Lipid nanogold particle compound and application thereof in delivering ERG and treating encephaledema disease

A technology of gold nanoparticles and composites, applied in gene therapy, genetic material components, pharmaceutical formulations, etc., can solve the problem of less brain edema, achieve the effects of treating brain edema, repairing the blood-brain barrier, and facilitating large-scale production

Pending Publication Date: 2022-04-05

QIANFOSHAN HOSPITAL OF SHANDONG

0 Cites 0 Cited by

AI-Extracted Technical Summary

Problems solved by technology

However, there are few studies on the ...

Abstract

The invention relates to the technical field of gene therapy, relates to a lipid gold nanoparticle compound and application of the lipid gold nanoparticle compound to ERG delivery and encephaledema disease treatment, and provides a lipid gold nanoparticle compound for treating encephaledema through a gene therapy method. The composite material comprises an Ang1-Lipo-AuNP-ERG compound, wherein the Ang1-Lipo-AuNP- The preparation method of the compound comprises the following specific steps: preparation of the AuNPs, preparation of the cation AuNPs, preparation of the AuNP-ERG, and preparation of the Ang1-Lipo-AuNP-ERG. The preparation method has the beneficial effects that the lipid gold nanoparticle compound Ang1-Lipo-AuNP-ERG serving as the gene carrier of the ERG has good biocompatibility and relatively high delivery efficiency. When Ang1-Lipo-AuNP-ERG is taken in by brain endothelial cells, ERG plasmids are expressed in the cells, so that the expression of Claudin-5, VE-Cadherin and ICAM-1 is regulated and controlled, and the effects of repairing the blood brain barrier and treating encephaledema can be achieved.

Application Domain

Genetic material ingredientsGene therapy

Technology Topic

CadherinGene carrier +8

Image

Examples

Experimental program(5)

Example Embodiment

[0028] The invention provides a lipid-gold nanoparticle complex for treating cerebral edema by gene therapy, the complex is an Ang1-Lipo-AuNP-ERG complex, and the preparation method includes:

[0029] Add sodium dodecyl sulfate to the AuNPs solution to carry out the reaction; then add β-mercaptoethylamine to carry out the reaction; carry out dialysis and purification on the reacted solution, and the dialysate is a weakly acidic citric acid solution; filter after dialysis to obtain cationic AuNPs solution;

[0030] The cationic AuNPs solution was added to the ERG plasmid solution and stirred overnight to obtain AuNP-ERG solution;

[0031] DSPE-PEG 2000 -MAL and Ang-1 are reacted in solution and purified by dialysis to obtain Ang-DP 2000;

[0032] Take soybean lecithin, cholesterol and 2-dioleoyl hydroxypropyl-3-N,N,N-trimethylammonium chloride DOTAP respectively, add them to methanol solution, heat to dissolve the three, mix well, cool to room temperature , adding the Ang-DP 2000 Mix well and evaporate methanol to obtain liposomes;

[0033] The AuNP-ERG solution and the liposome are subjected to shaking hydration, ultrasonic crushing, solid-liquid separation and filtration to obtain Ang1-Lipo-AuNP-ERG complex.

[0034] In some embodiments, the mass ratio of the sodium dodecyl sulfate to β-mercaptoethylamine is 1-5:3-7.

[0035] In some embodiments, the mass ratio of cationic AuNPs to ERG is 1:1.3 to 1:1.7.

[0036] In some embodiments, DSPE-PEG 2000 - The mass ratio of MAL and Ang-1 is 1.3:1 to 1.7:1.

[0037] In some embodiments, the mass ratio of soybean lecithin, cholesterol and 2-dioleoylhydroxypropyl-3-N,N,N-trimethylammonium chloride is 0.2-0.6:0.05-0.3:0.01-0.09.

[0038] In some embodiments, the specific conditions of the ultrasonic fragmentation are: a power of 80-120 W, a working time of 3-7 s, an intermittent time of 3-7 s, and a total of 3-7 min.

