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CN-122029204-A - Long-acting parathyroid hormone receptor agonist fusion protein and application thereof

CN122029204ACN 122029204 ACN122029204 ACN 122029204ACN-122029204-A

Abstract

A long-acting parathyroid hormone receptor agonist fusion protein and its application are provided. The fusion protein can inhibit the activity of RANKL and promote PTHR mediated bone anabolism, and comprises a first arm and a second arm, wherein the first arm comprises a functional region targeting the RANKL and an Fc region, and the second arm comprises a functional region capable of activating the PTHR and the Fc region. The long-acting parathyroid hormone receptor agonist fusion protein has multiple advantages of better efficacy, longer-lasting effect, better safety, higher structural stability, higher expression quantity and the like, and the comprehensive performance is superior to the existing or potential treatment means such as teriparatide, PEG-PTH, anti-RANKL monoclonal antibody, luo Moshan antibody or a control sample synPTH-alpha RANKL-1.

Inventors

  • SONG LIPING
  • ZHANG HUANHUAN
  • HUANG XIAOHUI
  • MU JUN
  • XU HONG
  • FAN YI
  • WANG HONGYAN

Assignees

  • 上海博生德生物科技有限公司

Dates

Publication Date
20260512
Application Date
20240911
Priority Date
20230911

Claims (18)

  1. The long-acting parathyroid hormone receptor agonist fusion protein is characterized by comprising a first arm and a second arm, wherein the first arm comprises a RANKL-targeting functional region and an Fc region, the RANKL-targeting functional region is Fab or ScFab, the RANKL-targeting functional region is positioned at the N end of the Fc region, the second arm comprises a PTHR-activatable functional region and the Fc region, and the C end of the PTHR-activatable functional region of the second arm is directly connected with the N end of the Fc region or is connected with the Fc region through a linker.
  2. The long acting parathyroid hormone receptor agonist fusion protein of claim 1 wherein the Fc region consists of FcA and FcB, the FcA and FcB hinge region-containing tandem CH2 and CH3 of human IgG, respectively, fcA and FcB Knob(s) mutations and Hole(s) mutations, respectively; Preferably, the human IgG is selected from the group consisting of IgG1, igG2, igG4, said IgG1 being a mutant with silent Fc immune effect function, said IgG2 comprising a mutant with normal Fc immune effect function or silent Fc immune effect function, said IgG4 comprising a mutant with silent Fc immune effect function and/or a mutant of S228P mutation that improves molecular stability.
  3. The long acting parathyroid hormone receptor agonist fusion protein of claim 2 wherein when said FcA comprises the amino acid sequence shown as SEQ ID No. 7, said FcB comprises the amino acid sequence shown as SEQ ID No. 8, when said FcA comprises the amino acid sequence shown as SEQ ID No. 8, said FcB comprises the amino acid sequence shown as SEQ ID No. 7, when said FcA comprises the amino acid sequence shown as SEQ ID No. 39, said FcB comprises the amino acid sequence shown as SEQ ID No. 40, and when said FcA comprises the amino acid sequence shown as SEQ ID No. 40, said FcB comprises the amino acid sequence shown as SEQ ID No. 39.
  4. The long acting parathyroid hormone receptor agonist fusion protein of any one of claims 1-3 wherein the PTHR-activatable functional region comprises a PTHR-targeting agonistic antigen-binding variable region or PTH functional region selected from the group consisting of mature human parathyroid hormone (PTH), PTH truncations, PTH mutants, mutants of PTH truncations, mature human parathyroid hormone-related protein (PTHrP) truncations and mutants of PTHrP truncations, preferably truncations of at least 11 amino acid residues from the N-terminus of mature human PTH, more preferably truncations of at least 34 amino acid residues from the N-terminus of mature human PTH, further preferably PTH functional regions of PTH (PTH 1-34), PTH (PTH 1-37), PTH (1-38), PTH (1-41), PTH (1-84), PTP (1-34), amino acid residues from the N-terminus of PTH, amino acid residues from PTH-34, further preferably PTH (PTH-34), amino acid residues from PTH-34, amino acid residues from PTH-33, amino acid residues from PTH-3, amino acid residues from amino acid sequence conservative substitutions such as shown in any one of the amino acid sequence; And/or the number of the groups of groups, The sequence of the linker is selected from (GGGGS) n, (GGGS) n, (GGS) n, (G) n, (GS) n, (EAAAK) n, or (XP) n, or a combination of the foregoing, n is a natural number, and the length of the linker is not more than 20 amino acids, preferably the sequence of the linker comprises an amino acid sequence selected from any one of SEQ ID NOs 9-12, more preferably the sequence of the linker comprises an amino acid sequence shown as SEQ ID NO 9.
