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

A manufacturing method and polymer technology, which can be applied to the manufacture of gene therapy compositions, other methods of inserting foreign genetic materials, and the use of microcapsules, etc., can solve problems such as insufficient introduction efficiency, and achieve excellent intake efficiency and blood. Excellent retention effect

Active Publication Date: 2016-03-30
THE JAPAN SCI & TECH AGENCY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The biggest problem in gene therapy is that the efficiency of introducing genes into target cells or tissues is not sufficient

Method used

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

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Embodiment

[0111] Hereinafter, although an Example etc. demonstrate this invention in more detail, this invention is not limited to these examples. In addition, all the following animal experiments were performed in accordance with the guidelines related to the management and use of experimental animals of the University of Tokyo, a national university corporation.

reference example 1

[0113] Using a block copolymer consisting of a PEG block and a PLys block, investigation of the degree of polymerization of the PLys block and the polymer micelle complex in a polymer micelle complex containing double-stranded DNA (pDNA) with a double helix structure shape relationship.

[0114] (1) PEG-PLys

[0115] α-Methoxy-ω-amino PEG (PEG, Mw=12kDa, M w / M n =1.05) as an initiator, and make N ε -N-formic anhydride (NCA) of trifluoroacetyl-L-lysine was subjected to ring-opening polymerization to produce PEG block-poly(ε-trifluoroacetyl-L-lysine) block (PEG -PLys(TFA)). At this time, three kinds of PEG-PLys(TFA) having different degrees of polymerization were produced by adjusting the ratio of the initiator to the NCA monomer. The trifluoroacetyl group (TFA group) of the three kinds of PEG-PLys (TFA) thus obtained was deprotected with sodium hydroxide to obtain three kinds of PEG-PLys having different degrees of polymerization ("n1" in the following chemical formula). ...

Embodiment 1

[0145] Composite of nucleic acid-encapsulating polymer microcapsules produced by the conventional method of directly encapsulating pDNA and nucleic acid-encapsulating polymer microcapsules produced by combining the dissociated double helix structure of pDNA with a block copolymer body, the shape, size, and density of block copolymers were compared.

[0146] (1) PEG-PAsp(DET)-Chole

[0147] Use α-methoxy-ω-amino PEG (PEG, Mw=12kDa, M w / M n =1.05) as an initiator, and the NCA of β-benzyl-L-aspartic acid ester (BLA) was subjected to ring-opening polymerization, thereby producing PEG block-poly(β-benzyl-L-aspartic acid Ester) block (PEG-PBLA). At this time, three kinds of PEG-PBLAs having different degrees of polymerization were produced by adjusting the ratio of the initiator to the NCA monomer.

[0148] In the presence of 10 times equivalents of dicyclohexylcarbodiimide and 2 times equivalents of 4-dimethylaminopyridine, the amino group at the end of the obtained PEG-PBLA a...

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Abstract

This nucleic acid-encapsulating polymer micelle complex is characterized in being formed of: a block copolymer containing an uncharged hydrophilic polymer chain block and a cationic polymer chain block; and two single-stranded DNA molecules comprising 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 the double helix structure has dissociated and taken on a single-stranded structure, or one single-stranded DNA molecule of 1000 or more bases in length.

Description

technical field [0001] The invention relates to a polymer microcapsule complex containing nucleic acid (DNA). More specifically, it involves a sufficiently small polymer microcapsule complex enclosing a relatively long DNA. [0002] This application claims priority based on Japanese Patent Application No. 2013-163106 for which it applied in Japan on August 6, 2013, and uses the content here. Background technique [0003] Gene therapy, which treats diseases by controlling gene expression, is expected to be a next-generation therapy. The biggest problem in gene therapy is that the efficiency of gene introduction into target cells or tissues is not sufficient. In particular, in order to realize gene therapy by systemic administration, it is necessary to allow the gene to circulate stably in the blood to accumulate in the target tissue, and to efficiently express the gene after invading the target cell. Therefore, in order to solve these problems, development of a gene carrie...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/107A61K31/711A61K31/713A61K47/34A61K48/00C12N15/115
CPCA61K47/34A61K48/00A61K31/711A61K31/713A61K9/0019A61K9/1075C12N15/88A61K48/0041A61K48/0091
Inventor 片冈一则长田健介赛奥弗里斯·阿格里斯·由佳利
Owner THE JAPAN SCI & TECH AGENCY
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