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KR-20260062935-A - Antibody binding to BTN3A1 and its uses

KR20260062935AKR 20260062935 AKR20260062935 AKR 20260062935AKR-20260062935-A

Abstract

An isolated monoclonal antibody that specifically binds to human BTN3A1, or its antigen-binding portion, is provided herein. A nucleic acid molecule encoding the antibody or its antigen-binding portion, an expression vector, a host cell, and a method for expressing the antibody or its antigen-binding portion are also provided. The present disclosure further provides a bispecific molecule and a pharmaceutical composition comprising the antibody or its antigen-binding portion, as well as a therapeutic method using the anti-BTN3A1 antibody or its antigen-binding portion of the present disclosure.

Inventors

  • 리 준
  • 펑 체위
  • 천 밍쥬

Assignees

  • 바이오션, 인코포레이티드

Dates

Publication Date
20260507
Application Date
20240830
Priority Date
20230831

Claims (17)

  1. An isolated monoclonal antibody capable of binding to BTN3A1 or its antigen-binding portion, comprising (ii) a heavy chain variable region including VH CDR1, VH CDR2 and VH CDR3 and (ii) a light chain variable region including VL CDR1, VL CDR2 and VL CDR3, wherein Here, VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 contain the following amino acid sequences: SEQ ID NO: 1, 2, 3, 4, 5 and 6 respectively; Here, X of SEQ ID NO: 1 is Y, X1 and X2 of SEQ ID NO: 2 are A and T respectively, X of SEQ ID NO: 3 is T, and X1 and X2 of SEQ ID NO: 4 are V and V respectively; Here, X of SEQ ID NO: 1 is Y, X1 and X2 of SEQ ID NO: 2 are A and T respectively, X of SEQ ID NO: 3 is T, and X1 and X2 of SEQ ID NO: 4 are A and A respectively; Here, X of SEQ ID NO: 1 is Y, X1 and X2 of SEQ ID NO: 2 are N and T respectively, X of SEQ ID NO: 3 is T, and X1 and X2 of SEQ ID NO: 4 are N and N respectively; Here, X of SEQ ID NO: 1 is Y, X1 and X2 of SEQ ID NO: 2 are N and T respectively, X of SEQ ID NO: 3 is T, and X1 and X2 of SEQ ID NO: 4 are A and A respectively; Here, X of SEQ ID NO: 1 is Y, X1 and X2 of SEQ ID NO: 2 are N and T respectively, X of SEQ ID NO: 3 is T, and X1 and X2 of SEQ ID NO: 4 are S and S respectively; Here, X of SEQ ID NO: 1 is Y, X1 and X2 of SEQ ID NO: 2 are N and T respectively, X of SEQ ID NO: 3 is T, and X1 and X2 of SEQ ID NO: 4 are V and V respectively; or Here, X of SEQ ID NO: 1 is Y, X1 and X2 of SEQ ID NO: 2 are N and A, respectively, X of SEQ ID NO: 3 is T, and X1 and X2 of SEQ ID NO: 4 are V and V, respectively. (2) SEQ ID NO: 1, 12, 3, 4, 5 and 6 respectively Here, X of SEQ ID NO: 1 is Y, X of SEQ ID NO: 3 is S, and X1 and X2 of SEQ ID NO: 4 are N and N, respectively. (3) SEQ ID NO: 1, 15, 3, 4, 5 and 6 respectively Here, X of SEQ ID NO: 1 is H, X of SEQ ID NO: 15 is G, X of SEQ ID NO: 3 is S, and X1 and X2 of SEQ ID NO: 4 are N and N, respectively; or Here, X of SEQ ID NO: 1 is H, X of SEQ ID NO: 15 is S, X of SEQ ID NO: 3 is S, and X1 and X2 of SEQ ID NO: 4 are N and N, respectively, Isolated monoclonal antibody or its antigen-binding portion.
