Engineered hepatitis B core polypeptide

Patent metadata

Field	Value
Publication number	US-11261216-B2
Application number	US-201916725626-A
Country	US
Kind code	B2
Filing date	Dec 23, 2019
Priority date	Dec 28, 2018
Publication date	Mar 1, 2022
Grant date	Mar 1, 2022

Abstract

Genetically modified HBc polypeptides are provided.

First claim

What is claimed is:

1. A hepatitis B core protein (HBc) polypeptide, comprising a hydrophobic cargo-loading domain at the carboxy terminus, wherein the cargo-loading domain is selected from EGFGEGFGEGF (SEQ ID NO:4); EGFGEGFGEGFC (SEQ ID NO:5); IGIGC (SEQ ID NO:6); and IGIGIC (SEQ ID NO:7).

2. The HBc polypeptide of claim 1 , wherein the cargo-loading domain comprises one cysteine residue.

3. The HBc polypeptide of claim 1 , further comprising a C-terminal poly-histidine tag before or after the cargo-loading domain.

4. The HBc polypeptide of claim 1 , wherein the polypeptide further comprises at least one unnatural amino acid.

5. The HBc polypeptide of claim 4 , wherein the unnatural amino acid provides a reactant group for Click chemistry reactions.

6. The HBc polypeptide of claim 5 , wherein the unnatural amino acid is azidohomoalanine, homopropargylglycine, p-acetyl-L-phenylalanine or p-azido-L-phenylalanine.

7. The HBc polypeptide of claim 6 , further comprising one or more additional moieties conjugated to the unnatural amino acid.

8. A VLP comprised of HBc polypeptides according to claim 1 .

9. The VLP of claim 8 , wherein the VLP comprises encapsulated cargo.

10. The VLP of claim 9 , wherein the cargo is ssRNA, dsRNA, ssDNA, dsDNA, protein, or small molecule drug by loading mediated by hydrophobic interaction directly to the cargo or to a hydrophobic moiety attached to the cargo.

11. The VLP of claim 9 , wherein the cargo is a chemotherapeutic drug.

12. The VLP of claim 9 , wherein the cargo is modified to enhance retention or uptake.

13. The VLP of claim 12 , wherein the cargo is modified by conjugation to cholesterol.

14. The VLP of claim 12 , wherein the cargo is modified to add a single sulfhydryl group.

15. The HBc polypeptide of claim 1 , wherein the cargo-loading domain comprises from one to three hydrophobic amino acids.

16. A method of assembling a cargo loaded VLP, the method comprising adding cargo to a solution comprising a polypeptide according to claim 1 , and increasing ionic strength of the solution with a solution comprising at least 10% of an organic solvent.

17. The method of claim 16 , wherein the organic solvent is DMSO.

Assignees

• Univ Leland Stanford Junior

Inventors

• Swartz James R
• Mathew Rinchu
• Rohovie Marcus John
• Nagasawa Maya

Classifications

• C07K14/02 Primary

  Hepadnaviridae, e.g. hepatitis B virus · CPC title

• C12N2730/10122

  New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes · CPC title

• C12N2730/10123

  Virus like particles [VLP] · CPC title

• C12N2730/10142

  virus or viral particle as vehicle, e.g. encapsulating small organic molecule · CPC title

• C12N7/00

  Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00) · CPC title