to BAFF antigen, comprising a heavy chain variable region B-VH and a light chain variable region B-VL, the B-VH comprising B-HCDR1, B- HCDR2 and B-HCDR3, the B-VL comprises B-LCDR1, B-LCDR2 and B-LCDR3, wherein the B-HCDR1, B-HCDR2 and B-HCDR3 respectively comprise Bv-HCDR1, Bv-HCDR1, Amino acid sequences of Bv-HCDR2 and Bv-HCDR3, And the B-LCDR1, B-LCDR2 and B-LCDR3 respectively comprise the amino acid sequences of Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 in SEQ ID NO: 64, wherein, SEQ ID NO: 63 is:

SEQ ID NO: 64 is:

Wherein, _X1 is selected from S, A, V or G, _X2 is selected from A, I, M or S, _X3 is selected from T or G, _X4 is selected from Q, E, G or K, _X5 is selected from N, G or Q, X ₆ is selected from L or P, X ₇ is selected from H, Q or D, X ₈ is selected from H or D, X ₉ is selected from A or G, X ₁₀ is selected from S or L, X ₁₁ selected from P or S, X ₁₂ selected from Q or H, X ₁₃ selected from D, A or N, X ₁₄ selected from S or I, X ₁₅ selected from K, R or T, X ₁₆ selected from S, W, T or D, X ₁₇ is selected from H, Y or S, X ₁₈ is selected from Y or R, X ₁₉ is selected from G or S, X ₂₀ is selected from A or D, X ₂₁ is selected from E or S, X ₂₂ is selected from S or A, X ₂₃ is selected from V, E, A, N or W, X ₂₄ is selected from G, K, H or R, X ₂₅ is selected from L or V, X ₂₆ is selected from G or C, X ₂₇ is selected from R or G, X ₂₈ is selected from G or C; And, the B-HCDR1, B-HCDR2, B-HCDR3, B-LCDR1, B-LCDR2 and B-LCDR3 do not contain the following CDR combination: B-HCDR1, B- HCDR2 and B-HCDR3 respectively comprise the amino acid sequences of Bv-HCDR1, Bv-HCDR2 and Bv-HCDR3 in SEQ ID NO: 1, and B-LCDR1, B-LCDR2 and B-LCDR3 respectively comprise SEQ ID NO: 2 Amino acid sequences of Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 in .

In some embodiments, the B-HCDR1, B-HCDR2, B-HCDR3, B-LCDR1, B-LCDR2, B-LCDR3, Bv-HCDR1, Bv-HCDR2, Bv-HCDR3, Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 are defined according to the same numbering convention selected from Kabat, IMGT, Chothia, AbM and Contact.

In some embodiments, the B-HCDR1, B-HCDR2, B-HCDR3, B-LCDR1, B-LCDR2, B-LCDR3, Bv-HCDR1, Bv-HCDR2, Bv-HCDR3, Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 are defined according to the Kabat numbering convention.

In some embodiments, the B-HCDR1, B-HCDR2, B-HCDR3, B-LCDR1, B-LCDR2, B-LCDR3, Bv-HCDR1, Bv-HCDR2, Bv-HCDR3, Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 are defined according to the IMGT numbering rules.

In some embodiments, the B-HCDR1, B-HCDR2, B-HCDR3, B-LCDR1, B-LCDR2, B-LCDR3, Bv-HCDR1, Bv-HCDR2, Bv-HCDR3, Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 are defined according to the Chothia numbering convention.

In some embodiments, the B-HCDR1, B-HCDR2, B-HCDR3, B-LCDR1, B-LCDR2, B-LCDR3, Bv-HCDR1, Bv-HCDR2, Bv-HCDR3, Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 are defined according to the AbM numbering convention.

In some embodiments, the B-HCDR1, B-HCDR2, B-HCDR3, B-LCDR1, B-LCDR2, B-LCDR3, Bv-HCDR1, Bv-HCDR2, Bv-HCDR3, Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 are defined according to the Contact Numbering Rules.

In some embodiments, the B-HCDR1, B-HCDR2, B-HCDR3, B-LCDR1, B-LCDR2, B-LCDR3 are defined according to the Kabat numbering rules, wherein the B-HCDR1 comprises the amino acid sequence NNAIN( SEQ ID NO: 18), B-HCDR2 contains the amino acid sequence X ₁ IX ₂ PMFGX ₃ AKYSX ₄ X ₅ FQG (SEQ ID NO: 65), B-HCDR3 contains the amino acid sequence SRDX ₆ LLFPX ₇ X ₈ X ₉ LX ₁₀ X ₁₁ (SEQ ID NO: 66), B-LCDR1 comprises the amino acid sequence X ₁₂ GX ₁₃ X ₁₄ LX ₁₅ X ₁₆ X ₁₇ X ₁₈ AS (SEQ ID NO: 67), B-LCDR2 comprises the amino acid sequence The sequence GKNNRPS (SEQ ID NO: 32) and B-LCDR3 comprise the amino acid sequence X ₁₉ SRX ₂₀ X ₂₁ X ₂₂ GX ₂₃ X ₂₄ WX ₂₅ (SEQ ID NO: 68), wherein X ₁ is selected from S, A, V or G, _X2 is selected from A, I, M or S, _X3 is selected from T or G, _X4 is selected from Q, E, G or K, _X5 is selected from N, G or Q, _X6 is selected from L or P, X ₇ is selected from H, Q or D, X ₈ is selected from H or D, X ₉ is selected from A or G, X ₁₀ is selected from S or L, X ₁₁ is selected from P or S, X ₁₂ is selected from Q or H, X ₁₃ is selected from D, A or N, X ₁₄ is selected from S or I, X ₁₅ is selected from K, R or T, X ₁₆ is selected from S, W, T or D, X ₁₇ is selected from H, Y or S, X ₁₈ is selected from Y or R, X ₁₉ is selected from G or S, X ₂₀ is selected from A or D, X ₂₁ is selected from E or S, X ₂₂ is selected from S or A, X ₂₃ is selected from V, E, A, N or W, X ₂₄ is selected from G, K, H or R, X ₂₅ is selected from L or V; and the B-HCDR1, B-HCDR2, B-HCDR3, B-LCDR1, B-LCDR2 and B-LCDR3 does not contain the following CDR combinations: B-HCDR1 contains the amino acid sequence of SEQ ID NO: 18, B-HCDR2 contains the amino acid sequence of SEQ ID NO: 19, and B-HCDR3 contains the amine of SEQ ID NO: 20 The amino acid sequence, B-LCDR1 comprises the amino acid sequence of SEQ ID NO:31, B-LCDR2 comprises the amino acid sequence of SEQ ID NO:32, and B-LCDR3 comprises the amino acid sequence of SEQ ID NO:33.

In some embodiments, the antigen-binding molecule that specifically binds to a BAFF antigen according to any one of the above, wherein (i) the B-HCDR1, B-HCDR2, and B-HCDR3 comprise Bv- in SEQ ID NO:47, respectively. Amino acid sequences of HCDR1, Bv-HCDR2 and Bv-HCDR3, and the B-LCDR1, B-LCDR2 and B-LCDR3 respectively comprise the amines of Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 in SEQ ID NO: 48 amino acid sequence, or (ii) the B-HCDR1, B-HCDR2 and B-HCDR3 respectively comprise the amino acid sequences of Bv-HCDR1, Bv-HCDR2 and Bv-HCDR3 in SEQ ID NO: 5, and the B- LCDR1, B-LCDR2 and B-LCDR3 respectively comprise the amino acid sequences of Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 in SEQ ID NO: 12, or (iii) the B-HCDR1, B-HCDR2 and B- HCDR3 respectively comprises the amino acid sequences of Bv-HCDR1, Bv-HCDR2 and Bv-HCDR3 in SEQ ID NO: 6, and the B-LCDR1, B-LCDR2 and B-LCDR3 respectively comprise Bv in SEQ ID NO: 13 - the amino acid sequence of LCDR1, Bv-LCDR2 and Bv-LCDR3, or (iv) the B-HCDR1, B-HCDR2 and B-HCDR3 respectively comprise Bv-HCDR1, Bv-HCDR2 and Bv in SEQ ID NO: 7 - the amino acid sequence of HCDR3, and the B-LCDR1, B-LCDR2 and B-LCDR3 respectively comprise the amino acid sequence of Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 in SEQ ID NO: 14, or (v ) The B-HCDR1, B-HCDR2 and B-HCDR3 respectively comprise the amino acid sequences of Bv-HCDR1, Bv-HCDR2 and Bv-HCDR3 in SEQ ID NO: 8, and the B-LCDR1, B-LCDR2 and B - LCDR3 respectively comprises the amino acid sequence of Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 in SEQ ID NO: 15, or (vi) the B-HCDR1, B-HCDR2 and B-HCDR3 respectively comprise SEQ ID NO: The amino acid sequences of Bv-HCDR1, Bv-HCDR2 and Bv-HCDR3 in 9, and the B-LCDR1, B-LCDR2 and B-LCDR3 respectively comprise SEQ ID NO: The amino acid sequences of Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 in 16, or (vii) the B-HCDR1, B-HCDR2 and B-HCDR3 respectively comprise Bv-HCDR1, Bv in SEQ ID NO: 10 - the amino acid sequences of HCDR2 and Bv-HCDR3, and the B-LCDR1, B-LCDR2 and B-LCDR3 respectively comprise the amino acid sequences of Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 in SEQ ID NO: 17 , or (viii) the B-HCDR1, B-HCDR2 and B-HCDR3 respectively comprise the amino acid sequences of Bv-HCDR1, Bv-HCDR2 and Bv-HCDR3 in SEQ ID NO: 11, and the B-LCDR1, B - LCDR2 and B-LCDR3 respectively comprise the amino acid sequences of Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 in SEQ ID NO: 2; and the B-HCDR1, B-HCDR2, B-HCDR3, B-LCDR1, B-LCDR2, B-LCDR3, Bv-HCDR1, Bv-HCDR2, Bv-HCDR3, Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 are defined according to the same numbering convention selected from Kabat, IMGT, Chothia, AbM and Contact of.

In some embodiments, the B-HCDR1, B-HCDR2, B-HCDR3, B-LCDR1, B-LCDR2, B-LCDR3, Bv-HCDR1, Bv-HCDR2, Bv-HCDR3, Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 are defined according to the Kabat numbering convention.

In some embodiments, the B-HCDR1, B-HCDR2, B-HCDR3, B-LCDR1, B-LCDR2, B-LCDR3, Bv-HCDR1, Bv-HCDR2, Bv-HCDR3, Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 are defined according to the IMGT numbering rules.

In some embodiments, the B-HCDR1, B-HCDR2, B-HCDR3, B-LCDR1, B-LCDR2, B-LCDR3, Bv-HCDR1, Bv-HCDR2, Bv-HCDR3, Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 are defined according to the Chothia numbering convention.

In some embodiments, the B-HCDR1, B-HCDR2, B-HCDR3, B-LCDR1, B-LCDR2, B-LCDR3, Bv-HCDR1, Bv-HCDR2, Bv-HCDR3, Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 are defined according to the AbM numbering convention.

In some embodiments, the B-HCDR1, B-HCDR2, B-HCDR3, B-LCDR1, B-LCDR2, B-LCDR3, Bv-HCDR1, Bv-HCDR2, Bv-HCDR3, Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 are defined according to the Contact Numbering Rules.

In some embodiments, wherein, the B-HCDR1, B-HCDR2, B-HCDR3, B-LCDR1, B-LCDR2, B-LCDR3 are defined according to the Kabat numbering rules, wherein (i) the B-HCDR1 comprises SEQ The amino acid sequence of ID NO: 18, the B-HCDR2 comprises the amino acid sequence of SEQ ID NO: 28, the B-HCDR3 comprises the amino acid sequence of SEQ ID NO: 29, and the B-LCDR1 comprises the amino acid sequence of SEQ ID The amino acid sequence of NO: 43, the B-LCDR2 comprises the amino acid sequence of SEQ ID NO: 32, and the B-LCDR3 comprises the amino acid sequence of SEQ ID NO: 44; or (ii) the B-HCDR1 comprising the amino acid sequence of SEQ ID NO: 18, the B-HCDR2 comprising the amino acid sequence of SEQ ID NO: 21, the B-HCDR3 comprising the amino acid sequence of SEQ ID NO: 20, and the B-LCDR1 comprising The amino acid sequence of SEQ ID NO: 34, the B-LCDR2 comprises the amino acid sequence of SEQ ID NO: 32, and the B-LCDR3 comprises the amino acid sequence of SEQ ID NO: 35; or (iii) the B -HCDR1 comprises the amino acid sequence of SEQ ID NO: 18, the B-HCDR2 comprises the amino acid sequence of SEQ ID NO: 22, the B-HCDR3 comprises the amino acid sequence of SEQ ID NO: 23, and the B- LCDR1 comprises the amino acid sequence of SEQ ID NO: 31, the B-LCDR2 comprises the amino acid sequence of SEQ ID NO: 32, and the B-LCDR3 comprises the amino acid sequence of SEQ ID NO: 36; or (iv) The B-HCDR1 comprises the amino acid sequence of SEQ ID NO: 18, The B-HCDR2 comprises the amino acid sequence of SEQ ID NO: 24, the B-HCDR3 comprises the amino acid sequence of SEQ ID NO: 20, and the B-LCDR1 comprises the amino acid sequence of SEQ ID NO: 37, the B-LCDR2 comprises the amino acid sequence of SEQ ID NO: 32, and the B-LCDR3 comprises the amino acid sequence of SEQ ID NO: 38; or (v) the B-HCDR1 comprises the amino acid sequence of SEQ ID NO: 18 Sequence, the B-HCDR2 comprises the amino acid sequence of SEQ ID NO: 25, the B-HCDR3 comprises the amino acid sequence of SEQ ID NO: 20, and the B-LCDR1 comprises the amino acid sequence of SEQ ID NO: 39 , the B-LCDR2 comprises the amino acid sequence of SEQ ID NO: 32, and the B-LCDR3 comprises the amino acid sequence of SEQ ID NO: 40; or (vi) the B-HCDR1 comprises the amine of SEQ ID NO: 18 Amino acid sequence, the B-HCDR2 comprises the amino acid sequence of SEQ ID NO: 26, the B-HCDR3 comprises the amino acid sequence of SEQ ID NO: 27, and the B-LCDR1 comprises the amino acid sequence of SEQ ID NO: 41 acid sequence, the B-LCDR2 comprises the amino acid sequence of SEQ ID NO: 32, and the B-LCDR3 comprises the amino acid sequence of SEQ ID NO: 42; or (vii) the B-HCDR1 comprises the amino acid sequence of SEQ ID NO: 18 The amino acid sequence of the B-HCDR2 comprises the amino acid sequence of SEQ ID NO: 30, the B-HCDR3 comprises the amino acid sequence of SEQ ID NO: 20, and the B-LCDR1 comprises the amino acid sequence of SEQ ID NO: 31 Amino acid sequence, the B-LCDR2 comprises the amino acid sequence of SEQ ID NO: 32, and the B-LCDR3 comprises the amino acid sequence of SEQ ID NO: 33.

In some embodiments, the antigen-binding molecule that specifically binds to a BAFF antigen according to any one of the above, wherein the B-VH comprises at least 90% (eg, at least 90%, 95%, 96%) of SEQ ID NO: 63 , 97%, 98% or 99%) sequence identity of the amino acid sequence, the B-VL comprising and SEQ ID NO: 64 has at least 90% (such as at least 90%, 95%, 96%, 97%, 98% or 99%) sequence identity amino acid sequence, wherein SEQ ID NO: 63 is:

SEQ ID NO: 64 is:

，其中，X₁選自S、A、V或G，X₂選自A、I、M或S，X₃選自T或G，X₄選自Q、E、G或K，X₅選自N、G或Q，X₆選自L或P，X₇選自H、Q或D，X₈選自H或D，X₉選自A或G，X₁₀選自S或L，X₁₁選自P或S，X₁₂選自Q或H，X₁₃選自D、A或N，X₁₄選自S或I，X₁₅選自K、R或T，X₁₆選自S、W、T或D，X₁₇選自H、Y或S，X₁₈選自Y或R，X₁₉選自G或S，X₂₀選自A或D，X₂₁選自E或S，X₂₂選自S或A，X₂₃選自V、E、A、N或W，X₂₄選自G、K、H或R，X₂₅選自L或V，X₂₆選自G或C，X₂₇選自R或G，X₂₈選自G或C；並且該B-VH和B-VL不包含如下可變區組合：B-VH包含SEQ ID NO：1的胺基酸序列，且B-VL包含SEQ ID NO：2的胺基酸序列。

, wherein, _X1 is selected from S, A, V or G, _X2 is selected from A, I, M or S, _X3 is selected from T or G, _X4 is selected from Q, E, G or K, _X5 is selected from From N, G or Q, X ₆ is selected from L or P, X ₇ is selected from H, Q or D, X ₈ is selected from H or D, X ₉ is selected from A or G, X ₁₀ is selected from S or L, X ₁₁ is selected from P or S, X ₁₂ is selected from Q or H, X ₁₃ is selected from D, A or N, X ₁₄ is selected from S or I, X ₁₅ is selected from K, R or T, X ₁₆ is selected from S, W , T or D, X ₁₇ is selected from H, Y or S, X ₁₈ is selected from Y or R, X ₁₉ is selected from G or S, X ₂₀ is selected from A or D, X ₂₁ is selected from E or S, X ₂₂ is selected from From S or A, X ₂₃ is selected from V, E, A, N or W, X ₂₄ is selected from G, K, H or R, X ₂₅ is selected from L or V, X ₂₆ is selected from G or C, X ₂₇ is selected from From R or G, X ₂₈ is selected from G or C; and the B-VH and B-VL do not comprise the following variable region combination: B-VH comprises the amino acid sequence of SEQ ID NO: 1, and B-VL comprises Amino acid sequence of SEQ ID NO:2.

