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CN-122011196-A - Bispecific antibodies against TL1A and TNF-alpha and uses thereof

CN122011196ACN 122011196 ACN122011196 ACN 122011196ACN-122011196-A

Abstract

The present disclosure relates to a bispecific antibody comprising a first domain that specifically binds TNF-like ligand 1A (TL 1A), and a second domain that specifically binds tumor necrosis factor alpha (TNF-a). The disclosure also relates to methods of treating or preventing diseases associated with TL1A and/or TNF- α by the bispecific antibodies.

Inventors

  • ZHOU JINGYUN
  • JIANG YANJING
  • WANG MENGMENG
  • MA RONG
  • XIE XIAOJUAN
  • LIU XUEWEI
  • ZHANG LILI
  • LI DONGXIA
  • LI ZIQIANG

Assignees

  • 北京伟德杰生物科技有限公司

Dates

Publication Date
20260512
Application Date
20250523
Priority Date
20241111

Claims (20)

  1. 1. A bispecific antibody comprising a first domain that specifically binds TNF-like ligand 1A (TL 1A) and a second domain that specifically binds tumor necrosis factor alpha (TNF-alpha).
  2. 2. The bispecific antibody according to claim 1, characterized in that the first domain is an antibody or a functional fragment thereof which specifically binds TL1A, and/or The second domain is an antibody or functional fragment thereof that specifically binds TNF- α.
  3. 3. The bispecific antibody according to claim 1 or 2, characterized in that the first domain comprises an antibody Fab fragment, fab 'fragment, F (ab') 2 fragment, fv fragment, scFv fragment, nanobody, heavy chain variable region (VH) fragment and/or light chain variable region (VL) fragment which specifically binds TL1A, and/or The second domain includes an antibody Fab fragment, fab 'fragment, F (ab') 2 fragment, fv fragment, scFv fragment, nanobody, heavy chain variable region (VH) fragment, and/or light chain variable region (VL) fragment that specifically binds TNF- α.
  4. 4. The bispecific antibody according to any one of claims 1 to 3, characterized in that the first domain and the second domain are linked directly or via a linker, Preferably, the linker has an amino acid sequence as shown in the general formula (G n S) m , n and m are integers of 1 to 10 respectively, more preferably n is an integer of 1 to 4, and m is an integer of 1 to 3, or an amino acid sequence having 1, 2 or 3 amino acid insertions, substitutions or deletions compared to the amino acid sequence shown in the general formula (G n S) m ).
  5. 5. The bispecific antibody of any one of claims 1 to 4, wherein the first domain comprises (1) the following 3 heavy chain variable region complementarity determining regions (HCDR): HCDR1 having an amino acid sequence of HCDR1 contained in the heavy chain variable region as shown in SEQ ID No. 1 or 3, or an amino acid sequence having substitution, deletion or addition of one or more amino acids as compared with the amino acid sequence of HCDR1 contained in the heavy chain variable region; HCDR2 having an amino acid sequence of HCDR2 contained in the heavy chain variable region as set forth in any one of SEQ ID NOs 1 or 3, or an amino acid sequence having a substitution, deletion or addition of one or several amino acids compared to the amino acid sequence of HCDR2 contained in the heavy chain variable region; HCDR3 having an amino acid sequence of HCDR3 contained in the heavy chain variable region as set forth in any one of SEQ ID NOs 1 or 3, or an amino acid sequence having substitution, deletion or addition of one or several amino acids as compared with the amino acid sequence of HCDR3 contained in the heavy chain variable region, and/or (2) The following 3 light chain variable region complementarity determining regions (LCDR): LCDR1, which has the amino acid sequence of LCDR1 contained in the light chain variable region as shown in SEQ ID NO. 2 or 4, or has one or several amino acid substitutions, deletions or additions compared to the amino acid sequence of LCDR1 contained in the light chain variable region; LCDR2 having the amino acid sequence of LCDR2 contained in the light chain variable region as shown in SEQ ID No. 2 or 4 or having one or more amino acid substitutions, deletions or additions