Search

RU-2026110935-A - LONG-ACTING PARATHYROID HORMONE RECEPTOR AGONIST FUSION PROTEIN AND ITS APPLICATIONS

RU2026110935ARU 2026110935 ARU2026110935 ARU 2026110935ARU-2026110935-A

Inventors

  • СУН, Липин
  • ЧЖАН, Хуаньхуань
  • ХУАН, Сяохуэй
  • МУ, Цзюнь
  • Сюй, Хун
  • Фань, И
  • ВАН, Хунянь

Assignees

  • ШАНХАЙ БиТиДи-БИО ТЕКНОЛОДЖИ КО., ЛТД.

Dates

Publication Date
20260505
Application Date
20240911
Priority Date
20230911

Claims (20)

  1. 1. A long-acting parathyroid hormone receptor agonist fusion protein comprising a first arm and a second arm; wherein the first arm comprises a functional region targeting RANKL and an Fc region, wherein the functional region targeting RANKL is a Fab or ScFab, and the functional region targeting RANKL is located at the N-terminus of the Fc region; the second arm comprises a functional region capable of activating PTHR (parathyroid hormone receptor 1) and an Fc region, and wherein the C-terminus of the functional region capable of activating PTHR in the second arm is connected to the N-terminus of the Fc region directly or via a linker.
  2. 2. The long-acting parathyroid hormone receptor agonist fusion protein of claim 1, wherein the Fc region consists of FcA and FcB, wherein both FcA and FcB are tandem CH2 and CH3 human IgG containing a hinge region, respectively; wherein FcA and FcB contain a knob(s) mutation and a pit(s) mutation, respectively;
  3. preferably, the human IgG is selected from IgG1, IgG2, IgG3 and IgG4; wherein IgG1 is a mutant type with an inactive immune effector function Fc, and IgG4 provides a mutant type with an inactive immune effector function Fc and/or a mutant type with the S228P mutation, which improves molecular stability.
  4. 3. The long-acting parathyroid hormone receptor agonist fusion protein of claim 2, wherein if FcA comprises the amino acid sequence of SEQ ID NO: 7, FcB comprises the amino acid sequence of SEQ ID NO: 8; if FcA comprises the amino acid sequence of SEQ ID NO: 8, FcB comprises the amino acid sequence of SEQ ID NO: 7; if FcA comprises the amino acid sequence of SEQ ID NO: 39, FcB comprises the amino acid sequence of SEQ ID NO: 40; and if FcA comprises the amino acid sequence of SEQ ID NO: 40, FcB comprises the amino acid sequence of SEQ ID NO: 39.
  5. 4. The long-acting parathyroid hormone receptor agonist fusion protein of any one of claims 1 to 3, wherein the functional region capable of activating PTHR comprises an agonistic antigen-binding variable region targeting PTHR, or a PTH functional region, wherein the agonistic antigen-binding variable region targeting PTHR is ScFv or SDA; the PTH functional region is selected from mature human parathyroid hormone (PTH), truncated PTH, a mutant variant of PTH, a mutant variant of truncated PTH, a truncated mature human parathyroid hormone-related protein (PTHrP), a mutant variant of truncated PTH; wherein the truncated PTH is preferably obtained by truncating at least 11 amino acid residues from the N-terminus of mature human PTH; and the truncated PTH is more preferably obtained by truncating at least 34 amino acid residues from the N-terminus of mature human PTH; even more preferably, the functional region of PTH is PTH(1-34), PTH(1-37), PTH(1-38), PTH(1-41), PTH(1-84), PTHrP(1-34), PTHrP(1-36); even more preferably, the functional region of PTH comprises an amino acid sequence presented by any one of SEQ ID NOs: 5 and 33-37, or an amino acid sequence obtained by adding, deleting, modifying and/or conservatively substituting at least one amino acid residue, such as 1, 2, 3, 4, 5 or 6 amino acid residues, in the amino acid sequences presented in the above-mentioned sequences;
  6. and/or
  7. the linker sequence is selected from (GGGGS)n, (GGGS)n, (GGS)n, (G)n, (GS)n, (EAAAK)n or (XP)n or a combination of the above sequences, wherein n is a natural number such as 1-20, and preferably the length of the linker is no more than 20 amino acids; preferably the linker sequence comprises an amino acid sequence selected from any of the sequences under SEQ ID NO: 9-12; more preferably the linker sequence comprises the amino acid sequence under SEQ ID NO: 9.