  5. The long acting parathyroid hormone receptor agonist fusion protein of any one of claims 1-4, wherein the first arm comprises a first arm a chain and a first arm B chain; the structure of the first arm A chain is VL-CL from the N end to the C end; The structure of the first arm B chain is VH-CH1-FcA from the N end to the C end; optionally, the VL and VH or CL and CH1 are interchanged by cross mab technology. And/or the number of the groups of groups, The structure of the second arm from the N end to the C end is a functional region-FcB capable of activating PTHR or a functional region-linker-FcB capable of activating PTHR;
  6. The long acting parathyroid hormone receptor agonist fusion protein of any of claims 1-5 wherein RANKL-targeting Fab or ScFab comprises 1) a VH comprising heavy chain complementarity determining regions HCDR1-3, 2) a VL comprising light chain complementarity determining regions LCDR1-3, 3) CL and 4) CH1, the amino acid sequences of said HCDR1-3 being shown as SEQ ID NOs 16-18 or 22-24, respectively, the amino acid sequences of LCDR1-3 being shown as SEQ ID NOs 19-21 or 25-27, respectively, optionally said Fab or ScFab exchanging VH and VL or exchanging CH1 and CL by cross mab technology.
  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 shown in SEQ ID No. 1 or 3 and the amino acid sequence of VL comprises the amino acid sequence shown in SEQ ID No. 2 or 4; And/or the amino acid sequence of the CH1 comprises an amino acid sequence shown as SEQ ID NO. 14, and the amino acid sequence of the CL comprises an amino acid sequence shown as SEQ ID NO. 15.
  8. A long-acting parathyroid hormone receptor agonist fusion protein, wherein the long-acting parathyroid hormone receptor agonist fusion protein comprises a first arm and a second arm, the first arm comprising a first arm a chain and a first arm B chain, wherein: When the amino acid sequence of the first arm A chain comprises SEQ ID NO. 2, SEQ ID NO. 15, the amino acid sequence of the first arm B chain comprises SEQ ID NO. 1, SEQ ID NO. 14, SEQ ID NO. 7 or SEQ ID NO. 1, SEQ ID NO. 14, SEQ ID NO. 8 or SEQ ID NO. 1, SEQ ID NO. 14, SEQ ID NO. 39 or SEQ ID NO. 1, SEQ ID NO. 14, SEQ ID NO. 40, or When the amino acid sequence of the first arm A chain comprises SEQ ID NO. 1, SEQ ID NO. 15, the amino acid sequence of the first arm B chain comprises SEQ ID NO. 2, SEQ ID NO. 14, SEQ ID NO. 7 or SEQ ID NO. 2, SEQ ID NO. 14, SEQ ID NO. 8 or SEQ ID NO. 2, SEQ ID NO. 14, SEQ ID NO. 39 or SEQ ID NO. 2, SEQ ID NO. 14, SEQ ID NO. 40, or When the amino acid sequence of the first arm A chain comprises SEQ ID NO. 2, SEQ ID NO. 14, the amino acid sequence of the first arm B chain comprises SEQ ID NO. 1, SEQ ID NO. 15, SEQ ID NO. 7 or SEQ ID NO. 1, SEQ ID NO. 15, SEQ ID NO. 8 or SEQ ID NO. 1, SEQ ID NO. 15, SEQ ID NO. 39 or SEQ ID NO. 1, SEQ ID NO. 15, SEQ ID NO. 40, or When the amino acid sequence of the first arm A chain comprises SEQ ID NO. 1, SEQ ID NO. 14, the amino acid sequence of the first arm B chain comprises