In some embodiments, the antigen-binding molecule that specifically binds to a BAFF antigen as described in any one of the above,

(i) the B-VH 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: 47, and the B-VL 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: 48, or

(ii) the B-VH 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: 5, and the B-VL 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: 12, or

(iii) the B-VH 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: 6, and the B-VL 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: 13, or

(iv) the B-VH 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: 7, and the B-VL 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: 14, or

(v) the B-VH 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: 8, and the B-VL 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: 15, or

(vi) the B-VH 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: 9, and the B-VL package 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: 16, or

(vii) the B-VH 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: 10, and the B-VL 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: 17, or

(viii) the B-VH 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: 11, and the B-VL 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: 2;

In some embodiments, the antigen-binding molecule that specifically binds a BAFF antigen as described in any one of the above, the B-VH comprises the amino acid sequence of SEQ ID NO: 47 and the B-VL comprises the amine of SEQ ID NO: 48 amino acid sequence, or the B-VH comprises the amino acid sequence of SEQ ID NO: 5, and the B-VL comprises the amino acid sequence of SEQ ID NO: 12; or the B-VH comprises the amino acid sequence of SEQ ID NO: 6 Amino acid sequence, and the B-VL comprises the amino acid sequence of SEQ ID NO: 13; or the B-VH comprises the amino acid sequence of SEQ ID NO: 7, and the B-VL comprises the amino acid sequence of SEQ ID NO: 14 or the B-VH comprises the amino acid sequence of SEQ ID NO: 8, and the B-VL comprises the amino acid sequence of SEQ ID NO: 15; or the B-VH comprises the amino acid sequence of SEQ ID NO: 9, and the B-VL comprises the amino acid sequence of SEQ ID NO: 16; or the B-VH comprises the amino acid sequence of SEQ ID NO: 10, and the B-VL comprises the amino acid sequence of SEQ ID NO : the amino acid sequence of 17; or the B-VH comprises the amino acid sequence of SEQ ID NO: 11, and the B-VL comprises the amino acid sequence of SEQ ID NO: 2.

In some embodiments, the antigen-binding molecule that specifically binds to a BAFF antigen as described in any one of the above includes a heavy chain constant region CH and a light chain constant region CL. In some embodiments, the heavy chain constant region CH is a human IgG heavy chain constant region and the light chain constant region CL is a human lambda or kappa light chain constant region. In some embodiments, the heavy chain constant region CH is a human IgG1 heavy chain constant region and the light chain constant region CL is a human lambda light chain constant region. In some embodiments, the heavy chain constant region CH is a human IgG1 heavy chain constant region, and the light chain constant region CL is a human kappa light chain constant region. In some embodiments, the heavy chain constant region CH comprises the amino acid sequence of SEQ ID NO:45, and the light chain constant region CL comprises the amino acid sequence of SEQ ID NO:46. In some embodiments, the antigen binding molecule that specifically binds a BAFF antigen is a full length antibody. In some embodiments, the heavy chain constant region of the antigen-binding molecule that specifically binds a BAFF antigen has one or more amino acid substitutions that reduce the binding of its Fc region to an Fc receptor. In some embodiments, the Fc region has YTE mutations (M252Y, S254T, and T256E), L234A, L235A mutations, and/or S228P mutations, the mutations are numbered according to the EU index.

In some embodiments, an antigen-binding molecule as described in any one of the above (for example, an antigen-binding molecule comprising an antigen-binding moiety 1 that specifically binds BAFF and an antigen-binding moiety 2 that specifically binds IL-12 and/or IL-23, Or an antigen-binding molecule that specifically binds to a BAFF antigen), the antigen-binding molecule has at least one of the following characteristics (for example, comprising any 1, 2, 3, 4, 5, 6 or 7 characteristics of the following A-G):

A. The antigen-binding molecule can block the binding of human BAFF to human BAFF-R; preferably, the IC50 value for blocking the binding of human BAFF to human BAFF-R is less than 11 nM (for example, less than 11 nM, less than or equal to 10 nM, less than or equal to Equal to 9nM, less than or equal to 8nM, less than or equal to 7 nM, less than or equal to 6nM, less than or equal to 5nM or less), the IC50 value is detected by the Elisa method; in some embodiments, the IC50 value is detected according to the method of Test Example 2 of the present disclosure;

B. The antigen-binding molecule can inhibit BAFF-induced B cell proliferation; preferably, the antigen-binding molecule can inhibit BAFF-induced B cell proliferation; , less than or equal to 0.10 nM, less than or equal to 0.09 nM, less than or equal to 0.08 nM, less than or equal to 0.07 nM, less than or equal to 0.06 nM or less) inhibits BAFF-induced B cell proliferation; in some embodiments , the IC50 value is detected according to the method of Test Example 3 disclosed herein;

C. The antigen binding molecule can be less than 9.99E-10M (such as less than 9.99E-10M, less than or equal to 9.00E-10M, less than or equal to 8.20E-10M, less than or equal to 7.40E-10M, less than or equal to 6.60E-10M, 10M, less than or equal to 5.20E-10M, less than or equal to 4.00E-10M, less than or equal to 3.00E-10M or less) combined with human BAFF, the KD value is detected by Biacore method; in some embodiments Among them, the KD value is detected according to the method of Test Example 5 of this disclosure;

D. The antigen-binding molecule can bind to cynomolgus monkey BAFF with a KD value of less than 5.00E-10M (for example, less than or equal to 4.00E-10M, less than or equal to 3.00E-10M or less), and the KD value is determined by Biacore Method detection; In some embodiments, the KD value is detected according to the method of Test Example 5 of the present disclosure;

E. The antigen-binding molecule can bind to mouse BAFF with a KD value of less than 2.20E-10M (for example, less than or equal to 2.00E-10M, less than or equal to 1.00E-10M or less), and the KD value is detected by Biacore method ; In some embodiments, the KD value is detected according to the method of Test Example 5 of the present disclosure;

F. The antigen-binding molecule can block the binding of human BAFF to human BCMA; preferably, the IC50 value for blocking the binding of human BAFF to human BCMA is less than 0.9nM (for example, less than or equal to 0.4nM, less than or equal to 0.35nM, less than or equal to 0.31nM or less), the IC50 value is detected by the Elisa method; in some embodiments, the IC50 value is detected according to the method of Test Example 2 of the present disclosure; or

G. The antigen-binding molecule can block the binding of human BAFF to human TACI; preferably, the IC50 value of blocking the binding of human BAFF to human TACI is less than 0.6nM (for example, less than or equal to 0.55nM, less than or equal to 0.4nM, less than or equal to 0.33nM, less than or equal to 0.23nM or less), the IC50 value is detected by the Elisa method; in some embodiments, the IC50 value is detected according to the method of Test Example 2 of the present disclosure.

In some embodiments, the antigen-binding molecule according to any one of the above, which comprises an antigen-binding moiety 1 that specifically binds BAFF and an antigen-binding moiety 2 that specifically binds IL-12 and/or IL-23, It has at least one of the following characteristics (for example, contains any 1, 2, 3, 4 or 5 characteristics of the following H-L):

H. The antigen-binding molecule can block the binding of human IL-12/23 p40 to human IL-12Rβ1; preferably, the IC50 value for blocking the binding of human IL-12/23 p40 to human IL-12Rβ1 is less than 3.6nM ( For example, less than or equal to 3.0nM, less than or equal to 2.0nM, less than or equal to 1.0nM, less than or equal to 0.5nM, less than or equal to 0.23nM or less), the IC50 value is detected by the Elisa method; in some embodiments, The IC50 value is detected according to the method of Test Example 2 of this disclosure;

I. The antigen-binding molecule can inhibit the secretion of IFNγ induced by IL-12; preferably, the IC50 value of the antigen-binding molecule inhibiting the secretion of IFNγ induced by IL-12 is less than 3.0nM (for example, less than or equal to 2.6nM, Less than or equal to 1.6nM or less); In some embodiments, the IC50 value is detected according to the method of Test Example 4 of the present disclosure;

J. The antigen-binding molecule can inhibit the secretion of IL-17 induced by IL-23; preferably, the IC50 value of the antigen-binding molecule inhibiting the secretion of IL-17 induced by IL-23 is less than 0.034nM (for example, less than or equal to 0.031nM, less than or equal to 0.030 nM or less); In some embodiments, the IC50 value is detected according to the method of Test Example 4 of the present disclosure;

K. The antigen-binding molecule can bind to cynomolgus monkey IL-12/23 p40 with a KD value of less than 2.7E-10M (for example, less than or equal to 2.0E-10M, less than or equal to 1.0E-10M or less), the KD The value is detected by the Biacore method; in some embodiments, the KD value is detected according to the method of Test Example 5 of the present disclosure; or

L. The antigen-binding molecule can bind to human IL-23 with a KD value of less than 6.4E-11M (for example, less than or equal to 6.3E-11M, less than or equal to 6.2E-11M or less), and the KD value is determined by the Biacore method Detection; In some embodiments, the KD value is detected according to the method of Test Example 5 of the present disclosure.

In another aspect, the present disclosure provides a pharmaceutical composition comprising: a therapeutically effective amount of any one of the above antigen-binding molecules, and one or more pharmaceutically acceptable carriers, diluents, buffers or excipients agent.

In another aspect, the present disclosure provides an isolated nucleic acid encoding an antigen-binding molecule according to any one of the above.

In another aspect, the present disclosure provides a host cell comprising the nucleic acid of any one of the above.

In another aspect, the present disclosure provides a method for treating a disease, the method comprising the step of administering the antigen-binding molecule or the pharmaceutical composition as described above to a subject.

In some embodiments, the disease is a B cell disorder or an autoimmune disease. In some embodiments, the B cell disorder or autoimmune disease is a disease or condition associated with BAFF expression. In some embodiments, 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 inflammation and psoriatic arthritis; the 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 gammaglobulinopathy. In some embodiments, the autoimmune disease is systemic lupus erythematosus.

On the other hand, the present disclosure also provides the use of the antigen-binding molecule or the pharmaceutical composition according to any one of the above in the preparation of a medicament for treating the aforementioned diseases.

In another aspect, the present disclosure also provides an antigen-binding molecule or a pharmaceutical composition as described above for use as a medicament.

Figure 1 is a schematic diagram of the structure of BU-1.

Figure 2 is a schematic diagram of the structure of BU-2.

Fig. 3 is a graph showing the results of an experiment on inhibiting TNFα secretion by a bispecific antibody specifically binding to BAFF and IL-12/23.

Fig. 4 is a graph showing the results of an experiment on inhibiting IgA secretion by a bispecific antibody specifically binding to BAFF and IL-12/23.

Fig. 5 is a diagram showing the results of an experiment on inhibiting IL-22 secretion by a bispecific antibody that specifically binds to BAFF and IL-12/23.

Fig. 6 is a diagram showing the results of an experiment of inhibiting IFNγ secretion by a bispecific antibody specifically binding to BAFF and IL-12/23.

the term

To facilitate understanding of the present disclosure, certain technical and scientific terms are described below. Unless defined otherwise herein, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art.

As used in the specification and claims, the singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise.

Unless the context clearly requires otherwise, in the patent specification and scope of claims, the words "comprising", "having", "comprising", etc. should be understood as "including but not limited to", rather than an exclusive or exhaustive meaning .

The term "cytokine" is a general term for proteins/polypeptides released by a population of cells that act as intercellular mediators on other cells. Examples of such cytokines include lymphokines, monokines, chemokines and traditional polypeptide hormones. Exemplary cytokines include: IL-2, IFN-γ, IL-6, TNFα, IL-17, and IL-5.

The term "and/or" means to include both "and" and "or". For example the phrase "A, B and/or C" is intended to cover each of the following: A, B and C; A, B or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone). In this disclosure, unless otherwise specified, "IL-12 and/or IL-23" or "IL-12/23" are meant to cover: "IL-12", "IL-23" and "IL-12 and IL-23" -23" each.

The three-letter and one-letter codes for amino acids used in this disclosure are as described in J.biol.chem, 243, p3558 (1968).

The term "amino acid" refers to naturally occurring and synthetic amino acids, as well as amino acid analogs and amino acid mimetics that function in a similar manner to naturally occurring amino acids. Naturally occurring amino acids are those encoded by the genetic code, as well as those that are later modified, eg, hydroxyproline, γ-carboxyglutamate, and O-phosphoserine. Amino acid analogs are compounds that have the same basic chemical structure (i.e., alpha carbon bonded to hydrogen, carboxyl, amine and R groups) as naturally occurring amino acids, such as homoserine, norleucine, Methionine imine, methionine methyl peroxide. 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.

The term "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 terminus and/or carboxyl terminus of the polypeptide chain. Specific amino acid mutations may be amino acid substitutions. In one embodiment, the amino acid mutation Mutations are non-conservative amino acid substitutions in which one amino acid is replaced with another amino acid having different structural and/or chemical properties. Amino acid substitutions include non-naturally occurring amino acids or derivatives of 20 natural amino acids (e.g., 4-hydroxyproline, 3-methylhistidine, ornithine, homoserine, 5 -Hydroxylysine) replacement. 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. Methods of altering amino acid side chain groups other than genetic engineering, such as chemical modification, may also be available. Various names may be used herein to refer to mutations of the same amino acid. Herein, the 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 position 366 is mutated from the original T to W.

The term "antigen-binding molecule" is used in the broadest sense and covers various molecules that specifically bind to an antigen, including but not limited to antibodies, other polypeptides with antigen-binding activity, and antibody fusion proteins fused thereto. Exemplarily, the antigen-binding molecules herein are bispecific antigen-binding molecules (eg, bispecific antibodies). The term "bispecific antigen binding molecule" refers to an antigen binding molecule capable of specifically binding to two different antigens or at least two different epitopes of the same antigen.

The term "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 proteins of approximately 150,000 Daltons, composed of two light chains and two heavy chains joined by disulfide bonds. Each heavy chain has a variable region from N to C-terminus (VH), also known as variable heavy domain, heavy chain variable region, followed by heavy chain constant region (CH), the native IgG heavy chain constant region usually contains three constant domains (CH1, CH2 and CH3). Similarly, from N to C-terminus, each light chain has a variable region (VL, also known as variable light domain, or light chain variable domain), followed by a constant light domain (light chain constant region, CL ). The terms "full-length antibody", "intact antibody" and "whole antibody" are used interchangeably herein to refer to an antibody having a structure substantially similar to that of a native antibody or having a heavy chain with an Fc region as defined herein. Natural whole 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 κ chain and λ chain; heavy chain includes variable region VH and constant region (CH1, CH2 and CH3), VH is at the amino terminal of the heavy chain (also known as N-terminal), the constant region is at the carboxy-terminal (also known as C-terminal), wherein CH3 is closest to the carboxyl-terminal 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.

The term "bispecific antibody" refers to an antibody (including an antibody or an antigen-binding fragment thereof, such as a single-chain antibody) capable of specifically binding to two different antigens or two different epitopes of the same antigen. Bispecific antibodies of various structures have been disclosed in the prior art. According to the integrity of the IgG molecule, it can be divided into IgG-like bispecific antibodies and antibody fragment bispecific antibodies. According to the number of antigen-binding regions, bispecific antibodies can be divided into bivalent, trivalent, tetravalent or more valent. According to whether the structure is symmetrical, it can be divided into symmetrical structure bispecific antibody and asymmetric structure bispecific antibody. Among them, fragment-type bispecific antibodies, such as Fab fragments lacking Fc fragments, which form bispecific antibodies by combining two or more Fab fragments in one molecule, have lower immunogenicity, and have a lower molecular weight. Small, with high tumor tissue permeability, typical antibody structures of this type are F(ab) ₂ , scFv-Fab, (scFv) ₂ -Fab, etc. IgG-like bispecific antibodies (for example, with Fc fragments), such antibodies have a relatively large molecular weight, and the Fc fragments are helpful for the purification of antibodies and improve their solubility and stability. The Fc part may also bind to the receptor FcRn, Increase antibody serum half-life. Typical structural models of bispecific antibodies such as KiH, CrossMAb, Triomab quadroma, FcΔAdp, ART-Ig, BiMAb, Biclonics, BEAT, DuoBody, Azymetric, XmAb, 2:1 TCBs, 1Fab-IgG TDB, FynomAb, two-in- one/DAF, scFv-Fab-IgG, DART-Fc, LP-DART, CODV-Fab-TL, HLE-BiTE, F(ab)2-CrossMAb, IgG-(scFv)2, Bs4Ab, DVD-Ig, Tetravalent - Bispecific antibodies such as DART-Fc, (scFv)4-Fc, CODV-Ig, mAb2, F(ab)4-CrossMAb (see Aran F. Labrijn et al., Nature Reviews Drug Discovery volume 18, pages 585-608 (2019 ); Chen S1 et al., J Immunol Res. 2019 Feb 11; 2019: 4516041).

The term "variable region" or "variable domain" refers to the antigen-binding domain of an antigen-binding molecule (eg, an antibody). Herein, the heavy chain variable region VH and the light chain variable region VL each comprise four conserved framework regions (FRs) and three complementarity determining regions (CDRs). Herein, the heavy chain variable region in the antigen-binding module that specifically binds BAFF is marked as B-VH, and the light chain variable region is marked as B-VL; the antigen-binding module that specifically binds IL-12/23 p40 The heavy chain variable region is denoted as P-VH and the light chain variable region as P-VL. The term "complementarity determining region" or "CDR" refers to the region within a variable domain that primarily contributes to antigen binding; "framework" or "FR" refers to the variable domain residues other than the CDR residues. 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 terminal (also called N-terminal) to the carboxy-terminal (also called C-terminal): FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. each VH and VL are composed of three CDRs and four FRs arranged in the following order from amino terminus to carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. A single VH or VL may be sufficient to confer antigen binding specificity. In this disclosure, the three CDR regions in B-VH are respectively marked as B-HCDR1, B-HCDR2 and B-HCDR3, or Bv-HCDR1, Bv-HCDR2 and Bv-HCDR3; the three CDR regions in B-VL Labeled as B-LCDR1, B-LCDR2, and B-LCDR3, or Bv-LCDR1, Bv-LCDR2, and Bv-LCDR3, respectively. In addition, the three CDR regions in P-VH are respectively marked as P-HCDR1, P-HCDR2 and P-HCDR3, or Pv-HCDR1, Pv-HCDR2 and Pv-HCDR3; the three CDR regions in P-VL are respectively marked as P-LCDR1, P-LCDR2, and P-LCDR3, or Pv-LCDR1, Pv-LCDR2, and Pv-LCDR3.

