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Preparation and application of gene and chemotherapeutic drug co-delivery anti-tumor nanoparticle of target SR-BI (Scavenger Receptor-BI)

A chemotherapeutic drug and nanoparticle technology, applied in gene therapy, antineoplastic drugs, drug combinations, etc., can solve the problems of difficult extraction, easy contamination of blood products and biological safety, and achieve the effect of improving encapsulation efficiency and efficient treatment

Inactive Publication Date: 2014-05-28
CHINA PHARM UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] At the same time, we should also note that there are some problems in the extraction of endogenous HDL from blood, such as the difficulty of extraction, and the biological safety caused by easy contamination of blood products, which limit the use of endogenous HDL as an anti-tumor drug carrier. application

Method used

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  • Preparation and application of gene and chemotherapeutic drug co-delivery anti-tumor nanoparticle of target SR-BI (Scavenger Receptor-BI)
  • Preparation and application of gene and chemotherapeutic drug co-delivery anti-tumor nanoparticle of target SR-BI (Scavenger Receptor-BI)
  • Preparation and application of gene and chemotherapeutic drug co-delivery anti-tumor nanoparticle of target SR-BI (Scavenger Receptor-BI)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: Preparation of rHDL drug-loaded nanoparticles

[0030]

[0031] Accurately measure the plasmid DNA to make a 0.5mg / ml solution, and dissolve DODAB in ethanol to make a 3.88mg / ml ethanol solution. According to the ratio of N / P ratio of 4, the DNA solution was injected into the DODAB ethanol solution to obtain the DODAB / DNA complex, vortexed for 30 s, and stood at room temperature for 30 min.

[0032] N / P=(m DODAB × M DNA ) / (m DNA × M DODAB )

[0033] where m DODAB is the mass of DODAB, m DNA is the mass of DNA, M DODAB is the relative molecular mass of DODAB, M DNA is the relative molecular mass of DNA.

[0034] Dissolve the prescribed amount of phospholipids, cholesterol, and cholesteryl ester in chloroform, add paclitaxel methanol solution, and dry the organic solvent by rotary evaporation at 37°C, and then put it in a vacuum desiccator overnight to remove the residual solvent. Add pH 7.4 PBS buffer containing DODAB / DNA, hydrate until a suspension...

Embodiment 2

[0035] Example 2: Research on the properties of rHDL drug-loaded nanoparticles

[0036] 2.1 Particle size and morphology of rHDL drug-loaded nanoparticles

[0037] The average particle size of the complex was measured by a laser particle size analyzer to be 184 nm. Transmission electron microscope results are as figure 1 As shown, the prepared rHDL drug-loaded nanoparticles are all spherical particles with good roundness and smooth appearance. The measured particle size is about 150nm, which is smaller than that measured by the laser particle size analyzer. The reason may be that the surface band There is an electric double layer around the nanoparticles with a certain negative charge, and the electric double layer can scatter light, resulting in larger results measured by the laser particle size analyzer. The protein on the surface of rHDL drug-loaded nanoparticles contains a large number of hydrophilic amino groups, which will bind more water molecules to form a thicker hy...

Embodiment 3

[0047] Example 3: Cytotoxicity Test (MTT)

[0048] 3.1 Cytotoxicity test of the carrier

[0049] MCF-7 cells in good growth state and in the logarithmic growth phase were taken, digested with 0.25% trypsin, and blown into a single cell suspension. Adjust the cell concentration to 1 x 10 5 1 / ml, 100 μl per well was inoculated in a 96-well plate, and 5 duplicate wells were set up. Place the cell plate at 37°C, 5% CO 2 cultured overnight in a cell culture incubator. Discard the medium, add 100 μl of fresh medium, then add 100 μl of blank NLC, blank rHDL, DODAB solution diluted in serum-free medium, continue to cultivate for 24 hours, suck off the medium, and add 20 μl of 0.5 mg / ml MTT solution to each well , 37°C, 5% CO 2 Incubate for 4 h in a cell culture incubator. The supernatant was discarded, and 150 μl of dimethyl sulfoxide (DMSO) solution was added to each well, and shaken for 10 min. The absorbance value of each well was measured at 570 nm with an enzyme-linked imm...

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Abstract

The invention relates to preparation and application of a gene and chemotherapeutic drug co-delivery anti-tumor nanoparticle of a target SR-BI (Scavenger Receptor-BI). The nanoparticle is a recombinant high density lipoprotein drug-loaded nanoparticle, comprises phospholipid, cholesterol, cholesteryl ester, apolipoprotein, cation lipid, a chemotherapeutic drug, a gene drug and the like, and is prepared by a film dispersion method. The invention aims at preparing the bionic tumor target drug-loaded nanoparticle, the preparation condition is mild, and the cost is low. The invention further provides the application of the gene and chemotherapeutic drug co-delivery anti-tumor nanoparticle. The gene and chemotherapeutic drug co-delivery anti-tumor nanoparticle has the advantages of spherical structure, good encapsulation efficiency, low cytotoxicity, good targeting, high safety, significant in-vitro anti-tumor effect and the like.

Description

technical field [0001] The invention relates to the field of pharmaceutical preparations, in particular to the preparation and application of co-delivery anti-tumor nanoparticles targeting SR-BI receptor genes and chemotherapeutic drugs. Background technique [0002] Cancer is one of the most serious diseases that have plagued human beings for centuries and endangers human health. Its mortality rate is second only to cardiovascular diseases. According to WHO statistics, among the more than 7 billion people in the world, an average of 6.9 million people die from malignant tumors each year, and 8.7 million new cases are reported, and the number is still increasing year by year. Therefore, governments, research institutes and pharmaceutical companies of various countries have long attached great importance to tumor research and anti-tumor drugs, and significant progress has been made in the research of anti-tumor drugs. [0003] Tumor drug therapy began in the 1940s, and there...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/14A61K48/00A61K47/42A61K47/20A61K47/18A61P35/00A61K31/704A61K31/337
Inventor 周建平王伟张洁蕾鲍秀丽李敏
Owner CHINA PHARM UNIV
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