Nucleic acid-encapsulating polymer micelle complex and method for producing same

Patent metadata

Field	Value
Publication number	US-10046065-B2
Application number	US-201414909162-A
Country	US
Kind code	B2
Filing date	Aug 5, 2014
Priority date	Aug 6, 2013
Publication date	Aug 14, 2018
Grant date	Aug 14, 2018

Abstract

A nucleic acid-encapsulating polymer micelle complex is formed of a block copolymer containing an uncharged hydrophilic polymer chain block and a cationic polymer chain block; and two single-stranded DNA molecules having mutually complementary base sequences of 1000 or more bases in length, double-stranded DNA of 1000 or more base pairs in length in which at least a part of a double helix structure is dissociated and forms a single-stranded structure, or one single-stranded DNA molecule of 1000 or more bases in length.

First claim

The invention claimed is:

1. A method for producing a nucleic acid-encapsulating polymer micelle complex which accommodates DNA, the method comprising:
a process of mixing a block copolymer containing an uncharged hydrophilic polymer chain block and a cationic polymer chain block with double-stranded DNA of 1000 or more base pairs in a state in which at least a part of the double-stranded DNA is denatured in an aqueous medium;
wherein the double-stranded DNA has been denatured at 80° C. or higher prior to the process of mixing with the block copolymer, and
the complex is spherical and has diameters having a ratio of a longest diameter to another diameter of 1:1.

2. The method for producing a nucleic acid-encapsulating polymer micelle complex according to claim 1 , wherein the double-stranded DNA is 2000 or more base pairs in length.

3. The method for producing a nucleic acid-encapsulating polymer micelle complex according to claim 1 , wherein the double-stranded DNA is linear.

4. The method for producing a nucleic acid-encapsulating polymer micelle complex according to claim 1 ,
wherein the average particle diameter thereof in an aqueous medium measured according to a dynamic light scattering method is 100 nm or less.

5. The method for producing a nucleic acid-encapsulating polymer micelle complex according to claim 1 ,
wherein the DNA and the cationic polymer chain block bonded to the DNA due to an electrostatic interaction form a core portion, and
the uncharged hydrophilic polymer chain block forms a shell portion.

6. The method for producing a nucleic acid-encapsulating polymer micelle complex according to claim 5 ,
wherein the average particle diameter of the core portion is 50 nm or less.

7. The method for producing a nucleic acid-encapsulating polymer micelle complex according to claim 1 ,
wherein at least a part of the block copolymer is mutually cross-linked.

8. The method for producing a nucleic acid-encapsulating polymer micelle complex according to claim 1 ,
wherein a hydrophobic group is covalently bonded to a main chain or a side chain of the cationic polymer chain block.

9. The method for producing a nucleic acid-encapsulating polymer micelle complex according to claim 1 ,
wherein the cationic polymer chain block has an ethylamine structure or a propylamine structure in the side chain thereof.

Assignees

• Japan Science & Tech Agency

Inventors

• Kataoka Kazunori
• Osada Kensuke
• TOCKARY Theofilus Agrios

Classifications

• A61K48/0091

  Purification or manufacturing processes for gene therapy compositions · CPC title

• A61K48/00

  Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy · CPC title

• A61K31/711

  Natural deoxyribonucleic acids, i.e. containing only 2'-deoxyriboses attached to adenine, guanine, cytosine or thymine and having 3'-5' phosphodiester links · CPC title

• A61K47/34 Primary

  Macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyesters, polyamino acids, polysiloxanes, polyphosphazines, copolymers of polyalkylene glycol or poloxamers (A61K47/10 takes precedence) · CPC title

• A61K31/713

  Double-stranded nucleic acids or oligonucleotides · CPC title