A method for constructing multifunctional gene therapy vectors based on gold nanoparticles

A gold nanoparticle and gene therapy technology, applied in gene therapy, preparations for in vivo experiments, pharmaceutical formulations, etc., can solve the problems of increased endocytosis, increased cytotoxicity, and increased transfection efficiency, achieving The effect of stable reaction, good storage stability and easy regulation

Active Publication Date: 2017-06-23
BEIJING UNIV OF CHEM TECH
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  • Abstract
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  • Claims
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Problems solved by technology

[0011] 3. When preparing a high-performance gold nanoparticle gene carrier, as the monomers are continuously grafted onto the gold nanoparticle skeleton, the molecular weight of the cationic gene carrier increases, the endocytosis of the cell increases, and the transfection efficiency increases. However, the toxicity of cells also increases, how to minimize the toxicity of gene carriers has become a problem to be solved
[0012] 4. The concentration of gold nanoparticles in the solution is directly related to the effect of CT imaging, but as the concentration of gold nanoparticles increases, the concentration of cationic polymers also increases, and the cytotoxicity will also increase. How to find a suitable The concentration of gold nanoparticles becomes a problem to be solved

Method used

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  • A method for constructing multifunctional gene therapy vectors based on gold nanoparticles
  • A method for constructing multifunctional gene therapy vectors based on gold nanoparticles
  • A method for constructing multifunctional gene therapy vectors based on gold nanoparticles

Examples

Experimental program
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Effect test

Embodiment 1

[0033] 1) Synthesis of gold nanoparticles:

[0034] Seeding solution: To cetyltrimethylammonium bromide solution (2.5 mL, 0.2 M) was added 74 μL of 0.024 M HAuCl 4 solution, then add 1.0458mLH 2 O, mix well; then add 0.5 mL of 10 mM NaBH kept in ice water 4 solution, stirred vigorously for 2 minutes; the solution turned brownish yellow, stored at 27°C for 1 hour before taking;

[0035] Growth solution: To cetyltrimethylammonium bromide solution (25 mL, 0.2 M) was sequentially added 1.4 mL of AgNO at a concentration of 4 mM 3 solution, add 1.5mL of 0.024M HAuCl 4 solution, then add 21mLH 2 O; each solution should be shaken and mixed evenly after adding; then add 620 μL ascorbic acid (0.0788M), the solution turns from dark yellow to colorless; finally add 500 μL seed solution, react at 27 ° C for 24 h; centrifuge at 2000 rpm for 10 min , then take the supernatant and continue to centrifuge at 12000rpm for 20min to obtain rod-shaped gold nanoparticles;

[0036] 2) Bovine se...

Embodiment 2

[0041] The reaction time of step 4) in Example 1 was 10 minutes, and the rest of the reaction conditions were the same as in Example 1. The obtained multifunctional gene therapy carrier based on gold nanoparticles was designated as Au-pDMA2.

Embodiment 3

[0043] The reaction time of step 4) in Example 1 was 30 minutes, and the rest of the reaction conditions were the same as in Example 1. The obtained multifunctional gene therapy carrier based on gold nanoparticles was designated as Au-pDMA3.

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Abstract

A method for constructing multifunctional gene therapy vectors based on gold nanoparticles. The invention belongs to the technical field of CT contrast agents and non-viral gene carriers, and specifically relates to gold nanoparticles as the skeleton, including spherical gold nanoparticles, rod-shaped gold nanoparticles (with different axis-to-diameter ratios), arrowhead-shaped gold nanoparticles, and octahedral gold nanoparticles. The morphology of gold nanoparticles, etc., using modified bovine serum albumin to wrap gold nanoparticles, and ATRP method to construct a multifunctional gene carrier. The polymerization reaction is stable and easy to control, and a variety of high-performance cationic gene carriers with different monomer contents can be prepared according to needs, and the storage stability is good, and the original performance can be maintained after being placed for several days or months. The cationic gene carrier has a higher transfection efficiency than the international "gold standard" PEI in HepG2, COS7, 293, C6 and other cells, and its cytotoxicity is much lower than that of PEI. It is simple to use and has commercial potential. And the material has a good CT imaging effect, which lays a good experimental foundation for the development of new contrast agents.

Description

technical field [0001] The invention belongs to the technical field of CT contrast agents and non-viral gene carriers, and specifically relates to gold nanoparticles as the skeleton, including spherical gold nanoparticles, rod-shaped gold nanoparticles (with different axis-to-diameter ratios), arrowhead-shaped gold nanoparticles, and octahedral gold nanoparticles. The morphology of gold nanoparticles, etc., using modified bovine serum albumin to wrap gold nanoparticles, and the ATRP method to construct a multifunctional gene therapy carrier. Background technique [0002] Gene therapy is to introduce normal genes into hematopoietic stem cells or other tissue cells, correct their specific genetic defects, disorders of gene structure or function, prevent the progression of lesions, or inhibit the replication and reproduction of viral genetic information by restoring or increasing gene expression. expression, so as to achieve the purpose of treatment. At present, gene therapy p...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K48/00A61K47/02A61K49/18A61K49/14
Inventor 徐福建赵娜娜闫彭
Owner BEIJING UNIV OF CHEM TECH
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