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US-20260125467-A1 - NOVEL ANTI-LILRB4 ANTIBODIES AND DERIVATIVE PRODUCTS

US20260125467A1US 20260125467 A1US20260125467 A1US 20260125467A1US-20260125467-A1

Abstract

The present disclosure provides bispecific antibodies or antigen-binding fragments capable of binding to LILRB4 and CD3, isolated polynucleotides encoding the same, pharmaceutical compositions comprising the same, and the uses thereof.

Inventors

  • An Song
  • J. Paul WOODARD
  • X. Charlene Liao
  • Tao Huang
  • Ryan Stafford
  • Maria Jose COSTA
  • Kyu Hee HONG
  • Caroline BONNANS
  • Jianhui Zhou
  • Li Zhou
  • Ji Li

Assignees

  • IMMUNE-ONC THERAPEUTICS, INC.

Dates

Publication Date
20260507
Application Date
20251026
Priority Date
20200312

Claims (20)

  1. 1 . A bispecific antibody or antigen-binding fragment thereof capable of binding to Leukocyte Immunoglobulin-like subfamily B member 4 (LILRB4) and CD3, comprising: (a) a first antigen-binding region capable of binding to LILRB4 comprising a first light chain variable (V L ) domain and a first heavy chain variable (V H ) domain; and (b) a second antigen-binding region capable of binding to CD3 comprising a second V L domain and a second V H domain, wherein (i) the first V H domain comprising a first HC-CDR1 having the amino acid sequence of SEQ ID NO: 5, a first HC-CDR2 having the amino acid sequence of SEQ ID NO: 6 and a first HC-CDR3 having the amino acid sequence of SEQ ID NO: 7; and (ii) a first V L domain comprising a first LC-CDR1 having the amino acid sequence of SEQ ID NO: 8 with a mutation at amino acid residues NS, a first LC-CDR2 having the amino acid sequence of SEQ ID NO: 9 and a first LC-CDR3 having the amino acid sequence of SEQ ID NO: 10.
  2. 2 . The bispecific antibody or antigen-binding fragment thereof of claim 1 , wherein the LC-CDR1 having the amino acid sequence of SEQ ID NO: 28.
  3. 3 . The bispecific antibody or antigen-binding fragment thereof of claim 1 , wherein the first V H domain has the amino acid sequence of SEQ ID NO: 1; and wherein the first V L domain has the amino acid sequence of SEQ ID NO: 27.
  4. 4 . The bispecific antibody or antigen-binding fragment of claim 1 , wherein the first V L domain and the first V H domain link to a first pair of constant domains, respectively, and wherein the second V L domain and the second V H domain link to a second pair of constant domains, respectively.
  5. 5 . The bispecific antibody or antigen-binding fragment of claim 4 , wherein (a) the first V L domain links to a first light chain constant (C L ) domain, and the first V H domain links to a first heavy chain constant domain 1 (C H 1), or (b) the first V L domain links to a first C H 1 domain, and the V H domain links to a second C L domain. (c) the first V L domain links to a T cell receptor (TCR) α chain constant domain, and the first V H domain links to a TCR β chain constant domain, or (d) the first V L domain links to a TCR β chain constant domain, and the first V H domain links to a TCR α chain constant domain.
  6. 6 . The bispecific antibody or antigen-binding fragment of claim 4 , wherein (a) the second V L domain links to a second CL domain, and the second V H domain links to a second C H 1 domain, or (b) the second V L domain links to a second C H 1 domain, and the second V H domain links to a second CL domain, or (c) the second V L domain links to a T cell receptor (TCR) α chain constant domain, and the second V H domain links to a TCR β chain constant domain, or (d) the second V L domain links to a TCR β chain constant domain, and the second V H domain links to a TCR α chain constant domain.
