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Preparation method of nano-particles

A nanoparticle and solution technology, applied in the fields of block polymer and its synthesis, and nanoparticle preparation, can solve the problems of affecting the adhesion of polyplex particles to diseased cells and poor targeting effect, so as to improve the circulation efficiency in the body and improve the targeting effect. effect of effect

Inactive Publication Date: 2014-03-26
SHANGHAI JIAO TONG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The first purpose of the present invention is to provide a block polymer to solve the problem that in the prior art, there are too many positive charges on the outer surface of the surface membrane of Polyplex particles, which affects the attachment of Polyplex particles to diseased cells, and the targeting effect is poor technical issues

Method used

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  • Preparation method of nano-particles

Examples

Experimental program
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Embodiment 1

[0040] Embodiment 1 The synthetic method of block polymer PEG-PCL-maltotriose-COOH

[0041] The synthetic route of block polymer PEG-PCL-maltotriose-COOH is as follows figure 1 shown. The whole reaction is carried out in an anhydrous and oxygen-free environment. Take a certain amount of PEG, polycaprolactone, and stannous octoate into a three-necked bottle, add fresh anhydrous toluene, and stir and react at 120°C for 24 hours. After completion, add diethyl ether to precipitate, then add dichloromethane to dissolve, and then diethyl ether to precipitate, repeat three times to obtain PEG-PCL block polymer.

[0042] Then PEG-PCL was dissolved in methylene chloride, excessive oxalyl chloride was added in the reaction flask, and the PEG-PCL solution was added dropwise in the reaction flask under the condition of anhydrous and oxygen-free ice bath, and returned to Stir at room temperature, remove the remaining oxalyl chloride under reduced pressure after 12 hours, then add dichlor...

Embodiment 2

[0045] Example 2 Preparation of Nanoparticles

[0046] Take a certain amount of polycationic polymer (take PEI as an example) and plasmid DNA, because of the investigation of nanoparticle structure and charge shielding, so the mass ratio of 1:5 (pDNA:PEI) is used to prepare polyplexes samples, and then add block height Molecules, fully compounded. See specific steps Figure 4 , 5 .

Embodiment 3

[0047] Example 3 Characterization of Nanoparticles

[0048] Nanoparticles prepared according to the above preparation method were characterized by fluorescence colocalization. The specific method was as follows: PEI and FITC were covalently bonded, and the hydrophobic PCL block of the block polymer was labeled with nile red at the same time. The prepared fluorescent particles were immobilized in PVA hydrogel and cross-linked and solidified through repeated "freezing-room temperature" cycles to limit the two-dimensional movement of the particles on the horizontal plane. Observation see Figure 6 , red (nile red) and green (FITC) appear and coincide at the same position, verifying the formation of nanoparticles.

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Abstract

The invention provides a preparation method of nano-particles. The preparation method comprises steps as follows: 1), a polycation polymer is dissolved in a polycation solution made from ultrapure water or RNase-free water, and a nucleic acid drug is dissolved in a nucleic acid solution made from ultrapure water or RNase-free water; 2), the polycation solution is added into the nucleic acid solution, the mixture is repeatedly blown and beaten to be uniform and cultivated at the room temperature, and polyplexes is obtained; 3), a block polymer is dissolved in a block polymer solution made from ultrapure water or RNase-free water, added into polyplexes particles prepared in the step 2) slowly, blown and beaten to be uniform and subjected to standing, so that the particles are fully wrapped, and the nano-particles wrapped with the block polymer can be prepared. Compared with the prior art, the prepared nano-particles can remove obstacles blocking circulation of cation particles in a body and improve the circulation efficiency of the particles in the body; and meanwhile, a targeted group can be grafted, so that diseased cells in the body can be targeted, and the targeting effect is effectively improved.

Description

[0001] This application is a divisional application of the Chinese invention patent application with the application number 201210163285.1, the title of the invention is "a block polymer and its synthesis method, and the preparation method of nanoparticles", and the application date is May 23, 2012. technical field [0002] The invention relates to the field of biotechnology, in particular to a block macromolecule, a synthesis method thereof, and a preparation method of nanoparticles. Background technique [0003] siRNA is a 21-25 base pair RNA molecule found in the mechanism of single-celled organisms to resist viral invasion. Single-cell organisms synthesize a complementary siRNA to the mRNA sequence of the invading virus, which actively binds to the mRNA, thereby blocking the replication of the virus. If this one-to-one interference strategy corresponding to pathogen genes is used to develop drugs for the treatment of human diseases, it will fundamentally change the curre...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K47/34A61K9/14
Inventor 袁伟恩金拓吴飞葛雪梅
Owner SHANGHAI JIAO TONG UNIV
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