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Preparation method of gene and hydrophobic drug co-supported PEG (polyethyleneglycol) nanoparticles

A hydrophobic drug and nanoparticle technology, which is applied in gene therapy, drug combination, pharmaceutical formulation and other directions, can solve problems such as affecting gene transfection efficiency, reducing the circulation time of gene delivery system, etc., and achieves simple and easy preparation method, easy operation, and easy loading capacity. The effect of adjusting and improving stability

Inactive Publication Date: 2012-10-03
ZHEJIANG UNIV
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Problems solved by technology

However, this kind of polycation / DNA assembly is easy to aggregate in the body, and is cleared out of the body by the reticuloendothelial system, which reduces the circulation time of the gene delivery system in the body and affects the efficiency of gene transfection

Method used

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  • Preparation method of gene and hydrophobic drug co-supported PEG (polyethyleneglycol) nanoparticles
  • Preparation method of gene and hydrophobic drug co-supported PEG (polyethyleneglycol) nanoparticles
  • Preparation method of gene and hydrophobic drug co-supported PEG (polyethyleneglycol) nanoparticles

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preparation example Construction

[0020] The steps of a method for preparing PEGylated nanoparticles co-loaded with genes and hydrophobic drugs are as follows:

[0021] 1) Prepare a β-cyclodextrin modified polycation solution with a concentration of 0.5-4 mg / mL;

[0022] 2) Prepare ferrocene-modified polyethylene glycol solution with a concentration of 0.5-4 mg / mL;

[0023] 3) Mix the solutions of step 1) and step 2) so that the molar ratio of β-cyclodextrin to ferrocene is 4:1-2:1, and then stand still after ultrasonication for 30 min;

[0024] 4) Prepare a hydrophobic drug solution with a concentration of 2 mg / mL;

[0025] 5) Add the hydrophobic drug solution prepared in step 4) dropwise to the solution in step 3), so that the molar ratio of β-cyclodextrin to hydrophobic drug is 1:1~1:4, stir in the dark for 24 hours, and dialyze for 8~ 24h, lyophilized to obtain β-cyclodextrin modified polycation / ferrocene modified polyethylene glycol / hydrophobic drug inclusion compound powder;

[0026] 6) Prepare β-cycl...

Embodiment 1

[0031] Prepare a β-cyclodextrin-modified polyethyleneimine (PEI-g-CD) solution with a concentration of 4 mg / mL and a ferrocene-modified polyethylene glycol (PEG-Fc) solution with a concentration of 4 mg / mL. The two solutions were mixed according to the molar ratio of β-CD and PEG-Fc in PEI-g-CD at a ratio of 2:1, sonicated for 30 min and left to stand for use. Add 2 mg / mL doxorubicin (DOX) dimethyl sulfoxide solution dropwise to the above mixed solution so that the molar ratio of CD and DOX in PEI-g-CD is 1:1, and stir in the dark for 24 h, then dialyzed for 8 h to 24 h, and freeze-dried to obtain PEI-g-CD / PEG-Fc / DOX clathrate powder. The PEI-g-CD / PEG-Fc / DOX clathrate powder prepared as above was used to measure the drug loading and encapsulation efficiency of doxorubicin (DOX) by ultraviolet-visible spectrophotometer (UV). The content was 9%, and the encapsulation efficiency was 57%, which indicated that β-cyclodextrin modified polyethyleneimine could effectively include the...

Embodiment 2

[0034] Prepare a solution of β-cyclodextrin-modified polylysine (PLL-g-CD) with a concentration of 2 mg / mL and a solution of ferrocene-modified polyethylene glycol (PEG-Fc) with a concentration of 2 mg / mL. The two solutions were mixed according to the molar ratio of β-CD and PEG-Fc in PLL-g-CD as 3:1, and then left to stand after ultrasonication for 30 min. Add 2 mg / mL paclitaxel (PTX) chloroform solution dropwise to the above mixed solution, so that the molar ratio of CD and PTX in PLL-g-CD is 1:2, stir for 24 h in the dark, and then dialyze for 8 h ~24 h, lyophilized to obtain PLL-g-CD / PEG-Fc / PTX clathrate powder. Prepare a PLL-g-CD / PEG-Fc / PTX solution with a concentration of 0.5 mg / mL and a p16 (multiple tumor suppressor 1 gene) solution with a concentration of 75 μg / mL. The above-mentioned PLL-g-CD / PEG-Fc / PTX solution was mixed with 250 μL DNA solution in equal volume by vortexing and allowed to stand for 30 min to prepare PEGylated nanoparticles co-loaded with genes and ...

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Abstract

The invention discloses a preparation method of gene and hydrophobic drug co-supported PEG (polyethyleneglycol) nanoparticles. The method comprises the following steps of: 1) preparing beta-cyclodextrin modified polycation solution; 2) preparing ferrocene modified polyethylene glycol solution; 3) mixing solutions in step 1) and step 2), standing after ultrasound is carried out; 4) preparing a hydrophobic drug solution; 5) dripping the hydrophobic drug solution prepared in step 4) into the solution in step 3), stirring away from light, dialyzing and lyophilizing to obtain inclusion powder; 6) preparing an inclusion solution; 7) preparing a DNA (deoxyribonucleic acid) solution; and 8) adding the solution in step 6) to isovolumetric solution in step 7), performing eddy mixing and standing to obtain the gene and hydrophobic drug co-supported PEG nanoparticles. The gene and hydrophobic drug co-supported PEG nanoparticles prepared by virtue of a host-guest effect; and the preparation method is simple and easy in operation, so that the stability of the gene and hydrophobic drug co-supported PEG nanoparticles in physiological salt solution can be remarkably improved.

Description

technical field [0001] The invention relates to a method for preparing PEGylated nanoparticles co-loaded with genes and hydrophobic drugs. It combines gene therapy and chemotherapy, and belongs to the preparation method and technology of a new type of high-efficiency non-viral gene delivery system in the field of gene therapy. Background technique [0002] Cancer (malignant tumor) has become an important disease affecting human health due to the lack of effective treatment methods, and tens of billions of dollars are directly used for tumor treatment every year. Cancer has become one of the most stubborn fortresses that scientists have not yet conquered, and it is also one of the most challenging topics in contemporary scientific research. Chemotherapy is one of the important means of comprehensive treatment of malignant tumors. However, due to the large amount of drugs used and the lack of specificity, it can cause serious toxic and side effects to normal human cells whil...

Claims

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Application Information

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IPC IPC(8): A61K9/14A61K47/40A61K47/34A61K48/00A61K31/704A61K31/337A61P35/00
Inventor 王幽香陈丽娜李文宇唐三
Owner ZHEJIANG UNIV
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