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JP-7856264-B2 - SAP FC fusion protein and method of use

JP7856264B2JP 7856264 B2JP7856264 B2JP 7856264B2JP-7856264-B2

Inventors

  • ジョーム・ポンズ
  • ジョナサン・エス・ウォール

Assignees

  • アトララス・インコーポレイテッド
  • ユニバーシティ オブ テネシー リサーチ ファウンデーション

Dates

Publication Date
20260511
Application Date
20211101
Priority Date
20201102

Claims (20)

  1. Starting from the N-terminus and moving towards the C-terminus, the following equation applies: SAP-Fc1-L1-Fc2 [During the ceremony, Fc1 is a first Fc domain sequence containing hinge-CH2-CH3, L1 is a linker, and Fc2 is a second Fc domain sequence containing hinge-CH2-CH3. SAP is a component protein of human serum amyloid-P (SAP). The aforementioned Fc1 and Fc2 each include amino acid substitutions in C226S and C229S according to EU numbering. A fusion protein comprising a structure represented by, The fusion protein comprises the amino acid sequence shown in SEQ ID NO: 1, which has or does not have a C-terminal lysine, or SEQ ID NO: 23, which has or does not have a C-terminal lysine.
  2. The fusion protein according to claim 1, wherein the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 1, with or without C-terminal lysine.
  3. The fusion protein according to claim 1, wherein the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 23, which may or may not have a C-terminal lysine.
  4. Starting from the N-terminus and moving towards the C-terminus, the following equation applies: SAP-Fc1-L1-Fc2 [During the ceremony, Fc1 is a first Fc domain sequence containing hinge-CH2-CH3, L1 is a linker, and Fc2 is a second Fc domain sequence containing hinge-CH2-CH3. SAP is a component protein of human serum amyloid-P (SAP). The Fc1 and Fc2 respectively contain amino acid substitutions in C226S and C229S according to EU numbering, The aforementioned human SAP contains the amino acid sequence shown in Sequence ID No. 20, [The Fc1 and Fc2 are human IgG1Fc domains .] A fusion protein containing the structure represented by .
  5. The fusion protein according to claim 4, wherein L1 comprises the amino acid sequence shown in SEQ ID NO: 19.
  6. The fusion protein according to claim 4 , wherein Fc1 and/or Fc2 comprises the amino acid sequence shown in SEQ ID NO: 18.
  7. The fusion protein according to claim 6, wherein the first and/or second Fc domain contains a mutation that reduces FcRn binding ability.
  8. The fusion protein according to claim 7 , wherein Fc1 and/or Fc2 comprises the amino acid sequence shown in SEQ ID NO: 21.
  9. The fusion protein according to any one of claims 1 to 8 , wherein the fusion protein forms a pentamer or a decamer.
  10. The fusion protein according to any one of claims 1 to 8 , wherein the fusion protein exhibits reduced aggregation compared to a fusion protein lacking one or more of the aforementioned amino acid substitutions.
  11. A pharmaceutical composition comprising a fusion protein according to any one of claims 1 to 8 , and a pharmaceutically acceptable carrier .
  12. The pharmaceutical composition according to claim 11, wherein at least 50% of the fusion protein is in the state of pentamers and/or decamers.
  13. A nucleic acid encoding a fusion protein according to any one of claims 1 to 8 .
  14. A vector comprising the nucleic acid described in claim 13.
  15. A host cell containing the nucleic acid described in claim 13.
  16. The host cell according to claim 15, wherein the host cell is a CHO cell or a 293 cell.
  17. A host cell comprising the vector according to claim 14.
  18. The host cell according to claim 17, wherein the host cell is a CHO cell or a 293 cell.
  19. A method for producing a fusion protein according to any one of claims 1 to 8 , comprising culturing a host cell containing a nucleic acid encoding the fusion protein under conditions for expressing the fusion protein.
  20. The method according to claim 19, wherein the host cell is a CHO cell or a 293 cell.