  2. In paragraph 1, The heavy chain variable region comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO: 7, 8, 9, 13, or 16, and Here, X1 and X2 of SEQ ID NO: 7 are A and T, N and T, or N and A, respectively. Here, X1, X2, X3, and X4 of SEQ ID NO: 9 are L, A, K, and F, respectively; L, T, K, and F, respectively; M, T, K, and F, respectively; L, T, T, and F, respectively; L, T, K, and Y, respectively; L, A, K, and F, respectively; or L, T, K, and F, respectively. Herein, X1, X2 and X3 of SEQ ID NO: 16 are isolated monoclonal antibodies or their antigen-binding portions, which are G, I and G; G, T and S; or E, T and S.
  3. In paragraph 1, The light chain variable region comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO: 10, 11, or 14, and Here, X1 and X2 of SEQ ID NO: 10 are V and V, respectively; A and A, respectively; N and N, respectively; or S and S, respectively. Here, X1, X2, and X3 of SEQ ID NO: 11 are V, L, and V, respectively; V, V, and L, respectively; or I, V, and L, respectively. Here, X of SEQ ID NO: 14 is T or Q, Isolated monoclonal antibody or its antigen-binding portion.
  4. In paragraph 2, The heavy chain variable region and the light chain variable region are SEQ ID NO: 7 and 10 respectively (where X1 and X2 of SEQ ID NO: 7 are A and T respectively, and X1 and X2 of SEQ ID NO: 10 are V and V respectively), SEQ ID NO: 8 and 11 respectively (where X1, X2, and X3 of SEQ ID NO: 11 are V, L, and V respectively), SEQ ID NO: 9 and 11 respectively (where X1, X2, X3, and X4 of SEQ ID NO: 9 are L, A, K, and F, respectively, and X1, X2, and X3 of SEQ ID NO: 11 are V, V, and L, respectively), SEQ ID NO: 9 and 11 respectively (where X1, X2, X3, and X4 of SEQ ID NO: 9 are L, T, K, and F, respectively, and X1, X2, and X3 of SEQ ID NO: 11 are V, V, and L, respectively), SEQ ID NO: 9 and 11 respectively (where X1, X2, X3, and X4 of SEQ ID NO: 9 are M, T, K, and F, respectively, and X1, X2, and X3 of SEQ ID NO: 11 are V, V, and L, respectively), SEQ ID NO: 9 and 11 respectively (where X1, X2, X3, and X4 of SEQ ID NO: 9 are L, T, T, and F respectively, and X1, X2, and X3 of SEQ ID NO: 11 are V, V, and L respectively), SEQ ID NO: 9 and 11 respectively (where X1, X2, X3, and X4 of SEQ ID NO: 9 are L, T, K, and Y, respectively, and X1, X2, and X3 of SEQ ID NO: 11 are V, V, and L, respectively), SEQ ID NO: 9 and 11 respectively (where X1, X2, X3, and X4 of SEQ ID NO: 9 are L, A, K, and F, respectively, and X1, X2, and X3 of SEQ ID NO: 11 are I, V, and L, respectively), SEQ ID NO: 9 and 11 respectively (where X1, X2, X3, and X4 of SEQ ID NO: 9 are L, T, K, and F, respectively, and X1, X2, and X3 of SEQ ID NO: 11 are I, V, and L, respectively), SEQ ID NO: 7 and 10 respectively (where X1 and X2 of SEQ ID NO: 7 are A and T respectively, and X1 and X2 of SEQ ID NO: 10 are A and A respectively), SEQ ID NO: 7 and 10 respectively (where X1 and X2 of SEQ ID NO: 7 are N and T respectively, and X1 and X2 of SEQ ID NO: 10 are N and N respectively), SEQ ID NO: 7 and 10 respectively (where X1 and X2 of SEQ ID NO: 7 are N and T respectively, and X1 and X2 of SEQ ID NO: 10 are A and A respectively), SEQ ID NO: 7 and 10 respectively (where X1 and X2 of SEQ ID NO: 7 are N and T respectively, and X1 and X2 of SEQ ID NO: 10 are S and S respectively), SEQ ID NO: 7 and 10 respectively (where X1 and X2 of SEQ ID NO: 7 are N and T respectively, and X1 and X2 of SEQ ID NO: 10 are V and V respectively), SEQ ID NO: 7 and 10 respectively (where X1 and X2 of SEQ ID NO: 7 are N and A respectively, and X1 and X2 of SEQ ID NO: 10 are