Search

US-12624112-B2 - Anti-ST2 antibody, encoding nucleic acid molecules thereof and methods of use thereof

US12624112B2US 12624112 B2US12624112 B2US 12624112B2US-12624112-B2

Abstract

Provided are an anti-ST2 antibody or a fragment thereof. The antibody or the fragment thereof can be specifically bound to human ST2, for inhibiting combination of IL-33 and human ST2, blocking an IL-33/ST2 intracellular signaling pathway, and inhibiting the promoting effect of different forms of IL-33 in cell-derived IL5, IL6 and 118 production. Compared with a known anti-ST2 antibody, this anti-ST2 antibody has higher biological activity, and can be used for preventing, treating or alleviating diseases related to ST2 expression or IL-33/ST2 pathway disorders.

Inventors

  • Xinhui XIONG
  • Tao Zhang
  • Kai Zhong
  • Wei Wu
  • Qianhui HUANG
  • Xing Li
  • Hong Pan

Assignees

  • Mabwell (shanghai) Bioscience Co., Ltd.

Dates

Publication Date
20260512
Application Date
20210121
Priority Date
20200121

Claims (20)

  1. 1 . An antibody or fragment thereof comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the heavy chain variable region and the light chain variable region comprise heavy chain CDRs and light chain CDRs shown in any one selected from the following combinations: (1) H-CDR1, H-CDR2, H-CDR3 as shown in SEQ ID NOs. 56, 63, 70 sequentially; and, L-CDR1, L-CDR2, L-CDR3 as shown in SEQ ID NOs. 85, 93, 97 sequentially; (II-1): H-CDR1, H-CDR2, H-CDR3 as shown in SEQ ID NOs. 57, 64, 71 sequentially; and, L-CDR1, L-CDR2, L-CDR3 as shown in SEQ ID NOs. 86, 93, 98 sequentially; (II-2): H-CDR1, H-CDR2, H-CDR3 as shown in SEQ ID NOs. 57, 64, 71 sequentially; and, L-CDR1, L-CDR2, L-CDR3 as shown in SEQ ID NOs. 87, 93, 98 sequentially; (III): H-CDR1, H-CDR2, H-CDR3 as shown in SEQ ID NOs. 58, 65, 72 sequentially; and, L-CDR1, L-CDR2, L-CDR3 as shown in SEQ ID NOs. 88, 94, 99 sequentially; (IV-1): H-CDR1, H-CDR2, H-CDR3 as shown in SEQ ID NOs. 59, 66, 73 sequentially; and, L-CDR1, L-CDR2, L-CDR3 as shown in SEQ ID NOs. 89, 95, 100 sequentially; (IV-2): H-CDR1, H-CDR2, H-CDR3 as shown in SEQ ID NOs. 59, 66, 74 sequentially; and, L-CDR1, L-CDR2, L-CDR3 as shown in SEQ ID NOs. 89, 95, 100 sequentially; (IV-3): H-CDR1, H-CDR2, H-CDR3 as shown in SEQ ID NOs. 59, 66, 75 sequentially; and, L-CDR1, L-CDR2, L-CDR3 as shown in SEQ ID NOs. 89, 95, 100 sequentially; (V-1): H-CDR1, H-CDR2, H-CDR3 as shown in SEQ ID NOs. 60, 67, 76 sequentially; and, L-CDR1, L-CDR2, L-CDR3 as shown in SEQ ID NOs. 90, 95, 101 sequentially; (V-2): H-CDR1, H-CDR2, H-CDR3 as shown in SEQ ID NOs. 60, 67, 76 sequentially; and, L-CDR1, L-CDR2, L-CDR3 as shown in SEQ ID NOs. 90, 95, 102 sequentially; (V-3): H-CDR1, H-CDR2, H-CDR3 as shown in SEQ ID NOs. 60, 67, 77 sequentially; and, L-CDR1, L-CDR2, L-CDR3 as shown in SEQ ID NOs. 90, 95, 101 sequentially; (V-4): H-CDR1, H-CDR2, H-CDR3 as shown in SEQ ID NOs. 60, 67, 77 sequentially; and, L-CDR1, L-CDR2, L-CDR3 as shown in SEQ ID NOs. 90, 95, 102 sequentially; (V-5): H-CDR1, H-CDR2, H-CDR3 as shown in SEQ ID NOs. 60, 67, 78 sequentially; and, L-CDR1, L-CDR2, L-CDR3 as shown in SEQ ID NOs. 90, 95, 101 sequentially; (V-6): H-CDR1, H-CDR2, H-CDR3 as shown in SEQ ID NOs. 60, 67, 78 sequentially; and, L-CDR1, L-CDR2, L-CDR3 as shown in SEQ ID NOs. 90, 95, 102 sequentially; (VI-1): H-CDR1, H-CDR2, H-CDR3 as shown in SEQ ID NOs. 61, 68, 79 sequentially; and, L-CDR1, L-CDR2, L-CDR3 as shown in SEQ ID NOs. 91, 95, 100 sequentially; (VI-2): H-CDR1, H-CDR2, H-CDR3 as shown in SEQ ID NOs. 61, 68, 80 sequentially; and, L-CDR1, L-CDR2, L-CDR3 as shown in SEQ ID NOs. 91, 95, 100 sequentially; (VI-3): H-CDR1, H-CDR2, H-CDR3 as shown in SEQ ID NOs. 61, 68, 81 sequentially; and, L-CDR1, L-CDR2, L-CDR3 as shown in SEQ ID NOs. 91, 95, 100 sequentially; (VII-1): H-CDR1, H-CDR2, H-CDR3 as shown in SEQ ID NOs. 62, 69, 82 sequentially; and, L-CDR1, L-CDR2, L-CDR3 as shown in SEQ ID NOs. 92, 96, 103 sequentially; (VII-2): H-CDR1, H-CDR2, H-CDR3 as shown in SEQ ID NOs. 62, 69, 83 sequentially; and, L-CDR1, L-CDR2, L-CDR3 as shown in SEQ ID NOs. 92, 96, 103 sequentially; and (VII-3): H-CDR1, H-CDR2, H-CDR3 as shown in SEQ ID NOs. 62, 69, 84 sequentially; and, L-CDR1, L-CDR2, L-CDR3) as shown in SEQ ID NOs. 92, 96, 103 sequentially.
