Recombinant self-assembling protein comprising target-oriented peptide and use thereof

Patent metadata

Field	Value
Publication number	US-9814907-B2
Application number	US-201514849379-A
Country	US
Kind code	B2
Filing date	Sep 9, 2015
Priority date	May 10, 2013
Publication date	Nov 14, 2017
Grant date	Nov 14, 2017

Abstract

The present invention relates to a recombinant self-assembled protein comprising a target-oriented peptide and a use thereof. The recombinant self-assembled protein according to the present invention, comprising a target-oriented peptide, does not require an additional process for providing target-orientedness, and is thus capable of delivering a desired drug to a target tissue or target cell without using additives, such as chemical binders or stabilizers; therefore, the protein can be used for photothermal therapy, drug delivery, imaging, or the like. In particular, according to the present invention, it is possible to prepare gold-protein nanoparticle fusions in which uniform high-density gold nanoparticles having target-orientedness are bound to protein surfaces, without an additional process of surface stabilization or process for providing target-orientedness. Compared with conventional gold nanoparticles, the gold-protein nanoparticle fusions according to the present invention show structural stability against pH variation and concentration variation, and also have excellent target-orientedness; therefore, the fusions can bring a dramatic enhancement to the utilization of gold nanoparticles in photothermal therapy.

First claim

The invention claimed is:

1. A recombinant self-assembled protein, comprising a target-oriented peptide fused to an HBV capsid protein as a self-assembled protein and a gold ion reducing peptide self-assembled.

2. The recombinant self-assembled protein of claim 1 , wherein the target-oriented peptide is introduced into a spike region of the recombinant HBV capsid protein.

3. The recombinant self-assembled protein of claim 1 , wherein the target-oriented peptide targets EGFR (epidermal growth factor receptor) or EDB (human fibronectin extra domain B).

4. The recombinant self-assembled protein of claim 1 , wherein the target-oriented peptide is located between two sequences of amino acids 1-78 and 81-149 on the recombinant HBV capsid protein.

5. The recombinant self-assembled protein of claim 1 , wherein the target-oriented peptide is inserted in two or more copies into the recombinant HBV capsid protein.

6. The recombinant self-assembled protein of claim 1 , wherein the gold ion reducing peptide comprising at least one selected from the group consisting of two or more tyrosine residues, two or more histidine residues, and two or more cysteine residues.

7. The recombinant self-assembled protein of claim 1 , wherein the gold ion reducing peptide is introduced into an N- or C-terminus of the recombinant self-assembled protein.

8. The recombinant self-assembled protein of claim 1 , further comprising a gold nanoparticle size-controlling peptide in a vicinity of the gold ion reducing peptide.

9. The recombinant self-assembled protein of claim 8 , wherein the gold nanoparticle size-controlling peptide is a biotinylated peptide.

10. The recombinant self-assembled protein of claim 8 , further comprising a linker peptide between the gold ion reducing peptide and the gold nanoparticle size-controlling peptide.

11. A recombinant self-assembled protein nanoparticle, comprising copies of the recombinant self-assembled protein of claim 1 .

12. A method for preparing a gold-protein particle fusion, comprising reacting the recombinant self-assembled protein nanoparticle of claim 11 with a gold precursor to form a gold nanoparticle on the recombinant self-assembled protein nanoparticle.

13. The method of claim 12 , wherein the gold precursor is chloro(trimethylphosphine)gold (AuClP(CH 3 ) 3 ), potassium tetrachloroaurate (III) (KAuCl 4 ), sodium chloroaurate (NaAuCl 4 ), chloroauric acid (HAuCl 4 ), sodium bromoaurate (NaAuBr 4 ), gold chloride (AuCl), gold chloride (III) (AuCl 3 ), or gold bromide (AuBr 3 ).

14. A gold-protein particle fusion in which a gold nanoparticle is formed on the recombinant capsid nanoparticle of claim 11 .

15. The gold-protein particle fusion of claim 14 which is used in preparing a medicine for photothermal therapy.

16. The gold-protein particle fusion of claim 15 , wherein the photothermal therapy is to treat cancer.

17. A method for preparing the recombinant self-assembled protein of claim 1 , comprising:
a) cloning a gene coding for a self-assembled protein;
b) cloning a gene including a nucleotide sequence coding for a target-oriented peptide fused to an HBV capsid protein for insertion into the self-assembled protein;
c) constructing an expression vector containing the clones by ligation; and
d) transforming the expression vector into a host to express the recombinant self-assembled protein.

Assignees

• Univ Korea Res & Bus Found

Inventors

• Lee Jeewon
• Kwon Koo-Chul

Classifications

• C12N15/09

  Recombinant DNA-technology · CPC title

• C12N2730/10133

  Use of viral protein as therapeutic agent other than vaccine, e.g. apoptosis inducing or anti-inflammatory · CPC title

• C07K2319/21

  containing a His-tag · CPC title

• A61N5/062 Primary

  Photodynamic therapy, i.e. excitation of an agent · CPC title

• B82Y5/00

  Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery · CPC title