[0039] In some embodiments, the preparation method of AuNPs is as follows: mixing gold tetrachloride solution and distilled water, heating to boiling; then adding trisodium citrate solution, continuing to heat, the solution changes from light yellow to wine red, and boiling for 15-30 min Then, the heating was stopped and cooled to room temperature to obtain gold nanoparticles.

Example Embodiment

[0044] Example 1

[0045] The lipid-gold nanoparticle complex for treating cerebral edema according to the present invention includes Ang1-Lipo-AuNP-ERG complex. The preparation steps of the compound are as follows:

[0046] (1) Preparation of AuNPs

[0047] The glass instruments used were soaked in aqua regia, washed three times with distilled water and dried for later use; add 30 μL gold tetrachloride solution with a mass fraction of 25.6% in a 100 mL two-neck bottle, and 60 mL distilled water, and heat to boiling. Then add 8 mL of 1% trisodium citrate solution, continue heating, the solution changes from light yellow to wine red, stop heating and cool to room temperature after boiling for 20 min, to obtain 13±2 nm gold nanoparticles. Store at 4°C in the dark.

[0048] (2) Preparation of cationic AuNPs

[0049] 0.3 g of sodium dodecyl sulfate SDS was added to the synthesized AuNPs solution and stirred for 2 h. Subsequently, 0.5 g of β-mercaptoethylamine was added and stirring was continued for 4 h. The solution was put into a dialysis bag with a cut-off of 3500 Da, and the dialysate was a weakly acidic citric acid solution (Ph=2-4). After two days of dialysis, the solution in the dialysis bag was filtered with a 0.22 μm membrane to obtain a cationic AuNPs solution, which was stored at 4°C in the dark.

[0050] (3) Preparation of AuNP-ERG

[0051] The ERG plasmid solution was added to the cationic AuNPs solution at a mass ratio of 1:1.5 (AuNPs:ERG), and stirred overnight to obtain an AuNP-ERG solution.

[0052] (4) Preparation of Ang1-Lipo-AuNP-ERG

[0053] First, DSPE-PEG 2000 -MAL and Ang-1 were dissolved in methanol solution at a ratio of 1.5:1, stirred at room temperature for 48h, and purified by dialysis to obtain Ang-DP 2000.

[0054] Take 0.4g of soybean lecithin, 0.1g of cholesterol and 0.05g of 2-dioleoyl hydroxypropyl-3-N,N,N-trimethylammonium chloride DOTAP respectively, put them in a 500mL eggplant-shaped bottle, add 200mL methanol solution, heat To 70 ~ 80 ℃ to dissolve the three, stir to mix. After cooling to room temperature, add 0.05g Ang-DP 2000 Stir well. Spin dry methanol on a rotary evaporator to form a uniform layer of liposomes on the bottle wall. The AuNP-ERG solution was added to the eggplant-shaped flask, shaken and hydrated for 2h. The large liposomes were broken up by ultrasonic crusher, 100W, working for 5s, intermittent for 5s, and a total of 5min. It was then purified by centrifugation. 10000rpm, 3min. The Ang1-Lipo-AuNP-ERG complex was obtained by filtering three times with a 0.22 μm filter membrane.

Example Embodiment

[0055] Example 2

[0056] The lipid-gold nanoparticle complex for treating cerebral edema according to the present invention includes Ang1-Lipo-AuNP-ERG complex. The preparation steps of the compound are as follows:

[0057] (1) Preparation of AuNPs

[0058] All glass instruments used were soaked in aqua regia, then washed three times with distilled water and dried. In a 100 mL two-necked bottle, add 10 μL of gold tetrachloride solution with a mass fraction of 25.6% and 40 mL of distilled water, and heat to boiling. Then add 6 mL of 1% trisodium citrate solution, continue heating, the solution changes from light yellow to wine red, stop heating and cool to room temperature after boiling for 15 min, to obtain 13±2 nm gold nanoparticles. Store at 4°C in the dark.