Description

Cross-reference of related applications This application claims priority under U.S. Provisional Application No. 63/108,799 filed on 2 November 2020 and U.S. Provisional Application No. 63/153,777 filed on 25 February 2021, the entire contents of each of those applications are incorporated herein by reference. Submission of Sequence Listings in ASCII Text Files By reference, the entire contents of the following submission in ASCII text file are incorporated herein by reference: Computer-Readable Format (CRF) of Sequence Listings (filename: 165992000200SEQLIST.TXT, date of submission: October 28, 2021, size: 91,399 bytes). This invention relates to a SAP-Fc fusion protein and a method for treating amyloid-related disorders by administering the SAP-Fc fusion protein. Amyloidosis is a broad group of diseases belonging to the protein conformational disease group, which includes other diseases such as Alzheimer's disease, transmissible spongiform encephalopathy, Huntington's disease, or type II diabetes mellitus. Amyloidosis is a rare disease characterized by the presence of insoluble protein deposits with abnormal fibrous conformations in tissues. The most common cause is fragments of serum precursor proteins. Many organs can be affected by these extracellular deposits, called "amyloid material." The main organs affected by amyloid deposits are the kidneys, heart, digestive tract, liver, skin, peripheral nerves, and eyes. The affected organs usually occupy a considerable volume. Ultimately, amyloidosis can affect all organs and the central nervous system, resulting in a wide variety of symptoms. Various methods have been attempted to treat amyloidosis. For example, chemotherapy with glucocorticoids (dexamethasone) and anti-mitotic drugs. The effectiveness of new anti-inflammatory drugs (anti-TNF, anti-IL-1) is currently under clinical evaluation. However, for the time being, amyloidosis remains incurable and fatal because there is no effective treatment that can eliminate the deposits more rapidly. DMSO, colchicine, and i-Dox anthracyclines have also been tried. Anti-SAP antibodies are another therapeutic agent being developed for amyloidosis. EOD001 is a monoclonal antibody that specifically targets amyloid-amyloid AL or AA. WO2015063728 discloses an SAP-Fc antibody fusion for the treatment of amyloidosis. Therefore, effective treatments for amyloidosis and amyloid-related diseases are needed. In one embodiment, the Specified Reference Indicators provide a fusion protein comprising a first polypeptide and a second polypeptide, wherein the first polypeptide comprises a human serum amyloid-P (SAP) component protein ligated to the N-terminus of a first human Fc domain, and the second polypeptide comprises a second human Fc domain but does not contain a human SAP component protein, the first and second Fc domains form a dimer, one of the two Fc domains comprises a knob mutation, and the other Fc domain comprises a hole mutation. In some embodiments, the first polypeptide comprises a knob mutation and the second polypeptide comprises a hole mutation. In some embodiments, the first polypeptide comprises an amino acid sequence shown in SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 8, or SEQ ID NO: 10, and the second polypeptide comprises an amino acid sequence shown in SEQ ID NO: 6, or SEQ ID NO: 9. In some embodiments, the first polypeptide comprises a hole mutation and the second polypeptide comprises a knob mutation. In some embodiments, the first polypeptide comprises an amino acid sequence shown in SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 14, or SEQ ID NO: 16, and the second polypeptide comprises an amino acid sequence shown in SEQ ID NO: 12, or SEQ ID NO: 15. In some embodiments, the human serum amyloid-P component protein contains an amino acid substitution at position N32 or N110 based on SEQ ID NO: 17. In some embodiments, the human serum amyloid-P component protein contains the amino acid sequence shown in SEQ ID NO: 20. In some embodiments, the first and second Fc domains contain an amino acid substitution at position C226 or C229 according to EU numbering. In some embodiments, the first and second Fc domains contain the amino acid substitution C226S or C229S according to EU numbering. In some embodiments, the first and second Fc domains contain an amino acid substitution at amino acid position 11 or 14, numbered based on the amino acid sequence of SEQ ID NO: 18. In some embodiments, the first and second Fc domains contain a serine residue at amino acid position 11 or 14, numbered based on the amino acid sequence of SEQ ID NO: 18. In some embodiments, the first and second Fc domains contain the amino acid sequence shown in SEQ ID NO: 18. In some embodiments, the first or second Fc domain contains a mutation that reduces FcRn binding. In some embodiments, the fusion protein forms a pentamer. In another embodiment, provided herein are fusion proteins comprising a structure represented by the following formula from N