The 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]. 2001) and ImMunoGenTics (IMGT ) numbering rules (Lefranc, M.P., etc., Dev.Comp.Immunol., 27, 55-77 (2003); Front Immunol.2018 Oct 16; 9: 2278), etc.; the corresponding relationship between various numbering systems is the technical field Exemplary ones that are well known to those of ordinary skill are shown in Table 1 below.

Table 1. Relationship between CDR numbering systems

Unless otherwise stated, the "Kabat" numbering convention is used for variable region and CDR sequences in this disclosure. Although in specific embodiments, the Kabat numbering rules are used to define amino acid residues, the corresponding technical solutions in other numbering systems will be regarded as equivalent technical solutions.

The term "antibody fragment" refers to a molecule other than an intact antibody that comprises a portion of an intact antibody that retains the antigen-binding ability of the intact antibody. Examples of 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 (scFv) , and multispecific antibodies formed from antibody fragments.

The term "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. In some embodiments, the Fc region comprises the same or different two subunits. In some embodiments, 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 with the antibodies herein include human IgG1, IgG2 (IgG2A, IgG2B), IgG3 and IgG4 Fc region. In some embodiments, 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 (according to Residues 446 and 447 of the EU numbering system). Unless otherwise stated, the numbering convention for the Fc region is the EU numbering system, also known as the EU index.

The term "chimeric" antibody refers to an antibody in which a portion of the heavy and/or light chains is from a particular source or species and the remaining portion of the heavy and/or light chains is from another, different source or species.

The term "humanized" antibody is an antibody that retains the reactivity of a non-human antibody while being less immunogenic in humans. Humanization 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).

The terms "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.

The term "affinity" refers to the overall strength of the non-covalent interaction between a single binding site of a molecule (eg, an antigen-binding molecule of the disclosure) and its binding partner (eg, an antigen). Unless otherwise indicated, as used herein, binding "affinity" refers to internal binding affinity, It 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.

As used 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. The term "KD" refers to the dissociation constant, which is obtained from the ratio of kd to ka (ie, kd/ka) and is expressed as a molar concentration (M). The KD value of an antibody can be determined using methods well known in the art. For example, use biosensing systems such as the system to measure surface plasmon resonance, or measure affinity in solution by solution equilibrium titration (SET). In some embodiments, the KD value is detected by Biacore.

The term "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 antibody isotype. Examples of 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 antibodies or members thereof that are substantially homogeneous, ie, the antibody molecules comprised in the population are identical in amino acid sequence, except for natural mutations that may be present in minor amounts. In contrast, 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. In some embodiments, the antibodies provided by the present disclosure are monoclonal antibodies.

The term "antigen" refers to a molecule or portion of a molecule capable of being selectively bound by an antigen-binding molecule (eg, an antibody). An antigen may have one or more epitopes capable of interacting with different antigen-binding molecules (eg, antibodies).

The term "epitope" refers to an area (area or region) on an antigen capable of specifically binding to an antibody or antigen-binding fragment thereof. An epitope may be formed from contiguous amino acids (linear epitope) or comprise non-contiguous amino acids (conformational epitope), for example, non-contiguous amino acids resulting from folding of the antigen (i.e., by tertiary folding of the antigen). close in space. 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. Antibodies that bind to a particular epitope (i.e., those that bind to the same epitope) can be screened using methods routine in the art, such as, but not limited to, alanine scanning, peptide blotting, peptide cleavage analysis, epitope excision, epitope extraction, antigenicity Chemical modification (see Prot. Sci. 9 (2000) 487-496), and cross-blocking.

The term "capable of specifically binding", "specifically binds" or "binds" means that an antigen-binding molecule (eg, an antibody) is capable of binding to a certain antigen or epitope thereof with higher affinity than to other antigens or epitopes. Typically, an antibody binds an antigen or an epitope thereof with an equilibrium dissociation constant (KD) of about 1×10 ⁻⁷ M or less (eg, about 1×10 ⁻⁸ M or less). In some embodiments, 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, such as by ^BIACORE® surface plasmon resonance assay. However, an antibody that specifically binds to an antigen or an epitope thereof does not exclude cross-reactivity to other related antigens, e.g. The corresponding antigens of cynomolgus, cyno), chimpanzee (Pan troglodytes) (chimpanzee, chimp)) or marmoset (Callithrix jacchus) (commonmarmoset, marmoset) are cross-reactive.

The term "antigen binding moiety" refers to a polypeptide molecule that specifically binds an antigen of interest. Antigen binding moieties include antibodies and fragments thereof as otherwise defined herein. In some embodiments, the antigen binding moiety comprises an antigen binding domain of an antibody comprising an antibody heavy chain variable region and an antibody light chain variable region. The term "antigen-binding moiety that specifically binds BAFF" refers to an antigen-binding moiety that is capable of binding BAFF with sufficient affinity. In certain embodiments, the antigen binding moiety that specifically binds human BAFF has an equilibrium dissociation constant (KD) of less than 10.00E-10M (e.g., less than 9.99E-10M, less than or equal to 9.00E-10M, less than or equal to 8.20 E-10M, less than or equal to 7.40E-10M, less than or equal to 6.60E-10M, less than or equal to 5.20E-10M, less than or equal to 4.00E-10M, less than or equal to 3.00E-10M or less), which is Measured by Biacore method. In certain embodiments, an antigen binding moiety that specifically binds BAFF binds a conserved epitope in BAFF from a different species. The term "antigen-binding moiety that specifically binds IL-12/23" refers to an antigen-binding moiety capable of binding IL-12/23 with sufficient affinity. In certain embodiments, the antigen binding moiety that specifically binds IL-12/23 has an equilibrium dissociation constant (KD) of less than 6.4E-11M (e.g., less than or equal to 6.3E-11M, less than or equal to 6.2E-11M, 11M or less) for binding to human IL-23, which was detected by the Biacore method. In certain embodiments, the antigen binding moiety that specifically binds IL-12/23 binds a conserved epitope in IL-12/23 from a different species. Antigen binding moieties include antibody fragments as defined herein, eg Fab or scFv.

In this context, the ordinal numbers in "antigen-binding module 1" or "antigen-binding module 2" are only used to distinguish different technical features and chemical entities; they do not limit any order, level or quantity.

The term "linker", "Linker" or "linker" refers to a linking unit that joins two polypeptide fragments. Herein, 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. When "-" appears in the structural formula, it means that the units on both sides are directly linked by covalent bonds. When the term "bond" appears in a structural unit, it means that the unit has no amino acids, and the units on either side of the unit are directly connected.

The terms "antibody-dependent cellular cytotoxicity", "antibody-dependent cell-mediated cytotoxicity" or "ADCC" are mechanisms for inducing cell death that rely on the interaction of antibody-coated target cells with lytically active effector cells (such as natural killer cells (NK), monocytes, macrophages, and neutrophils) via Fcγ receptors (FcγRs) expressed on effector cells. For example, NK cells express FcyRIIIa, while 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 markers released from lysed cells, such as radioacceptors, fluorescent dyes, or native intracellular proteins.

The term "antibody-dependent cellular phagocytosis" ("ADCP") refers to the mechanism by which antibody-coated target cells are eliminated by internalization by phagocytes, such as macrophages or dendritic cells.

The term "complement-dependent cytotoxicity" or "CDC" refers to a mechanism that induces cell death in which the Fc effector domains of bound antibodies on target cells bind and activate complement component C1q, which in turn activates the complement cascade, resulting in target cell die. 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.

The term "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 a reference nucleic acid, and operating in a manner similar to Metabolized in the manner of reference nucleotides. Examples of such analogs include, but are not limited to, phosphorothioate, phosphoroamidate, methylphosphonate, chiral-methylphosphonate, 2-O-methylribonucleotide, peptide-nucleic acid ( PNA).

An "isolated" 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 an antigen-binding molecule refers to one or more nucleic acid molecules encoding an antigen-binding molecule, including such one or more nucleic acid molecules in a single vector or in separate vectors, and one or more nucleic acid molecules present in a host cell. Such one or more nucleic acid molecules at more positions. Unless otherwise indicated, 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. Specifically, as detailed below, degenerate codon substitutions can be obtained by generating sequences in which In the sequence, the third position of one or more selected (or all) codons is replaced by a degenerate base and/or a deoxyinosine residue.

The terms "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 the corresponding artificial chemical mimetic of a naturally occurring amino acid, as well as to naturally occurring amino acid polymers and non-naturally occurring amino acid polymer. Unless otherwise stated, a particular polypeptide sequence also implicitly encompasses conservatively modified variants thereof.

The term sequence "identity" refers to the degree (percentage) to which the amino acids/nucleic acids of two sequences are identical at equivalent positions when the two sequences are optimally aligned; Gaps are allowed to achieve the maximum percent sequence identity, and any conservative substitutions are not considered part of the sequence identity. To determine percent sequence identity, 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 of ordinary skill 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 terms "fused" or "linked" refer to the joining of components (eg, B-VH and B-VL) by covalent bonds, either directly or via one or more linkers.

The term "vector" means a polynucleotide molecule capable of transporting another polynucleotide to which it has been linked. One type of vector is a "plasmid," which refers to a circular double-stranded DNA loop into which additional DNA segments can be ligated. Another type of vector is a viral vector, such as an adeno-associated viral vector (AAV or AAV2), in which additional DNA segments can be ligated into the viral genome. Certain vectors are capable of autonomous replication in the host cells into which they are introduced (e.g., with bacterial vectors and episomal mammalian vectors with a bacterial origin of replication). Other vectors (eg, non-episomal mammalian vectors) can integrate into the genome of the host cell after introduction into the host cell, thereby replicating along with the host genome. The term "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. A vector of nucleic acid sequences. 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.

The terms "host cell", "host cell line" and "host cell culture" are used interchangeably and refer 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. Herein, the term includes mutant progeny, including cells having the same function or biological activity as those screened or selected in the primary transformed cells. 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, rat, 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 salictaria, Pichia guercuum, Pichia pijperi, Pichia Pichia stiptis, Pichia methanolica, Pichia sp., Saccharomyces cerevisiae, Saccharomyces cerevisiae, Hansenula polymorpha, Kluyveromyces gram Kluyveromyces lactis, Candida albicans, Aspergillus nidulans, Aspergillus niger, Aspergillus oryzae, Trichoderma reesei, Lucker's Chrysosporium lucknowense, Fusarium sp., Fusarium gramineum, Fusarium venenatum, Physcomitrella patens and Neurospora crassa. Pichia, any Saccharomyces, Hansenula polymorpha, any Kluyveromyces, Candida albicans, any Aspergillus, Trichoderma reesei, Luke Mold (Chrysosporium lucknowense), any Fusarium species, Yarrowia lipolytica (Yarrowia lipolytica) and Neurospora crassa (Neurospora crassa).

The host cell cannot develop into a plant or individual animal.

"Optional" or "optionally" means that the subsequently described feature may but need not occur, and the indication includes instances where the feature occurs or does not occur.

The term "pharmaceutical composition" means a mixture comprising one or more antigen binding molecules described herein and other chemical components such as physiological/pharmaceutical acceptable carriers and excipients.

The term "pharmaceutically acceptable carrier (vehicle), diluent, buffer or excipient" refers to an ingredient in a pharmaceutical formulation (formulation) that is different from the active ingredient and is non-toxic to the subject.

The term "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. The terms "patient" or "subject" are used interchangeably herein unless otherwise indicated. As used herein, the term "cyno" or "cynomolgus" refers to Macaca fascicularis. In certain embodiments, the individual or subject is a human.

"Administration" or "administration", when applied to animals, humans, experimental subjects, cells, tissues, organs or biological fluids, refers to the interaction of exogenous drugs, therapeutic agents, diagnostic agents or compositions with animals, humans , subjects, cells, tissues, organs or biological fluids.

The term "sample" refers to a collection (such as a fluid, cell, or tissue) isolated from a subject, as well as a fluid, cell, or tissue present in a subject. Exemplary samples are biological fluids such as blood, serum and serosal fluids, plasma, lymph, urine, saliva, cyst fluid, tears, faeces, sputum, mucosal secretions of secretory tissues or 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 culture medium (including conditioned medium), lavage fluid, etc. , tissue biopsy sample, fine needle aspiration, surgically removed tissue, organ culture, or cell culture.

"Treatment" and "treatment" (and grammatical variants thereof) refer to clinical intervention applied to the individual being treated, and may be for prophylactic purposes, or in clinical pathology implemented during the learning process. 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 improved prognosis . In some embodiments, the molecules of the present 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 those symptoms and/or their 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 amount. In some embodiments, 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 retard, delay or reverse the progression of the disease state or any other undesirable symptom associated with the disease quantity.

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. Thus, 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.

target molecule

"BAFF" should be broadly understood and is intended to cover various forms of molecules of BAFF in various stages of mammalian body, such as but not limited to BAFF gene in amplification, replication, transduction molecules produced during transcription, splicing, processing, translation, modification (e.g., precursor BCMA, mature BAFF, membrane-expressed BAFF, BAFF splice variants, modified BAFF, or fragments thereof); the term also covers artificially produced or BAFF expressed in vitro.

"IL-12" should be broadly understood and is intended to cover various forms of IL-12 molecules in various stages in mammals, such as but not limited to IL-12 gene in amplification, replication, transcription, splicing, processing, Molecules produced during translation, modification (e.g. precursor IL-12, mature IL-12, membrane expressed IL-12, IL-12 splice variants, modified IL-12, or fragments, subunits thereof); The term also encompasses artificially produced or in vitro expressed IL-12.

"IL-23" should be broadly understood and is intended to cover various forms of IL-23 molecules in various stages of mammalian body, such as but not limited to IL-23 gene in amplification, replication, transcription, splicing, processing, Molecules produced during translation, modification (e.g. precursor IL-23, mature IL-23, membrane expressed IL-23, IL-23 splice variants, modified IL-23, or fragments, subunits thereof); The term also encompasses artificially produced or in vitro expressed IL-23.

Antigen binding molecules of the present disclosure

In one aspect, the disclosure obtains an antigen-binding molecule that specifically binds BAFF by modifying Belimumab. In addition, antigen-binding molecules that specifically bind BAFF and IL-12/23 were also constructed. In some embodiments, the antigen-binding molecules of the present disclosure have many favorable properties, such as binding to antigen with high affinity, blocking the binding of BAFF to its receptors (such as BAFF-R, BCMA and/or TACI), inhibiting BAFF-induced B cell proliferation, blocking IL-12/23 binding to its receptor, inhibiting IL-12-induced IFNγ secretion, inhibiting IL-23-induced IL-17 secretion, pharmacokinetic properties and/or druggability, etc.

In some embodiments, the present disclosure provides an antigen binding molecule that specifically binds BAFF, which is an anti-BAFF antibody. In some embodiments, the antibody is a full-length antibody or an antigen-binding fragment thereof (e.g., Fv, Fab, Fab', Fab'-SH, F(ab ' )2, single domain antibody, single chain Fab (scFab), diabody , linear antibody, single-chain antibody (scFv)), which has one or more of the following functional activities:

A. The antigen-binding molecule can block the binding of human BAFF to human BAFF-R; preferably, the IC50 value for blocking the binding of human BAFF to human BAFF-R is less than 11 nM (for example, less than 11 nM, less than or equal to 10 nM, less than or equal to Equal to 9nM, less than or equal to 8nM, less than or equal to 7nM, less than or equal to 6nM, less than or equal to 5nM or less), the IC50 value is detected by the Elisa method; in some embodiments, the IC50 value is according to the disclosed test example 2 method detection;

B. The antigen-binding molecule can inhibit BAFF-induced B cell proliferation; preferably, the antigen-binding molecule can inhibit BAFF-induced B cell proliferation; , less than or equal to 0.10 nM, less than or equal to 0.09 nM, less than or equal to 0.08 nM, less than or equal to 0.07 nM, less than or equal to 0.06 nM or less) inhibits BAFF-induced B cell proliferation; in some embodiments , the IC50 value is detected according to the method of Test Example 3 disclosed herein;

C. The antigen binding molecule can be less than 9.99E-10M (such as less than 9.99E-10M, less than or equal to 9.00E-10M, less than or equal to 8.20E-10M, less than or equal to 7.40E-10M, less than or equal to 6.60E-10M, 10M, less than or equal to 5.20E-10M, less than or equal to 4.00E-10M, less than or equal to 3.00E-10M or less) combined with human BAFF, the KD value is detected by Biacore method; in some embodiments Among them, the KD value is detected according to the method of Test Example 5 of this disclosure;

D. The antigen-binding molecule can bind to cynomolgus monkey BAFF with a KD value of less than 5.00E-10M (for example, less than or equal to 4.00E-10M, less than or equal to 3.00E-10M or less), and the KD value is determined by Biacore Method detection; In some embodiments, the KD value is detected according to the method of Test Example 5 of the present disclosure;

E. The antigen-binding molecule can bind to mouse BAFF with a KD value of less than 2.20E-10M (for example, less than or equal to 2.00E-10M, less than or equal to 1.00E-10M or less), and the KD value is detected by Biacore method ; In some embodiments, the KD value is detected according to the method of Test Example 5 of the present disclosure;

F. The antigen-binding molecule can block the binding of human BAFF to human BCMA; preferably, the IC50 value for blocking the binding of human BAFF to human BCMA is less than 0.9nM (for example, less than or equal to 0.4nM, less than or equal to 0.35nM, less than or equal to 0.31nM or less), the IC50 value is detected by the Elisa method; in some embodiments, the IC50 value is detected according to the method of Test Example 2 of the present disclosure; or

G. The antigen-binding molecule can block the binding of human BAFF to human TACI; preferably, the IC50 value of blocking the binding of human BAFF to human TACI is less than 0.6nM (for example, less than or equal to 0.55nM, less than or equal to 0.4nM, less than or equal to 0.33nM or less), the IC50 value is detected by the Elisa method; in some embodiments, the IC50 value is detected according to the method of Test Example 2 of the present disclosure.

