Method of promoting nucleic acid transfer

• US 20040266004A1
• Filed: 12/19/2003
• Published: 12/30/2004
• Est. Priority Date: 06/20/2001
• Status: Active Grant

• Current Assignee: National Cancer Center, Sumitomo Pharma Co Ltd (Sumitomo Chemical Company Limited)
• Original Assignee: Sumitomo Dainippon Pharma Company Limited (Sumitomo Chemical Company Limited)
• Inventors: Sano, Akihiko, Terada, Masaaki, Ochiya, Takahiro, Nagahara, Shunji, Aso, Yu, Honma, Kimi
• Granted Patent: US 8,742,091 B2
• Time in Patent Office: Days
• US Class Current: 435/455
• CPC Class Codes: A61K 47/6435 the peptide or protein in the drug conjugate being a connective tissue peptide, e.g. collagen, fibronectin or gelatin
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61K 48/0008 characterised by an aspect of the 'non-active' part of the composition delivered, e.g. wherein such 'non-active' part is not delivered simultaneously with the 'active' part of the composition
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C12N 15/87 Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation

Abstract

Means for transferring efficiently a desired nucleic acid into a cell is provided.

The present invention comprises using a complex comprising a collagen or a collagen derivative, and a desired nucleic acid.

Claims

1. A preparation for facilitating the transfer of a nucleic acid into a target cell, which comprises a collagen or a collagen derivative.

2. A preparation according to claim 1 for facilitating the transfer of a nucleic acid into a target cell, which comprises a collagen or a collagen derivative that is made complex with a desired nucleic acid.

3. A preparation according to claim 2 for facilitating the transfer of a nucleic acid, wherein the complex is in a form of particle.

4. A preparation according to claim 2 for facilitating the transfer of a nucleic acid, wherein the major axis of the particle is 300 nm to 300 μ

m.

5. A preparation according to any one of claims 2 to 4 for facilitating the transfer of a nucleic acid, wherein the desired nucleic acid is a plasmid DNA.

6. A preparation according to claim 5 for facilitating the transfer of a nucleic acid, wherein the ratio of the number of a collagen molecule or a collagen derivative molecule to the number of a nucleotide monomer of the plasmid DNA in the complex is 1:

20 to 1;

the number of a nucleotide monomer of the plasmid DNA.

7. A preparation according to any one of claims 2 to 4 for facilitating the transfer of a nucleic acid, wherein the desired nucleic acid is an oligonucleotide.

8. A preparation according to claim 7 for facilitating the transfer of a nucleic acid, wherein the ratio of the number of a collagen molecule or a collagen derivative molecule to the number of a nucleotide monomer of the oligonucleotide in the complex is 1:

1 to 1;

200.

9. A particle of the complex comprising a collagen or a collagen derivative and a desired nucleic acid.

10. A particle of the complex according to claim 9, wherein the major axis is 300 nm to 300 μ

m.

11. A particle of the complex according to claim 9 or 10, wherein the desired nucleic acid is an plasmid DNA.

12. A particle of the complex according to claim 11, wherein the ratio of the number of a collagen molecule or a collagen derivative molecule to the number of a nucleotide monomer of the plasmid DNA in the complex is 1:

20 to 1;

the number of a nucleotide monomer of the plasmid DNA.

13. A particle of the complex according to claim 9 or 10, wherein the desired nucleic acid is an oligonucleotide.

14. A particle of the complex according to claim 13, wherein the ratio of the number of a collagen molecule or a collagen derivative molecule to the number of a nucleotide monomer of the oligonucleotide in the complex is 1:

1 to 1;

200.

15. A process for preparing a particle of the complex according to claim 9, which comprises mixing a collagen or a collagen derivative and a desired nucleic acid in a solution comprising an agent that inhibits the formation of collagen association body.

16. A medical instrument, of which the surface is coated with a particle of the complex according to claim 9.

17. A cell culture instrument, of which the surface is coated with a particle of the complex according to claim 9.

18. A process for transferring a desired nucleic acid into a target cell, which comprises using a particle of the complex according to claim 9.

19. A process for improving the expression level of a desired nucleic acid in a target cell, which comprises using a particle of the complex according to claim 9.

20. A process for examining the function of a gene or a protein in a target cell, which comprises coating a solid surface with a particle of the complex according to claim 9 that comprises the gene, a gene encoding the protein, or a nucleic acid inhibiting the expression of the gene or the protein in a cell;

culturing the target cell on the solid surface; and

examining the expression level of the nucleic acid or the expression level of the gene or the protein in the target cell, or the proliferation ratio or the phenotype of the cell.

21. A process for screening for a nucleic acid that treats a disease, which comprises coating a solid surface with a particle of the complex according to claim 9 that comprises a nucleic acid candidate that inhibits the expression of a gene associated with the disease in a cell;

culturing the cell presenting the condition of the disease on the solid surface; and

examining the expression level of the gene to be inhibited with each of the nucleic acid candidate, or the proliferation ratio or the phenotype of the cell.