compared to the amino acid sequence of LCDR2 contained in said light chain variable region; LCDR3 having an amino acid sequence of LCDR3 contained in a light chain variable region as shown in SEQ ID No. 2 or 4 or having a substitution, deletion or addition of one or more amino acids compared to the amino acid sequence of LCDR3 contained in said light chain variable region; Preferably, the first domain comprises: HCDR1, HCDR2 and HCDR3 contained in the heavy chain variable region as shown in SEQ ID No. 1, and LCDR1, LCDR2 and LCDR3 contained in the light chain variable region as shown in SEQ ID No. 2, or HCDR1, HCDR2 and HCDR3 contained in the heavy chain variable region as shown in SEQ ID No. 3, and LCDR1, LCDR2 and LCDR3 contained in the light chain variable region as shown in SEQ ID No. 4; Preferably, said HCDR1-3 and/or said LCDR1-3 are defined by an IMGT numbering system, a Kabat numbering system, a Chothia numbering system, a Contact numbering system or a combination thereof.
  6. 6. The bispecific antibody of any one of claims 1-5, wherein the first domain comprises: (1) Heavy chain variable regions and/or light chain variable regions wherein HCDR1-3 and/or LCDR1-3 are defined by the IMGT numbering system: (a) A heavy chain variable region comprising 3 HCDRs of HCDR1 of SEQ ID NO. 7, HCDR2 of SEQ ID NO. 8, HCDR3 of SEQ ID NO. 9, and/or a light chain variable region comprising 3 LCDRs of LCDR1 of SEQ ID NO. 10, LCDR2 of YAT, LCDR3 of SEQ ID NO. 11, or (B) A heavy chain variable region comprising 3 HCDRs, a HCDR1 of SEQ ID NO. 25, a HCDR2 of SEQ ID NO. 26, a HCDR3 of SEQ ID NO. 27, and/or a light chain variable region comprising 3 LCDRs, a LCDR1 of SEQ ID NO. 28, a LCDR2 of DAS, a LCDR3 of SEQ ID NO. 29; Or alternatively (2) Heavy chain variable regions and/or light chain variable regions wherein HCDR1-3 and/or LCDR1-3 are defined according to the Kabat numbering system: (a) A heavy chain variable region comprising 3 HCDRs of HCDR1 sequence SEQ ID NO. 12, HCDR2 sequence SEQ ID NO. 13, HCDR3 sequence SEQ ID NO. 14, and/or a light chain variable region comprising 3 LCDRs of LCDR1 sequence SEQ ID NO. 15, LCDR2 sequence SEQ ID NO. 16, LCDR3 sequence SEQ ID NO. 11, or (B) A heavy chain variable region comprising 3 HCDRs of HCDR1 sequence SEQ ID NO. 30, HCDR2 sequence SEQ ID NO. 31, HCDR3 sequence SEQ ID NO. 32, and/or a light chain variable region comprising 3 LCDRs of LCDR1 sequence SEQ ID NO. 33, LCDR2 sequence SEQ ID NO. 34, LCDR3 sequence SEQ ID NO. 29; Or alternatively (3) Heavy chain variable regions and/or light chain variable regions wherein HCDR1-3 and/or LCDR1-3 are defined by the Chothia numbering system: (a) Comprising 3 HCDR heavy chain variable regions of HCDR1 of SEQ ID NO. 17, HCDR2 of SEQ ID NO. 18, HCDR3 of SEQ ID NO. 14, and/or comprising 3 LCDR light chain variable regions of LCDR1 of SEQ ID NO. 15, LCDR2 of SEQ ID NO. 16, LCDR3 of SEQ ID NO. 11, or (B) A heavy chain variable region comprising 3 HCDRs of HCDR1 sequence SEQ ID NO. 35, HCDR2 sequence SEQ ID NO. 36, HCDR3 sequence SEQ ID NO. 32, and/or a light chain variable region comprising 3 LCDRs of LCDR1 sequence SEQ ID NO. 33, LCDR2 sequence SEQ ID NO. 34, LCDR3 sequence SEQ ID NO. 29; Or alternatively (4) Heavy chain variable regions and/or light chain variable regions, wherein HCDR1-3 and/or LCDR1-3 are defined by the Contact numbering system: (a) Comprising 3 HCDR heavy chain variable regions of HCDR1 of SEQ ID NO. 19, HCDR2 of SEQ ID NO. 20, HCDR3 of SEQ ID NO. 21, and/or comprising 3 LCDR light chain variable regions of LCDR1 of SEQ ID NO. 22, LCDR2 of SEQ ID NO. 23, LCDR3 of SEQ ID NO. 24, or (B) Comprising 3 heavy chain variable regions of HCDR1 of SEQ ID NO. 37, HCDR2 of SEQ ID NO. 38, HCDR3 of SEQ ID NO. 39, and/or light chain variable regions of LCDR1 of SEQ ID NO. 40, LCDR2 of SEQ ID NO. 41, LCDR3 of SEQ ID NO. 42.