In some embodiments, the present disclosure provides an antigen binding molecule that specifically binds BAFF and IL-12/23. In some embodiments, the antigen binding molecule is a bispecific antibody that specifically binds BAFF and IL-12/23 P40 subunits. In some embodiments, the antigen binding molecule has one or more of the following functional activities:

A. The antigen-binding molecule can block the binding of human BAFF to human BAFF-R; preferably, the IC50 value for blocking the binding of human BAFF to human BAFF-R is less than 11 nM (for example, less than 11 nM, less than or equal to 10 nM, less than or equal to Equal to 9nM, less than or equal to 8nM, less than or equal to 7nM, less than or equal to 6nM, less than or equal to 5nM or less), the IC50 value is detected by the Elisa method; in some embodiments, the IC50 value is according to the disclosed test example 2 method detection;

B. The antigen-binding molecule can inhibit BAFF-induced B cell proliferation; preferably, the antigen-binding molecule can inhibit BAFF-induced B cell proliferation; , less than or equal to 0.10 nM, less than or equal to 0.09 nM, less than or equal to 0.08 nM, less than or equal to 0.07 nM, less than or equal to 0.06 nM or less) inhibits BAFF-induced B cell proliferation; in some embodiments , the IC50 value is detected according to the method of Test Example 3 disclosed herein;

C. The antigen binding molecule can be less than 9.99E-10M (such as less than 9.99E-10M, less than or equal to 9.00E-10M, less than or equal to 8.20E-10M, less than or equal to 7.40E-10M, less than or equal to 6.60E-10M, 10M, less than or equal to 5.20E-10M, less than or equal to 4.00E-10M, less than or equal to 3.00E-10M or less) combined with human BAFF, the KD value is detected by Biacore method; in some embodiments Among them, the KD value is detected according to the method of Test Example 5 of this disclosure;

D. The antigen-binding molecule can bind to cynomolgus monkey BAFF with a KD value of less than 5.00E-10M (for example, less than or equal to 4.00E-10M, less than or equal to 3.00E-10M or less), and the KD value is determined by Biacore Method detection; In some embodiments, the KD value is detected according to the method of Test Example 5 of the present disclosure;

E. The antigen-binding molecule can bind to mouse BAFF with a KD value of less than 2.20E-10M (for example, less than or equal to 2.00E-10M, less than or equal to 1.00E-10M or less), and the KD value is detected by Biacore method ; In some embodiments, the KD value is detected according to the method of Test Example 5 of the present disclosure;

F. The antigen-binding molecule can block the binding of human BAFF to human BCMA; preferably, the IC50 value for blocking the binding of human BAFF to human BCMA is less than 0.9nM (for example, less than or equal to 0.4nM, less than or equal to 0.35nM, less than or equal to 0.31nM or less), the IC50 value is detected by the Elisa method; in some embodiments, the IC50 value is detected according to the method of Test Example 2 of the present disclosure;

G. The antigen-binding molecule can block the binding of human BAFF to human TACI; preferably, the IC50 value of blocking the binding of human BAFF to human TACI is less than 0.6nM (for example, less than or equal to 0.55nM, less than or equal to 0.4nM, less than or equal to 0.33nM or less), the IC50 value is detected by the Elisa method; in some embodiments, the IC50 value is detected according to the method of Test Example 2 of the present disclosure;

H. The antigen-binding molecule can block the binding of human IL-12/23 p40 to human IL-12Rβ1; preferably, the IC50 value for blocking the binding of human IL-12/23 p40 to human IL-12Rβ1 is less than 3.6nM ( For example, less than or equal to 3.0nM, less than or equal to 2.0nM, less than or equal to 1.0nM, less than or equal to 0.5nM, less than or equal to 0.23nM or less), the IC50 value is detected by the Elisa method; in some embodiments, The IC50 value is detected according to the method of Test Example 2 of this disclosure;

I. Antigen-binding molecules can inhibit IL-12-induced IFNγ secretion; preferably, the IC50 value of antigen-binding molecules inhibiting IL-12-induced IFNγ secretion is less than 3.0nM (for example, less than or equal to 2.6nM, less than or equal to 1.6nM or less ); In some embodiments, the IC50 value is detected according to the method of Test Example 4 of the present disclosure;

J. The antigen-binding molecule can inhibit the secretion of IL-17 induced by IL-23; preferably, the IC50 value of the antigen-binding molecule inhibiting the secretion of IL-17 induced by IL-23 is less than 0.034nM (for example, less than or equal to 0.031nM, less than or equal to 0.030 nM or less); In some embodiments, the IC50 value is detected according to the method of Test Example 4 of the present disclosure;

K. The antigen-binding molecule can bind to cynomolgus monkey IL-12/23 p40 with a KD value of less than 2.7E-10M (for example, less than or equal to 2.0E-10M, less than or equal to 1.0E-10M or less), the KD The value is detected by the Biacore method; in some embodiments, the KD value is detected according to the method of Test Example 5 of the present disclosure; or

L. The antigen-binding molecule can bind to human IL-23 with a KD value of less than 6.4E-11M (for example, less than or equal to 6.3E-11M, less than or equal to 6.2E-11M or less), and the KD value is determined by the Biacore method Detection; In some embodiments, the KD value is detected according to the method of Test Example 5 of the present disclosure.

In some embodiments, the antigen-binding molecule of any one of the above, which comprises a heavy chain variable region B-VH and a light chain variable region B-VL, the B-VH comprises B-HCDR1, B-HCDR2 and B-HCDR3, the B-VL comprises B-LCDR1, B-LCDR2 and B-LCDR3, wherein the amino acid sequences of the B-HCDR1, B-HCDR2 and B-HCDR3 are respectively the same as those of Bv in SEQ ID NO: 63 - the amino acid sequences of HCDR1, Bv-HCDR2 and Bv-HCDR3 are the same, and the amino acid sequences of B-LCDR1, B-LCDR2 and B-LCDR3 are respectively the same as those of Bv-LCDR1 and Bv-LCDR1 in SEQ ID NO: 64 The amino acid sequences of LCDR2 and Bv-LCDR3 are identical. In some embodiments, the B-HCDR1, B-HCDR2, B-HCDR3, B-LCDR1, B-LCDR2, B-LCDR3, Bv-HCDR1, Bv-HCDR2, Bv-HCDR3, Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 are defined according to the same numbering convention selected from Kabat, IMGT, Chothia, AbM and Contact.

In some embodiments, the antigen-binding molecule of any one of the above, the B-HCDR1, B-HCDR2, B-HCDR3, B-LCDR1, B-LCDR2, B-LCDR3 are defined according to the Kabat numbering rules, wherein , the amino acid sequence of B-HCDR1 is NNAIN (SEQ ID NO: 18), the amino acid sequence of B-HCDR2 is X ₁ IX ₂ PMFGX ₃ AKYSX ₄ X ₅ FQG (SEQ ID NO: 65), B-HCDR3 _{The amino} acid sequence _of B-LCDR1 is _{X 12} GX _{13 X 14} LX ₁₅ X ₁₆ X _{17 X 18} AS (SEQ ID NO: 67), the amino acid sequence of B-LCDR2 is GKNNRPS (SEQ ID NO: 32) and the amino acid sequence of B-LCDR3 is X ₁₉ SRX ₂₀ X ₂₁ X ₂₂ GX ₂₃ X ₂₄ WX ₂₅ (SEQ ID NO: 68), wherein, X ₁ is selected from S, A, V or G, X ₂ is selected from A, I, M or S, X ₃ is selected from T or G, X ₄ is selected from Q, E, G or K, _X5 is selected from N, G or Q, _X6 is selected from L or P, _X7 is selected from H, Q or D, _X8 is selected from H or D, _X9 is selected from A or G, _X10 Be selected from S or L, X ₁₁ is selected from P or S, X ₁₂ is selected from Q or H, X ₁₃ is selected from D, A or N, X ₁₄ is selected from S or I, X ₁₅ is selected from K, R or T, X ₁₆ is selected from S, W, T or D, X ₁₇ is selected from H, Y or S, X ₁₈ is selected from Y or R, X ₁₉ is selected from G or S, X ₂₀ is selected from A or D, X ₂₁ is selected from E or S, _X22 is selected from S or A, _X23 is selected from V, E, A, N or W, _X24 is selected from G, K, H or R, _X25 is selected from L or V. Preferably, _X1 to _X25 are not selected from the following combinations: _X1 is G, _X2 is I, _X3 is T, _X4 is Q, _X5 is N, _X6 is L, _X7 is H, X ₈ is H, X ₉ is A, X ₁₀ is S, X 11 is P, X ₁₂ is Q, X ₁₃ is _D , X ₁₄ is S, X ₁₅ is R, X ₁₆ is S, X ₁₇ is Y, X ₁₈ is Y, X ₁₉ is S, X ₂₀ is D, X ₂₁ is S, X ₂₂ is S, X ₂₃ is N, X ₂₄ is H, and X ₂₅ is V.

In some embodiments, (i) the B-HCDR1 is set forth in SEQ ID NO: 18, the B-HCDR2 is set forth in SEQ ID NO: 28, the B-HCDR3 is set forth in SEQ ID NO: 29, and the B-LCDR1 as shown in SEQ ID NO: 43, the B-LCDR2 as shown in SEQ ID NO: 32, and the B-LCDR3 as shown in SEQ ID NO: 44; or (ii) the B-HCDR1 as shown in SEQ ID NO: 18, the B-HCDR2 shown in SEQ ID NO: 21, the B-HCDR3 shown in SEQ ID NO: 20, and the B-LCDR1 shown in SEQ ID NO: 34, the B-LCDR2 as shown in SEQ ID NO: 32, and the B-LCDR3 as shown in SEQ ID NO: 35; or (iii) the B-HCDR1 as shown in SEQ ID NO: 18, the B-HCDR2 as shown in SEQ ID NO: 22 As shown, the B-HCDR3 is shown in SEQ ID NO: 23, and the B-LCDR1 is shown in SEQ ID NO: 31, the B-LCDR2 is shown in SEQ ID NO: 32, and the B-LCDR3 is shown in SEQ ID NO: 32. ID NO: 36; or (iv) the B-HCDR1 is shown in SEQ ID NO: 18, the B-HCDR2 is shown in SEQ ID NO: 24, the B-HCDR3 is shown in SEQ ID NO: 20, and the B-LCDR1 is shown in SEQ ID NO: 37, the B-LCDR2 is shown in SEQ ID NO: 32, and the B-LCDR3 is shown in SEQ ID NO: 38; or (v) the B-HCDR1 is shown in As shown in SEQ ID NO: 18, the B-HCDR2 is shown in SEQ ID NO: 25, the B-HCDR3 is shown in SEQ ID NO: 20, and the B-LCDR1 is shown in SEQ ID NO: 39, the B - LCDR2 as shown in SEQ ID NO: 32, and the B-LCDR3 as shown in SEQ ID NO: 40; or (vi) the B-HCDR1 as shown in SEQ ID NO: 18, the B-HCDR2 as shown in SEQ ID NO : 26, the B-HCDR3 is shown in SEQ ID NO: 27, and the B-LCDR1 is shown in SEQ ID NO: 41, the B-LCDR2 is shown in SEQ ID NO: 32, and the B-LCDR3 As shown in SEQ ID NO: 42; or (vii) the B-HCDR1 is shown in SEQ ID NO: 18, the B-HCDR2 as shown in SEQ ID NO: 30, the B-HCDR3 as shown in SEQ ID NO: 20, and the B-LCDR1 as shown in SEQ ID NO: 31, the B-LCDR2 as shown in SEQ ID NO: 32, and The B-LCDR3 is represented by SEQ ID NO:33.

In some embodiments, the antigen-binding molecule of any one of the above, which comprises B-VH and B-VL, the B-VH is shown in SEQ ID NO: 47, and the B-VL is shown in SEQ ID NO: 48. In some embodiments, the B-VH is shown in SEQ ID NO: 5, and the B-VL is shown in SEQ ID NO: 12; or the B-VH is shown in SEQ ID NO: 6, and the B-VL is shown in SEQ ID NO: 6, and the B-VL is shown in SEQ ID NO: 12; -VL as shown in SEQ ID NO: 13; or the B-VH as shown in SEQ ID NO: 7, and the B-VL as shown in SEQ ID NO: 14; or the B-VH as shown in SEQ ID NO: 8 shown, and the B-VL is shown in SEQ ID NO: 15; or the B-VH is shown in SEQ ID NO: 9, and the B-VL is shown in SEQ ID NO: 16; or the B-VH As shown in SEQ ID NO: 10, and the B-VL as shown in SEQ ID NO: 17; or the B-VH as shown in SEQ ID NO: 11, and the B-VL as shown in SEQ ID NO: 2 .

In some embodiments, the antigen-binding molecule that specifically binds BAFF and IL-12/23 comprises an antigen-binding moiety 1 that specifically binds BAFF and an antigen-binding moiety 2 that specifically binds IL-12 and/or IL-23 ; wherein the antigen-binding module 2 comprises a heavy chain variable region P-VH and a light chain variable region P-VL, the P-VH comprises P-HCDR1, P-HCDR2 and P-HCDR3, and the P-VL comprises P- LCDR1, P-LCDR2 and P-LCDR3; the amino acid sequences of P-HCDR1, P-HCDR2 and P-HCDR3 are respectively the same as those of Pv-HCDR1, Pv-HCDR2 and Pv-HCDR3 in SEQ ID NO: 51 The acid sequence is the same, and the amino acid sequence of the P-LCDR1, P-LCDR2 and P-LCDR3 is respectively identical to the amino acid sequence of Pv-LCDR1, Pv-LCDR2 and Pv-LCDR3 in SEQ ID NO: 52; the P-HCDR1, P- HCDR2, P-HCDR3, P-LCDR1, P-LCDR2, P-LCDR3, Pv-HCDR1, Pv-HCDR2, Pv-HCDR3, Pv-LCDR1, Pv-LCDR2 and Pv-LCDR3 are selected from Kabat, IMGT, Chothia , AbM and Contact are defined by the same numbering rules. In some embodiments, the antigen-binding molecule of any one of the above, the P-HCDR1, P-HCDR2, P-HCDR3, P-LCDR1, P-LCDR2, P-LCDR3 are defined according to the Kabat numbering rules, wherein , the P-HCDR1 is shown in SEQ ID NO: 53, the P-HCDR2 is shown in SEQ ID NO: 54, the P-HCDR3 is shown in SEQ ID NO: 55, and the P-LCDR1 is shown in SEQ ID NO: 56, the P-LCDR2 is shown in SEQ ID NO: 57, and the P-LCDR3 is shown in SEQ ID NO: 58. In some embodiments, the P-VH of the heavy chain variable region is shown in SEQ ID NO:51, and the P-VL of the light chain variable region is shown in SEQ ID NO:52.

Variants of Antigen Binding Molecules

In certain embodiments, amino acid sequence variants of the antigen binding molecules provided herein are contemplated. For example, it may be desirable to improve the binding affinity and/or other biological properties of the antibody. 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 antigen-binding molecule. 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.

Variants containing substitutions, insertions, and deletions

In certain embodiments, antigen binding molecule 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 2 under the heading "Preferred Substitutions". More substantial changes are listed in Table 2 under "Exemplary Substitutions" are provided under the heading of , and are further described below with reference to the amino acid side chain category. 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.

Table 2. Amino acid substitutions

According to common side chain properties, amino acids can be grouped as follows:

(1) Hydrophobic: Nle, Met, Ala, Val, Leu, Ile;

(2) Neutral, hydrophilic: Cys, Ser, Thr, Asn, Gln;

(3) acidic: Asp, Glu;

(4) Basic: His, Lys, Arg;

(5) Residues affecting chain orientation: Gly, Pro;

(6) Aromatic: Trp, Tyr, Phe.

Non-conservative substitutions refer to the substitution of a member of one class for a member of another class.

One type of substitutional variant involves substituting one or more CDR residues of a parent antibody (eg, a humanized or human antibody). Generally, the resulting variants selected for further study will have altered (e.g. improved) certain biological properties (e.g. increased affinity, reduced immunogenicity) relative to the parental antibody, and/or will be substantially Certain 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 (eg, substitutions) can be made to the CDRs, eg, to improve antibody affinity. Such changes can be made to CDR "hotspots" (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. In some embodiments of 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 . Then, create secondary libraries. The library is then screened to identify any antibody variants with the desired affinity. Another method of introducing diversity involves a CDR-directed approach, in which several CDR residues (eg, 4-6 residues at a time) are randomized. CDR residues involved in antigen binding can be specifically identified, for example, using alanine scanning mutagenesis or modeling. In particular, HCDR3 and LCDR3 are frequently targeted.

In certain embodiments, 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. For example, conservative changes (eg, conservative substitutions, as provided herein) can be made to the CDRs that do not substantially reduce binding affinity. Such changes do not occur at antigen contact residues.