SEQ ID NO. 2, SEQ ID NO. 15, SEQ ID NO. 7 or SEQ ID NO. 2, SEQ ID NO. 15, SEQ ID NO. 8 or SEQ ID NO. 2, SEQ ID NO. 15, SEQ ID NO. 39 or SEQ ID NO. 2, SEQ ID NO. 15, SEQ ID NO. 40, or When the amino acid sequence of the first arm A chain comprises SEQ ID NO. 4, SEQ ID NO. 15, the amino acid sequence of the first arm B chain comprises SEQ ID NO. 3, SEQ ID NO. 14, SEQ ID NO. 7, or SEQ ID NO. 3, SEQ ID NO. 14, SEQ ID NO. 8, or SEQ ID NO. 3, SEQ ID NO. 14, SEQ ID NO. 39, or SEQ ID NO. 3, SEQ ID NO. 14, SEQ ID NO. 40, or When the amino acid sequence of the first arm A chain comprises SEQ ID NO 3, SEQ ID NO 15, the amino acid sequence of the first arm B chain comprises SEQ ID NO 4, SEQ ID NO 14, SEQ ID NO 7, or SEQ ID NO 4, SEQ ID NO 14, SEQ ID NO 8, or SEQ ID NO 4, SEQ ID NO 14, SEQ ID NO 39, or SEQ ID NO 4, SEQ ID NO 14, SEQ ID NO 40, or When the amino acid sequence of the first arm A chain comprises SEQ ID NO. 4, SEQ ID NO. 14, the amino acid sequence of the first arm B chain comprises SEQ ID NO. 3, SEQ ID NO. 15, SEQ ID NO. 7, or SEQ ID NO. 3, SEQ ID NO. 15, SEQ ID NO. 8, or SEQ ID NO. 3, SEQ ID NO. 15, SEQ ID NO. 39, or SEQ ID NO. 3, SEQ ID NO. 15, SEQ ID NO. 40, or When the amino acid sequence of the first arm A chain comprises SEQ ID NO 3, SEQ ID NO 14 connected in sequence, the amino acid sequence of the first arm B chain comprises SEQ ID NO 4, SEQ ID NO 15, SEQ ID NO 7 or SEQ ID NO 4, SEQ ID NO 15, SEQ ID NO 8 connected in sequence or SEQ ID NO 4, SEQ ID NO 15, SEQ ID NO 39 or SEQ ID NO 4, SEQ ID NO 15, SEQ ID NO 40 connected in sequence; Preferably, the method comprises the steps of, When the B-chain amino acid sequence of the first arm comprises SEQ ID NO. 7, the amino acid sequence of the second arm comprises a sequence linked to a PTH functional region selected from the group consisting of SEQ ID NO. 5, 33-37, a linker selected from the group consisting of SEQ ID NO. 9-12, and an Fc region having the sequence SEQ ID NO. 8, or When the B-chain amino acid sequence of the first arm comprises SEQ ID NO. 8, the amino acid sequence of the second arm comprises a sequence linked to a PTH functional region selected from the group consisting of SEQ ID NO. 5, 33-37, a linker selected from the group consisting of SEQ ID NO. 9-12, and an Fc region having the sequence SEQ ID NO. 7, or When the B-chain amino acid sequence of the first arm comprises SEQ ID NO:39, the amino acid sequence of the second arm comprises a sequence of a sequence linked to a PTH functional region selected from the group consisting of SEQ ID NO:5, 33-37, a linker selected from the group consisting of SEQ ID NO:9-12, and an Fc region having the sequence SEQ ID NO:40, or When the B-chain amino acid sequence of the first arm comprises SEQ ID NO. 40, the amino acid sequence of the second arm comprises a sequence linked to a PTH functional region selected from the group consisting of SEQ ID NO. 5, 33-37, a linker selected from the group consisting of SEQ ID NO. 9-12, and an Fc region sequence of SEQ ID NO. 39.
