Gene drug co-carrier system using cation liposome to establish gold nano-spherical shell and preparation method

A gold nanosphere shell and cation technology, applied in gene therapy, drug combination, pharmaceutical formulation, etc., can solve the problems of difficult removal of impurities, gene instability, gene decomposition, etc.

Inactive Publication Date: 2012-12-05
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Nowadays, many methods of constructing drug-gene co-loading are to connect drugs and genes together using chemical methods. The chemical reaction process is relatively complicated, and the impurities are not easy to remove after the reaction, and the gene is very unstable. The operation process is a little careless. , will cause the decomposition of genes; some researchers have achieved the co-loading of drugs and genes, but when the carrier carries the drugs and genes into the cells, the drugs and genes cannot be released into the cytoplasm, and the drugs and genes must be able to Therefore, a simple and quick method is needed to construct a co-carrying system of genes and drugs, and the release of drugs and genes can be achieved well

Method used

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  • Gene drug co-carrier system using cation liposome to establish gold nano-spherical shell and preparation method
  • Gene drug co-carrier system using cation liposome to establish gold nano-spherical shell and preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] a. Preparation of cationic liposomes: DPPC, MPPC, and DPPE-PEG2000 were dissolved in PBS solution (pH=7.4) according to the weight ratio of 40:40:10 to form a 60nM lipid solution, and doxorubicin hydrochloride was added, The concentration of doxorubicin hydrochloride was 5nM, and the solution was prepared into liposomes according to the standard cycle freeze-thaw method, and extruded through a 100nm polycarbonate membrane to form liposomes. The prepared liposomes were dialyzed in PBS solution to remove excess doxorubicin hydrochloride.

[0023] b. Formation of gold nanoshells: Take 1mL of the liposome solution prepared above, add 18uL 100nM chloroauric acid solution and 27uL 500nM ascorbic acid solution, the formed gold nanoshells have an absorption peak of 1064nm; In PBS solution, dialyze for 12h.

[0024] c. The connection between siRNA and the gold nanosphere shell: the selected siRNA is green fluorescent protein silencing gene, and the nucleotide sequence of the se...

Embodiment 2

[0026] a. Preparation of cationic liposomes: DPPC, MPPC, DPPE-PEG2000 weight ratio is 90:0:10, add PBS solution, add doxorubicin hydrochloride so that the concentration of doxorubicin hydrochloride is 10nM, and the remaining implementation steps and implementation The preparation process of the cationic liposomes in Example 1 was the same.

[0027] b. The formation of gold nanoshells: the concentrations of chloroauric acid solution and ascorbic acid solution are different, the thickness and particle size of the formed gold shells are all different, and the absorption peaks produced by the plasmon resonance effect are also different. The gold nano shell formed in Example 1 has an absorption peak of 1064nm. In this example, 7uL chloroauric acid solution and 10.5uL ascorbic acid solution are added to form a gold nano shell with an absorption peak of 655nm. The prepared samples were dialyzed again in PBS solution for 12 hours.

[0028] c. The connection between siRNA and the gold...

Embodiment 3

[0030] a. Preparation of cationic liposomes: DPPC, MPPC, DPPE-PEG2000 weight ratio is 90:0:10, and the rest of the implementation steps are the same as the preparation process of cationic liposomes in Example 1.

[0031] b. Formation of gold nanoshells: add 18uL 100nM chloroauric acid solution and 27uL 500nM ascorbic acid solution, and the formed gold nanoshell has an absorption peak of 1064nm; add 7uL chloroauric acid solution and 10.5uL ascorbic acid solution to form gold nanospheres The shell absorption peak is 655nm. The prepared samples were dialyzed again in PBS solution for 12 hours.

[0032]c. Connection of gene and gold nanoshell: Dilute SH-miR-21 (20 μmol / L) with TE buffer (10 mmol / L, Tris-HCL with pH=8.0, 1 mmol / L EDTA) to a certain concentration, The concentration of miR-21 with sulfhydryl groups was 2 μmol / L. According to the pre-designed volume ratio (64:1, 16:1, 2:1), the same amount of SH-miR-21 and gold nanoshells were mixed in TE buffer Mix in the solution ...

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Abstract

The invention relates to a gene drug co-carrier system using cation liposome to establish a gold nano-spherical shell and a preparation method. The method comprises: using phosphatides such as DPPC, MPPC and DPPE-PEG and the derivatives thereof to form the cation liposome via membrane extrusion; carrying the hydrophilic drug hydrochloric acid adriacin into the liposome in the forming process; forming the drug-carried cation liposome; adding chlorauric acid solution into cation liposome solution; adsorbing on the surface of the cation liposome; adding ascorbic acid solution for reduction; forming the gold nano-spherical shell structure; reducing sulfhydryl out of the single chain of green fluorescent protein silent gene with a disulfide bond, grafting onto the surface of the gold nano-spherical shell; and forming the gene drug co-carrier system. The gold nano-spherical shell has low toxicity and unique optical properties. The gene drug co-carrier system using cation liposome to establish the gold nano-spherical shell can provide carrier for the co-treatment of drugs and genes, can select light to illuminate the gold nano-spherical shell, and utilize light-heat conversion property to realize the co-release of the drugs and genes.

Description

technical field [0001] The invention relates to the formation of a gold nano-shell structure using a cationic liposome as a template, and using the gold nano-shell structure as a carrier to construct a drug and gene co-carrying system. Because siRNA etc. Background technique [0002] Cationic liposomes are widely used in drug carriers and gene carriers, but liposomes are metastable structures, not stable enough in body fluids, and have a relatively short circulation time in the body. Cationic liposomes can be modified to improve their stability. [0003] Gold nanomaterials have relatively unique physical and chemical properties. The gold nanosphere shell structure has an absorption peak in the near-infrared region due to the surface plasmon resonance effect. Researchers have used liposomes as templates to deposit gold on the surface to form hollow nanosphere structures, and used liposomes to carry anticancer drugs such as doxorubicin hydrochloride, and irradiated gold nano...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K31/704A61K9/10A61K47/48A61K47/34A61K47/02A61P35/00A61K47/24A61K47/69
Inventor 高立章刘贵高原续波何芳
Owner TIANJIN UNIV
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