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". In this approach, a residue or group of residues (e.g. charged residues such as Arg, Asp, His, Lys and Glu) are identified and neutral or negatively charged amino acids (e.g. Ala or polyalanine) to determine whether the antibody-antigen interaction was affected. Further substitutions can be introduced at amino acid positions showing functional sensitivity to the initial substitution. In addition, 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: 1 residue fused at the amino and/or carboxy terminus or polypeptides with a length of 100 or more residues, and intrasequence insertions of single or multiple amino acid residues. Examples of terminal insertions include antibodies with an N-terminal methionyl residue. Other insertional variants of antibody molecules include fusions with enzymes (or polypeptides that extend the serum half-life of antibodies) at the N- or C-terminus of the antibody.

Modification of the Fc region

In one aspect, the Fc region of an antigen binding molecule of the disclosure comprises one or more amino acid substitutions that reduce its binding to an Fc receptor, e.g., its binding to an Fcγ receptor, And reduce or eliminate effector function. A native IgG Fc region, specifically an IgG ₁ Fc region or an IgG ₄ Fc region, may result in targeting of an antigen-binding molecule of the present disclosure to cells expressing Fc receptors, rather than cells expressing antigen. The engineered Fc regions of the present disclosure exhibit reduced binding affinity to Fc receptors and/or reduced effector functions. In some embodiments, the engineered Fc region has a reduced binding affinity for Fc receptors by more than 50%, 80%, 90% or more than 95% compared to a native Fc region. In some embodiments, the Fc receptor is an Fc gamma receptor. In some embodiments, the Fc receptor is a human Fc gamma receptor, eg, FcγRI, FcγRIIa, FcγRIIB, FcγRIIIa. In some embodiments, the engineered Fc region has reduced binding affinity for complement (eg, C1q) compared to a native Fc region. In some embodiments, the engineered Fc region has no reduced binding affinity for neonatal Fc receptor (FcRn) compared to a native Fc region. In some embodiments, 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 Binding, reduced binding to macrophages, reduced binding to monocytes, reduced binding to polymorphonuclear cells, reduced direct signaling-induced apoptosis, reduced dendritic cell maturation, or reduced T cell priming.

For the IgG ₁ Fc region, amino acid residue substitutions at positions 238, 265, 269, 270, 297, 327, and 329 may reduce effector function. In some embodiments, the Fc region is a human IgG ₁ Fc region, and the amino acid residues at positions 234 and 235 are A, numbered according to the EU index. For the IgG ₄ Fc region, substitution of amino acid residues at positions such as 228 may reduce effector function.

Antigen binding molecules may also comprise disulfide bond engineering, eg, 354C of the first subunit and 349C of the second subunit.

Undesired homodimerization may result when the antigen binding molecule comprises a binding module fused to two subunits of the Fc region. In order to increase yield and purity, it would therefore be advantageous to introduce modifications in the Fc region of the antigen binding molecules of the present disclosure that promote heterodimerization. In some embodiments, the Fc region of the present disclosure comprises a modification according to the knob-into-hole (KIH) technique, which involves introducing a knob at the interface of one subunit and a knob at the interface of the other subunit. A hole structure (hole) is introduced at the interface. This allows the protrusion to be positioned in the pore structure, promoting the formation of heterodimers and inhibiting the generation of homodimers. Protrusion structures are constructed by replacing small amino acid side chains from the interface of one subunit with larger side chains such as tyrosine or tryptophan. Instead, the pore structure is created in the interface of another subunit by replacing a large amino acid side chain with a smaller amino acid side chain, 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 3 below:

Table 3. KIH mutation combinations

In addition to the knob-and-hole technique, other techniques for modifying the CH3 domain of a heavy chain to achieve heterodimerization are known in the art, for example WO96/27011, WO98/050431, EP1870459, WO2007/110205, WO007/147901, WO2009/089004, WO2010/129304, WO2011/90754, WO2011/143545, WO2012/058768, WO2013/15795 4 and WO 013/096291.

The C-terminus of the Fc region can be a complete C-terminus ending with the amino acid residue PGK; it can also be a truncated C-terminus, for example, one or two C-terminal amino acids have been removed from the truncated C-terminus Residues. In a preferred aspect, the C-terminus of the Fc region is a shortened C-terminus ending with PG. Thus, in some embodiments, a composition of intact antibodies can include a population of antibodies from which all K447 residues and/or G446+K447 residues have been removed. In some embodiments, the composition of intact antibodies can include a population of antibodies without removal of the K447 residue and/or the G446+K447 residues. In some embodiments, the composition of whole antibodies has a population of antibodies with and without the K447 residue and/or the antibody mixture of G446+K447 residues.

Recombination method

Antigen binding molecules (eg, antibodies) can be produced using recombinant methods. For these methods, one or more isolated nucleic acids encoding the antigen binding molecule are provided.

In one embodiment, the present disclosure provides an isolated nucleic acid encoding an antigen binding molecule as previously described. Such nucleic acids may independently encode any of the aforementioned polypeptide chains. In another aspect, the present disclosure provides one or more vectors (eg, expression vectors) comprising such nucleic acids. In another aspect, the present disclosure provides host cells comprising such nucleic acids. In one embodiment, there is provided a method of making an antigen-binding molecule, wherein the method comprises, under conditions suitable for expression, culturing a host cell comprising a nucleic acid encoding the antigen-binding molecule, as provided above, and optionally from The host cell (or host cell culture medium) recovers the antigen-binding molecule.

For recombinant production of antigen-binding molecules, nucleic acid encoding the protein is isolated and inserted into one or more vectors for further cloning and/or expression in host cells. Such 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 antigen-binding molecules include prokaryotic or eukaryotic cells as 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.

In addition to prokaryotes, eukaryotic microorganisms such as filamentous fungi or yeast are suitable strains or expression hosts for vectors encoding antigen-binding molecules, including fungal and yeast strains. Suitable host cells suitable for expression of antigen binding molecules 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, especially for the transfection of Spodoptera frugiperda cells; plant cell cultures can also be used as hosts, e.g. US5959177, US 6040498, US6420548, US7125978 and US6417429; vertebrate cells can also be used as hosts, eg mammalian cell lines adapted for growth in suspension. Other examples of 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 (buffalo rat) liver cells ( BRL3A); human lung cells (W138); human hepatocytes (Hep G2); mouse breast tumor (MMT 060562); TRI cells; MRC 5 cells; and FS4 cells. Other 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. For a review of certain mammalian host cell lines suitable for antibody production see, e.g., Yazaki, P. and Wu, AM, Methods in Molecular Biology , Vol. 248, Lo, BKC (eds.), Humana Press, Totowa, NJ (2004) , pp. 255-268.

Detection and Diagnosis

Antigen binding molecules provided herein can be identified, screened or characterized for their physical/chemical characteristics and/or biological activity by a variety of assays known in the art. In one aspect, the activity of the antigen-binding molecules of the present disclosure is tested, for example, by known methods such as ELISA, Western blotting, and the like.

In certain embodiments, the antigen binding molecules provided by the present disclosure can be used to detect the presence or level of BAFF or IL-12/23 in a biological sample. As used herein, the term "detection" encompasses quantitative or qualitative detection. In certain embodiments, the biological sample comprises cells or tissue, such as tumor tissue.

In one embodiment, an antigen binding molecule for use in a diagnostic or detection method is provided. In one aspect, methods of detecting the presence of BAFF or IL-12/23 in a biological sample are provided. In certain embodiments, the method comprises contacting a biological sample with an antigen-binding molecule under suitable conditions, and detecting whether a complex is formed between the detection reagent and the antigen. Such methods can be in vitro or in vivo methods. In one embodiment, antigen binding molecules are used to select subjects suitable for treatment, for example BAFF or IL-12/23 are biomarkers for patient selection.

Exemplary disorders that can be diagnosed using the antigen binding molecules of the disclosure, such as autoimmune diseases, for example: systemic lupus erythematosus, myasthenia gravis, multiple sclerosis, insulin Dependent diabetes mellitus, Crohn's disease, rheumatoid arthritis, polyarticular juvenile rheumatoid arthritis, or psoriatic arthritis; or B-cell disorders such as tumors, chronic leukaemia, multiple myeloma, Non-Hodgkin's lymphoma, post-transplant lymphoproliferative disease, or light chain gamma globulinopathy.

In certain embodiments, labeled antigen binding molecules are provided. Labels include, but are not limited to, markers or moieties for direct detection (such as fluorescent, chromogenic, electron-dense, chemiluminescent, and radioactive labels), and moieties for indirect detection (e.g., indirect detection via enzymatic reactions or molecular interactions). modules, such as enzymes or ligands).

Methods of treatment and routes of administration

Any of the antigen binding molecules (eg, antibodies) provided herein can be used in methods of treatment. In yet another aspect, the present disclosure provides the use of an antigen-binding molecule in the manufacture or preparation of a medicament. In some embodiments, the B cell disorder or autoimmune disease is a disease or condition associated with BAFF or IL-12/23. In some embodiments, 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 inflammation and psoriatic arthritis; the 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 gammaglobulinopathy. In some embodiments, the autoimmune disease is systemic lupus erythematosus. In one such embodiment, the use further comprises administering to the subject an effective amount of at least one additional therapeutic agent (eg, one, two, three, four, five, or six additional therapeutic agents).

A "subject" according to any of the above embodiments may be a human. When used for therapeutic purposes, a subject is an individual who has, or is suspected of having, the disease of interest. When used for prophylactic purposes, the subject is an individual susceptible to the disease of interest.

In yet another aspect, there is provided a pharmaceutical composition comprising the antigen binding molecule, eg, for any of the above pharmaceutical uses or methods of treatment. In one embodiment, a pharmaceutical composition comprises any of the antigen binding molecules provided herein and a pharmaceutically acceptable carrier. In another embodiment, the pharmaceutical composition further comprises at least one additional therapeutic agent.

The antigen binding molecules of the present disclosure can be used alone or in combination with other agents for therapy. For example, an antibody of the present disclosure can be administered in combination (simultaneously, or sequentially) with at least one additional therapeutic agent.

The antigen binding molecules of the disclosure (and any additional therapeutic agents) can be administered by any suitable means, including parenteral, intrapulmonary, and intranasal, and, if local treatment is desired, intralesional. 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.

Antigen binding molecules of the present disclosure will be formulated, dosed and administered in a manner consistent with good medical practice (eg, GOOD MEDICAL PRACTICE Guidelines, GMP). 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. factor. Antigen binding molecules may or may not be formulated with one or more agents currently used to prevent or treat the disorder. Effective amounts 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 by any route empirically/clinically determined to be suitable.

For the prophylaxis or treatment of disease, appropriate dosages of the antigen binding molecules of the present disclosure (when used alone or in combination with one or more other additional therapeutic agents) will depend on the type of disease to be treated, the type of therapeutic molecule , the severity and course of the disease, whether administered for prophylactic or therapeutic purposes, previous therapy, the subject's clinical history and response to the therapeutic molecule, and the judgment of the attending physician. The therapeutic molecule is suitably administered to a subject at one time or over a series of treatments. For example, a daily dosage might range from about 1 μg/kg to 100 mg/kg, depending on the factors mentioned above.

products

In another aspect of the present disclosure, an article of manufacture comprising materials useful for the treatment, prevention and/or diagnosis of the disorders described above is provided. 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 port (e.g., the container may be a vein with a stopper pierceable by a hypodermic needle) inner solution bag or vial). At least one active agent in the composition is an antigen binding molecule of the present disclosure. The label or package insert indicates that the composition is used to treat the condition of choice. In addition, the article of manufacture may comprise: (a) a first container having a composition therein, wherein the composition comprises an antigen binding molecule of the present disclosure; and (b) a second container having the composition therein device, wherein the composition comprises an additional cytotoxic or other therapeutic agent. The articles 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. Alternatively, or in addition, 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. As an example, the article of manufacture is prepared in the form of a kit.

Example and test case

The present disclosure is further described below in conjunction with examples and test examples, but these examples and test examples do not limit the scope of the present disclosure. For the experimental methods not specified in the examples and test examples of this disclosure, conventional conditions are usually followed, such as Cold Spring Harbor Antibody Technology Experiment Manual, Molecular Breeding Manual; or the conditions suggested by raw material or commodity manufacturers. Reagents without specific sources indicated are conventional reagents purchased in the market.

Example 1: Belimumab mutant antibody and its activity detection

Belimumab was modified by phage display technology. By designing mutation primers, the CDRs of the light and heavy chains of Belimumab were mutated separately to construct a mutation library. The mutant library was enriched and screened using biotinylated BAFF protein (Sino biological, 10056-HNCH) to obtain candidate strains. The amino acid sequences of the light and heavy chain variable regions of the strains were determined by sequencing, and the light/heavy chains of the candidate strains were The variable regions are fused with the antibody light/heavy chain constant regions respectively to construct full-length antibodies. in,

The sequence information of the Belimumab antibody is as follows:

>Amino acid sequence of B-VH0 (Belimumab heavy chain variable region):

SEQ ID NO：1

SEQ ID NO: 1

>Amino acid sequence of B-VL0 (Belimumab light chain variable region):

SEQ ID NO：2

SEQ ID NO: 2

>Amino acid sequence of Belimumab heavy chain:

SEQ ID NO：3

SEQ ID NO: 3

>Amino acid sequence of Belimumab light chain:

SEQ ID NO：4

SEQ ID NO: 4

Belimumab modified heavy chain variable region and light chain variable region:

>Amino acid sequence of B-VH1:

SEQ ID NO：5

SEQ ID NO: 5

>Amino acid sequence of B-VH2

SEQ ID NO：6

SEQ ID NO: 6

>Amino acid sequence of B-VH3:

SEQ ID NO：7

SEQ ID NO: 7

>Amino acid sequence of B-VH4:

SEQ ID NO：8

SEQ ID NO: 8

>Amino acid sequence of B-VH5:

SEQ ID NO：9

SEQ ID NO: 9

>Amino acid sequence of B-VH6:

SEQ ID NO：10

SEQ ID NO: 10

>Amino acid sequence of B-VH7:

SEQ ID NO：11

SEQ ID NO: 11

>Amino acid sequence of B-VL1:

SEQ ID NO：12

SEQ ID NO: 12

>Amino acid sequence of B-VL2:

SEQ ID NO：13

SEQ ID NO: 13

>Amino acid sequence of B-VL3:

SEQ ID NO：14

SEQ ID NO: 14

>Amino acid sequence of B-VL4:

SEQ ID NO：15

SEQ ID NO: 15

>Amino acid sequence of B-VL5:

SEQ ID NO：16

SEQ ID NO: 16

>Amino acid sequence of B-VL6:

SEQ ID NO：17。

SEQ ID NO:17.

Remarks: Among the above variable region sequences, the sequence is FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4, the underlined part is the CDR region (determined according to the Kabat numbering system), and the italic part is the mutated amino acid residue, The ununderlined part is the framework region.

Table 4. Heavy chain CDR sequence list

Table 5. List of light chain CDR sequences

The above heavy chain variable region and light chain variable region are fused with the antibody heavy chain constant region and light chain constant region, respectively, to construct a full-length antibody. Exemplarily, the above-mentioned heavy chain variable region is fused with the heavy chain constant region of Belimumab (sequence shown in SEQ ID NO: 45), and the light chain variable region of the antibody is fused with the light chain constant region of Belimumab (sequence shown in SEQ ID NO: 45). : shown in 46) fusion to construct anti-BAFF antibodies: B1 to B7, the variable region sequences of the obtained antibodies are shown in Table 6.

>Amino acid sequence of heavy chain constant region of Belimumab antibody

SEQ ID NO：45

SEQ ID NO: 45

>Amino acid sequence of light chain constant region of Belimumab antibody

SEQ ID NO：46。

SEQ ID NO:46.

Table 6. Variable region sequences of anti-BAFF antibodies

Through in vitro activity experiments, the activity of the constructed antibodies B1 to B7 to block the binding of human BAFF to BAFF-R (see Test Example 2 of the present disclosure) and the activity of inhibiting BAFF-induced B cell proliferation (see Test Example 2 of the present disclosure) were detected. 3) and the binding affinity to human BAFF (see test example 5 of the present disclosure). Experimental results show that the disclosed B1 to B7 antibodies can be Binding to human BAFF with high affinity, compared with the parent antibody Belimumab, it has better activity in blocking the binding ability of BAFF to BAFF-R and inhibiting BAFF-induced B cell proliferation.

In addition, the framework regions of the above-mentioned anti-BAFF antibodies were modified. Exemplarily, the 44th amino acid residue in the heavy chain variable region of B6 (the site determined according to the Kabat numbering system) is mutated to C (ie 44C), and the 100th amino acid residue in the light chain variable region of B6 is mutated (According to the position determined by the Kabat numbering system) the amino acid residue is mutated to C (ie 100C) in order to increase the interchain disulfide bond between VH and VL. The sequence of the variable region of antibody B61 after modification of the B6 framework region is as follows :

>Amino acid sequence of heavy chain variable region of B61

SEQ ID NO：47

SEQ ID NO: 47

>Amino acid sequence of light chain variable region of B61

SEQ ID NO：48。

SEQ ID NO:48.

Example 2: Construction of bispecific antibodies specifically binding to BAFF and IL-12/23

The anti-BAFF antibody obtained in Example 1 and the known anti-IL-12/23 antibody were used to construct a bispecific antibody specifically binding to BAFF and IL-12/23. Antibodies against IL-12/23 can It can be derived from any suitable antibody, for example, anti-IL-12/23 p40 subunit antibody Ustekinumab (abbreviated as Umab), the sequence of Ustekinumab is as follows:

>Amino acid sequence of Ustekinumab heavy chain

SEQ ID NO：49

SEQ ID NO: 49

>Amino acid sequence of Ustekinumab light chain

SEQ ID NO：50

SEQ ID NO: 50

>Amino acid sequence of heavy chain variable region of Ustekinumab

SEQ ID NO：51

SEQ ID NO: 51

>Amino acid sequence of Ustekinumab light chain variable region

SEQ ID NO：52，

SEQ ID NO: 52,

Remarks: Among the above sequences, the double underlined part is the variable region sequence, and the single underlined part is the CDR region sequence. The variable region and CDR are identified according to the Kabat numbering system.