  7. 7. The bispecific antibody of any one of claims 1-6, wherein the first domain comprises: A heavy chain variable region having an amino acid sequence as shown in SEQ ID NO. 1 or 3, an amino acid sequence having one or more amino acid substitutions, deletions or additions as compared with it, or an amino acid sequence having at least 80% sequence identity thereto, and A light chain variable region having an amino acid sequence as shown in SEQ ID NO. 2 or 4, an amino acid sequence having one or more amino acid substitutions, deletions or additions as compared to it, or an amino acid sequence having at least 80% sequence identity thereto; Preferably, the first domain comprises a heavy chain variable region having the amino acid sequence shown in SEQ ID NO. 1 and a light chain variable region having the amino acid sequence shown in SEQ ID NO. 2; Preferably, the first domain comprises a heavy chain variable region having the amino acid sequence shown in SEQ ID NO.3 and a light chain variable region having the amino acid sequence shown in SEQ ID NO. 4.
  8. 8. The bispecific antibody of any one of claims 1-7, wherein the second domain comprises the following 3 heavy chain variable region complementarity determining regions (HCDR): HCDR1 having an amino acid sequence of HCDR1 contained in the heavy chain variable region as shown in SEQ ID No. 5 or an amino acid sequence having one or more amino acid substitutions, deletions or additions compared to the amino acid sequence of HCDR1 contained in the heavy chain variable region; HCDR2 having the amino acid sequence of HCDR2 contained in the heavy chain variable region as shown in SEQ ID No. 5 or having one or more amino acid substitutions, deletions or additions compared to the amino acid sequence of HCDR2 contained in the heavy chain variable region; HCDR3 having an amino acid sequence of HCDR3 contained in the heavy chain variable region as shown in SEQ ID NO. 5 or an amino acid sequence having substitution, deletion or addition of one or several amino acids as compared with the amino acid sequence of HCDR3 contained in the heavy chain variable region, and/or The following 3 light chain variable region complementarity determining regions (LCDR): LCDR1, which has the amino acid sequence of LCDR1 contained in the light chain variable region as shown in SEQ ID NO. 6, or an amino acid sequence having substitution, deletion or addition of one or several amino acids as compared with the amino acid sequence of LCDR1 contained in the light chain variable region; LCDR2, which has the amino acid sequence of LCDR2 contained in the light chain variable region as shown in SEQ ID NO. 6, or an amino acid sequence having substitution, deletion or addition of one or several amino acids as compared with the amino acid sequence of LCDR2 contained in the light chain variable region; LCDR3, which has the amino acid sequence of LCDR3 contained in the light chain variable region as shown in SEQ ID NO. 6, or an amino acid sequence having substitution, deletion or addition of one or several amino acids as compared with the amino acid sequence of LCDR3 contained in the light chain variable region; Preferably, said HCDR1-3 and/or said LCDR1-3 are defined by an IMGT numbering system, a Kabat numbering system, a Chothia numbering system, a Contact numbering system or a combination thereof.
  9. 9. The bispecific antibody of any one of claims 1-8, wherein the second domain comprises: (1) Heavy chain variable regions and/or light chain variable regions wherein HCDR1-3 and/or LCDR1-3 are defined by the IMGT numbering system: A heavy chain variable region comprising 3 HCDRs, a HCDR1 having the sequence SEQ ID NO. 43, a HCDR2 having the sequence SEQ ID NO. 44, a HCDR3 having the sequence SEQ ID NO. 45, and/or a light chain variable region comprising 3 LCDRs, a LCDR1 having the sequence SEQ ID NO. 46, a LCDR2 having the sequence DAS, a LCDR3 having the sequence SEQ ID NO. 47; Or alternatively (2) Heavy chain variable regions and/or light chain variable regions wherein HCDR1-3 and/or LCDR1-3 are defined according to the Kabat numbering system: a heavy chain variable region comprising 3 HCDRs of HCDR1 sequence SEQ ID NO. 48, HCDR2 sequence SEQ ID NO. 49, HCDR3 sequence SEQ ID NO. 50, and/or a light chain variable region comprising 3 LCDRs of LCDR1 sequence SEQ ID NO. 51, LCDR2 sequence SEQ ID NO. 52, LCDR3 sequence SEQ ID NO. 47; Or alternatively (3) Heavy chain variable regions and/or light chain variable regions wherein