  2. 2 . The antibody or fragment thereof according to claim 1 , wherein the heavy chain variable region comprises an amino acid sequence as shown in any one of SEQ ID NO. 1 to SEQ ID NO. 28 or an amino acid sequence having at least 75% identity to the amino acid sequence as shown; and/or, the light chain variable region comprises an amino acid sequence as shown in any one of SEQ ID NO. 29 to SEQ ID NO. 53 or an amino acid sequence having at least 75% identity to the amino acid sequence as shown.
  3. 3 . The antibody or fragment thereof according to claim 1 , wherein the antibody or fragment thereof comprises a heavy chain variable region and a light chain variable region as shown in any one selected from the following combinations: (I-1): the heavy chain variable region as shown in SEQ ID NO. 1 and the light chain variable region as shown in SEQ ID NO. 29; (I-2): the heavy chain variable region as shown in SEQ ID NO. 2 and the light chain variable region as shown in SEQ ID NO. 30; (I-3): the heavy chain variable region as shown in SEQ ID NO. 2 and the light chain variable region as shown in SEQ ID NO. 31; (I-4): the heavy chain variable region as shown in SEQ ID NO. 3 and the light chain variable region as shown in SEQ ID NO. 30; (I-5): the heavy chain variable region as shown in SEQ ID NO. 3 and the light chain variable region as shown in SEQ ID NO. 31; (II-1): the heavy chain variable region as shown in SEQ ID NO. 4 and the light chain variable region as shown in SEQ ID NO. 32; (II-2): the heavy chain variable region as shown in SEQ ID NO. 5 and the light chain variable region as shown in SEQ ID NO. 33; (II-3): the heavy chain variable region as shown in SEQ ID NO. 5 and the light chain variable region as shown in SEQ ID NO. 34; (II-4): the heavy chain variable region as shown in SEQ ID NO. 5 and the light chain variable region as shown in SEQ ID NO. 35; (II-5): the heavy chain variable region as shown in SEQ ID NO. 6 and the light chain variable region as shown in SEQ ID NO. 33; (II-6): the heavy chain variable region as shown in SEQ ID NO. 6 and the light chain variable region as shown in SEQ ID NO. 34; (II-7): the heavy chain variable region as shown in SEQ ID NO. 6 and the light chain variable region as shown in SEQ ID NO. 35; (III-1): the heavy chain variable region as shown in SEQ ID NO. 7 and the light chain variable region as shown in SEQ ID NO. 36; (III-2): the heavy chain variable region as shown in SEQ ID NO. 8 and the light chain variable region as shown in SEQ ID NO. 37; (III-3): the heavy chain variable region as shown in SEQ ID NO. 8 and the light chain variable region as shown in SEQ ID NO. 38; (III-4): the heavy chain variable region as shown in SEQ ID NO. 9 and the light chain variable region as shown in SEQ ID NO. 37; (III-5): the heavy chain variable region as shown in SEQ ID NO. 9 and the light chain variable region as shown in SEQ ID NO. 38; (IV-1): the heavy chain variable region as shown in SEQ ID NO. 10 and the light chain variable region as shown in SEQ ID NO. 39; (IV-2): the heavy chain variable region as shown in SEQ ID NO. 11 and the light chain variable region as shown in SEQ ID NO. 40; (IV-3): the heavy chain variable region as shown in SEQ ID NO. 11 and the light chain variable region as shown in SEQ ID NO. 41; (IV-4): the heavy chain variable region as shown in SEQ ID