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Method for loading siRNA (small interfering Ribonucleic Acid) by using mesoporous silicon dioxide nanoparticles

A technology of mesoporous silica and nanoparticles, which is applied in the direction of pharmaceutical formulations, medical preparations of non-active ingredients, gene therapy, etc., can solve the problems of high specific surface area and failure to exert the channel structure of mesoporous nanoparticles, and achieve Avoid chemical modification, easy-to-handle effect

Active Publication Date: 2012-01-25
SHANGHAI MAG GENE NANOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the above works only used the modified outer surface of mesoporous silica nanoparticles to load siRNA, which failed to take advantage of the advantages of mesoporous nanoparticle pore structure, high specific surface area, and large pore volume; A large number of amino groups are used to adsorb siRNA molecules, making the particle surface unable to be further functionalized such as polyethylene glycol, which limits the application of particles in vivo

Method used

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  • Method for loading siRNA (small interfering Ribonucleic Acid) by using mesoporous silicon dioxide nanoparticles
  • Method for loading siRNA (small interfering Ribonucleic Acid) by using mesoporous silicon dioxide nanoparticles
  • Method for loading siRNA (small interfering Ribonucleic Acid) by using mesoporous silicon dioxide nanoparticles

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example 1

[0026] 1. First add 0.2mg of M-MSNs into a 2mL centrifuge tube, then add 0.03mL of pure ethanol to it, and ultrasonically oscillate for 30s at 100w to disperse the particles evenly in the solution.

[0027] 2. Add 0.01 mL of high-salt solution (the composition of this high-salt solution is: 0.04 mmol of guanidine hydrochloride, 0.0025 mL of isopropanol, 0.0075 mL of sterilized water, pH 5) to the final solution obtained in step 1 and mix uniform.

[0028]3. Add 0.01 mL of siRNA aqueous solution (solvent is deionized water) with a concentration of 0.6 mg / mL to the solution obtained in step 2, and mix well again. In this embodiment, ethanol: high salt solution: siRNA aqueous solution three The volume ratio between the two was 3:1:1, the mixture was placed in an oven at 25°C for 4 hours, and the M-MSNs adsorbed by siRNA were separated from the solution by centrifugation at 10,000 rpm. At this time, the unit loading capacity of M-MSNs to siRNA was 8.8mg / g ( figure 2 shown).

...

example 2

[0034] 1. Keep the other conditions of steps 1 to 3 in Example 1 unchanged, change the volume ratio of ethanol: high salt solution: siRNA aqueous solution (solvent is double distilled water) to 2: 1: 1 or 4: 1 : 1.

[0035] 2. After the above-mentioned finally obtained particles are separated from the adsorption solution, the unit loading capacity of M-MSNs to siRNA is respectively: when the ratio is 2:1:1, the loading amount is 5.3 mg / g, and the ratio is 4: 1:1 loading capacity is 14.5mg / g ( figure 2 shown).

[0036] 3. Wash the separated particles once with 0.06mL pure ethanol, then add 0.06mL pure ethanol, and use 100w ultrasonic vibration to disperse the particles evenly.

[0037] 4. Add 0.06mL of pure ethanol solution containing polyethyleneimine to the final solution obtained in step 3 (the concentration of polyethyleneimine in pure ethanol solution is 1.5mg / mL), and the mixture is reacted under 100w ultrasonic vibration conditions 20 minutes.

[0038] 5. Separate t...

example 3

[0040] 1. First add 0.2mg of M-MSNs to a 2mL centrifuge tube, then add 0.04mL of pure ethanol to it, and ultrasonically oscillate for 30s at 100w to disperse the particles evenly in the solution.

[0041] 2. Add 0.01 mL of high-salt solution (the composition of this high-salt solution is: 0.04 mmol of guanidine hydrochloride, 0.0025 mL of isopropanol, 0.0075 mL of sterilized water, pH 5) to the final solution obtained in step 1 and mix uniform.

[0042] 3. Add 0.01 mL of siRNA aqueous solution (solvent is sterilized water) with a concentration of 0.5 mg / mL or 2.5 mg / mL to the solution obtained in step 2 and mix the solution evenly. In this embodiment, ethanol: high salt The volume ratio between the solution: siRNA aqueous solution was 4:1:1, the mixture was placed in an oven at 25°C for 4h, and the M-MSNs adsorbed by siRNA were separated from the solution by centrifugation at 10000rpm. The unit loading of siRNA is: when adding the siRNA aqueous solution with a concentration o...

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Abstract

The invention discloses a method for loading siRNA (small interfering Ribonucleic Acid) by using mesoporous silicon dioxide nanoparticles. In the method, the siRNA is induced into a pore cannel of the mesoporous silicon dioxide nanoparticles, so that loading of the siRNA is realized, and the loading capacity of the siRNA in the particles can be adjusted. The invention has the advantages: siRNA molecules are prevented from being separated from the pole canal under the condition of physiological environment; the surfaces of the loaded nanoparticles can be further decorated; and the method has a good application prospect.

Description

technical field [0001] The invention relates to a method for loading siRNA by a mesoporous silicon dioxide material under specific solution conditions. Background technique [0002] With the advancement of cell and molecular biology technology, RNA interference technology has been widely used to explore the gene function and the field of gene therapy for infectious diseases and malignant tumors (Nat.Rev.Mol.Cell Biol.4 Volume, 457 pages, 2003 year). RNA interference refers to the phenomenon of specific post-transcriptional gene silencing induced by double-stranded RNA homologous to the target gene. siRNA complementarily binds to homologous target RNA, and specifically enzymatically degrades the target RNA, thereby inhibiting and down-regulating gene expression. In 1998, after Fire et al. (Nature, Vol. 391, Page 806, 1998) discovered the phenomenon of RNA interference in nematodes, RNA interference technology, as a method for specifically inhibiting gene expression, has gra...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K47/04A61K47/32A61K47/34A61K47/36
Inventor 古宏晨李旭
Owner SHANGHAI MAG GENE NANOTECH CO LTD
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