HCDR1-3 and/or LCDR1-3 are defined by the Chothia numbering system: A heavy chain variable region comprising 3 HCDRs of HCDR1 sequence SEQ ID NO. 53, HCDR2 sequence SEQ ID NO. 54, HCDR3 sequence SEQ ID NO. 50, and/or a light chain variable region comprising 3 LCDRs of LCDR1 sequence SEQ ID NO. 51, LCDR2 sequence SEQ ID NO. 52, LCDR3 sequence SEQ ID NO. 47; Or alternatively (4) Heavy chain variable regions and/or light chain variable regions, wherein HCDR1-3 and/or LCDR1-3 are defined by the Contact numbering system: Comprising 3 heavy chain variable regions of HCDR1 of SEQ ID NO:55, HCDR2 of SEQ ID NO:56, HCDR3 of SEQ ID NO:57, and/or comprising 3 light chain variable regions of LCDR1 of SEQ ID NO:58, LCDR2 of SEQ ID NO:59, LCDR3 of SEQ ID NO: 60.
  10. 10. The bispecific antibody of any one of claims 1-9, wherein the second domain comprises: A heavy chain variable region having an amino acid sequence as shown in SEQ ID NO. 5, an amino acid sequence having one or more amino acid substitutions, deletions or additions thereto or an amino acid sequence having at least 80% sequence identity thereto, and A light chain variable region having an amino acid sequence as shown in SEQ ID NO. 6, an amino acid sequence having one or more amino acid substitutions, deletions or additions as compared to it, or an amino acid sequence having at least 80% sequence identity thereto.
  11. 11. An isolated nucleic acid molecule encoding the bispecific antibody of any one of claims 1 to 10.
  12. 12. An expression vector comprising the isolated nucleic acid of claim 10.
  13. 13. A host cell comprising the isolated nucleic acid molecule of claim 11 or the expression vector of claim 12.
  14. 14. A chimeric antigen receptor comprising an extracellular antigen-binding fragment, a transmembrane domain, and an intracellular signaling domain, wherein the extracellular antigen-binding fragment comprises the bispecific antibody of any one of claims 1-10.
  15. 15. A conjugate comprising the bispecific antibody of any one of claims 1 to 10, and a coupling moiety; Preferably, the coupling moiety is selected from the group consisting of detectable labels, radioisotopes, fluorescent substances, luminescent substances, colored substances, enzymes, polyethylene glycol (PEG), nuclides, nucleic acids, small molecule toxins, polypeptides having binding activity, proteins, receptors, ligands, and other active substances that inhibit tumor cell growth, promote tumor cell apoptosis or necrosis.
  16. 16. A pharmaceutical composition comprising the bispecific antibody of any one of claims 1 to 10, the isolated nucleic acid molecule of claim 11, or the expression vector of claim 12, and a pharmaceutically acceptable carrier; Preferably, the pharmaceutical composition is selected from the form of a tablet, powder, granule, pill, injection, suspension, powder, emulsion, aerosol, gel, eye drop, sustained release agent or sustained release implant.
  17. 17. A kit comprising the pharmaceutical composition of claim 16, packaged in a container, Preferably, the container is selected from the group consisting of a glass ampoule, a glass bottle, a plastic ampoule, a plastic bottle, a plastic bag, and a prefilled syringe.
  18. 18. A kit comprising the bispecific antibody of any one of claims 1 to 10, and optionally, instructions for use.
  19. 19. Use of a bispecific antibody according to any one of claims 1 to 10, an isolated nucleic acid molecule according to claim 11, an expression vector according to claim 12, a chimeric antigen receptor according to claim 14, a conjugate according to claim 15 or a pharmaceutical composition according to claim 16 for the preparation of a medicament for the prevention and/or treatment of TL1A and/or TNF-a related diseases.
  20. 20. A method of treating or preventing a disorder associated with TL1A and/or TNF-a comprising administering to a subject a therapeutically effective amount of the bispecific antibody of any one of claims 1 to 10, the isolated nucleic acid molecule of claim 11, the expression vector of claim 12, the chimeric antigen receptor of claim 14, the conjugate of claim 15, or the pharmaceutical composition of claim 16.