  7. 7 . The bispecific antibody or antigen-binding fragment of claim 6 , wherein the TCR α chain constant domain has a S91A mutation.
  8. 8 . The bispecific antibody or antigen-binding fragment of claim 1 , wherein the first antigen-binding region and/or the second antigen-binding region is a single chain variable fragment (scFv).
  9. 9 . The bispecific antibody or antigen-binding fragment of claim 1 , further comprising a third antigen-binding region comprising a third V L domain and a third V H domain, wherein the third antigen-binding region is capable of binding to LILRB4 or CD3.
  10. 10 . The bispecific antibody or antigen-binding fragment of claim 9 , wherein the third V L domain and the third heavy chain variable domain link to a first pair of constant domains, respectively.
  11. 11 . The bispecific antibody or antigen-binding fragment of claim 10 , wherein (a) the third V L domain links to a third CL domain, and the third V H domain links to a third C H 1 domain, or (b) the third V L domain links to a third C H 1 domain, and the third V H domain links to a third C L domain, or (c) the third V L domain links to a TCR α chain constant domain, and the third V H domain links to a TCR β chain constant domain, or (d) the third V L domain links to a TCR β chain constant domain, and the third V H domain links to a TCR α chain constant domain.
  12. 12 . The bispecific antibody or antigen-binding fragment thereof of claim 1 , which is linked to one or more conjugate moieties.
  13. 13 . The bispecific antibody or antigen-binding fragment thereof of claim 12 , wherein the conjugate moiety comprises a clearance-modifying agent, a toxin, a detectable label, a chemotherapeutic agent, a cytokine, or purification moiety.
  14. 14 . A pharmaceutical composition comprising the bispecific antibody or antigen-binding fragment thereof of claim 1 , and a pharmaceutically acceptable carrier.
  15. 15 . An isolated polynucleotide encoding the bispecific antibody or antigen-binding fragment thereof of claim 1 .
  16. 16 . A vector comprising the isolated polynucleotide of claim 15 .
  17. 17 . A host cell comprising the vector of claim 16 .
  18. 18 . A method of expressing a bispecific antibody or antigen-binding fragment thereof capable of binding to LILRB4 and CD3, comprising culturing the host cell of claim 17 under the condition at which the bispecific antibody or antigen-binding fragment is expressed.
  19. 19 . A method of treating or ameliorating the effect of a cancer in a subject, comprising administering to the subject a therapeutically effective amount of the bispecific antibody or antigen-binding fragment thereof of claim 1 .
  20. 20 . The method of claim 19 , wherein the cancer is selected from the group consisting of multiple myeloma (MM), blastic plasmacytoid dendritic cell neoplasm (BPDCN), lymphoma, lymphocytic leukemia, Hodgkin's Disease, acute myeloid leukemia (AML), acute lymphocytic/lymphoblastic leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia (CML), myelodysplastic syndrome (MDS), myeloproliferative neoplasms, and chronic myelomonocytic leukemia (CMML).

