Self assembling protein nanoparticles as carrier molecules

Patent metadata

Field	Value
Publication number	US-11911482-B2
Application number	US-202017002571-A
Country	US
Kind code	B2
Filing date	Aug 25, 2020
Priority date	Aug 25, 2020
Publication date	Feb 27, 2024
Grant date	Feb 27, 2024

Abstract

Disclosed herein are self-assembling protein nanoparticles comprising passenger molecules of interest.

First claim

What is claimed is:

1. A polypeptide comprising (a) a scaffolding protein consisting of an amino acid sequence having 100% sequence identity to SEQ ID NO: 64, (b) a passenger peptide covalently linked thereto, wherein (i) the passenger peptide is a peptide tag, a covalent peptide tag, a protein tag, an enzyme, or an enzyme substrate, and the passenger peptide neither comprises nor consists of SEQ ID NO: 41 or SEQ ID NO: 42, and/or (ii) the passenger peptide is covalently linked to an amino acid selected from the group consisting of: residues 19-26, 131-141, 245-257, 300-304, and 350-357 of SEQ ID NO: 64 or replaces one or more amino acids selected from the group consisting of: residues 19-26, 131-141, 245-257, 300-304, and 350-357 of SEQ ID NO: 64; and (c) optionally, a peptide having a sequence selected from the group consisting of SEQ ID NO: 45, SEQ ID NO: 52, and SEQ ID NO: 56 covalently attached to a terminal end of the scaffolding protein.

2. The polypeptide according to claim 1 , wherein the polypeptide comprises the peptide having SEQ ID NO: 45 covalently attached to the terminal end of the scaffolding protein.

3. The polypeptide according to claim 1 , wherein the polypeptide does not contain the peptide having SEQ ID NO: 45 covalently attached to the terminal end of the scaffolding protein.

4. The polypeptide according to claim 1 , wherein the polypeptide comprises the peptide having SEQ ID NO: 52 covalently attached to the terminal end of the scaffolding protein.

5. The polypeptide according to claim 1 , wherein the polypeptide comprises the peptide having SEQ ID NO: 56 covalently attached to the terminal end of the scaffolding protein.

6. The polypeptide according to claim 1 , wherein the passenger peptide is covalently linked to the amino acid selected from the group consisting of: residues 19-26, 131-141, 245-257, 300-304, and 350-357 of SEQ ID NO: 64.

7. The polypeptide according to claim 1 , wherein the passenger peptide replaces one or more amino acids selected from the group consisting of: residues 19-26, 131-141, 245-257, 300-304, and 350-357 of SEQ ID NO: 64.

8. The polypeptide according to claim 1 , wherein the passenger peptide is covalently linked to residue 19, 20, 21, 22, 23, 24, 25, 26, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 245, 246, 247, 248, 249, 253, 254, 255, 256, 257, 300, 301, 302, 303, 304, 350, 351, 352, 353, 354, 355, 356, or 357 of SEQ ID NO: 64 or replaces one or more amino acids selected from the group consisting of: residues 19, 20, 21, 22, 23, 24, 25, 26, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 245, 246, 247, 248, 249, 253, 254, 255, 256, 257, 300, 301, 302, 303, 304, 350, 351, 352, 353, 354, 355, 356, and 357 of SEQ ID NO: 64.

9. The polypeptide according to claim 1 , wherein the passenger peptide is covalently linked to residue residues 22, 23, 24, 25, 139, 238, 244, 245, 246, 301, 302, 303, 352, 353, 354, 355, or 356 or replaces one or more amino acids selected from the group consisting of: residues 22, 23, 24, 25, 139, 238, 244, 245, 246, 301, 302, 303, 352, 353, 354, 355, and 356 of SEQ ID NO: 64.

10. The polypeptide according to claim 1 , wherein the polypeptide comprises the passenger peptide which is a peptide tag, a covalent peptide tag, a protein tag, an enzyme, or an enzyme substrate.

11. The polypeptide according to claim 1 , wherein the passenger peptide binds another molecule or is cleaved by an enzyme.

12. The polypeptide according to claim 1 , wherein the passenger peptide is an enzyme that is a narrow-specificity protease, whose recognition sequence depends on two or more amino acid positions.

13. The polypeptide according to claim 1 , wherein the passenger peptide is an enzyme that is a protease such as thrombin or TEV protease.

14. The polypeptide according to claim 1 , wherein the passenger peptide comprises or consists of LVPRGSG (SEQ ID NO: 53), GSENLYFQGGS (SEQ ID NO: 54), LPXTG (SEQ ID NO: 65), AHIVMVDAYKPTK (SEQ ID NO: 66), ATHIKFSKRD (SEQ ID NO: 67), CCPGCC (SEQ ID NO: 68), DIPATYEFTDGKHYITNEPIPPK (SEQ ID NO: 69), DLYDDDDK (SEQ ID NO: 70), DPIVMIDNDKPIT (SEQ ID NO: 71), DYKDDDDK (SEQ ID NO: 72), EEEEEE (SEQ ID NO: 73), EQKLISEEDL (SEQ ID NO: 74), EVHTNQDPLD (SEQ ID NO: 75), GAPVPYPDPLEPR (SEQ ID NO: 76), GKPIPNPLLGLDST (SEQ ID NO: 77), GLNDIFEAQKIEWHE (SEQ ID NO: 78), KETAAAKFERQHMDS (SEQ ID NO: 79), KLGDIEFIKVNK (SEQ ID NO: 80), KLGSIEFIKVNK (SEQ ID NO: 81), KRRWKKNFIAVSAANRFKKISSSGAL (SEQ ID NO: 82), MASMTGGQQMG (SEQ ID NO: 83), MDEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP (SEQ ID NO: 84), PDRVRAVSHWSS (SEQ ID NO: 85), SLAELLNAGLGGS (SEQ ID NO: 86), SRLEEELRRRLTE (SEQ ID NO: 87), TDKDMTITFTNKKDAE (SEQ ID NO: 88), TETSQVAPA (SEQ ID NO: 89), TKENPRSNQEESYDDNES (SEQ ID NO: 90), TQDPSRVG (SEQ ID NO: 91), VPTIVMVDAYKRYK (SEQ ID NO: 92), WSHPQFEK (SEQ ID NO: 93), YPYDVPDYA (SEQ ID NO: 94), or YTDIEMNRLGK (SEQ ID NO: 95).

15. The polypeptide according to claim 1 , wherein the passenger peptide neither comprises nor consists of SEQ ID NO: 41 or SEQ ID NO: 42.

Assignees

• Univ California

Inventors

• Yeates Todd
• Lai Yen-Ting
• Miller Justin
• Nguyen Phillip
• Srinivasan Yashes
• Dharmaraj Nithin
• Taylor Scott
• Liu Andrew

Classifications

• A61K38/00

  Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00) · CPC title

• C12Y304/22044

  Nuclear-inclusion-a endopeptidase (3.4.22.44) · CPC title

• C07K16/44

  against material not provided for elsewhere {, e.g. haptens, metals, DNA, RNA, amino acids} · CPC title

• C07K14/47

  from mammals · CPC title

• C12N9/50

  Proteinases {, e.g. Endopeptidases (3.4.21-3.4.25)} · CPC title