V and V respectively), SEQ ID NO: 13 and 14 respectively (where X of SEQ ID NO: 14 is T), SEQ ID NO: 16 and 14 respectively (where X1, X2, and X3 of SEQ ID NO: 16 are G, I, and G respectively, and X of SEQ ID NO: 14 is Q), SEQ ID NO: 16 and 14 respectively (where X1, X2, and X3 of SEQ ID NO: 16 are G, T, and S respectively, and X of SEQ ID NO: 14 is Q), or Each comprising SEQ ID NO: 16 and 14 (where X1, X2, and X3 of SEQ ID NO: 16 are E, T, and S, respectively, and X of SEQ ID NO: 14 is Q) and amino acid sequences having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, Isolated monoclonal antibody or its antigen-binding portion.
  5. In paragraph 1, An isolated monoclonal antibody or its antigen-binding portion that is an IgG1, IgG2, or IgG4 isoform.
  6. In paragraph 1, An isolated monoclonal antibody or its antigen-binding portion comprising a heavy chain constant region having an amino acid sequence of SEQ ID NOs: 17, 18, or 20 connected to a heavy chain variable region, and/or a light chain constant region having an amino acid sequence of SEQ ID NO: 23 or 25 connected to a light chain variable region.
  7. In paragraph 1, (a) able to bind to human BTN3A1; (b) able to bind to synomolgus monkey BTN3A1; (c) able to bind to rhesus macaque BTN3A1; (d) able to bind to human BTN3A2; (e) able to bind to human BTN3A3; (f) able to induce apoptosis of target cells by γδ T cells, or (f) an isolated monoclonal antibody or its antigen-binding portion having in vivo anti-tumor activity.
  8. In paragraph 1, A monoclonal antibody or its antigen-binding portion, which is a mouse, chimeric, or humanized antibody.
  9. A nucleic acid molecule encoding the isolated monoclonal antibody according to claim 1 or the antigen-binding portion thereof.
  10. An expression vector comprising a nucleic acid molecule according to claim 9.
  11. A host cell having an expression vector according to claim 10 or a nucleic acid molecule according to claim 9 integrated into the genome.
  12. A pharmaceutical composition comprising an isolated monoclonal antibody or its antigen-binding portion according to any one of claims 1 to 8, a nucleic acid molecule of claim 9, an expression vector of claim 10, or a host cell of claim 11, and a pharmaceutically acceptable carrier.
  13. A method for treating a disease beneficial to a subject requiring BTN3A-mediated T cell activation, comprising the step of administering a therapeutically effective amount of the pharmaceutical composition of claim 12 to a subject.
  14. In Paragraph 13, The disease is BTN3A + cancer, method.
  15. In Paragraph 14, Cancer is colorectal cancer, breast cancer, lung cancer, prostate cancer, head and neck cancer, pancreatic cancer, bladder cancer, rectal cancer, cervical cancer, ovarian cancer, liver cancer, esophageal cancer, thyroid cancer, kidney cancer, stomach cancer, chronic lymphocytic leukemia (CLL), non-Hodgkin lymphoma (NHL), lymphoma neoplasm, small lymphocytic lymphoma (SLL), mantle cell lymphoma (MCL), or myeloid cell lineage neoplasm.
  16. In Paragraph 13, The disease is an infectious disease, method.
  17. A method for killing BTN3A + cells in a subject requiring this, comprising the step of administering a therapeutically effective amount of a pharmaceutical composition according to claim 12 to the subject.