Description

CROSS-REFERENCE TO RELATED APPLICATIONS This application is a continuation application of U.S. application Ser. No. 17/905,943, which is the national phase of PCT/US2021/022029 filed on Mar. 12, 2021, which claims priority to U.S. provisional patent application No. 62/988,892, filed Mar. 12, 2020, the disclosure of which is incorporated herein by reference. SEQUENCE LISTING The sequence listing that is contained in the file named “066564-8013WO01_ST25”, which is 150,794 bytes (as measured in Microsoft Windows) and was created on Mar. 12, 2021, is filed herewith by electronic submission and is incorporated by reference herein. FIELD OF THE INVENTION The present disclosure relates generally to the fields of medicine, oncology, and immunology. More particular, the disclosure relates to antibodies that bind to LILRB4. BACKGROUND Human Leukocyte Immunoglobulin-Like Receptor subfamily B member 4 (LILRB4), also known as Immunoglobulin-like transcript 3 (ILT3 or ILT-3), Leukocyte Immunoglobulin-like Receptor 5 (LIR5 or LIR-5), and CD85k or CD85K, is a type I membrane protein that contains cytoplasmic immunoreceptor tyrosine-based inhibition motif (ITIM) and involves in negative regulation of immune cell activation. LILRB4 is expressed on monocytes, macrophages and dendritic cells and can inhibit innate immunity in a cell-autonomous manner as well as suppress T cell activation through an indirect mechanism. LILRB4 is a specific marker for monocytic acute myeloid leukemia (AML) including refractory and relapsed disease. It has been shown that LILRB4 supports tumor cell infiltration into tissues and suppresses T cell activity via a signaling pathway that involves APOE, LILRB4, SHP-2, uPAR and ARG1 in AML cells (Deng M. et al., Nature (2018) 562:605-09). There is a significant need for novel anti-LILRB4 antibodies. BRIEF SUMMARY OF THE INVENTION The present disclosure provides anti-LILRB4 antibodies and antigen-binding fragment thereof, amino acid and nucleotide sequences thereof, anti-LILRB4 chimeric antigen receptors, and uses thereof. In one aspect, the present disclosure provides an isolated anti-LILRB4 antibody or an antigen-binding fragment thereof. In some embodiments, the anti-LILRB4 antibody or an antigen-binding fragment comprises: (a) a heavy chain variable region comprising a heavy chain complementarity determining region (HC-CDR) 1 having an amino acid sequence of SEQ ID NO: 5, an HC-CDR2 having an amino acid sequence of SEQ ID NO: 6 and an HC-CDR3 having an amino acid sequence of SEQ ID NO: 7; and (b) a light chain variable region comprising a light chain complementarity determining region (LC-CDR) 1 having an amino acid sequence of SEQ ID NO: 8 with a mutation at amino acid residues NS, an LC-CDR2 having an amino acid sequence of SEQ ID NO: 9 and an LC-CDR3 having an amino acid sequence of SEQ ID NO: 10. In certain embodiments, the LC-CDR1 has an amino acid sequence of SEQ ID NO: 28. In certain embodiments, the heavy chain variable region has an amino acid sequence at least about 90% identical to SEQ ID NO: 1; and wherein the light chain variable region has an amino acid sequence at least about 90% identical to SEQ ID NO: 27. In certain embodiments, the heavy chain variable region has an amino acid sequence of SEQ ID NO: 1; and wherein the light chain variable region has an amino acid sequence of SEQ ID NO: 27. In certain embodiments, the antibody or the antigen-binding fragment further comprises an immunoglobulin constant region, optionally a constant region of Ig, or optionally a constant region of human IgG. In certain embodiments, the antibody described herein is of the IgG1, IgG2, IgG3 or IgG4 isotype. In certain embodiments, the antibody or the antigen-binding fragment is humanized. In certain embodiments, the antigen-binding fragment is a camelized single domain antibody, a diabody, a ds (disulfide-stabilized) diabody or ds diabody, a scFv, a scFv dimer, a BsFv, a dsFv, a (dsFv)2, a dsFv-dsFv′, an Fv fragment, a Fab, a Fab′, a F(ab′)2, a bispecific antibody, a nanobody, a domain antibody, or a bivalent antibody. In certain embodiments, the anti-LILRB4 antibody described herein is a bispecific antibody. In some embodiments, the anti-LILRB4 bispecific antibody is against a T-cell receptor such as CD3. In some embodiments, the anti-LILRB4 bispecific antibody is against an NK-cell receptor such as CD16A. Therefore, the present disclosure in another aspect provides a bispecific antibody or antigen-binding fragment capable of binding to LILRB4 and CD3. In certain embodiments, the bispecific antibody or antigen-binding fragment provided herein comprises: (a) a first antigen-binding region comprising a first light chain variable (VL) domain and a first heavy chain variable (VH) domain; and (b) a second antigen-binding region comprising a second VL domain and a second VH domain, wherein the first antigen-binding region is capable of binding to LILRB4 and the second antigen-binding region is capable of bind