NO. 12 and the light chain variable region as shown in SEQ ID NO. 40; (IV-5): the heavy chain variable region as shown in SEQ ID NO. 12 and the light chain variable region as shown in SEQ ID NO. 41; (IV-6): the heavy chain variable region as shown in SEQ ID NO. 13 and the light chain variable region as shown in SEQ ID NO. 40; (IV-7): the heavy chain variable region as shown in SEQ ID NO. 13 and the light chain variable region as shown in SEQ ID NO. 41; (IV-8): the heavy chain variable region as shown in SEQ ID NO. 14 and the light chain variable region as shown in SEQ ID NO. 40; (IV-9): the heavy chain variable region as shown in SEQ ID NO. 14 and the light chain variable region as shown in SEQ ID NO. 41; (V-1): the heavy chain variable region as shown in SEQ ID NO. 15 and the light chain variable region as shown in SEQ ID NO. 42; (V-2): the heavy chain variable region as shown in SEQ ID NO. 16 and the light chain variable region as shown in SEQ ID NO. 43; (V-3): the heavy chain variable region as shown in SEQ ID NO. 16 and the light chain variable region as shown in SEQ ID NO. 44; (V-4): the heavy chain variable region as shown in SEQ ID NO. 16 and the light chain variable region as shown in SEQ ID NO. 45; (V-5): the heavy chain variable region as shown in SEQ ID NO. 17 and the light chain variable region as shown in SEQ ID NO. 43; (V-6): the heavy chain variable region as shown in SEQ ID NO. 17 and the light chain variable region as shown in SEQ ID NO. 44; (V-7): the heavy chain variable region as shown in SEQ ID NO. 17 and the light chain variable region as shown in SEQ ID NO. 45; (V-8): the heavy chain variable region as shown in SEQ ID NO. 18 and the light chain variable region as shown in SEQ ID NO. 43; (V-9): the heavy chain variable region as shown in SEQ ID NO. 18 and the light chain variable region as shown in SEQ ID NO. 44; (V-10): the heavy chain variable region as shown in SEQ ID NO. 18 and the light chain variable region as shown in SEQ ID NO. 45; (V-11): the heavy chain variable region as shown in SEQ ID NO. 19 and the light chain variable region as shown in SEQ ID NO. 43; (V-12): the heavy chain variable region as shown in SEQ ID NO. 19 and the light chain variable region as shown in SEQ ID NO. 44; (V-13): the heavy chain variable region as shown in SEQ ID NO. 19 and the light chain variable region as shown in SEQ ID NO. 45; (VI-1): the heavy chain variable region as shown in SEQ ID NO. 20 and the light chain variable region as shown in SEQ ID NO. 46; (VI-2): the heavy chain variable region shown as SEQ ID NO. 21 and the light chain variable region shown as SEQ ID NO. 47; (VI-3): the heavy chain variable region as shown in SEQ ID NO. 21 and the light chain variable region as shown in SEQ ID NO. 48; (VI-4): the heavy chain variable region as shown in SEQ ID NO. 22 and the light chain variable region as shown in SEQ ID NO. 47; (VI-5): the heavy chain variable region as shown in SEQ ID NO. 22 and the light chain variable region as shown in SEQ ID NO. 48; (VI-6): the heavy chain variable region as shown in SEQ ID NO. 23 and the light chain variable region as shown in SEQ ID NO. 47; (VI-7): the heavy chain variable region as shown in SEQ ID NO. 23 and the light chain variable region as shown in SEQ ID NO. 48; (VI-8): the heavy chain variable region as shown in SEQ ID NO. 24 and the light chain variable region as shown in SEQ ID NO. 47; (VI-9): the heavy chain variable region as shown in SEQ ID NO. 24 and the light chain variable region as shown in SEQ ID NO. 48; (VII-1): the heavy chain variable region as shown in SEQ ID NO. 25 and the light chain variable region as shown in SEQ ID NO. 49; (VII-2): the heavy chain variable region as shown in SEQ ID NO. 26 and the light chain variable region as shown in SEQ ID NO. 52; (VII-3): the heavy chain