  9. The long acting parathyroid hormone receptor agonist fusion protein of claim 8, When the amino acid sequence of the first arm A chain comprises SEQ ID NO. 2, SEQ ID NO. 15, the amino acid sequence of the first arm B chain comprises SEQ ID NO. 1, SEQ ID NO. 14, SEQ ID NO. 7, or the amino acid sequence of the first arm A chain comprises SEQ ID NO. 2, SEQ ID NO. 14, the amino acid sequence of the first arm B chain comprises SEQ ID NO. 1, SEQ ID NO. 15, SEQ ID NO. 7, or the amino acid sequence of the first arm A chain comprises SEQ ID NO. 1, SEQ ID NO. 14, the amino acid sequence of the first arm B chain comprises SEQ ID NO. 2, SEQ ID NO. 15, SEQ ID NO. 7, or the amino acid sequence of the first arm A chain comprises SEQ ID NO. 1, SEQ ID NO. 15, the amino acid sequence of the first arm B chain comprises SEQ ID NO. 2, SEQ ID NO. 14, SEQ ID NO. 7; The amino acid sequence of the second arm comprises SEQ ID NO. 5, SEQ ID NO. 9, SEQ ID NO. 8, or SEQ ID NO. 5, SEQ ID NO. 10, SEQ ID NO. 8, or SEQ ID NO. 5, SEQ ID NO. 11, SEQ ID NO. 8, or SEQ ID NO. 5, SEQ ID NO. 12, SEQ ID NO. 8, or SEQ ID NO. 33, SEQ ID NO. 9, SEQ ID NO. 8, or SEQ ID NO. 33, SEQ ID NO. 10, SEQ ID NO. 8, or SEQ ID NO. 33, SEQ ID NO. 11, SEQ ID NO. 8, or SEQ ID NO. 33, SEQ ID NO. 12, SEQ ID NO. 8, or SEQ ID NO. 34, SEQ ID NO. 9, SEQ ID NO. 8, or SEQ ID NO. 34, SEQ ID NO. 10, SEQ ID NO. 8, or SEQ ID NO. 34, SEQ ID NO. 11, SEQ ID NO. 8, or SEQ ID NO. 34, SEQ ID NO. 12, SEQ ID NO. 8, or SEQ ID NO. 35, SEQ ID NO. 9, SEQ ID NO. 8, or SEQ ID NO. 35, SEQ ID NO. 10, SEQ ID NO. 8, or SEQ ID NO. 35, SEQ ID NO. 11, SEQ ID NO. 8, or SEQ ID NO. 35, SEQ ID NO. 12, SEQ ID NO. 8, or SEQ ID NO. 36, SEQ ID NO. 9, SEQ ID NO. 8, or SEQ ID NO. 36, SEQ ID NO. 10, SEQ ID NO. 8, or SEQ ID NO. 36, SEQ ID NO. 11, SEQ ID NO. 8, or SEQ ID NO. 36, SEQ ID NO. 12, SEQ ID NO. 8, or SEQ ID NO. 37, SEQ ID NO. 9, SEQ ID NO. 8, or SEQ ID NO. 37, SEQ ID NO. 10, SEQ ID NO. 8, or SEQ ID NO. 37, SEQ ID NO. 11, SEQ ID NO. 8, or SEQ ID NO. 37, SEQ ID NO. 12, SEQ ID NO. 8; When the amino acid sequence of the first arm A chain comprises SEQ ID NO. 2, SEQ ID NO. 15, the amino acid sequence of the first arm B chain comprises SEQ ID NO. 1, SEQ ID NO. 14, SEQ ID NO. 8, or the amino acid sequence of the first arm A chain comprises SEQ ID NO. 2, SEQ ID NO. 14, the amino acid sequence of the first arm B chain comprises SEQ ID NO. 1, SEQ ID NO. 15, SEQ ID NO. 8, or the amino acid sequence of the first arm A chain comprises SEQ ID NO. 1, SEQ ID NO. 14, the amino acid sequence of the first arm B chain comprises SEQ ID NO. 2, SEQ ID NO. 15, SEQ ID NO. 8, or the amino acid sequence of the first arm A chain comprises SEQ ID NO. 1, SEQ ID NO. 15, the amino acid sequence of the first arm B chain comprises SEQ ID NO. 2, SEQ ID NO. 14, SEQ ID NO. 8; The amino acid sequence of the second arm comprises SEQ ID NO. 5, SEQ ID