The CDR sequence of Ustekinumab is shown in Table 7 below:

Table 7. CDR sequences of Ustekinumab

Exemplarily, the human IgG1 heavy chain constant region (sequence shown in SEQ ID NO: 59), human kappa light chain constant region (sequence shown in SEQ ID NO: 60), B61 antibody heavy chain variable region and Light chain variable region (sequence as shown in SEQ ID NO: 47,48) and Ustekinumab antibody heavy chain variable region and light chain variable region (sequence as shown in SEQ ID NO: 51,52 shown), to construct a bispecific antibody specifically binding to BAFF and IL-12/23. Among them, the C-terminal of the variable region of the light chain of the Ustekinumab antibody is fused with the N-terminal of the constant region of the human κ light chain to form the second chain; the C-terminal of the variable region of the heavy chain of the Ustekinumab antibody is fused with the N-terminal of the constant region of the human IgG1 heavy chain, The C-terminus of the human IgG1 heavy chain constant region is fused to the N-terminus of the scFv (constructed from the heavy and light chain variable regions of the B61 antibody) to form the first chain. Finally, bispecific antibodies specifically binding to BAFF and IL-12/23 were constructed: BU-1 and BU-2. Both BU-1 and BU-2 are composed of 4 chains (including two identical first chains and two identical second chains), the structural diagram of BU-1 is shown in Figure 1, and the structural diagram of BU-2 is shown in Figure 1 2.

>Amino acid sequence of human IgG1 heavy chain constant region

SEQ ID NO：59

SEQ ID NO: 59

>Amino acid sequence of human κ light chain constant region

SEQ ID NO：60

SEQ ID NO: 60

>Amino acid sequence of the first strand of BU-1

SEQ ID NO：61；

SEQ ID NO: 61;

>The amino acid sequence of the second chain of BU-1 is the same as that of the light chain of Ustekinumab, and the sequence is shown in SEQ ID NO:50.

>Amino acid sequence of the first strand of BU-2

SEQ ID NO：62；

SEQ ID NO: 62;

>The amino acid sequence of the second chain of BU-2 is the same as that of the light chain of Ustekinumab, and the sequence is shown in SEQ ID NO:50.

Remarks: In the first chain of BU-1 and BU-2 above, the wavy part is the heavy chain variable region of the Ustekinumab antibody, the underlined part is the human IgG1 heavy chain constant region, and the double underlined part is the B61 antibody The variable region of the heavy chain, the underlined part is the variable region of the light chain of the B61 antibody, and the part in italics is the linker sequence.

Test Example 1: Elisa Binding Experiment

The binding activity of the antibody to BAFF and IL-12/23 p40 was detected by the Elisa method. The specific method is as follows:

The sample to be tested was diluted to 2 μg/mL with pH 7.4 PBS (B320) buffer, added to a 96-well microplate plate (Corning, 3590) at a volume of 100 μL/well, and incubated overnight at 4°C. After the liquid was discarded, 300 μL of 5% skimmed milk (BD, 232100) diluted with PBS was added to each well for blocking, and incubated at 37°C for 2 hours. After blocking, the blocking solution was discarded, and the plate was washed 3 times with PBST buffer (pH7.4 PBS containing 0.1% tween-20), and 100 μL of serially diluted BAFF (ACROBiosystems, BAF-H5248) or IL-12 was added to each well. /23 p40 (Sino Biological, 10052-H08H) solution, incubated at 37°C for 1 hour. After incubation, the plate was washed 3 times with PBST. Add 100 μL Anti-His-HRP (Sino biological, 105327-MM02T-H, 1:2000 dilution) to each well. After washing the plate 3 times with PBST, add 100 μL TMB chromogenic substrate (KPL, 5120-0077) to each well, incubate at room temperature for 10 to 15 minutes, add 50 μL 1M H ₂ SO ₄ to each well to terminate the reaction, and read with a microplate reader Take the absorbance at 450nm, use software to fit the binding curve of antibody and antigen, and calculate the EC50 value. The experimental results are shown in Table 8 and Table 9 below. The experimental results show that the bispecific antibodies specifically binding to BAFF and IL-12/23 constructed in this disclosure can effectively bind to BAFF and IL-12/23 p40.

Table 8. The results of antibody binding experiments with IL-12/23 p40

Table 9. Results of antibody binding experiment with BAFF

Test Example 2: Ligand and Receptor Blocking Experiment

The blocking activity of the antibody against the following receptors and ligands was detected by Elisa method: human BAFF and human BAFF-R, human BAFF and human BCMA, human BAFF and human TACI, and human IL-12/23 p40 and human IL- 12Rβ1. The specific method is as follows:

The receptor protein was diluted to 2 μg/mL with pH 7.4 PBS (B320) buffer, added to a 96-well microtiter plate (Corning, 3590) at a volume of 100 μL/well, and incubated overnight at 4°C. After the liquid was discarded, 200 μL of 1% Casein blocking solution (Thermo, 37528) was added to each well for blocking, and incubated at 37°C for 2 hours. After 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. A fixed concentration of biotin-labeled ligand protein was mixed with a gradiently diluted antibody, 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 μL streptavidin-HRP (Invitrogen, 434323, diluted 1:4000) was added to each well, and incubated at 37° C. for 1 hour. Remove the supernatant, wash the plate 3 times with PBST, add 100 μL TMB chromogenic substrate (KPL, 5120-0077) to each well, incubate at room temperature for 10 to 15 minutes, add 50 μL 1M H ₂ SO ₄ to each well to terminate the reaction, and use enzyme The absorbance value at 450nm was read by the standard instrument, and the curve of inhibiting ligand and receptor binding was fitted by software, and the IC50 value was calculated. The sources of receptors and ligand proteins used in this test example are as follows: human BAFF (Sino biological, 10056-HNCH), human IL-12/23 p40 (Sino biological, 10052-H08H), human BAFF-R (Sino biological , 16079-H02H), human BCMA (Sino biological, 10620-H02H), human TACI (ACROBiosystems, TAI-H5256), human IL-12Rβ1 (ACROBiosystems, ILB-H5255).

The experimental results are shown in Table 10, Table 11, and Table 12. The experimental results show that the antibody specifically binding to BAFF constructed in this disclosure can effectively block the binding of human BAFF and human BAFF-R, and the IC50 value is lower than that of Belimumab. In addition, the disclosed bispecific antibody specifically binding to BAFF and IL-12/23 can effectively block human BAFF and human BAFF-R, human BAFF and human BCMA, human BAFF and human TACI, and the IC50 value is less than that of Belimumab ; Can also effectively block the binding of human IL-12/23 p40 and human IL-12Rβ1.

Table 10. Antibody Blocking Experimental Results of Human BAFF and Human BAFF-R

備註：“-”表示未檢測 Table 11. Antibody blocking human BAFF and its receptor binding experiment results blocking IL-12/23 p40 and its receptor

Note: "-" means not detected

Table 12. Experimental results of bispecific antibodies blocking the binding of human BAFF to human BAFF-R

Test Example 3: B cell proliferation experiment

B cell proliferation assay was used to detect whether the antibody could inhibit BAFF-induced B cell proliferation. The experimental method is as follows:

The mouse spleen was taken for grinding, centrifuged at 4°C for 5 minutes to collect the cells in the lower layer, washed once with washing solution (PBS+2% FBS+2mM EDTA) and centrifuged, then added RBC lysis buffer (Invitrogen, 00-4333- 57), stand at room temperature for 5 minutes until the red blood cells are completely lysed. Centrifuge again and resuspend cells for counting. The cell suspension was sorted with a 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 μM2 - Mercaptoethanol (5igma-Aldrich, M6250) was resuspended and counted, and the cells were plated in 96-well cell plates (Costar, 3903) for later use. Dilute the BAFF (R&D Systems, 7537-BF) protein to a fixed concentration, and add the diluted antibody to mix well, pre-incubate at 37°C for 30 minutes, add it to a 96-well cell plate, and culture it in a cell culture incubator at 37°C for 48 hours. Take out the cell culture plate, add 50 μL Celltiter Glo detection solution (Promega, G7573) to each well, and incubate at room temperature for 10 Minutes, use a microplate reader to detect the bioluminescent signal, use software to fit the detection results to an inhibition curve, and calculate the IC50 value.

The experimental results are shown in Table 13 and Table 14 below. The experimental results show that the disclosed antibody specifically binding to BAFF and the bispecific antibody specifically binding to BAFF and IL-12/23 can effectively inhibit BAFF-induced B cell proliferation, and Belimumab inhibits The IC50 value of BAFF-induced B cell proliferation is more than 2 times that of the disclosed antibody specifically binding to BAFF, while ustekinumab has no function of inhibiting BAFF-induced B cell proliferation.

Table 13. Results of anti-BAFF antibody inhibition of BAFF-induced B cell proliferation

Table 14. Experimental results of bispecific antibodies inhibiting BAFF-induced B cell proliferation

Test Example 4: Experimental Detection of IFNγ and IL-17 Secretion

Whether the antibody can inhibit T cell differentiation induced by IL-12 and IL-23 was detected by IFNγ and IL-17 secretion assay. The experimental method is as follows:

Add 100 μL of 2 μg/mL anti-mouse CD3 antibody (BioLegend, 100238) and 2 μg/mL anti-mouse CD28 antibody (BioLegend, 102116) to each well of a 96-well plate (Corning, 3599), incubate at 37°C for 1 hour, and wash with PBS for 2 hours. For later use. The mouse spleen was taken for grinding, centrifuged at 4°C for 5 minutes to collect the cells in the lower layer, washed once with washing solution (PBS+2% FBS+2mM EDTA) and centrifuged, then added RBC lysis buffer (Invitrogen, 00-4333- 57), stand at room temperature for 5 minutes until the red blood cells are completely lysed. Centrifuge again and resuspend cells for counting. The cell suspension was sorted with a mouse CD4 cell kit (Invitrogen, 11415D), and the isolated CD4+ T cells were resuspended in RPMI 1640 medium (Gibco, 11875119) + 10% FBS (Gibco, 10099-141) and counted for later use .

When detecting IL-12-induced T cell differentiation, 20 μg/mL anti-mouse IL-4 (BioLegend, 504122) was added to T cells, and the cell suspension was spread in a coated 96-well plate. A fixed concentration of chimeric IL-12 (human p40 fused with mouse p35) protein was mixed with serially diluted antibodies, pre-incubated at 37°C for 1 hour, added to a 96-well plate, and incubated in a 37°C cell culture incubator for 48 hours. The 96-well plate was taken out, centrifuged at 1000 rpm for 3 minutes, and the supernatant was collected, and the content of IFNγ in the supernatant was detected with a mouse IFN-gamma DuoSet ELISA Kit (R&D Systems, DY485).

When detecting the differentiation of T cells induced by IL-23, the cell suspension was spread in a well-coated 96-well plate, and a fixed concentration of IL-23 (R&D Systems, 1290-IL-010) was mixed with a serially diluted antibody for pretreatment. After incubation for 1 hour, add it to a 96-well plate and culture it in a cell culture incubator at 37°C for 48 Hour. The 96-well plate was taken out, centrifuged at 1000 rpm for 3 minutes, and the supernatant was collected, and the IL-17 content in the supernatant was detected with a mouse IL-17 DuoSet ELISA Kit (R&D Systems, DY421).

The experimental results are shown in Table 15 and Table 16 below. The experimental results show that the disclosed bispecific antibody specifically binding to BAFF and IL-12/23 can effectively inhibit IL-23-induced IL-17 secretion, and can effectively inhibit IL-12 Induced IFNγ secretion, while Belimumab could neither inhibit IL-23-induced IL-17 secretion nor IL-12-induced IFNγ secretion.

Table 15. Results of Antibody Inhibition of IFNγ Secretion Induced by IL-12

Table 16. Experimental results of antibodies inhibiting IL-23-induced IL-17 secretion

Test Example 5: Affinity Test

Use biosensor 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 dissociation curve. After each cycle of dissociation, the biochip was washed and regenerated with 10 mM glycine-hydrochloric acid solution pH 1.5 (GE, BR-1003-54). The experimental data was fitted with BIA evaluation version 4.1 and GE software with a 1:1 model to obtain the affinity value. The relevant antigenic proteins used in this test are as follows: Human IL-23 (CT048-H08H, Sino biological), human BAFF (10056-HNCH, Sino biological), cynomolgus IL-12/23 P40 (10215-CL, R&D Systems), cynomolgus BAFF (BAF-CM412B , Kactus), mouse BAFF (BAF-M521y, Acro Biosystems).

The experimental results are shown in Table 17 and Table 18 below. The experimental results show that, compared with Belimumab, the anti-BAFF antibody constructed in this disclosure can bind to human BAFF with a smaller KD value; in addition, the construction of this disclosure specifically binds to BAFF and IL-12 The /23 bispecific antibody can bind to human IL-23, human BAFF, cynomolgus monkey IL-12/23 P40, cynomolgus monkey BAFF, and mouse BAFF with high affinity.

Table 17. Antibody and human BAFF affinity test results

Table 18. Antibody and related antigen affinity test results

Test Example 6: In Vivo Activity Evaluation

Simultaneously stimulate mice with chimeric IL-12 (human p40 fused with mouse p35), human IL-23 and human BAFF to induce the production of cytokines such as IFNγ, TNFα, IL-22 and IgA in mice. The levels of these cytokines were used to evaluate the in vivo activity of antibodies.

SPF-grade female C57BL/6 mice (Beijing Weitong Lihua Experimental Animal Technology Co., Ltd., 8 weeks old), the mice were randomly divided into 5 groups, and chimeric IL-12 (2 μg/hour) was mixed and injected intraperitoneally. Rat), human IL-23 (2 μg/mouse) and human BAFF (1 mg/kg) were injected once a day for four days. The sample to be tested (Belimumab 8mpk, BU-1 10.7mpk, or BU-1 5.35mpk) was injected intraperitoneally one hour before the protein injection on the first day and the third day respectively. On the fifth day, the plasma samples of mice in each group were collected, and the levels of IFNγ, TNFα, IL-22 and IgA were detected respectively. The sources of the detection kits used in this test case are as follows: Mouse IFN-gamma Quantikine ELISA Kit (R&D Systems, MIF00), 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). Belimumab was used as a positive control, and PBS was used as a negative control. BU-1 10.7mpk has the same molar drug concentration as Belimumab 8mpk.

The experimental results are shown in Figures 3 to 6. The experimental results show that the disclosed bispecific antibody that specifically binds to BAFF and IL-12/23 can significantly inhibit the secretion of TNFα, IFNγ, and IL-22, while Belimumab cannot inhibit the secretion of TNFα and IFNγ secretion. In addition, BU-1 can significantly inhibit the secretion of IgA at two doses of 10.7mpk and 5.35mpk, and the inhibitory activity is stronger than that of Belimumab.

Test Example 7. In vivo pharmacokinetic experiment in rats

In vivo pharmacokinetic tests were performed with SD rats. Male SD rats (Zhejiang Weitong Lihua Experimental Animal Technology Co., Ltd.) were divided into random groups, 4 in each group, administered intravenously, and the administration group was administered 5 minutes, 8 hours, 24 hours, and 48 hours , 84 hours, 9 days, 10 days, 14 days, 21 days, 28 days collected 0.2mL of whole blood, without anticoagulation, placed at 4°C for 30 minutes after blood collection, centrifuged at 1000g for 15 minutes, and the supernatant serum was placed in EP tube and stored at -80°C. The blood drug concentration in serum was detected by ELISA method, and the pharmacokinetic parameters and in vivo half-life of the tested drug were calculated by Winnolin software.

The experimental results are shown in Table 19 below. The experimental results show that the disclosed bispecific antibody specifically binding to BAFF and IL-12/23 has a good half-life in rats, and the anti-IL-12/23 p40 half-life reaches 14 days. The half-life is longer than that of Ustekinumab.