[0059] (2) Preparation of cationic AuNPs

[0060] 0.1 g of sodium dodecyl sulfate SDS was added to the synthesized AuNPs solution and stirred for 1 h. Subsequently, 0.3 g of β-mercaptoethylamine was added and stirring was continued for 2 h. Put the solution into a dialysis bag with an interception volume of 3000-4000 Da, and the dialysate is a weakly acidic citric acid solution. After two days of dialysis, the solution in the dialysis bag was filtered with a 0.22 μm membrane to obtain a cationic AuNPs solution, which was stored at 4°C in the dark.

[0061] (3) Preparation of AuNP-ERG

[0062] The ERG plasmid solution was added to the cationic AuNPs solution at a mass ratio of 1:1.3 (AuNPs:ERG), and stirred overnight to obtain an AuNP-ERG solution.

[0063] (4) Preparation of Ang1-Lipo-AuNP-ERG

[0064] First, DSPE-PEG 2000 -MAL and Ang-1 were dissolved in methanol solution at a ratio of 1.3:1, stirred at room temperature for 24 hours, and purified by dialysis to obtain Ang-DP 2000.

[0065] Take 0.2g of soybean lecithin, 0.05g of cholesterol and 0.01g of 2-dioleoyl hydroxypropyl-3-N,N,N-trimethylammonium chloride DOTAP respectively, put them in a 500mL eggplant-shaped bottle, add 100mL methanol solution, heat Dissolve the three at 60°C and stir to mix. After cooling to room temperature, add a small amount of Ang-DP 2000 Stir well. Spin dry methanol on a rotary evaporator to form a uniform layer of liposomes on the bottle wall. The AuNP-ERG solution was added to the eggplant-shaped flask, shaken and hydrated for 1 h. The large liposomes were broken up by ultrasonic disruptor, 80W, working for 3s, intermittent for 3s, and a total of 3min. It was then purified by centrifugation. 8000rpm, 2min. The Ang1-Lipo-AuNP-ERG complex was obtained by filtering three times with a 0.22 μm filter membrane.

PUM

Description & Claims & Application Information

We can also present the details of the Description, Claims and Application information to help users get a comprehensive understanding of the technical details of the patent, such as background art, summary of invention, brief description of drawings, description of embodiments, and other original content. On the other hand, users can also determine the specific scope of protection of the technology through the list of claims; as well as understand the changes in the life cycle of the technology with the presentation of the patent timeline. Login to view more.

Classification and recommendation of technical efficacy words

Improve delivery efficiency
good biocompatibility

Lipid nanogold particle compound and application thereof in delivering ERG and treating encephaledema disease

AI-Extracted Technical Summary

Problems solved by technology

Abstract

Image

Examples

Example Embodiment

Example Embodiment

Example Embodiment

PUM

Description & Claims & Application Information

Similar technology patents

Composite nano sliver polymer antibacterial micelle and preparation method thereof

Method for preparing three-dimensional porous stent composite layer

A kind of synthetic method of trihydrate 3-aminopropylamine ethylthiophosphoric acid

Preparation method of magnetic fluorescent dual-function nano particle with nuclear shell structure

Method for preparing structural-color contact lens

Classification and recommendation of technical efficacy words

Meal delivery robot applied to intelligent cafeteria and intelligent cafeteria

Database deployment method and device

Full-automatic express delivery trolley

Bucket type conveyor

Straw crushed aggregate conveying and lifting transport system

Implantable particles for urinary incontinence

Polymer with tumor organization pH responsiveness and micelle thereof

Hollow core-shell nanometer mesoporous medicament carrying system with magnetism and luminescent performance, preparation method and application thereof

Preparation method of calcium silicate/calcium alginate hybrid material with mesoporous silica gel on surface

Preparation method of silk fibroin/metal nanoparticle composite system