NO. 9, SEQ ID NO. 7, or SEQ ID NO. 5, SEQ ID NO. 10, SEQ ID NO. 7, or SEQ ID NO. 5, SEQ ID NO. 11, SEQ ID NO. 7, or SEQ ID NO. 5, SEQ ID NO. 12, SEQ ID NO. 7, or SEQ ID NO. 33, SEQ ID NO. 9, SEQ ID NO. 7, or SEQ ID NO. 33, SEQ ID NO. 10, SEQ ID NO. 7, or SEQ ID NO. 33, SEQ ID NO. 11, SEQ ID NO. 7, or SEQ ID NO. 33, SEQ ID NO. 12, SEQ ID NO. 7, or SEQ ID NO. 34, SEQ ID NO. 9, SEQ ID NO. 7, or SEQ ID NO. 34, SEQ ID NO. 10, SEQ ID NO. 7, or SEQ ID NO. 34, SEQ ID NO. 11, SEQ ID NO. 7, or SEQ ID NO. 34, SEQ ID NO. 12, SEQ ID NO. 7, or SEQ ID NO. 35, SEQ ID NO. 9, SEQ ID NO. 7, or SEQ ID NO. 35, SEQ ID NO. 10, SEQ ID NO. 7, or SEQ ID NO. 35, SEQ ID NO. 11, SEQ ID NO. 7, or SEQ ID NO. 35, SEQ ID NO. 12, SEQ ID NO. 7, or SEQ ID NO. 36, SEQ ID NO. 9, SEQ ID NO. 7, or SEQ ID NO. 36, SEQ ID NO. 10, SEQ ID NO. 7, or SEQ ID NO. 36, SEQ ID NO. 11, SEQ ID NO. 7, or SEQ ID NO. 36, SEQ ID NO. 12, SEQ ID NO. 7, or SEQ ID NO. 37, SEQ ID NO. 9, SEQ ID NO. 7, or SEQ ID NO. 37, SEQ ID NO. 10, SEQ ID NO. 7, or SEQ ID NO. 37, SEQ ID NO. 11, SEQ ID NO. 7, or SEQ ID NO. 37, SEQ ID NO. 12, SEQ ID NO. 7; When the amino acid sequence of the first arm A chain comprises SEQ ID NO. 2, SEQ ID NO. 15, the amino acid sequence of the first arm B chain comprises SEQ ID NO. 1, SEQ ID NO. 14, SEQ ID NO. 39, or the amino acid sequence of the first arm A chain comprises SEQ ID NO. 2, SEQ ID NO. 14, the amino acid sequence of the first arm B chain comprises SEQ ID NO. 1, SEQ ID NO. 15, SEQ ID NO. 39, or the amino acid sequence of the first arm A chain comprises SEQ ID NO. 1, SEQ ID NO. 14, the amino acid sequence of the first arm B chain comprises SEQ ID NO. 2, SEQ ID NO. 15, SEQ ID NO. 39, or the amino acid sequence of the first arm A chain comprises SEQ ID NO. 1, SEQ ID NO. 15, SEQ ID NO. 2, 14, SEQ ID NO. 39; The amino acid sequence of the second arm comprises SEQ ID NO. 5, SEQ ID NO. 9, SEQ ID NO. 40, or SEQ ID NO. 5, SEQ ID NO. 10, SEQ ID NO. 40, or SEQ ID NO. 5, SEQ ID NO. 11, SEQ ID NO. 40, or SEQ ID NO. 5, SEQ ID NO. 12, SEQ ID NO. 40, or SEQ ID NO. 33, SEQ ID NO. 9, SEQ ID NO. 40, or SEQ ID NO. 33, SEQ ID NO. 10, SEQ ID NO. 40, or SEQ ID NO. 33, SEQ ID NO. 11, SEQ ID NO. 40, or SEQ ID NO. 33, SEQ ID NO. 40, or SEQ ID NO. 40, connected in sequence, SEQ ID NO. 12, SEQ ID NO. 40, or SEQ ID NO. 34, SEQ ID NO. 40 connected in sequence, or SEQ ID NO. 34, SEQ ID NO. 9, SEQ ID NO. 40 connected in sequence, or SEQ ID NO. 34, SEQ ID NO. 10, SEQ ID NO. 40 connected in sequence, or SEQ ID NO. 34, SEQ ID NO. 11, SEQ ID NO. 40 connected in sequence, or SEQ ID NO. 34 connected in sequence, SEQ ID NO. 12, SEQ ID NO. 40, or SEQ ID NO. 35, SEQ ID NO. 9, SEQ ID NO. 40, or SEQ ID NO. 35, SEQ ID NO. 10, SEQ ID NO. 40, or SEQ ID NO. 35, SEQ ID NO. 11, SEQ ID NO. 40, or SEQ ID NO. 35, SEQ ID NO. 35, SEQ ID NO. 12, SEQ ID NO. 40, or SEQ ID NO. 36, SEQ ID NO. 9, SEQ ID NO. 40, or