variable region shown as SEQ ID NO. 26 and the light chain variable region shown as SEQ ID NO. 53; (VII-4): the heavy chain variable region as shown in SEQ ID NO. 26 and the light chain variable region as shown in SEQ ID NO. 50; (VII-5): the heavy chain variable region shown as SEQ ID NO. 26 and the light chain variable region shown as SEQ ID NO. 51; (VII-6): the heavy chain variable region shown as SEQ ID NO. 27 and the light chain variable region shown as SEQ ID NO. 52; (VII-7): the heavy chain variable region shown as SEQ ID NO. 27 and the light chain variable region shown as SEQ ID NO. 53; (VII-8): the heavy chain variable region shown as SEQ ID NO. 27 and the light chain variable region shown as SEQ ID NO. 50; (VII-9): the heavy chain variable region shown as SEQ ID NO. 27 and the light chain variable region shown as SEQ ID NO. 51; (VII-10): the heavy chain variable region as shown in SEQ ID NO. 28 and the light chain variable region as shown in SEQ ID NO. 52; (VII-11): the heavy chain variable region as shown in SEQ ID NO. 28 and the light chain variable region as shown in SEQ ID NO. 53; (VII-12): the heavy chain variable region as shown in SEQ ID NO. 28 and the light chain variable region as shown in SEQ ID NO. 50; and (VII-13): the heavy chain variable region as shown in SEQ ID NO. 28 and the light chain variable region as shown in SEQ ID NO. 51.
  4. 4 . The antibody or fragment thereof according to claim 1 , wherein the antibody or fragment thereof binds to human ST2.
  5. 5 . The antibody or fragment thereof according to claim 1 , wherein the antibody is a monoclonal antibody, a single chain antibody, a diabody, a single domain antibody, a nanobody, a fully or partially humanized antibody, or a chimeric antibody; and the fragment is a functionally active fragment of the antibody which is capable of specifically binding to ST2 or any portion thereof.
  6. 6 . A conjugate or fusion protein comprising the antibody or fragment thereof according to claim 1 .
  7. 7 . A composition comprising the antibody or fragment thereof according to claim 1 , a conjugate or fusion protein comprising the antibody or fragment thereof, a nucleic acid molecule comprising a nucleotide sequence encoding a heavy chain CDR, a light chain CDR, a heavy chain variable region, a light chain variable region, a heavy chain or a light chain comprised in the antibody or fragment thereof, a vector comprising the nucleic acid molecule, and/or a host cell comprising the nucleic acid molecule and/or the vector, or transformed or transfected with the nucleic acid molecule and/or the vector.
  8. 8 . The composition according to claim 7 manufactured to be a medicament for reducing the likelihood of treating, or ameliorating a disease associated with expression of ST2 or dysregulation of IL-33/ST2 pathway.
  9. 9 . The composition according to claim 7 wherein the composition is a pharmaceutical composition.
  10. 10 . The composition according to claim 8 , wherein the disease is an inflammatory disease or an autoimmune disease.
  11. 11 . The composition according to claim 8 , wherein the disease is heart failure, allergic rhinitis, nasal polyps, atopic dermatitis, chronic obstructive pulmonary disease, asthma, pulmonary fibrosis, sepsis, inflammatory bowel disease, systemic lupus erythematosus, rheumatoid arthritis, systemic sclerosis, wegener's granulomatosis, or chemotherapy-associated diarrhea.
  12. 12 . A pharmaceutical combination comprising the antibody or fragment thereof according to claim 1 , a conjugate or fusion protein comprising the antibody or fragment thereof, and/or a composition comprising the antibody or fragment thereof or the conjugate or fusion protein, and optionally an additional drug.
