Preparation method of a cationic lipid-mesoporous silicon composite nanocarrier co-loading siRNA and chemotherapy drugs

A technology of cationic lipids and chemotherapeutic drugs, applied in drug combinations, pharmaceutical formulas, genetic material components, etc., can solve the problem of poor treatment effect of malignant tumors

Active Publication Date: 2021-09-14
HENAN UNIVERSITY OF TECHNOLOGY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0002] Although tumor chemotherapy has made great progress in recent years, it is still not effective in the treatment of most malignant tumors. One of the important reasons is the generation of multidrug resistance (MDR) of tumor cells.

Method used

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  • Preparation method of a cationic lipid-mesoporous silicon composite nanocarrier co-loading siRNA and chemotherapy drugs
  • Preparation method of a cationic lipid-mesoporous silicon composite nanocarrier co-loading siRNA and chemotherapy drugs
  • Preparation method of a cationic lipid-mesoporous silicon composite nanocarrier co-loading siRNA and chemotherapy drugs

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Experimental program
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Embodiment 1

[0031] (1) Preparation of mesoporous silicon nanoparticles: Add a magnet rotor and 1.0 g of CTAB into a three-neck flask, then add 100 ml of deionized water, and stir at 95° C. for 30 min. Add 160μL DEA solution to the bottle and continue to stir for 15min. Take 8ml of TEOS and add it dropwise slowly (dropping for 1 hour). After the dropwise addition is completed, reflux for 3 hours, centrifuge, and discard the supernatant. The template was removed by cross-washing three times with methanol-HCl solution (4:1) and deionized water. Dry in a drying oven at 50° C. for 12 hours, and then vacuum-dry for 12 hours. That is, mesoporous silicon nanoparticles (MSNs) with an average particle diameter of about 80 nm were obtained.

[0032] (2) Take mesoporous silicon particles in a 10ml centrifuge tube, add an appropriate amount of PBS to disperse, add dropwise 5mg / ml doxorubicin (DOX) solution, mix and make up to volume. Stir for 24 hours, centrifuge, and wash with PBS until the washin...

Embodiment 2

[0039] (1) Chemically synthesize two complementary single-stranded DNAs corresponding to siRNA 1, with Hind III and BglII restriction sites at both ends. After annealing, it was ligated with the double-enzyme-digested plasmid pDual, and the product was transformed into Escherichia coli competent DH5a, single-clonal colonies were picked for amplification and culture, and the plasmid was extracted and sequenced for identification.

[0040] (2) Loading of siRNA 1: Centrifuge the dry powdered siRNA for 10 minutes, dissolve it in 375 μl of DEOC water, mix the obtained solution with the above-mentioned CLMSNs, and let it stand still, and record it as siRNA-CLMSNS. The particle size, PDI and zeta potential of siRNA-CLMSNS were measured with a Malvern particle size analyzer. The results are shown in Table 1. Compared with CLMSNS, the particle size of siRNA-CLMSNS was significantly larger, and the zeta potential was significantly lower. This is due to the negatively charged The siRNA n...

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Abstract

The invention discloses a preparation method of a cationic lipid-mesoporous silicon composite nanocarrier co-carrying siRNA and chemotherapy drugs. The method comprises the following steps: using a template method to synthesize mesoporous silicon nanoparticles, and synthesizing MDR1-siRNA; Mix mesoporous silicon and chemotherapeutic drugs in an organic solvent, wash and dry to obtain drug-loaded mesoporous silicon; disperse drug-loaded mesoporous silicon in a chloroform solution dissolved in lipid materials, evaporate the solvent to form a film, and then add the dispersed The medium is ultrasonically dispersed until completely dispersed, sterilized by filtration, and repeatedly extruded to adjust the particle size; the siRNA is dissolved, mixed with liposome solution, and left to stand. The composite carrier can enhance the cytotoxicity of the drug, induce cell apoptosis, and play a synergistic anti-tumor effect on the basis of inhibiting MDR.

Description

technical field [0001] The invention belongs to the technical field of chemical pharmacy, and relates to a preparation method of a cationic lipid-mesoporous silicon composite nanocarrier co-carrying siRNA and chemotherapeutic drugs. Background technique [0002] Although tumor chemotherapy has made great progress in recent years, it is still ineffective in the treatment of most malignant tumors. One of the important reasons is the generation of multidrug resistance (MDR) of tumor cells. MDR is ubiquitous in various tumor cells and is the main reason for the failure of chemotherapy. Therefore, inhibiting or reversing MDR has important theoretical and clinical significance for tumor therapy. MDR is generally considered to be a self-protection defense mechanism of tumor cells against chemotherapy drug toxicity, and its occurrence is related to many factors. Aiming at the common drug resistance mechanism, people have designed a variety of methods to reverse MDR, including chem...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K9/51A61K47/04A61K47/28A61K31/7105A61K31/704A61K48/00A61P35/00
CPCA61K9/5115A61K9/5123A61K9/5192A61K31/704A61K31/7105A61K48/005A61P35/00A61K2300/00
Inventor 杨硕晔宋世博韩凯硕刘艳萍陈凌志吴兴旺王金水张贝贝
Owner HENAN UNIVERSITY OF TECHNOLOGY
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