SEQ ID NO. 36, SEQ ID NO. 10, SEQ ID NO. 40, or SEQ ID NO. 36, SEQ ID NO. 11, SEQ ID NO. 40, or SEQ ID NO. 36, SEQ ID NO. 12, SEQ ID NO. 40, or SEQ ID NO. 37, SEQ ID NO. 40 connected in sequence, or SEQ ID NO. 37, SEQ ID NO. 9, SEQ ID NO. 40 connected in sequence, or SEQ ID NO. 37, SEQ ID NO. 10, SEQ ID NO. 40 connected in sequence, or SEQ ID NO. 37, SEQ ID NO. 11, SEQ ID NO. 40 connected in sequence, or SEQ ID NO. 37 connected in sequence, SEQ ID NO. 12, SEQ ID NO. 40; When the amino acid sequence of the first arm A chain comprises SEQ ID NO. 2, SEQ ID NO. 15, the amino acid sequence of the first arm B chain comprises SEQ ID NO. 1, SEQ ID NO. 14, SEQ ID NO. 40, or the amino acid sequence of the first arm A chain comprises SEQ ID NO. 2, SEQ ID NO. 14, the amino acid sequence of the first arm B chain comprises SEQ ID NO. 1, SEQ ID NO. 15, SEQ ID NO. 40, or the amino acid sequence of the first arm A chain comprises SEQ ID NO. 1, SEQ ID NO. 14, the amino acid sequence of the first arm B chain comprises SEQ ID NO. 2, SEQ ID NO. 15, SEQ ID NO. 40, or the amino acid sequence of the first arm A chain comprises SEQ ID NO. 1, SEQ ID NO. 15, the amino acid sequence of the first arm B chain comprises SEQ ID NO. 2, SEQ ID NO. 14, SEQ ID NO. 40; The amino acid sequence of the second arm comprises SEQ ID NO. 5, SEQ ID NO. 9, SEQ ID NO. 39, or SEQ ID NO. 5, SEQ ID NO. 10, SEQ ID NO. 39, or SEQ ID NO. 5, SEQ ID NO. 11, SEQ ID NO. 39, or SEQ ID NO. 5, SEQ ID NO. 12, SEQ ID NO. 39, or SEQ ID NO. 33, SEQ ID NO. 9, SEQ ID NO. 39, or SEQ ID NO. 33, SEQ ID NO. 10, SEQ ID NO. 39, or SEQ ID NO. 33, SEQ ID NO. 11, SEQ ID NO. 39, or SEQ ID NO. 33, SEQ ID NO. 33, SEQ ID NO. 12, SEQ ID NO. 39, or SEQ ID NO. 34, SEQ ID NO. 39 connected in sequence, or SEQ ID NO. 34, SEQ ID NO. 9, SEQ ID NO. 39 connected in sequence, or SEQ ID NO. 34, SEQ ID NO. 10, SEQ ID NO. 39 connected in sequence, or SEQ ID NO. 34, SEQ ID NO. 11, SEQ ID NO. 39 connected in sequence, or SEQ ID NO. 34 connected in sequence, SEQ ID NO. 12, SEQ ID NO. 39, or SEQ ID NO. 35, SEQ ID NO. 9, SEQ ID NO. 39, or SEQ ID NO. 35, SEQ ID NO. 10, SEQ ID NO. 39, or SEQ ID NO. 35, SEQ ID NO. 11, SEQ ID NO. 39, or SEQ ID NO. 35, SEQ ID NO. 12, SEQ ID NO. 39, or SEQ ID NO. 36, SEQ ID NO. 9, SEQ ID NO. 39, or SEQ ID NO. 36, SEQ ID NO. 10, SEQ ID NO. 39, or SEQ ID NO. 36, SEQ ID NO. 11, SEQ ID NO. 39, or SEQ ID NO. 36, SEQ ID NO. 36, SEQ ID NO. 12, SEQ ID NO. 39, or SEQ ID NO. 37, SEQ ID NO. 39 linked in sequence, or SEQ ID NO. 37, SEQ ID NO. 9, SEQ ID NO. 39 linked in sequence, or SEQ ID NO. 37, SEQ ID NO. 10, SEQ ID NO. 39 linked in sequence, or SEQ ID NO. 37, SEQ ID NO. 11, SEQ ID NO. 39 linked in sequence, or SEQ ID NO. 37, SEQ ID NO. 12, SEQ ID NO. 39.
  10. An isolated nucleic acid molecule encoding the long acting parathyroid hormone receptor agonist fusion protein of any one of claims 1-9.