  8. 5. A long-acting parathyroid hormone receptor agonist fusion protein according to any one of claims 1-4, wherein the first arm comprises an A-chain of the first arm and a B-chain of the first arm;
  9. the structure of the A-chain of the first arm from the N-terminus to the C-terminus is VL-CL;
  10. the structure of the B chain of the first arm from the N-terminus to the C-terminus is VH-CH1-FcA;
  11. optionally VL and VH or CL and CH1 are interchangeable via CrossMab technology;
  12. and/or
  13. The structure of the second arm from the N-terminus to the C-terminus represents the functional region capable of activating PTHR-FcB or the functional region capable of activating PTHR-linker-FcB.
  14. 6. The long-acting parathyroid hormone receptor agonist fusion protein of any one of claims 1 to 5, wherein the Fab or ScFab targeting RANKL comprises 1) VH comprising the heavy chain complementarity determining regions HCDR1-3, 2) VL comprising the light chain complementarity determining regions LCDR1-3, 3) CL and 4) CH1, wherein the amino acid sequences of HCDR1-3 are presented under SEQ ID NO: 16-18 or SEQ ID NO: 22-24, respectively, and the amino acid sequences of LCDR1-3 are presented under SEQ ID NO: 19-21 or SEQ ID NO: 25-27, respectively; and optionally, the Fab or ScFab has been exchanged between VH and VL or has been exchanged between CH1 and CL using CrossMab technology.
  15. 7. The long-acting parathyroid hormone receptor agonist fusion protein of claim 5 or 6, wherein the amino acid sequence of VH comprises the amino acid sequence presented under SEQ ID NO: 1 or 3, and the amino acid sequence of VL comprises the amino acid sequence presented under SEQ ID NO: 2 or 4;
  16. and/or
  17. the amino acid sequence of CH1 comprises the amino acid sequence of SEQ ID NO: 14, and the amino acid sequence of CL comprises the amino acid sequence of SEQ ID NO: 15.
  18. 8. A long-acting parathyroid hormone receptor agonist fusion protein comprising a first arm and a second arm, wherein the first arm comprises the A chain of the first arm and the B chain of the first arm; wherein
  19. if the amino acid sequence of the A chain of the first arm comprises the sequence of SEQ ID NO: 2 and the sequence of SEQ ID NO: 15, connected in series, the amino acid sequence of the B chain of the first arm comprises the sequence of SEQ ID NO: 1, the sequence of SEQ ID NO: 14 and the sequence of SEQ ID NO: 6, connected in series, or the sequence of SEQ ID NO: 1, the sequence of SEQ ID NO: 14 and the sequence of SEQ ID NO: 7, connected in series, or the sequence of SEQ ID NO: 1, the sequence of SEQ ID NO: 14 and the sequence of SEQ ID NO: 8, connected in series, or the sequence of SEQ ID NO: 1, the sequence of SEQ ID NO: 14 and the sequence of SEQ ID NO: 39, connected in series, or the sequence of SEQ ID NO: 1, the sequence of SEQ ID NO: 14 and the sequence of SEQ ID NO: 40, connected in series; or
  20. if the amino acid sequence of the A chain of the first arm comprises the sequence of SEQ ID NO: 1 and the sequence of SEQ ID NO: 15, connected in series, the amino acid sequence of the B chain of the first arm comprises the sequence of SEQ ID NO: 2, the sequence of SEQ ID NO: 14 and the sequence of SEQ ID NO: 6, connected in series, or the sequence of SEQ ID NO: 2, the sequence of SEQ ID NO: 14 and the sequence of SEQ ID NO: 7, connected in series, or the sequence of SEQ ID NO: 2, the sequence of SEQ ID NO: 14 and the sequence of SEQ ID NO: 8, connected in series, or the sequence of SEQ ID NO: 2, the sequence of SEQ ID NO: 14 and the sequence of SEQ ID NO: 39, connected in series, or the sequence of SEQ ID NO: 2, the sequence of SEQ ID NO: 14 and the sequence of SEQ ID NO: 40, connected in series; or