  13. 13 . A kit comprising the antibody or fragment thereof according to claim 1 , a conjugate or fusion protein comprising the antibody or fragment thereof, a nucleic acid molecule comprising a nucleotide sequence encoding a heavy chain CDR, a light chain CDR, a heavy chain variable region, a light chain variable region, a heavy chain or a light chain comprised in the antibody or fragment thereof, a vector comprising the nucleic acid molecule, and/or a host cell comprising the nucleic acid molecule and/or the vector, or transformed or transfected with the nucleic acid molecule and/or the vector, and/or a composition comprising the antibody or fragment thereof, the conjugate or fusion protein, the nucleic acid molecule, the vector, and/or the host cell.
  14. 14 . A nucleic acid molecule comprising a nucleotide sequence encoding a heavy chain CDR, a light chain CDR, a heavy chain variable region, a light chain variable region, a heavy chain or a light chain comprised in the antibody or fragment thereof according to claim 1 .
  15. 15 . A vector comprising the nucleic acid molecule according to claim 14 .
  16. 16 . An isolated host cell comprising the nucleic acid molecule according to claim 14 and/or a vector comprising the nucleic acid molecule, or transformed or transfected with the nucleic acid molecule and/or the vector.
  17. 17 . A method for detecting a disease associated with expression of ST2 or dysregulation of IL-33/ST2 pathway, including contacting the antibody or fragment thereof according to claim 1 , a conjugate or fusion protein comprising the antibody or fragment thereof, and/or a composition comprising the antibody or fragment thereof or the conjugate or fusion protein with a sample from a subject.
  18. 18 . The method according to claim 17 , wherein the disease is an inflammatory disease or an autoimmune disease.
  19. 19 . The method according to claim 17 , wherein the disease is heart failure, allergic rhinitis, nasal polyps, atopic dermatitis, chronic obstructive pulmonary disease, asthma, pulmonary fibrosis, sepsis, inflammatory bowel disease, systemic lupus erythematosus, rheumatoid arthritis, systemic sclerosis, wegener's granulomatosis, or chemotherapy-associated diarrhea.
  20. 20 . A method for reducing the likelihood of, treating or ameliorating a disease associated with expression of ST2 or dysregulation of IL-33/ST2 pathway, including administering to a subject in need thereof the antibody or fragment thereof according to claim 1 , a conjugate or fusion protein comprising the antibody or fragment thereof, and/or a composition comprising the antibody or fragment thereof or the conjugate or fusion protein, and optionally an additional drug.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS The present application is a U.S. National Phase Patent Application of International Application Number PCT/CN2021/073009, filed on Jan. 21, 2021, which claims the priority benefit of Chinese Patent Application No. CN202010072085.X filed on 21 Jan. 2020, the entire content of each of which is hereby incorporated by reference. SEQUENCE LISTING The instant application contains a Sequence Listing submitted in ASCII format via EFS-web, and the entire content of the electronic submission of the sequence listing is incorporated by reference in its entirety for all purposes. The ASCII file is named “224785_seq_list_rev_ST25,” was last modified on Jan. 23, 2023, and is 77,419 bytes in size. TECHNICAL FIELD The present invention relates to the field of antibody drugs, in particular to an antibody against human ST2 and use thereof in preparing a medicament. BACKGROUND OF THE INVENTION Interleukin 33 (IL-33) is a cytokine associated with IL-1 and IL-18, also known as NF-HEV or IL-1F11. ST2 (ST2L, IL-1RL1, T1, Fit-1, DER-4, IL-1R4 or ST2 alpha) is a binding receptor for IL-33, and is a member of Toll/IL-1 receptor family, expressed