Description

Bispecific antibodies against TL1A and TNF-alpha and uses thereof Technical Field The present disclosure relates to bispecific antibodies directed against TL1A and TNF- α and uses thereof. Background TNF-like ligand 1A (TL 1A, also known as TNF superfamily member 15 (TNFSF 15)) is one of the tumor necrosis factor superfamily members, a type II membrane protein, which is expressed by antigen presenting cells (including dendritic cells, B cells and macrophages), cd4+ and cd8+ T cells and endothelial cells and can be expressed on the cell surface or secreted in the form of a soluble cytokine. TL1A is associated with the development and severity of Inflammatory Bowel Disease (IBD), the major types of IBD being Ulcerative Colitis (UC) and Crohn's Disease (CD). These diseases are very common and the number of available therapies is limited and some patients either do not respond to the initial treatment or over time the response disappears. Currently, the only therapy for patients who do not respond to first line therapy is surgery. Surgical treatment of IBD is invasive, and about one third of patients present a post-operative risk after receiving surgery. The pathogenesis of IBD is thought to involve an uncontrolled immune response, and heterogeneity of disease pathogenesis and clinical course suggests that targeted therapeutic approaches to treat these diseases are ideal therapeutic strategies. Thus, novel therapeutic approaches that specifically target the pathogenesis of IBD, drugs that inhibit TL1A activity are hot targets for developing therapies for IBD. Tumor necrosis factor alpha (TNF- α) is a homotrimeric protein consisting of 157 amino acids, produced mainly by activated macrophages, T lymphocytes and natural killer cells, which play an important role in a number of physiological processes such as immune response, inflammatory response and apoptosis. TNF receptor family members include TNFR1 and TNFR2.TNFR1 and TNFR2 are receptor proteins on the surface of cell membranes, which bind to TNF- α and transmit cellular signals, and TNFR1 plays a major proinflammatory role. When cells are subjected to inflammatory stimuli or stress, TNFR1 expression increases, and tumor necrosis factor alpha (TNF-alpha) bound thereto activates a series of signaling pathways, which trigger biological processes such as apoptosis and inflammatory reactions. Currently, blocking TNF-TNFR1 by binding TNFR1 and ligand TNF is one of the important therapeutic strategies for the treatment of inflammation. Currently, there are a variety of TNF- α inhibitors on the market, which are largely classified into fusion proteins and monoclonal antibodies. Adalimumab is a TNF-alpha inhibitor biopharmaceutical with the greatest global sales compliance and is excellent in the treatment of various diseases such as rheumatoid arthritis, ankylosing spondylitis, psoriasis and the like. However, at present, no similar double-target medicines are approved to be marketed at home and abroad. Disclosure of Invention In order to solve one of the technical problems existing in the prior art, the disclosure provides a bispecific antibody which can better maintain the activity of each monoclonal antibody, can specifically bind to two targets of TNF-like ligand 1A (TL 1A) and tumor necrosis factor alpha (TNF-alpha) at the same time, and has similar or even better biological activity as the monoclonal antibody. One aspect of the present disclosure provides a bispecific antibody comprising a first domain that specifically binds TNF-like ligand 1A (TL 1A), and a second domain that specifically binds tumor necrosis factor alpha (TNF-a). In some embodiments, the first domain is an antibody or functional fragment thereof that specifically binds TL 1A. In some embodiments, the second domain is an antibody or functional fragment thereof that specifically binds TNF- α. In some embodiments, the first domain comprises an antibody Fab fragment, fab 'fragment, F (ab') 2 fragment, fv fragment, scFv fragment, nanobody, heavy chain variable region (VH) fragment, and/or light chain variable region (VL) fragment that specifically binds to TL 1A. In some embodiments, the second domain comprises an antibody Fab fragment, fab 'fragment, F (ab') 2 fragment, fv fragment, scFv fragment, nanobody, heavy chain variable region (VH) fragment, and/or light chain variable region (VL) fragment that specifically binds TNF-a. In some embodiments, the first domain and the second domain are linked directly or through a linker. In some embodiments, the linker has an amino acid sequence as shown in formula (G nS)m, n, m are each integers from 1 to 10, more preferably n is an integer from 1 to 4, m is an integer from 1 to 3, or an amino acid sequence having 1,2, or 3 amino acid insertions, substitutions, or deletions compared to the amino acid sequence shown in formula (G nS)m). In some embodiments, the bispecific antibody comprises a first domain and a second domain linked in any manner. In some