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EP-4737479-A1 - 4-1BB-BINDING PROTEIN

EP4737479A1EP 4737479 A1EP4737479 A1EP 4737479A1EP-4737479-A1

Abstract

Provided is a 4-1BB-binding protein that can specifically bind to 4-1BB and does not block the binding of 4-1BB to 4-1BBL. The protein can effectively activate NF-κB signaling and T cells under cross-linking conditions.

Inventors

  • LIU, GUOSHENG
  • CHEN, Caiyan
  • LIN, QIXING
  • LI, CHUANG
  • ZHANG, HUI
  • LI, Jiaping
  • CHEN, Junyou
  • WANG, ZHIWEI
  • HUANG, XIANMING

Assignees

  • Bio-Thera Solutions, Ltd.

Dates

Publication Date
20260506
Application Date
20240628

Claims (12)

  1. A 4-1BB-binding protein, comprising a heavy chain variable region, wherein the heavy chain variable region comprises one or more of the following complementarity determining regions (CDRs): an HCDR1 set forth in SEQ ID NO: 7, an HCDR2 set forth in any one of SEQ ID NOs: 8-13 or SEQ ID NOs: 31-36, and an HCDR3 set forth in any one of SEQ ID NOs: 14-19 or SEQ ID NOs: 37-42.
  2. The 4-1BB-binding protein according to claim 1, wherein the heavy chain variable region comprises an HCDR1 set forth in SEQ ID NO: 7, an HCDR2 set forth in SEQ ID NOs: 8-13, and an HCDR3 set forth in any one of SEQ ID NOs: 14-19; or the heavy chain variable region comprises an HCDR1 set forth in SEQ ID NO: 7, an HCDR2 set forth in SEQ ID NO: 8, and an HCDR3 set forth in any one of SEQ ID NOs: 14-17; or the heavy chain variable region comprises an HCDR1 set forth in SEQ ID NO: 7, an HCDR2 set forth in SEQ ID NO: 9, and an HCDR3 set forth in SEQ ID NO: 14 or 19; or the heavy chain variable region comprises an HCDR1 set forth in SEQ ID NO: 7, an HCDR2 set forth in SEQ ID NO: 10, and an HCDR3 set forth in SEQ ID NO: 14 or 16; or the heavy chain variable region comprises an HCDR1 set forth in SEQ ID NO: 7, an HCDR2 set forth in SEQ ID NO: 11 or 12, and an HCDR3 set forth in SEQ ID NO: 16; or the heavy chain variable region comprises an HCDR1 set forth in SEQ ID NO: 7, an HCDR2 set forth in SEQ ID NO: 13, and an HCDR3 set forth in SEQ ID NO: 18 or 19; or wherein the heavy chain variable region comprises an HCDR1 set forth in SEQ ID NO: 7, an HCDR2 set forth in SEQ ID NOs: 31-36, and an HCDR3 set forth in any one of SEQ ID NOs: 37-42; or the heavy chain variable region comprises an HCDR1 set forth in SEQ ID NO: 7, an HCDR2 set forth in SEQ ID NO: 31, and an HCDR3 set forth in any one of SEQ ID NOs: 37-40; or the heavy chain variable region comprises an HCDR1 set forth in SEQ ID NO: 7, an HCDR2 set forth in SEQ ID NO: 32, and an HCDR3 set forth in SEQ ID NO: 37 or 42; or the heavy chain variable region comprises an HCDR1 set forth in SEQ ID NO: 7, an HCDR2 set forth in SEQ ID NO: 33, and an HCDR3 set forth in SEQ ID NO: 37 or 39; or the heavy chain variable region comprises an HCDR1 set forth in SEQ ID NO: 7, an HCDR2 set forth in SEQ ID NO: 34 or 35, and an HCDR3 set forth in SEQ ID NO: 39; or the heavy chain variable region comprises an HCDR1 set forth in SEQ ID NO: 7, an HCDR2 set forth in SEQ ID NO: 36, and an HCDR3 set forth in SEQ ID NO: 41 or 42.
