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KR-102962913-B1 - Antibody-decorated self-assembling protein nanocages (SAPNA) and parts thereof

KR102962913B1KR 102962913 B1KR102962913 B1KR 102962913B1KR-102962913-B1

Abstract

The present invention provides "antibody-decorated self-assembled protein nanoparticles" (SAPNAs), which are chimeric protein assemblies comprising a protein cage polypeptide (or scaffolding protein) capable of forming or useful for forming a hollow tetrahedral pyramidal structure, and (a) one or more antibodies and (b) a protein cage polypeptide providing a scaffold for arranging antibodies. In some embodiments, the antibody may specifically bind to a pathogenic biological agent, or a part thereof.

Inventors

  • 호지 쿠르티스 디
  • 후라 그레고리 엘
  • 예츠 토드 오

Assignees

  • 더 리젠츠 오브 더 유니버시티 오브 캘리포니아

Dates

Publication Date
20260508
Application Date
20200427
Priority Date
20190425

Claims (20)

  1. As a protein cage polypeptide or scaffolding protein capable of forming or useful for forming a hollow tetrahedral pyramidal structure, The above protein cage polypeptide, or scaffolding protein (i) an amino acid sequence comprising at least 90% amino acid identity for any one of SEQ ID NOs: 20 to 39; and (ii) INSERT A comprising an amino acid sequence capable of specifically binding to an antibody or a part thereof, wherein the amino acid sequence comprises DCAWHLGELVWCT (SEQN: 41) or GCDCAWHLGELVWCTCG (SEQN: 42); or INSERT B comprising an amino acid sequence capable of specifically binding to an antibody or a part thereof, wherein the amino acid sequence comprises DCAWHLGELVWCT (SEQN: 41) or GCDCAWHLGELVWCTCG (SEQN: 42). A protein cage polypeptide, or scaffolding protein comprising
  2. The protein cage polypeptide or scaffolding protein of claim 1, wherein the protein cage polypeptide or scaffolding protein comprises a structure: polypeptide 1—AHL—polypeptide 2—INSERT A—polypeptide 3—INSERT B—polypeptide 4; wherein AHL is an “alpha helical linker”.
  3. A protein cage polypeptide or scaffolding protein according to paragraph 2, wherein INSERT A has a length of 17 to 25 amino acids and INSERT B has a length of 28 to 85 amino acids.
  4. A protein cage polypeptide according to claim 2 or 3, wherein polypeptide 1 comprises an amino acid sequence having at least 90% amino acid identity with respect to an amino acid sequence up to a maximum AQEAQKQK sequence from the N-terminus of any one of SEQ NOs: 20 to 39.
  5. A protein cage polypeptide according to claim 4, wherein polypeptide 1 comprises an amino acid sequence comprising YGTAR, TDD, LXENLGTR, IDV, TGXRT, and/or SA; wherein X in the sequence LXENLGTR is D, E, or K, and X in the sequence TGXRT is D or E.
  6. In paragraph 4, the protein cage polypeptide wherein polypeptide 1 comprises 278 to 303 amino acid residues.
  7. A protein cage polypeptide according to claim 2 or 3, wherein the AHL comprises an amino acid sequence including AQEAQKQK.
  8. A protein cage polypeptide according to claim 7, wherein the AHL comprises 5, 6, 7, 8, 9, 10, or 11 amino acid residues.
  9. A protein cage polypeptide according to claim 2 or 3, wherein polypeptide 2 comprises an amino acid sequence having at least 90% amino acid identity with respect to the amino acid sequence from the C-end of any one of SEQEAQKQK sequences 20, 22, 23, 24, 25, 37, 28, or 39 to the N-end of INSERT A.
  10. A protein cage polypeptide according to claim 9, wherein the polypeptide 2 comprises an amino acid sequence including LTEVETYVLS (SEQ ID: 43).
  11. A protein cage polypeptide according to claim 9, wherein polypeptide 2 comprises 30 to 36 amino acid residues.
  12. A protein cage polypeptide according to claim 11, wherein polypeptide 2 comprises 33 amino acid residues.
  13. A protein cage polypeptide according to claim 2 or 3, wherein the polypeptide 3 comprises an amino acid sequence having at least 90% amino acid identity with respect to the amino acid sequence from the C-end of INSERT A represented by SEQ ID NO: 20, 22, 23, 24, 25, 37, 28, or 39 to the N-end of INSERT B represented by SEQ ID NO: 21, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36.
  14. A protein cage polypeptide according to claim 13, wherein the polypeptide 3 comprises an amino acid sequence comprising FTLTVPSERGLQR (SEQ No. 44) and/or CATCEQIAD (SEQ No. 45).
  15. In claim 13, the protein cage polypeptide wherein the polypeptide 3 comprises 110 to 130 amino acid residues.
  16. In claim 15, the protein cage polypeptide wherein the polypeptide 3 comprises 121 amino acid residues.
  17. A protein cage polypeptide according to claim 2 or 3, wherein polypeptide 4 comprises an amino acid sequence having at least 90% amino acid identity with respect to the amino acid sequence from the C-end of INSERT B represented by SEQ ID NO: 21, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36.
  18. In claim 17, the protein cage polypeptide wherein the polypeptide 4 comprises an amino acid sequence including EHHHHHH.
  19. A protein cage polypeptide according to claim 17, wherein the polypeptide 4 comprises 5 to 13 amino acid residues.
  20. In claim 19, a protein cage polypeptide wherein the polypeptide 4 comprises eight amino acid residues.

