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A kind of nanocomposite and its preparation method and application

A nano-composite and complex technology, applied in the fields of polymer chemistry and biomedical engineering, can solve the problem of low nucleic acid delivery efficiency, and achieve the effects of enhancing and nucleic acid delivery efficiency, reducing usage, and rationally designing preparation methods.

Active Publication Date: 2020-02-21
NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, a common problem in the cationic polymer carrier system is that once the amount of polymer and nucleic acid molecules is reduced, it will lead to the adverse result of severely low nucleic acid delivery efficiency

Method used

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  • A kind of nanocomposite and its preparation method and application
  • A kind of nanocomposite and its preparation method and application
  • A kind of nanocomposite and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0114] The preparation of embodiment 1 cationic polythiophene

[0115] 1. Cationic linear polythiophene

[0116] The structural formula of cationic linear polythiophene of the present invention is:

[0117]

[0118] The preparation method comprises the following steps:

[0119] (1) Synthesis of fat-soluble linear polythiophene (LPT or LPTM)

[0120] Under nitrogen protection, anhydrous ferric chloride (1.05g, 6.5mmol) was dissolved in anhydrous nitromethane (25mL), and under nitrogen protection, ethyl 3-thiopheneacetate (0.50g, 3.34mmol) or Diethyl 3-thiophene malonate (0.80 g, 3.31 mmol) was dissolved in anhydrous CHCl 3 (25mL); under ice bath, FeCl 3 The nitromethane solution was added dropwise to the reaction system, the system was heated to 35°C, and stirred in the dark for 2 hours. After the reaction was stopped, the system was washed once with 0.01M hydrochloric acid, and then washed with triethanolamine (5%, w / v) and Wash three times with a mixed solution of sod...

Embodiment 2

[0136] Example 2 Cationic linear / branched polythiophene multi-component nanocomposite

[0137] Composition of raw materials: the complex includes degradable cationic star-shaped polyasparagine carrier, DNA and cationic linear / branched polythiophene; among them, cationic linear / branched polythiophene and degradable cationic star-shaped polyasparagine carrier The ratio of the sum of the charges to the negative charge of DNA is 2.5:1, and the ratio of the positive charge of the cationic linear / branched polythiophene to the degradable cationic star polyasparagine carrier is 2.5% or 5%.

[0138] Preparation method: under the condition that the ratio of the number of amine groups (including 1-4 grade amines) contained in the polymer material to the number of phosphate bonds contained in the nucleic acid molecule is 5 (that is, N / P=5, since only half of the amine groups Protonation occurs under neutral conditions, at this time the positive and negative charge ratio is 2.5:1), first d...

Embodiment 3

[0161] Embodiment 3 Cationic polyphenylene acetylene and its preparation

[0162] The structural formula of cationic polyphenylene acetylene of the present invention is:

[0163]

[0164] Among them, the synthetic route of cPPE-DET is as follows:

[0165]

[0166] Concrete preparation method is as follows:

[0167] (1) Synthesis of ethyl 2,5-diiodo-1,4-diphenylacetate: Dissolve 2,5-diiodo-1,4-diphenylacetic acid (4.45g, 10mmol) in absolute ethanol (100mL ), was added dropwise thionyl chloride (1mL), stirred at room temperature for 24h, and the solvent was removed by rotary evaporation, and a light yellow solid (4.0g) was obtained after separation on a silica gel column;

[0168] (2) Synthesis of PPE-Et: Dissolve ethyl 2,5-diiodo-1,4-diphenylacetate (0.5g, 1mmol) in anhydrous tetrahydrofuran (15mL) and triethylamine (5mL), after degassing Adding catalytic amount of Pd(PPh 3 ) 4 and CuI, blow nitrogen for 10min, add trimethylsilaneacetylene (0.27mL, 2mmol), react at r...

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Abstract

The invention discloses a nano-composite, a preparation method of the nano-composite and application of the nano-composite. The nano-composite is prepared from cationic polymer carriers, nucleic acid molecules and conjugated polyelectrolyte. The preparation method comprises the following steps of mixing the cationic polymer carriers with the nucleic acid molecules, adding the conjugated polyelectrolyte, and standing for 10 to 60min so as to obtain a multi-component nano-composite solution. The nano-composite is applied to enhancing the transportation of nucleic acid. The nano-composite has the advantages that the surface of the nano-composite is functionally transformed by adopting a non-covalent assembly way, so that the nano-composite is not only strong in stability, but also is applied to the transportation of the nucleic acid, therefore the permeability of a lysosome membrane in an illumination or non-illumination condition can be enhanced, the lysosome escape of the nano-composite is realized, a cell can be prevented from being damaged obviously, the gene delivery efficiency and nucleic acid transportation efficiency are increased, and the requirements of safety and high efficiency on gene delivery are met; meanwhile, the preparation method is reasonably designed, the preparation process is simple, the usage amount of a carrier material is greatly reduced, and a wide application prospect is obtained.

Description

technical field [0001] The invention belongs to the field of polymer chemistry and biomedical engineering, and in particular relates to a nanocomposite and its preparation method and application. Background technique [0002] Cationic polymer carriers are an important class of non-viral carriers. These materials can combine with negatively charged nucleic acid molecules through electrostatic interactions to ensure the integrity of nucleic acid molecules during delivery. Compared with viral vectors, one of the main challenges in the development of non-viral vectors is how to improve transfection efficiency. [0003] Lysosomal escape is a key point for the enhanced efficiency of cationic carrier-based nucleic acid delivery. In general, if endocytosed nanoparticles are transported into lysosomes, they will be at risk of being degraded by lysosomal hydrolases. Therefore, if polyplexes are internalized and transported into lysosomes, it is necessary to try to break through the ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08G61/12C08G61/02C08G61/10C08L77/02C08L77/00C08L5/00C08L65/00C12N15/87
CPCC08G61/02C08G61/10C08G61/126C08G2261/312C08G2261/3142C08G2261/3223C08L5/00C08L77/00C08L77/02C12N15/87C08L65/00
Inventor 冯福德张雅洁李志良
Owner NANJING UNIV
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