  3. The 4-1BB-binding protein according to claim 1 or 2, wherein the heavy chain variable region comprises an amino acid sequence set forth in any one of SEQ ID NOs: 1-6 and 29-30, or an amino acid sequence having at least 80% identity compared to the amino acid sequence set forth in any one of SEQ ID NOs: 1-6 and 29-30, or an amino acid sequence having one or more conservative amino acid substitutions compared to the amino acid sequence set forth in any one of SEQ ID NOs: 1-6 and 29-30.
  4. A 4-1BB-binding protein, comprising an amino acid sequence set forth in any one of SEQ ID NOs: 1-6 and 29-30.
  5. The 4-1BB-binding protein according to any one of claims 1-4, wherein the 4-1BB-binding protein is a single-domain antibody, a heavy-chain antibody, a bispecific antibody, a multispecific antibody, a Fab, a Fab', a F(ab')2, an Fv, or an scFv.
  6. The 4-1BB-binding protein according to any one of claims 1-4, wherein the 4-1BB-binding protein is a fusion protein and further comprises an Fc region; preferably, the Fc is an Fc of an immunoglobulin, such as an IgG1 Fc; or the Fc is a variant Fc having one or more amino acid modifications, such as substitutions, deletions, or insertions.
  7. The binding protein according to claim 6, wherein in the fusion protein, the C-terminus of the heavy chain variable region and the N-terminus of the Fc region are further linked by a peptide linker.
  8. The binding protein according to claim 7, wherein a sequence of the peptide linker is (G m S) n , wherein each m is independently 2, 3, 4, or 5, and n is independently 1, 2, 3, 4, 5, or 6.
  9. A 4-1BB-binding protein, comprising an amino acid sequence set forth in any one of SEQ ID NOs: 22-27.
  10. A biomaterial, wherein the biomaterial is 1) a nucleic acid molecule encoding the 4-1BB-binding protein according to any one of claims 1-9; 2) an expression vector comprising the nucleic acid molecule encoding the 4-1BB-binding protein according to any one of claims 1-9; or 3) a host cell comprising the nucleic acid molecule or the expression vector encoding the 4-1BB-binding protein according to any one of claims 1-9.
  11. A pharmaceutical composition, comprising the 4-1BB-binding protein according to any one of claims 1-9 and a pharmaceutically acceptable carrier.
  12. Use of the 4-1BB-binding protein according to any one of claims 1-9, the biomaterial according to claim 10, or the pharmaceutical composition according to claim 11 in the preparation of a medicament for treating or preventing a disease, wherein preferably, the disease is a tumor, an autoimmune disease, or an inflammatory disease.