Table 19. In vivo pharmacokinetic test results of antibody in rats

Claims (16)

An antigen-binding molecule comprising an antigen-binding module 1 that specifically binds BAFF and an antigen-binding module 2 that specifically binds IL-12 and/or IL-23, wherein the antigen-binding module 1 comprises a heavy chain variable region B- VH and light chain variable region B-VL, the B-VH comprising B-HCDR1, B-HCDR2 and B-HCDR3, the B-VL comprising B-LCDR1, B-LCDR2 and B-LCDR3, wherein, The B-HCDR1, B-HCDR2 and B-HCDR3 respectively comprise the amino acid sequences of Bv-HCDR1, Bv-HCDR2 and Bv-HCDR3 in SEQ ID NO: 63, and The B-LCDR1, B-LCDR2 and B-LCDR3 respectively comprise the amino acid sequences of Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 in SEQ ID NO: 64, wherein, SEQ ID NO: 63 is: QVQLQQSGAEVKKPGSSVRVSCKASGGTFNNNAINWVRQAPGQX ₂₆ LEWMGX ₁ IX ₂ PMFGX ₃ AKYSX ₄ X ₅ FQGRVAITADESTGTASMELSSLRSEDTAVYYCARSRDX ₆ LLFPX ₇ X ₈ X ₉ LX ₁₀ X ₁₁ WG _{X 27} GTMVTVSS, SEQ ID NO: 64 is: SSELTQDPAVSVALGQTVRVTCX ₁₂ GX ₁₃ X ₁₄ LX ₁₅ X ₁₆ X ₁₇ X ₁₈ ASWYQQKPGQAPVLVIYGKNNRPSGIPDRFSGSSSGNTASLTITGAQAEDEADYYCX ₁₉ SRX ₂₀ X ₂₁ X ₂₂ GX ₂₃ X ₂₄ WX ₂₅ FGX ₂₈ GTELTVL, Wherein, _X1 is selected from S, A, V or G, _X2 is selected from A, I, M or S, _X3 is selected from T or G, _X4 is selected from Q, E, G or K, _X5 is selected from N, G or Q, X ₆ is selected from L or P, X ₇ is selected from H, Q or D, X ₈ is selected from H or D, X ₉ is selected from A or G, X ₁₀ is selected from S or L, X ₁₁ selected from P or S, X ₁₂ selected from Q or H, X ₁₃ selected from D, A or N, X ₁₄ selected from S or I, X ₁₅ selected from K, R or T, X ₁₆ selected from S, W, T or D, X ₁₇ is selected from H, Y or S, X ₁₈ is selected from Y or R, X ₁₉ is selected from G or S, X ₂₀ is selected from A or D, X ₂₁ is selected from E or S, X ₂₂ is selected from S or A, X ₂₃ is selected from V, E, A, N or W, X ₂₄ is selected from G, K, H or R, X ₂₅ is selected from L or V, X ₂₆ is selected from G or C, X ₂₇ is selected from R or G, X ₂₈ is selected from G or C; Moreover, the B-HCDR1, B-HCDR2, B-HCDR3, B-LCDR1, B-LCDR2 and B-LCDR3 do not contain the following CDR combination: B-HCDR1, B-HCDR2 and B-HCDR3 respectively contain SEQ ID NO: 1 The amino acid sequences of Bv-HCDR1, Bv-HCDR2 and Bv-HCDR3 in, and B-LCDR1, B-LCDR2 and B-LCDR3 respectively comprise Bv-LCDR1, Bv-LCDR2 and Bv-LCDR1 in SEQ ID NO: 2 The amino acid sequence of LCDR3; Preferably, The B-HCDR1, B-HCDR2, B-HCDR3, B-LCDR1, B-LCDR2 and B-LCDR3 are defined according to the Kabat numbering rules, wherein, B-HCDR1 comprises the amino acid sequence NNAIN (SEQ ID NO: 18), B-HCDR2 comprises the amino acid sequence X ₁ IX ₂ PMFGX ₃ AKYSX ₄ X ₅ FQG (SEQ ID NO: 65), B-HCDR3 comprises the amino acid sequence SRDX ₆ LLFPX ₇ X ₈ X ₉ LX ₁₀ X ₁₁ (SEQ ID NO: 66), B-LCDR1 comprises the amino acid sequence X ₁₂ GX ₁₃ X ₁₄ LX ₁₅ X ₁₆ X ₁₇ X ₁₈ AS (SEQ ID NO: 67), B-LCDR2 comprises the amino acid sequence GKNNRPS (SEQ ID NO: 32) and B-LCDR3 comprises the amino acid sequence X ₁₉ SRX ₂₀ X ₂₁ X ₂₂ GX ₂₃ X ₂₄ WX ₂₅ (SEQ ID NO: 68), Wherein, _X1 is selected from S, A, V or G, _X2 is selected from A, I, M or S, _X3 is selected from T or G, _X4 is selected from Q, E, G or K, _X5 is selected from N, G or Q, X ₆ is selected from L or P, X ₇ is selected from H, Q or D, X ₈ is selected from H or D, X ₉ is selected from A or G, X ₁₀ is selected from S or L, X ₁₁ selected from P or S, X ₁₂ selected from Q or H, X ₁₃ selected from D, A or N, X ₁₄ selected from S or I, X ₁₅ selected from K, R or T, X ₁₆ selected from S, W, T or D, X ₁₇ is selected from H, Y or S, X ₁₈ is selected from Y or R, X ₁₉ is selected from G or S, X ₂₀ is selected from A or D, X ₂₁ is selected from E or S, X ₂₂ is selected from S or A, X ₂₃ is selected from V, E, A, N or W, X ₂₄ is selected from G, K, H or R, X ₂₅ is selected from L or V; And, the B-HCDR1, B-HCDR2, B-HCDR3, B-LCDR1, B-LCDR2 and B-LCDR3 do not contain the following CDR combination: B-HCDR1 contains the amino acid sequence of SEQ ID NO: 18, B-HCDR2 Contains the amino acid sequence of SEQ ID NO: 19, B-HCDR3 contains the amino acid sequence of SEQ ID NO: 20, B-LCDR1 contains the amino acid sequence of SEQ ID NO: 31, and B-LCDR2 contains the amino acid sequence of SEQ ID NO: 32, and B-LCDR3 comprises the amino acid sequence of SEQ ID NO:33. The antigen-binding molecule as claimed in item 1, wherein, (i) the B-HCDR1, B-HCDR2 and B-HCDR3 respectively comprise the amino acid sequences of Bv-HCDR1, Bv-HCDR2 and Bv-HCDR3 in SEQ ID NO: 47, and the B-LCDR1, B-LCDR2 and B-LCDR3 respectively comprising the amino acid sequences of Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 in SEQ ID NO: 48, or (ii) the B-HCDR1, B-HCDR2 and B-HCDR3 respectively comprise the amino acid sequences of Bv-HCDR1, Bv-HCDR2 and Bv-HCDR3 in SEQ ID NO: 5, and the B-LCDR1, B-LCDR2 and B-LCDR3 respectively comprising the amino acid sequences of Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 in SEQ ID NO: 12, or (iii) the B-HCDR1, B-HCDR2 and B-HCDR3 respectively comprise the amino acid sequences of Bv-HCDR1, Bv-HCDR2 and Bv-HCDR3 in SEQ ID NO: 6, and the B-LCDR1, B-LCDR2 and B-LCDR3 respectively comprising the amino acid sequences of Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 in SEQ ID NO: 13, or (iv) the B-HCDR1, B-HCDR2 and B-HCDR3 respectively comprise the amino acid sequences of Bv-HCDR1, Bv-HCDR2 and Bv-HCDR3 in SEQ ID NO: 7, and the B-LCDR1, B-LCDR2 and B-LCDR3 respectively comprising the amino acid sequences of Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 in SEQ ID NO: 14, or (v) the B-HCDR1, B-HCDR2 and B-HCDR3 respectively comprise the amino acid sequences of Bv-HCDR1, Bv-HCDR2 and Bv-HCDR3 in SEQ ID NO: 8, and the B-LCDR1, B-LCDR2 and B-LCDR3 respectively comprising the amino acid sequences of Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 in SEQ ID NO: 15, or (vi) The B-HCDR1, B-HCDR2 and B-HCDR3 respectively comprise the amino acid sequences of Bv-HCDR1, Bv-HCDR2 and Bv-HCDR3 in SEQ ID NO: 9, and the B-LCDR1, B-LCDR2 and B-LCDR3 respectively comprising the amino acid sequences of Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 in SEQ ID NO: 16, or (vii) the B-HCDR1, B-HCDR2 and B-HCDR3 respectively comprise the amino acid sequences of Bv-HCDR1, Bv-HCDR2 and Bv-HCDR3 in SEQ ID NO: 10, and the B-LCDR1, B-LCDR2 and B-LCDR3 respectively comprising the amino acid sequences of Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 in SEQ ID NO: 17, or (viii) the B-HCDR1, B-HCDR2 and B-HCDR3 respectively comprise the amino acid sequences of Bv-HCDR1, Bv-HCDR2 and Bv-HCDR3 in SEQ ID NO: 11, and the B- LCDR1, B-LCDR2 and B-LCDR3 respectively comprise the amino acid sequences of Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 in SEQ ID NO: 2; Preferably, The B-HCDR1, B-HCDR2, B-HCDR3, B-LCDR1, B-LCDR2, B-LCDR3 are defined according to the Kabat numbering rules, wherein, (i) The B-HCDR1 comprises the amino acid sequence of SEQ ID NO: 18, the B-HCDR2 comprises the amino acid sequence of SEQ ID NO: 28, and the B-HCDR3 comprises the amino acid sequence of SEQ ID NO: 29 , and the B-LCDR1 comprises the amino acid sequence of SEQ ID NO: 43, the B-LCDR2 comprises the amino acid sequence of SEQ ID NO: 32, and the B-LCDR3 comprises the amino acid sequence of SEQ ID NO: 44 sequence; or (ii) The B-HCDR1 comprises the amino acid sequence of SEQ ID NO: 18, the B-HCDR2 comprises the amino acid sequence of SEQ ID NO: 21, and the B-HCDR3 comprises the amino acid sequence of SEQ ID NO: 20 , and the B-LCDR1 comprises the amino acid sequence of SEQ ID NO: 34, the B-LCDR2 comprises the amino acid sequence of SEQ ID NO: 32, and the B-LCDR3 comprises the amino acid sequence of SEQ ID NO: 35 ;or (iii) The B-HCDR1 comprises the amino acid sequence of SEQ ID NO: 18, the B-HCDR2 comprises the amino acid sequence of SEQ ID NO: 22, and the B-HCDR3 comprises the amino acid sequence of SEQ ID NO: 23 , and the B-LCDR1 comprises the amino acid sequence of SEQ ID NO: 31, the B-LCDR2 comprises the amino acid sequence of SEQ ID NO: 32, and the B-LCDR3 comprises the amino acid sequence of SEQ ID NO: 36 ;or (iv) The B-HCDR1 comprises the amino acid sequence of SEQ ID NO: 18, the B-HCDR2 comprises the amino acid sequence of SEQ ID NO: 24, and the B-HCDR3 comprises the amino acid sequence of SEQ ID NO: 20 The amino acid sequence of, and the B-LCDR1 comprises the amino acid sequence of SEQ ID NO: 37, the B-LCDR2 comprises the amino acid sequence of SEQ ID NO: 32, and the B-LCDR3 comprises the amino acid sequence of SEQ ID NO: 38 the amino acid sequence of ; or (v) The B-HCDR1 comprises the amino acid sequence of SEQ ID NO: 18, the B-HCDR2 comprises the amino acid sequence of SEQ ID NO: 25, and the B-HCDR3 comprises the amino acid sequence of SEQ ID NO: 20 , and the B-LCDR1 comprises the amino acid sequence of SEQ ID NO: 39, the B-LCDR2 comprises the amino acid sequence of SEQ ID NO: 32, and the B-LCDR3 comprises the amino acid sequence of SEQ ID NO: 40 ;or (vi) The B-HCDR1 comprises the amino acid sequence of SEQ ID NO: 18, the B-HCDR2 comprises the amino acid sequence of SEQ ID NO: 26, and the B-HCDR3 comprises the amino acid sequence of SEQ ID NO: 27 , and the B-LCDR1 comprises the amino acid sequence of SEQ ID NO: 41, the B-LCDR2 comprises the amino acid sequence of SEQ ID NO: 32, and the B-LCDR3 comprises the amino acid sequence of SEQ ID NO: 42 ;or (vii) The B-HCDR1 comprises the amino acid sequence of SEQ ID NO: 18, the B-HCDR2 comprises the amino acid sequence of SEQ ID NO: 30, and the B-HCDR3 comprises the amino acid sequence of SEQ ID NO: 20 , and the B-LCDR1 comprises the amino acid sequence of SEQ ID NO: 31, the B-LCDR2 comprises the amino acid sequence of SEQ ID NO: 32, and the B-LCDR3 comprises the amino acid sequence of SEQ ID NO: 33 . The antigen-binding molecule according to claim 1 or 2, wherein the B-VH comprises an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 63, and the B-VL comprises an amino acid sequence identical to SEQ ID NO: 64 Amino acid sequences having at least 90% sequence identity, wherein SEQ ID NO: 63 is: QVQLQQSGAEVKKPGSSVRVSCKASGGTFNNNAINWVRQAPGQX ₂₆ LEWMGX ₁ IX ₂ PMFGX ₃ AKYSX ₄ X ₅ FQGRVAITADESTGTASMELSSLRSEDTAVYYCARSRDX ₆ LLFPX ₇ X ₈ X ₉ LX ₁₀ X ₁₁ WGX ₂₇ GTMVTVSS, SEQ ID NO: 64 is: SSELTQDPAVSVALGQTVRVTCX ₁₂ GX ₁₃ X ₁₄ LX ₁₅ X ₁₆ X ₁₇ X ₁₈ ASWYQQKPGQAPVLVIYGKNNRPSGIPDRFSGSSSGNTASLTITGAQAEDEADYYCX ₁₉ SRX ₂₀ X ₂₁ X ₂₂ GX ₂₃ X ₂₄ WX ₂₅ FGX ₂₈ GT ELTVL, Wherein, _X1 is selected from S, A, V or G, _X2 is selected from A, I, M or S, _X3 is selected from T or G, _X4 is selected from Q, E, G or K, _X5 is selected from N, G or Q, X ₆ is selected from L or P, X ₇ is selected from H, Q or D, X ₈ is selected from H or D, X ₉ is selected from A or G, X ₁₀ is selected from S or L, X ₁₁ selected from P or S, X ₁₂ selected from Q or H, X ₁₃ selected from D, A or N, X ₁₄ selected from S or I, X ₁₅ selected from K, R or T, X ₁₆ selected from S, W, T or D, X ₁₇ is selected from H, Y or S, X ₁₈ is selected from Y or R, X ₁₉ is selected from G or S, X ₂₀ is selected from A or D, X ₂₁ is selected from E or S, X ₂₂ is selected from S or A, X ₂₃ is selected from V, E, A, N or W, X ₂₄ is selected from G, K, H or R, X ₂₅ is selected from L or V, X ₂₆ is selected from G or C, X ₂₇ is selected from R or G, X ₂₈ is selected from G or C; And, the B-VH and B-VL do not comprise the following variable region combination: B-VH comprises the amino acid sequence of SEQ ID NO: 1, and B-VL comprises the amino acid sequence of SEQ ID NO: 2; Preferably, (i) the B-VH comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO:47, and the B-VL comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO:48 sequence, or (ii) the B-VH comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO:5, and the B-VL comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO:12 sequence, or (iii) the B-VH comprises an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 6, and the B-VL comprises an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 13 sequence, or (iv) the B-VH comprises an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 7, and the B-VL comprises an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 14 sequence, or (v) the B-VH comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO:8, and the B-VL comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO:15 sequence, or (vi) the B-VH comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 9, and the B-VL comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 16 sequence, or (vii) the B-VH comprises an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 10, and the B-VL comprises an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 17 sequence, or (viii) the B-VH comprises an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 11, and the B-VL comprises an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 2 sequence; Better yet, The B-VH comprises the amino acid sequence of SEQ ID NO:47, and the B-VL comprises the amino acid sequence of SEQ ID NO:48. The antigen-binding molecule according to any one of claims 1 to 3, wherein the antigen-binding module 2 comprises a heavy chain variable region P-VH and a light chain variable region P-VL, the P-VH comprising P-HCDR1 , P-HCDR2 and P-HCDR3, the P-VL comprises P-LCDR1, P-LCDR2 and P-LCDR3; The P-HCDR1, P-HCDR2 and P-HCDR3 respectively comprise the amino acid sequences of Pv-HCDR1, Pv-HCDR2 and Pv-HCDR3 in SEQ ID NO: 51, and the P-LCDR1, P-LCDR2 and P- LCDR3 respectively comprises the amino acid sequences of Pv-LCDR1, Pv-LCDR2 and Pv-LCDR3 in SEQ ID NO:52; Preferably, the P-HCDR1, P-HCDR2, P-HCDR3, P-LCDR1, P-LCDR2 and P-LCDR3 are defined according to the Kabat numbering rules, and the P-HCDR1 comprises the amino acid of SEQ ID NO: 53 Sequence, the P-HCDR2 comprises the amino acid sequence of SEQ ID NO: 54, the P-HCDR3 comprises the amino acid sequence of SEQ ID NO: 55, the P-LCDR1 comprises the amino acid sequence of SEQ ID NO: 56, The P-LCDR2 comprises the amino acid sequence of SEQ ID NO:57, and the P-LCDR3 comprises the amino acid sequence of SEQ ID NO:58; More preferably, the heavy chain variable region P-VH comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 51, and the light chain variable region P-VL comprises an amino acid sequence identical to SEQ ID NO: 52 Amino acid sequences having at least 90% sequence identity; Optimally, the heavy chain variable region P-VH comprises the amino acid sequence of SEQ ID NO:51, and the light chain variable region P-VL comprises the amino acid sequence of SEQ ID NO:52. The antigen-binding molecule according to any one of claims 1 to 4, comprising a heavy chain constant region CH and a light chain constant region CL; Preferably, the heavy chain constant region CH is a human IgG1 heavy chain constant region, and the light chain constant region CL is a human kappa light chain constant region; More preferably, the heavy chain constant region CH comprises the amino acid sequence of SEQ ID NO:59, and the light chain constant region CL comprises the amino acid sequence of SEQ ID NO:60. The antigen-binding molecule according to claim 5, wherein the antigen-binding module 1 is a scFv, and the antigen-binding module 2 comprises a heavy chain variable region P-VH, a light chain variable region P-VL, and a heavy chain constant region A full-length antibody of CH and light chain constant region CL, the antigen-binding module 1 is fused directly or via a linker to the N-terminal of the variable region or the C-terminal of the constant region of the antigen-binding module 2; Preferably, the antigen-binding molecule comprises: A first chain having a structure shown in formula (a) and a second chain having a structure shown in formula (b); or A first chain having a structure shown in formula (c) and a second chain having a structure shown in formula (b), Formula (a) [P-VH]-[CH]-[Linker 1]-[B-VH]-[Linker 2]-[B-VL], Formula (b) [P-VL]-[CL], Formula (c) [P-VH]-[CH]-[Linker 1]-[B-VL]-[Linker 2]-[B-VH], More preferably, in formula (a) and formula (c), linker 1 and linker 2 are the same or different peptide linkers; Optimally, in formula (a) and formula (c), the amino acid sequence of linker 1 is shown in SEQ ID NO:69, and the amino acid sequence of linker 2 is shown in SEQ ID NO:70. The antigen-binding molecule as claimed in item 6, wherein, The first strand comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO:61, and the second strand comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO:50, or The first strand comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO:62, and the second strand comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO:50; Preferably, the antigen-binding molecule has: (i) two first strands comprising the amino acid sequence of SEQ ID NO: 61 and two second strands comprising the amino acid sequence of SEQ ID NO: 50, or (ii) Two first strands comprising the amino acid sequence of SEQ ID NO:62 and two second strands comprising the amino acid sequence of SEQ ID NO:50. An antigen-binding molecule that specifically binds to a BAFF antigen comprising: a heavy chain variable region B-VH and a light chain variable region B-VL, The B-VH comprises B-HCDR1, B-HCDR2 and B-HCDR3, The B-VL includes B-LCDR1, B-LCDR2 and B-LCDR3, wherein, The B-HCDR1, B-HCDR2 and B-HCDR3 respectively comprise the amino acid sequences of Bv-HCDR1, Bv-HCDR2 and Bv-HCDR3 in SEQ ID NO: 63, and the B-LCDR1, B-LCDR2 and B- LCDR3 respectively comprises the amino acid sequences of Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 in SEQ ID NO: 64, wherein, SEQ ID NO: 63 is:

