Nanoparticles comprising fusion protein of single-chain variable fragment and ferritin, and use thereof

Patent metadata

Field	Value
Publication number	US-2025075003-A1
Application number	US-202218559548-A
Country	US
Kind code	A1
Filing date	May 9, 2022
Priority date	May 7, 2021
Publication date	Mar 6, 2025
Grant date	—

Abstract

The present invention provides: ferritin; a single-chain variable fragment bound to the N-terminus of the ferritin; and a fusion protein self-assembling nanoparticle structure in which an N-terminus RNA-interaction domain (RID) in a human-derived lysyl-tRNA synthetase is bound to the N-terminus of the single-chain variable fragment by means of a novel fusion partner. Therefore, the present invention provides: a fusion protein having enhanced water solubility; nanoparticles; a vector coding same; a host cell transformed by means of the vector, and a pharmaceutical composition, for treating a disease, using the same.

First claim

1 . A fusion protein, comprising:
a ferritin protein; a single-chain variable fragment which is bound to an N-terminus of the ferritin protein; and an N-terminus domain (hLysRS N-terminal appended RNA interacting domain; hRID) which is isolated from a human-derived lysyl tRNA synthetase that is bound to an N-terminus of the single-chain variable fragment.

2 . The fusion protein of claim 1 , wherein the single-chain variable fragment is derived from any one antibody selected from the group consisting of Trastuzumab, Bevacizumab and Pertuzumab.

3 . The fusion protein of claim 1 , wherein the hRID is represented by the amino acid sequence of SEQ ID NO: 1.

4 . The fusion protein of claim 1 , wherein the ferritin protein is represented by the amino acid sequence of SEQ ID NO: 4.

5 . The fusion protein of claim 1 , wherein the fusion protein further comprises a linker protein between the single-chain variable fragment (scFv) and the ferritin protein.

6 . The fusion protein of claim 5 , wherein the linker protein has an amino acid sequence represented by any one of the amino acid sequences of SEQ ID NOs: 8 to 10.

7 . Nanoparticles, which are formed by self-assembly of 2 to 24 pieces of the fusion protein of claim 1 as a monomer.

8 . A polynucleotide, which encodes the fusion protein of claim 1 .

9 . A recombination expression vector, comprising the polynucleotide of claim 8 .

10 . A host cell, which is transformed by the expression vector according to claim 9 .

11 . The host cell of claim 10 , wherein the host cell is selected from the group consisting of Escherichia bacteria, Bacillus bacteria, Pseudomonas bacteria, lactic acid bacteria, yeast, animal cells and insect cells.

12 . A method for preparing nanoparticles, comprising the steps of:
(a) preparing an expression vector comprising a polynucleotide encoding a fusion protein, which consists of a ferritin protein; a single-chain variable fragment which is bound to an N-terminus of the ferritin protein; and an N-terminus domain (hLysRS N-terminal appended RNA interacting domain; hRID) which is isolated from a human-derived lysyl tRNA synthetase that is bound to an N-terminus of the single-chain variable fragment; (b) preparing a transformant by introducing the expression vector into a host cell; (c) culturing the transformant to induce expression of the fusion protein; and (d) purifying nanoparticles formed by self-assembly of the expressed fusion protein.

13 . The method of claim 12 , wherein the nanoparticles are formed by self-assembly of 2 to 24 pieces of the fusion protein monomer.

14 . The method of claim 12 , wherein the single-chain variable fragment is derived from any one antibody selected from the group consisting of Trastuzumab, Bevacizumab and Pertuzumab.

15 . A pharmaceutical composition for ameliorating or treating a disease, comprising the fusion protein of claim 1 .

16 . Use of nanoparticles in the treatment of a target disease, wherein the nanoparticles are formed by self-assembly of 2 to 24 pieces of a fusion protein, which comprises a ferritin protein; a single-chain variable fragment which is bound to an N-terminus of the ferritin protein; and an N-terminus domain (hLysRS N-terminal appended RNA interacting domain; hRID) which is isolated from a human-derived lysyl tRNA synthetase that is bound to an N-terminus of the single-chain variable fragment, as a monomer.

Assignees

• Uif Univ Industry Foundation Yonsei Univ
• Research & Business Found Sungkyunkwan Univ

Inventors

• Seong Baik Lin
• Chung Sangjeon
• Kim Min Jin
• Kang Hyo Jin
• Kim Ga Hyeon
• Kim Young Seok
• Lee Je Ho
• Sohn Myunghyun
• Kim Tae Hoon

Classifications

• A61K2039/505

  comprising antibodies · CPC title

• C07K2317/76

  Antagonist effect on antigen, e.g. neutralization or inhibition of binding · CPC title

• C07K2317/24

  containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered · CPC title

• C07K2317/622

  Single chain antibody (scFv) · CPC title

• C07K2317/92

  Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value · CPC title