Description

TECHNICAL FIELD The present disclosure pertains to the field of bio-pharmaceuticals and relates to a 4-1BB-binding protein, such as a single-domain antibody, a heavy-chain antibody, and/or a fusion protein binding to 4-1BB, and use thereof. BACKGROUND 4-1BB, also known as CD137, belongs to the tumor necrosis factor receptor superfamily and plays an important role in T cell activation-driven immune responses. It mainly promotes T cell proliferation, enables T cells to acquire effector functions, promotes immune memory, and inhibits activation-induced cell death. The earliest 4-1BB-targeting agonistic antibodies to enter clinical trials include urelumab, developed by Bristol-Myers Squibb (BMS), and utomilumab, developed by Pfizer. They have shown potential as monotherapies or a combination therapy in mouse tumor models. However, due to toxicity or lack of efficacy in clinical cases (Chester C, Sanmamed MF, Wang J, Melero I. Immunotherapy targeting 4-1BB: mechanistic rationale, clinical results, and future strategies. Blood. 2018;131(1):49-57.), there is a great clinical need for safe and effective 4-1BB-targeting novel therapies. SUMMARY The present disclosure provides a 4-1BB-binding protein that can specifically bind to 4-1BB and does not block the binding of 4-1BB to 4-1BBL (a 4-1BB ligand). The protein can effectively activate NF-κB signaling and T cells under cross-linking conditions. In one aspect, the present disclosure provides a 4-1BB-binding protein, comprising a heavy chain variable region, wherein the heavy chain variable region comprises one or more of the following complementarity determining regions (CDRs): an HCDR1 set forth in SEQ ID NO: 7, an HCDR2 set forth in any one of SEQ ID NOs: 8-13 or SEQ ID NOs: 31-36, and an HCDR3 set forth in any one of SEQ ID NOs: 14-19 or SEQ ID NOs: 37-42. In some embodiments, the heavy chain variable region comprises an HCDR1 set forth in SEQ ID NO: 7, an HCDR2 set forth in SEQ ID NOs: 8-13, and an HCDR3 set forth in any one of SEQ ID NOs: 14-19. In some embodiments, the heavy chain variable region comprises an HCDR1 set forth in SEQ ID NO: 7, an HCDR2 set forth in SEQ ID NO: 8, and an HCDR3 set forth in any one of SEQ ID NOs: 14-17. In some embodiments, the heavy chain variable region comprises an HCDR1 set forth in SEQ ID NO: 7, an HCDR2 set forth in SEQ ID NO: 9, and an HCDR3 set forth in SEQ ID NO: 14 or 19. In some embodiments, the heavy chain variable region comprises an HCDR1 set forth in SEQ ID NO: 7, an HCDR2 set forth in SEQ ID NO: 10, and an HCDR3 set forth in SEQ ID NO: 14 or 16. In some embodiments, the heavy chain variable region comprises an HCDR1 set forth in SEQ ID NO: 7, an HCDR2 set forth in SEQ ID NO: 11 or 12, and an HCDR3 set forth in SEQ ID NO: 16. In some embodiments, the heavy chain variable region comprises an HCDR1 set forth in SEQ ID NO: 7, an HCDR2 set forth in SEQ ID NO: 13, and an HCDR3 set forth in SEQ ID NO: 18 or 19. In some embodiments, the heavy chain variable region comprises an HCDR1 set forth in SEQ ID NO: 7, an HCDR2 set forth in SEQ ID NOs: 31-36, and an HCDR3 set forth in any one of SEQ ID NOs: 37-42. In some embodiments, the heavy chain variable region comprises an HCDR1 set forth in SEQ ID NO: 7, an HCDR2 set forth in SEQ ID NO: 31, and an HCDR3 set forth in any one of SEQ ID NOs: 37-40. In some embodiments, the heavy chain variable region comprises an HCDR1 set forth in SEQ ID NO: 7, an HCDR2 set forth in SEQ ID NO: 32, and an HCDR3 set forth in SEQ ID NO: 37 or 42. In some embodiments, the heavy chain variable region comprises an HCDR1 set forth in SEQ ID NO: 7, an HCDR2 set forth in SEQ ID NO: 33, and an HCDR3 set forth in SEQ ID NO: 37 or 39. In some embodiments, the heavy chain variable region comprises an HCDR1 set forth in SEQ ID NO: 7, an HCDR2 set forth in SEQ ID NO: 34 or 35, and an HCDR3 set forth in SEQ ID NO: 39. In some embodiments, the heavy chain variable region comprises an HCDR1 set forth in SEQ ID NO: 7, an HCDR2 set forth in SEQ ID NO: 36, and an HCDR3 set forth in SEQ ID NO: 41 or 42. In some embodiments, the heavy chain variable region comprises an amino acid sequence set forth in any one of SEQ ID NOs: 1-6 and 29-30, or an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity compared to the amino acid sequence set forth in any one of SEQ ID NOs: 1-6 and 29-30, or an amino acid sequence having one or more conservative amino acid substitutions compared to the amino acid sequence set forth in any one of SEQ ID NOs: 1-6 and 29-30. In some embodiments, the heavy chain variable region comprises an amino acid sequence set forth in any one of SEQ ID NOs: 1-6 and 29-30. In some embodiments, the 4-1BB-binding protein is an antibody or an antigen-binding fragment. In some embodiments, the 4-1BB-binding protein is in the form of a single-domain antibody (e.g., a nanobody), a heavy-chain antibody, a bispecific antib