Description

Antibody-decorated self-assembling protein nanocages (SAPNA) and parts thereof Related patent application This application claims priority to U.S. provisional application serial number 62/838,826 filed on April 25, 2019, the full text of which is incorporated herein by reference. Government Support Statement The present invention was made with government support under Contract No. DE-AC02-05CH11231 granted by the U.S. Department of Energy. The government holds specific rights to the present invention. Field of invention The present invention relates to the field of protein-based antibody scaffold production. Therapeutic monoclonal antibodies exert a huge influence in the biopharmaceutical industry, but cancer immunotherapy is emerging as a field of intensive research. According to Ecker et al. (5), in 2013, monoclonal antibody products amounted to nearly $75 billion, accounting for approximately half of the total sales volume of all biopharmaceutical products. In this regard, only 47 monoclonal antibody products were approved for use in the United States or Europe as of the end of 2014. Therefore, there is much room for growth, and it is likely that most large pharmaceutical companies have antibody development programs. In fact, approximately 70 new monoclonal antibody products are expected to enter the market by 2020, reaching nearly $125 billion in the global market (5). U.S. Patent No. 6,756,039 (Yeates, Padilla, and Colovos) discloses a fusion protein capable of self-assembling into a regular structure, wherein the fusion protein comprises at least two oligomerization domains that are rigidly linked together, for example, through an alpha helical linker. U.S. Patent No. 7,608,681 (Dennis, Lowman and DeLano) discloses a peptide ligand having an affinity for IgG or serum albumin. U.S. Patent No. 8,969,521 (Baker, King, Sheffler and, Yeates) discloses a general method for designing self-assembling protein nanomaterials and an isolated polypeptide comprising a specific 184-amino acid sequence capable of forming a multimeric assembly. U.S. Patent Application Publication No. 20070218547 (Yeates, Padilla, Yoshida, and Colovos) discloses a self-assembling protein for producing an extended substance, comprising a fusion protein comprising a first oligomerization domain that naturally associates to a homomeric structure and a second oligomerization domain that naturally associates to a homotemeric structure, wherein the first and second oligomerization domains are rigidly connected to each other. The present invention provides a protein cage polypeptide (or scaffolding protein) capable of forming or useful for forming a hollow tetrahedral pyramidal structure, wherein the protein cage polypeptide or scaffolding protein can specifically bind to an antibody or part thereof, or any chimeric protein, molecule, or compound comprising an antibody or part thereof. In some embodiments, the antibody is an IgG antibody. In some embodiments, part of the antibody is the Fc region of an antibody such as an IgG, IgA, IgD, IgE, or IgM antibody. In some embodiments, the antibody is a human, chicken, mouse, rabbit, sheep, or goat antibody. In some embodiments, the antibody is a humanized antibody. In some embodiments, the IgG antibody is a human IgG antibody. In some embodiments, the antibody is part of a chimeric protein, molecule, or compound containing the antibody or part thereof. In some embodiments, the chimeric protein, or other molecule, or compound contains the Fc region of the antibody. In some embodiments, the antibody, or part thereof, is covalently bound to the chimeric protein, molecule, or compound. In some embodiments, the binding affinity K a of the protein cage polypeptide, or scaffolding protein to the antibody or part thereof is 10⁷ M⁻¹ , 10⁸ M⁻¹ , or 10⁹ M⁻¹ or greater. In some embodiments, the protein cage polypeptide comprises a polypeptide of about 400 to about 700 amino acid residues. In some embodiments, the protein cage polypeptide comprises a polypeptide of about 450 to about 650 amino acid residues. In some embodiments, the protein cage polypeptide comprises an amino acid sequence having the following structure: polypeptide 1―AHL―polypeptide 2―INSERT A―polypeptide 3―INSERT B―polypeptide 4 (chemical structure I); Here, AHL is an "alpha helical linker," and INSERT A and/or INSERT B can each independently bind specifically to an antibody or a part thereof. In some embodiments, INSERT A has a length of about 17 to about 25 amino acids. In some embodiments, INSERT B has a length of about 28 to about 85 amino acids. In some embodiments, the binding affinity K a of INSERT A and/or INSERT to the antibody or part thereof is each independently 10⁷ M⁻¹ , 10⁸ M⁻¹ , or 10⁹ M⁻¹ or greater. In some embodiments, INSERT A and/or INSERT B each independently comprises the amino acid sequence DCAWHLGELVWCT (SEQN: 41) or GCDCAWHLGELVWCTCG (SEQN: 42). In some embodiments, the protein cage polypeptide comprises an amino acid