on the cell surface of a variety of immune cells, including lymphocytes, especially helper T cells, natural killer (NK) cells and natural killer-T (NKT) cells expressing IL-5 and IL-13, as well as many so-called innate immune cells, such as mast cells, basophils, eosinophils, macrophages and innate helper cells (also known as new immune cells (nuocytes) (Neill, Wong et al, 2010)). ST2 can down-regulate the responsiveness of Toll-like receptors TLR2, TLR4 and TLR9, and can induce the release of Type 2 cytokines via activation by its ligand IL-33 and association with accessory protein IL-1RAcP. Relevant literatures have proposed models of the interaction between ST2, IL-33 and IL-1RAcP, and the interaction between IL-1R1 and IL-1RAcP(Lingel et al, Cell 17: 1398-1410, 2009; Wang et al, Nat Immunol, 11: 905-11, 2010). IL-33 has been described as an “Alarmin” because it is present in the nuclei of epithelial and endothelial cells in its full-length form during homeostasis, but can be cleaved and released during cellular necrosis. Examples of IL-33-induced cellular responses include the production of inflammatory cytokines such as IL-5, IL-6, IL-13, TNF, IFN-7, and GM-CSF, and the production of chemokines such as CXCL8, CCL17, and CCL24. IL-33 has also been shown to enhance acute allergic reactions by potentiating mast cell and basophil activation triggered by IgE receptor signaling or other mast cell and basophil activators. IL-33 also enhances the recruitment, survival and adhesion properties of ST2-expressing immune cells and is therefore of great importance in the excitation and maintenance of cellular inflammation in local tissues. Dysregulation of IL-33/ST2 pathway has been shown to be associated with a variety of immune-mediated diseases, including asthma, rheumatoid arthritis, inflammatory bowel disease, atopic dermatitis, allergic rhinitis, nasal polyps, and systemic sclerosis. Thus, therapeutic blockade of the IL-33/ST2 pathway may help to overcome hyperimmune responses. Inhibitors of this pathway mainly include IL33 antibodies (e.g., MEDI3506, ANB020, REGN3500, MT-2990, LY-3375880, PF-06817024) and ST2 antibodies (e.g., CNTO7160, AMG-282), being developed at clinical stages 1 and 2 for indications including allergic rhinitis, atopic dermatitis, chronic obstructive pulmonary disease, asthma, etc. Presently, incidence rates of allergic inflammation and respiratory disease are gradually increasing, and medicines available on the market still are mainly glucocorticoids and β2 receptor agonists. Antibodies reported to date, although all are able to block the interaction of ST2 with its ligand, differ in biological activities produced. Differences in biological activity may lead to differences in clinical efficacy and dosage of the antibodies, and therefore there is still a need in the art for ST2 antibodies that provide high affinity, high stability, and high biological activity. SUMMARY OF THE INVENTION The technical problem to be solved by the present invention is to obtain a new high affinity antibody which binds ST2 and is suitable for the treatment of diseases or any indications related to the IL-33/ST2 pathway, by immunizing mice with human ST2 as immunogen, obtaining murine antibodies through B cell panning, and further antibody engineering and humanization techniques. For the above technical problem, an object of the present invention is to provide an antibody or functional fragment thereof that specifically binds to ST2, and to provide uses thereof. Technical solutions of the present invention are as follows. As described herein, a “fragment” of an antibody as described in the present invention encompasses various functional or active fragments of the antibody, e.g., an antigen-binding portion thereof, such as Fab, F (ab′) 2, or scFv. In one a