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Polyethylene imine nanometer gel and its uses

A technology of polyethyleneimine and nanogel, which is applied in the direction of introducing foreign genetic material, gene therapy, and genetic material components by using carriers. Inhomogeneity and other problems, to achieve the effect of low cytotoxicity and immunogenicity, narrow particle size distribution, and fast reaction time

Inactive Publication Date: 2007-05-23
SHANGHAI INST OF APPLIED PHYSICS - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the molecular weight of PEI currently used as a gene transfection carrier is mostly 0.6-25kDa, and its structure [CH 2 -CH 2 -NH n ] does not contain double bonds or unstable rings, it is difficult to initiate its continuous polymerization with conventional chemical methods, and because the existing PEI synthesis technology is difficult to control the reaction to end on demand, the molecular weight and particle size of the obtained PEI are difficult to control. Wide distribution range, uneven particle size, and poor dispersion, such as the PCS (photon correlation spectroscopy, Chinese translation called photon correlation spectroscopy) of PEI prepolymer produced by Tokyo Chemical Industry Co., Ltd., which is currently used as a gene transfection carrier The results suggest a multimodal distribution with a wide range of particle size distributions (as shown in Figure 1)

Method used

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  • Polyethylene imine nanometer gel and its uses
  • Polyethylene imine nanometer gel and its uses
  • Polyethylene imine nanometer gel and its uses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] In the 50ml quartz flask, add the following various components to form the reaction system, the concentration of each component is in order: the concentration of PEI prepolymer is 1.15% (w / w), the concentration of iron carboxylate as nucleating agent is 1.54mg / ml, add a very small amount of dilute hydrochloric acid solution (0.5M) to adjust the pH value to 9.5. Control the stirring speed to be about 140 rev / min, pass nitrogen gas to remove oxygen for 30 minutes, then add 3% hydrogen peroxide (hydrogen peroxide) so that the concentration in the reaction solution is 0.057% (v / v). In the case of continuous nitrogen flow (nitrogen flow rate: 80ml / min), use an ultraviolet low-pressure mercury lamp with a light intensity of 18mw / cm 2 Illumination for 1h (hour) initiates the reaction. Control the reaction conditions as mentioned above to carry out three repeated experiments to obtain No. 1, No. 2, and No. 3 samples, and use a 0.45 μm needle filter (ANPEL product) and a dialys...

Embodiment 2

[0049] The PEI prepolymer concentration is 4.8% (w / w), the ferrous chloride concentration is 3.21 mg / ml, and a very small amount of dilute hydrochloric acid solution is added to adjust the pH value of the solution to 10.0. Add 3% hydrogen peroxide so that its concentration in the reaction solution is 0.12% (v / v), and the remainder is the same as in Example 1.

[0050] The polyethylenimine nanogel sample after the purification is measured with the laser particle size scattering instrument PCS method, all shows unimodal distribution (figure is not shown), and the particle diameter mean value of this polyethylenimine nanogel is 172.9nm, more than The dispersion coefficient was 0.355. After being placed at 4°C for 21 days, it was tested again, and the average particle size was 171.7nm, and the polydispersity coefficient was 0.415. The results show that the polyethyleneimine nanogel samples are basically stable during the storage process, and the particle size does not change sign...

Embodiment 3

[0052] The concentration of PEI was 1.15% (w / w), the concentration of iron carboxylate was 1.54 mg / ml, and the pH value was adjusted to 8.0 by adding dilute hydrochloric acid solution. The stirring speed was controlled at about 140 rpm, and after 30 minutes of deoxygenation by nitrogen gas, 3% hydrogen peroxide was added to make the concentration in the reaction solution 0.11% (v / v). Under the condition of continuous feeding of nitrogen (nitrogen flow rate: 80ml / min), the intensity of ultraviolet light is 18mw / cm 2 Light for 1 h to initiate the reaction. Control the above-mentioned reaction conditions and repeat the test every other month to obtain three samples No. 4, No. 5, and No. 6. After the samples are filtered through a 0.45 μm needle filter and purified by dialysis three times, the three nanocondensate samples are measured by PCS. The particle diameter of glue sample is each three times, and the result shows unimodal distribution, and particle diameter distribution is...

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Abstract

The invention discloses a PEI nano gel, which is prepared by the prepolymer solution of polyethylene imine on basic condition with pH value not more than 11 and radiation of ultraviolet light. The photon correlation spectrometry shows unimodal distribution for its particle size with dispesion coefficient 0.1-0.7. it also disloses its application as gene transfection carrier. The product has controllable particle size to decrease cytotoxicity and immunogenicity and increase its security and efficiency as carrier in biological medicine field.

Description

technical field [0001] The invention relates to a polyethyleneimine nano gel and its application in gene transfection in vivo and in vitro. Background technique [0002] Many diseases and cancers have been proven to be caused by mutations and deletions of single genes. Gene therapy is to introduce normal genes into the body to make up for the deficiency of missing and / or mutated genes. At present, the human genome project has been completed, and the post-genome project is being implemented. The development of new functional gene drugs is gradually becoming one of the hotspots of scientific research in the world, and as an important link in the development of gene drugs - the research and development of vectors is also attracting more and more researchers. [0003] The vectors used to mediate the binding of DNA or oligonucleotides mainly include viral vectors and non-viral vectors. Viral vectors use recombinant viruses to transfect therapeutic genes, such as: retroviral vec...

Claims

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Application Information

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IPC IPC(8): C08J3/075C08L39/02C12N15/64A61K48/00
Inventor 姚思德徐冬梅刘永彪乔向利余家会孙汉文盛康龙
Owner SHANGHAI INST OF APPLIED PHYSICS - CHINESE ACAD OF SCI
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