SEQ ID NO: 64 is:

Wherein, _X1 is selected from S, A, V or G, _X2 is selected from A, I, M or S, _X3 is selected from T or G, _X4 is selected from Q, E, G or K, _X5 is selected from N, G or Q, X ₆ is selected from L or P, X ₇ is selected from H, Q or D, X ₈ is selected from H or D, X ₉ is selected from A or G, X ₁₀ is selected from S or L, X ₁₁ selected from P or S, X ₁₂ selected from Q or H, X ₁₃ selected from D, A or N, X ₁₄ selected from S or I, X ₁₅ selected from K, R or T, X ₁₆ selected from S, W, T or D, X ₁₇ is selected from H, Y or S, X ₁₈ is selected from Y or R, X ₁₉ is selected from G or S, X ₂₀ is selected from A or D, X ₂₁ is selected from E or S, X ₂₂ is selected from S or A, X ₂₃ is selected from V, E, A, N or W, X ₂₄ is selected from G, K, H or R, X ₂₅ is selected from L or V, X ₂₆ is selected from G or C, X ₂₇ is selected from R or G, X ₂₈ is selected from G or C; Moreover, the B-HCDR1, B-HCDR2, B-HCDR3, B-LCDR1, B-LCDR2 and B-LCDR3 do not contain the following CDR combination: B-HCDR1, B-HCDR2 and B-HCDR3 respectively contain SEQ ID NO: 1 The amino acid sequences of Bv-HCDR1, Bv-HCDR2 and Bv-HCDR3 in, and B-LCDR1, B-LCDR2 and B-LCDR3 respectively comprise B-LCDR1, B-LCDR2 and B-LCDR1 in SEQ ID NO:2 LCDR3; Preferably, The B-HCDR1, B-HCDR2, B-HCDR3, B-LCDR1, B-LCDR2 and B-LCDR3 are defined according to the Kabat numbering rules, wherein, B-HCDR1 comprises the amino acid sequence NNAIN (SEQ ID NO: 18), B-HCDR2 comprises the amino acid sequence X ₁ IX ₂ PMFGX ₃ AKYSX ₄ X ₅ FQG (SEQ ID NO: 65), B-HCDR3 comprises the amino acid sequence SRDX ₆ LLFPX ₇ X ₈ X ₉ LX ₁₀ X ₁₁ (SEQ ID NO: 66), B-LCDR1 comprises the amino acid sequence X ₁₂ GX ₁₃ X ₁₄ LX ₁₅ X ₁₆ X ₁₇ X ₁₈ AS (SEQ ID NO: 67), B-LCDR2 comprises the amino acid sequence GKNNRPS (SEQ ID NO: 32) and B-LCDR3 comprises the amino acid sequence X ₁₉ SRX ₂₀ X ₂₁ X ₂₂ GX ₂₃ X ₂₄ WX ₂₅ (SEQ ID NO: 68), Wherein, _X1 is selected from S, A, V or G, _X2 is selected from A, I, M or S, _X3 is selected from T or G, _X4 is selected from Q, E, G or K, _X5 is selected from N, G or Q, X ₆ is selected from L or P, X ₇ is selected from H, Q or D, X ₈ is selected from H or D, X ₉ is selected from A or G, X ₁₀ is selected from S or L, X ₁₁ selected from P or S, X ₁₂ selected from Q or H, X ₁₃ selected from D, A or N, X ₁₄ selected from S or I, X ₁₅ selected from K, R or T, X ₁₆ selected from S, W, T or D, X ₁₇ is selected from H, Y or S, X ₁₈ is selected from Y or R, X ₁₉ is selected from G or S, X ₂₀ is selected from A or D, X ₂₁ is selected from E or S, X ₂₂ is selected from S or A, X ₂₃ is selected from V, E, A, N or W, X ₂₄ is selected from G, K, H or R, X ₂₅ is selected from L or V; And, the B-HCDR1, B-HCDR2, B-HCDR3, B-LCDR1, B-LCDR2 and B-LCDR3 do not contain the following CDR combination: B-HCDR1 contains the amino acid sequence of SEQ ID NO: 18, B-HCDR2 Comprising the amino acid sequence of SEQ ID NO: 19, B-HCDR3 comprising the amino acid sequence of SEQ ID NO: 20, B-LCDR1 comprising the amino acid sequence of SEQ ID NO: 31 Amino acid sequence, B-LCDR2 comprises the amino acid sequence of SEQ ID NO:32, and B-LCDR3 comprises the amino acid sequence of SEQ ID NO:33. The antigen-binding molecule as claimed in item 8, wherein, (i) the B-HCDR1, B-HCDR2 and B-HCDR3 respectively comprise the amino acid sequences of Bv-HCDR1, Bv-HCDR2 and Bv-HCDR3 in SEQ ID NO: 47, and the B-LCDR1, B-LCDR2 and B-LCDR3 respectively comprising the amino acid sequences of Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 in SEQ ID NO: 48, or (ii) the B-HCDR1, B-HCDR2 and B-HCDR3 respectively comprise the amino acid sequences of Bv-HCDR1, Bv-HCDR2 and Bv-HCDR3 in SEQ ID NO: 5, and the B-LCDR1, B-LCDR2 and B-LCDR3 respectively comprising the amino acid sequences of Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 in SEQ ID NO: 12, or (iii) the B-HCDR1, B-HCDR2 and B-HCDR3 respectively comprise the amino acid sequences of Bv-HCDR1, Bv-HCDR2 and Bv-HCDR3 in SEQ ID NO: 6, and the B-LCDR1, B-LCDR2 and B-LCDR3 respectively comprising the amino acid sequences of Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 in SEQ ID NO: 13, or (iv) the B-HCDR1, B-HCDR2 and B-HCDR3 respectively comprise the amino acid sequences of Bv-HCDR1, Bv-HCDR2 and Bv-HCDR3 in SEQ ID NO: 7, and the B-LCDR1, B-LCDR2 and B-LCDR3 respectively comprising the amino acid sequences of Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 in SEQ ID NO: 14, or (v) the B-HCDR1, B-HCDR2 and B-HCDR3 respectively comprise the amino acid sequences of Bv-HCDR1, Bv-HCDR2 and Bv-HCDR3 in SEQ ID NO: 8, and the B- LCDR1, B-LCDR2 and B-LCDR3 respectively comprise the amino acid sequences of Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 in SEQ ID NO: 15, or (vi) The B-HCDR1, B-HCDR2 and B-HCDR3 respectively comprise the amino acid sequences of Bv-HCDR1, Bv-HCDR2 and Bv-HCDR3 in SEQ ID NO: 9, and the B-LCDR1, B-LCDR2 and B-LCDR3 respectively comprising the amino acid sequences of Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 in SEQ ID NO: 16, or (vii) the B-HCDR1, B-HCDR2 and B-HCDR3 respectively comprise the amino acid sequences of Bv-HCDR1, Bv-HCDR2 and Bv-HCDR3 in SEQ ID NO: 10, and the B-LCDR1, B-LCDR2 and B-LCDR3 respectively comprising the amino acid sequences of Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 in SEQ ID NO: 17, or (viii) the B-HCDR1, B-HCDR2 and B-HCDR3 respectively comprise the amino acid sequences of Bv-HCDR1, Bv-HCDR2 and Bv-HCDR3 in SEQ ID NO: 11, and the B-LCDR1, B-LCDR2 and B-LCDR3 respectively comprise the amino acid sequences of Bv-LCDR1, Bv-LCDR2 and Bv-LCDR3 in SEQ ID NO: 2; Preferably, The B-HCDR1, B-HCDR2, B-HCDR3, B-LCDR1, B-LCDR2 and B-LCDR3 are defined according to the Kabat numbering rules, and, (i) The B-HCDR1 comprises the amino acid sequence of SEQ ID NO: 18, the B-HCDR2 comprises the amino acid sequence of SEQ ID NO: 28, and the B-HCDR3 comprises the amino acid sequence of SEQ ID NO: 29 , and the B-LCDR1 comprises the amino acid sequence of SEQ ID NO: 43, the B-LCDR2 comprises the amino acid sequence of SEQ ID NO: 32, and the B-LCDR3 comprises the amino acid sequence of SEQ ID NO: 44 ;or (ii) The B-HCDR1 comprises the amino acid sequence of SEQ ID NO: 18, the B-HCDR2 comprises the amino acid sequence of SEQ ID NO: 21, and the B-HCDR3 comprises the amino acid sequence of SEQ ID NO: 20 , and the B-LCDR1 comprises the amino acid sequence of SEQ ID NO: 34, the B-LCDR2 comprises the amino acid sequence of SEQ ID NO: 32, and the B-LCDR3 comprises the amino acid sequence of SEQ ID NO: 35 ;or (iii) The B-HCDR1 comprises the amino acid sequence of SEQ ID NO: 18, the B-HCDR2 comprises the amino acid sequence of SEQ ID NO: 22, and the B-HCDR3 comprises the amino acid sequence of SEQ ID NO: 23 , and the B-LCDR1 comprises the amino acid sequence of SEQ ID NO: 31, the B-LCDR2 comprises the amino acid sequence of SEQ ID NO: 32, and the B-LCDR3 comprises the amino acid sequence of SEQ ID NO: 36 ;or (iv) The B-HCDR1 comprises the amino acid sequence of SEQ ID NO: 18, the B-HCDR2 comprises the amino acid sequence of SEQ ID NO: 24, and the B-HCDR3 comprises the amino acid sequence of SEQ ID NO: 20 , and the B-LCDR1 comprises the amino acid sequence of SEQ ID NO: 37, the B-LCDR2 comprises the amino acid sequence of SEQ ID NO: 32, and the B-LCDR3 comprises the amino acid sequence of SEQ ID NO: 38 ;or (v) The B-HCDR1 comprises the amino acid sequence of SEQ ID NO: 18, the B-HCDR2 comprises the amino acid sequence of SEQ ID NO: 25, and the B-HCDR3 comprises the amino acid sequence of SEQ ID NO: 20 , and the B-LCDR1 comprises the amino acid sequence of SEQ ID NO: 39, the B-LCDR2 comprises the amino acid sequence of SEQ ID NO: 32, and the B-LCDR3 comprises the amino acid sequence of SEQ ID NO: 40 ;or (vi) The B-HCDR1 comprises the amino acid sequence of SEQ ID NO: 18, the B-HCDR2 comprises the amino acid sequence of SEQ ID NO: 26, and the B-HCDR3 comprises the amino acid sequence of SEQ ID NO: 27 The amino acid sequence of, and the B-LCDR1 comprises the amino acid sequence of SEQ ID NO: 41, the B-LCDR2 comprises the amino acid sequence of SEQ ID NO: 32, and the B-LCDR3 comprises the amino acid sequence of SEQ ID NO: 42 the amino acid sequence of ; or (vii) The B-HCDR1 comprises the amino acid sequence of SEQ ID NO: 18, the B-HCDR2 comprises the amino acid sequence of SEQ ID NO: 30, and the B-HCDR3 comprises the amino acid sequence of SEQ ID NO: 20 , and the B-LCDR1 comprises the amino acid sequence of SEQ ID NO: 31, the B-LCDR2 comprises the amino acid sequence of SEQ ID NO: 32, and the B-LCDR3 comprises the amino acid sequence of SEQ ID NO: 33 . The antigen-binding molecule according to claim 8 or 9, wherein the B-VH comprises an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 63, and the B-VL comprises an amino acid sequence identical to SEQ ID NO: 64 Amino acid sequences having at least 90% sequence identity, wherein SEQ ID NO: 63 is:

SEQ ID NO: 64 is:

Wherein, _X1 is selected from S, A, V or G, _X2 is selected from A, I, M or S, _X3 is selected from T or G, _X4 is selected from Q, E, G or K, _X5 is selected from N, G or Q, X ₆ is selected from L or P, X ₇ is selected from H, Q or D, X ₈ is selected from H or D, X ₉ is selected from A or G, X ₁₀ is selected from S or L, X ₁₁ selected from P or S, X ₁₂ selected from Q or H, X ₁₃ selected from D, A or N, X ₁₄ selected from S or I, X ₁₅ selected from K, R or T, X ₁₆ selected from S, W, T or D, X ₁₇ is selected from H, Y or S, X ₁₈ is selected from Y or R, X ₁₉ is selected from G or S, X ₂₀ is selected from A or D, X ₂₁ is selected from E or S, X ₂₂ is selected from S or A, X ₂₃ is selected from V, E, A, N or W, X ₂₄ is selected from G, K, H or R, X ₂₅ is selected from L or V, X ₂₆ is selected from G or C, X ₂₇ is selected from R or G, X ₂₈ is selected from G or C; And, the B-VH and B-VL do not comprise the following variable region combination: B-VH comprises the amino acid sequence of SEQ ID NO: 1, and B-VL comprises the amino acid sequence of SEQ ID NO: 2; Preferably, (i) the B-VH comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO:47, and the B-VL comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO:48 sequence, or (ii) the B-VH comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO:5, and the B-VL comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO:12 sequence, or (iii) the B-VH comprises an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 6, and the B-VL comprises an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 13 sequence, or (iv) the B-VH comprises an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 7, and the B-VL comprises an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 14 sequence, or (v) the B-VH comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO:8, and the B-VL comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO:15 sequence, or (vi) the B-VH comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 9, and the B-VL comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 16 sequence, or (vii) the B-VH comprises an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 10, and the B-VL comprises an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 17 sequence, or (viii) the B-VH comprises an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 11, and the B-VL comprises an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 2 sequence; Better yet, The B-VH comprises the amino acid sequence of SEQ ID NO: 47, and the B-VL comprises the amino acid sequence of SEQ ID NO: 48, or The B-VH comprises the amino acid sequence of SEQ ID NO: 5, and the B-VL comprises the amino acid sequence of SEQ ID NO: 12, or The B-VH comprises the amino acid sequence of SEQ ID NO: 6, and the B-VL comprises the amino acid sequence of SEQ ID NO: 13, or The B-VH comprises the amino acid sequence of SEQ ID NO: 7, and the B-VL comprises the amino acid sequence of SEQ ID NO: 14, or The B-VH comprises the amino acid sequence of SEQ ID NO: 8, and the B-VL comprises the amino acid sequence of SEQ ID NO: 15, or The B-VH comprises the amino acid sequence of SEQ ID NO: 9, and the B-VL comprises the amino acid sequence of SEQ ID NO: 16, or The B-VH comprises the amino acid sequence of SEQ ID NO: 10, and the B-VL comprises the amino acid sequence of SEQ ID NO: 17, or The B-VH comprises the amino acid sequence of SEQ ID NO:11, and the B-VL comprises the amino acid sequence of SEQ ID NO:2. The antigen-binding molecule according to any one of claims 8 to 10, comprising a heavy chain constant region CH and a light chain constant region CL; Preferably, the heavy chain constant region CH is a human IgG1 heavy chain constant region, and the light chain constant region CL is a human lambda light chain constant region; More preferably, the heavy chain constant region CH comprises the amino acid sequence of SEQ ID NO:45, and the light chain constant region CL comprises the amino acid sequence of SEQ ID NO:46. The antigen-binding molecule according to any one of claims 1 to 11, wherein the antigen-binding molecule has one or more of the following characteristics: A. The antigen-binding molecule can block the binding of human BAFF and human BAFF-R; preferably, the IC50 value of blocking the binding of human BAFF and human BAFF-R is less than 11 nM, and the IC50 value is detected by Elisa method; B. The antigen-binding molecule can inhibit BAFF-induced B cell proliferation; preferably, the antigen-binding molecule can inhibit BAFF-induced B cell proliferation with an IC50 value of less than 0.2nM; C. The antigen-binding molecule can bind to human BAFF with a KD value of less than 9.99E-10M, and the KD value is detected by a Biacore method; D. The antigen-binding molecule can bind to cynomolgus BAFF with a KD value less than 5.00E-10M, and the KD value is detected by Biacore method; E. The antigen-binding molecule can bind to mouse BAFF with a KD value less than 2.20E-10M, and the KD value is detected by the Biacore method; or G. The antigen-binding molecule can block the binding of human BAFF and human TACI; preferably, the IC50 value of blocking the binding of human BAFF and human TACI is less than 0.6nM, and the IC50 value is detected by Elisa method. A pharmaceutical composition, which contains: A therapeutically effective amount of the antigen-binding molecule of any one of claims 1 to 12, and One or more pharmaceutically acceptable carriers, diluents, buffers or excipients. An isolated nucleic acid encoding the antigen-binding molecule of any one of claims 1-12. A host cell comprising the isolated nucleic acid as described in claim 14. A method of treating a disease, the method comprising the step of administering the antigen-binding molecule according to any one of claims 1 to 12 or the pharmaceutical composition according to claim 13 to a subject; Preferably, the disease is a B cell disorder or an autoimmune disease; More preferably, 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; The 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 gamma globulinopathy; Optimally, the disease is systemic lupus erythematosus.