  11. A recombinant expression vector comprising the nucleic acid molecule of claim 10.
  12. An engineered cell comprising the recombinant expression vector of claim 11, preferably the host cell of the engineered cell is a eukaryotic cell, more preferably the eukaryotic cell is a yeast cell or a mammalian cell, even more preferably the mammalian cell is a CHO cell or a HEK293 cell.
  13. A pharmaceutical composition comprising the long-acting parathyroid hormone receptor agonist fusion protein of any one of claims 1-9, the nucleic acid molecule of claim 10, the recombinant expression vector of claim 11, and/or the engineered cell of claim 12; Optionally, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier and/or adjuvant.
  14. Use of a long-acting parathyroid hormone receptor agonist fusion protein of any one of claims 1-9, a nucleic acid molecule of claim 10, a recombinant expression vector of claim 11, an engineered cell of claim 12, or a pharmaceutical composition of claim 13 in the manufacture of a medicament for the treatment, prevention, co-treatment, or neoadjuvant treatment of a bone disorder in need of treatment, prevention, co-treatment, or neoadjuvant treatment by modulating bone formation and/or bone resorption at a site associated with a lesion; Preferably, the bone disorder is osteoporosis, osteopenia, osteogenesis imperfecta, bone loss associated with transplantation, autoimmune induced bone loss, disuse induced bone loss, bone injury, degenerative lumbar spondylolisthesis or degenerative disc disease, the bone injury is preferably a fracture, bone fissure, cartilage injury, bone necrosis, bone injury due to arthritis or tumors, the fracture is preferably an osteoporotic fracture, fracture disunion or delayed union of the fracture, the arthritis comprises arthritis due to degeneration, trauma, strain, infection, autoimmune diseases, such as rheumatoid arthritis, osteoarthritis, infectious arthritis, the tumors comprise bone tumors and tumor-bone metastases.
  15. A method of making a long acting parathyroid hormone receptor agonist fusion protein comprising the step of culturing the engineered cell of claim 12.
  16. A method of treating, preventing, co-treating or neoadjuvant treating a bone disorder by modulating bone formation or/and bone resorption at a site associated with a lesion, comprising administering to a subject in need thereof an effective amount of a long acting parathyroid hormone receptor agonist fusion protein of any one of claims 1-9, a nucleic acid molecule of claim 10, a recombinant expression vector of claim 11, an engineered cell of claim 12 or a pharmaceutical composition of claim 13; Preferably, the bone disorder is osteoporosis, osteopenia, osteogenesis imperfecta, bone loss associated with transplantation, autoimmune induced bone loss, disuse induced bone loss, bone injury, degenerative lumbar spondylolisthesis, or degenerative disc disease, preferably bone fracture, bone fissure, cartilage injury, osteonecrosis, bone injury from arthritis or tumors, preferably osteoporotic fracture, non-union of bone fracture, or delayed union of bone fracture, arthritis including arthritis caused by degeneration, trauma, strain, infection, autoimmune disease, or inflammation, such as rheumatoid arthritis, osteoarthritis, infectious arthritis, tumors including bone tumor and tumor bone metastasis; And/or the method of administration is selected from oral administration (e.g. by oral delivery device) or injection, preferably intravenous, subcutaneous, intramuscular, or topical injection.
  17. Use of a long acting parathyroid hormone receptor agonist fusion protein of any one of claims 1-9, a nucleic acid molecule of claim 10, a recombinant expression vector of claim 11, an engineered cell of claim 12 or a pharmaceutical composition of claim 13 as a medicament.
  18. A long acting parathyroid hormone receptor agonist fusion protein of any one of claims 1-9, a nucleic acid molecule of claim 10, a recombinant expression vector of claim 11, an engineered cell of claim 12, or a pharmaceutical composition of claim 13 for use in the treatment, prevention, adjuvant treatment, or neoadjuvant treatment of a bone disorder; Preferably, the bone disorder is osteoporosis, osteopenia, osteogenesis imperfecta, bone loss associated with transplantation, autoimmune induced bone loss, disuse induced bone loss, bone injury, degenerative lumbar spondylolisthesis or degenerative disc disease, the bone injury is preferably a fracture, bone fissure, cartilage injury, bone necrosis, bone injury due to arthritis or a tumor, the fracture is preferably an osteoporotic fracture, fracture disunion or delayed union of the fracture, the arthritis comprises arthritis due to degeneration, trauma, strain, infection, autoimmune disease or inflammation, such as rheumatoid arthritis, osteoarthritis, infectious arthritis, the tumor comprises bone tumor and tumor bone metastasis.