Description

Antibody binding to BTN3A1 and its uses The present disclosure relates to isolated monoclonal antibodies that bind to BTN3A1 with generally high affinity and activity, particularly mouse, chimeric, or humanized monoclonal antibodies, or their antigen-binding portions. Nucleic acid molecules encoding the antibody or its antigen-binding portion, expression vectors, host cells, and methods for expressing the antibody or its antigen-binding portion are also provided. The present disclosure further provides bispecific molecules, oncolytic viruses, and pharmaceutical compositions that may comprise the antibody or its antigen-binding portion, as well as therapeutic methods using the antibody or its antigen-binding portion of the present disclosure. The present application claims priority to U.S. provisional application serial number 63/535,618 filed on August 31, 2023. In humans, approximately 1% to 10% of CD3 + T cells are γδ T cells, and these cells can recognize conserved risk-related determinants that are upregulated after inflammation, infection, or cell transformation (Saura-Esteller J et al ., (2022) Front Immunol . 13:915837; Harly C et al ., (2012) Blood . 120:2269-2279). Vγ9Vδ2 T cells constitute the dominant γδ T cell subpopulation (95%) in peripheral blood and exhibit significant functional plasticity, such as differentiation into antigen-presenting or phagocytic cells and the regulation of immune responses through interactions with other immune cells (Hinz T et al., (1997) Int Immunol 9(8):1065-1072). Vγ9Vδ2 T cells can detect and respond to small phosphorylated metabolites (phosphoantigens or pAg) accumulated in cells with microbial infections or abnormal metabolism (including certain tumor cells) through a protein called BTN3A. The BTN3A protein, also known as CD277, has three isoforms: BTN3A1, BTN3A2, and BTN3A3. These three isoforms share 95% homology in the extracellular domain, each possessing a proximal IgC domain connected to a single transmembrane-transmembrane domain and an extracellular N-terminal IgV domain, respectively. However, their intracellular domains differ significantly; BTN3A1 and BTN3A3 possess the B30.2 domain, whereas this domain is absent in the short cytoplasmic tail of BTN3A2. The BTN3A protein is widely expressed and overexpression has been found in a number of solid tumors (e.g., bladder cancer, colorectal cancer, melanoma, ovarian cancer, pancreatic cancer, lung cancer) and hematological cancers (e.g., leukemia, lymphoma) (Gassart AD et al ., (2021) Sci Transl Med . 13(616): eabj0835). When cellular pAg levels increase under stress conditions, it binds to the B30.2 domain of BTN3A1, inducing structural changes, redistribution, and fixation of BTN3A1. It can also bind to BTN2A1, which directly interacts with the Vγ9 chain of Vγ9Vδ2 T cells, thereby activating Vγ9Vδ2 T cells and targeting and killing cells with high pAg levels (Saura-Esteller J et al ., (2022), cited supra ; Chen S et al ., (2021) J Cancer . 12(15):4505-4512). BTN3A2 and BTN3A3 have been reported to promote Vγ9Vδ2 T cell activation by increasing BTN3A1 levels, and their extracellular domains can undergo structural changes similar to those of BTN3A1 when treated with anti-BTN3A antibodies (Wang H et al ., (2019) J Immunol . 203:607-626). Therapeutics targeting BTN3A have been developed or are under development. ICT01 is a humanized anti-BTN3A monoclonal antibody developed by Imcheck that binds to all three BTN3A isoforms to induce Vγ9Vδ2 T cell activation and enhance cytotoxicity against BTN3A + tumor cell lines of various origins (Harly C et al ., (2012), cited supra ). Despite advancements in this field, patient choices remain limited, and additional monoclonal antibodies with improved pharmacological properties are needed. The citation or identification of any literature in this application does not constitute an acknowledgment that such literature is available as prior art for the present invention. The present disclosure provides an isolated monoclonal antibody (e.g., mouse, chimeric, or humanized antibody) or its antigen-binding portion that can specifically bind to BTN3A1 (e.g., human BTN3A1 and monkey BTN3A1) and, compared to a prior art anti-BTN3A1 antibody such as ICT01, i) has similar, if not higher, binding affinity/binding ability to human and monkey BTN3A1 proteins; ii) has similar, if not higher, binding ability to cells expressing human BTN3A1; iii) exhibits similar, if not higher, cross-reactivity to human BTN3A2 and BTN3A3 proteins; iv) induces targeted cell death by γδ T cells similarly, if not superiorly; or v) has similar, if not higher, in vivo antitumor efficacy. The inventors of this application believe that when the antibody of the present disclosure or its antigen-binding portion binds to a BTN3A protein (e.g., BTN3A1, BTN3A2, or BTN3A3), it enables structural changes in the extracellular domain of the BTN3A protein, particularly BTN3A1 and possibly BTN3A2 and/or BTN3A3, thereby in