Description

Long-acting parathyroid hormone receptor agonist fusion protein and application thereof The present application claims priority from chinese patent application 2023111664518 with application date 2023/9/11 and chinese patent application 2024112526007 with application date 2024/9/6. The present application incorporates the entirety of the above-mentioned chinese patent application. Technical Field The invention belongs to the field of biological medicine, and in particular relates to a long-acting parathyroid hormone receptor agonist fusion protein and application thereof. Background Osteoporosis is a common degenerative bone disease characterized by bone loss and bone structure deterioration, and is induced by an imbalance between osteoblast bone formation and osteoclast bone resorption (Dirckx N,Van Hul M,Maes C.Osteoblast recruitment to sites of bone formation in skeletal development,homeostasis,and regeneration.Birth Defects Res C Embryo Today.2013;99(3):170-91.), when the bone resorption rate exceeds bone formation, resulting in decreased bone density and increased risk of body fracture, thereby inducing osteoporosis (Egermann M,Schneider E,Evans C H,et al,The potential of gene therapy for fracture healing in osteoporosis[J].Osteoporos Int,2005,16(Suppl 2):S120-S128.). Osteoporosis has been listed by the world health organization as the second next disease to cardiovascular disease that is the second most damaging to human health. The first sign of the disease is a fracture, because of the lack of warning signals. In the population over 50 years, the proportion of osteoporotic fractures occur is as high as 19.2%. Within 1 year after hip fracture, 20% of all complications die, while about 50% of the surviving population is disabled. Osteoporotic fracture (also called brittle fracture) is the most serious consequence of osteoporosis, and not only affects the quality of life of patients, but also is one of the main causes of disability and death for elderly patients, resulting in heavy household and social burden. The current common medicines for treating osteoporosis clinically comprise calcitonin, bisphosphonate, estrogen medicines and monoclonal antibodies of anti-nuclear factor kappa B receptor activator ligand (Receptor activator of nuclear factor kappa-B ligand, RANKL), and although the medicines have certain curative effects, the medicines can only inhibit bone absorption and can not promote bone formation, so the medicines can only delay the development of diseases and can not cure the diseases. Most of the elderly patients with osteoporosis have low self bone metabolism and turnover rate, and many medicines for treating osteoporosis inhibit osteoclast, and have an effect of inhibiting bone metabolism or bone turnover. In terms of bone remodeling and regeneration, more drugs for promoting bone formation should be used in female patients with osteoporotic vertebral fractures of 65 years or older and BMD below-2.5 SD, in patients with multiple vertebral/hip fractures after menopause, in patients with osteoporotic fractures and multiple osteoporotic fractures after administration of drugs for inhibiting bone resorption. The two purposes are that firstly, the bone strength is effectively improved, the next fracture is prevented, and secondly, the bone quality is improved, because a part of patients need to do surgery, and the quality of the bone can influence the effect of surgery treatment. In recent years, bone formation promoters have received widespread attention. Parathyroid hormone (parathyroid hormone, PTH) is a peptide of 84 amino acids that plays an important role in bone remodeling. Binding of PTH to parathyroid hormone receptor (parathyroid hormone receptor, PTHR) directly induces bone formation by activating cAMP and classical wnt signaling pathways. Teriparatide (TERIPARATIDE, rhPTH (1-34)) is a commercially available therapeutic PTH peptide, an N-terminal 34 amino acid fragment of mature human PTH, which has been shown to increase bone formation activity in osteoporotic patients, approved by the FDA for the treatment of primary and hypogonadal osteoporosis, postmenopausal osteoporosis. Intermittent small doses of teriparatide may exhibit the effect of promoting bone formation and increasing bone mass, but if the drug is subcutaneously injected daily, it causes a great deal of inconvenience to the patient, and thus patient compliance is poor. In order to solve the problem of poor compliance of teriparatide, a research institution has constructed PEG-PTH 1-34 in order to prolong the half-life of teriparatide, and in vitro research has shown that PEG-PTH 1-34 has activity equivalent to teriparatide, but in vivo efficacy data show that PEG-PTH 1-34 has significantly deteriorated effect of promoting bone formation (Valderrama P,Jung R.E.,Thoma D.S.,et al.Evaluation of parathyroid hormone bound to a synthetic matrix for guided bone regeneration around dental implants:a histomorphometric study i