Exosome-nucleic acid aptamer liposome composite drug carrier system as well as preparation method and application thereof

A nucleic acid aptamer and drug-carrying system technology, applied in the field of cancer targeted drug delivery systems, can solve the problems of lack of function of natural liposomes, cumbersome synthesis process, and increased cost, so as to reduce drug resistance and improve drug resistance. Efficacy, good biocompatibility

Inactive Publication Date: 2018-04-17
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, natural liposomes lack functions, while artificially synthesized liposomes are convenient for mo

Method used

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  • Exosome-nucleic acid aptamer liposome composite drug carrier system as well as preparation method and application thereof
  • Exosome-nucleic acid aptamer liposome composite drug carrier system as well as preparation method and application thereof
  • Exosome-nucleic acid aptamer liposome composite drug carrier system as well as preparation method and application thereof

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Embodiment 1

[0038] A preparation method of the exosome-nucleic acid aptamer liposome composite drug delivery system of the present invention, comprising the following steps:

[0039] (1) Culture of exosome cells

[0040] A549 cells were incubated at 37°C, 5% CO 2 cultured at 75cm 2 In the culture flask, the serum concentration is 10%. When the cell confluence reaches 50%-60%, replace it with exosome-free serum and cultivate for 2-3 days. Collect the cell culture medium and remove it by centrifugation at 300g for 10min at 4°C. cell;

[0041] (2) Extraction of exosomes

[0042] Centrifuge the cell culture medium at 2000g (centrifugal force) for 10min to remove dead cells; take the supernatant after centrifugation and place it in a 15mL centrifuge tube, and centrifuge at 20000g for 30 minutes to remove cell debris and microvesicles; take the supernatant with a 0.22μm Filter the filter to further remove impurities; use PBS buffer to balance the centrifuge tube, and centrifuge at 120,000g ...

Embodiment 2

[0051] A preparation method of the exosome-nucleic acid aptamer liposome composite drug delivery system of the present invention, comprising the following steps:

[0052] (1) Culture of exosome cells

[0053] MCF7 cells were incubated at 37°C, 5% CO 2 cultured at 75cm 2 In the culture flask, the serum concentration is 10%. When the cell confluence reaches 50%-60%, replace it with exosome-free serum and cultivate for 2-3 days. Collect the cell culture medium and remove it by centrifugation at 300g for 10min at 4°C. cell;

[0054] (2) Extraction of exosomes

[0055] Centrifuge the cell culture medium at 2000g (centrifugal force) for 10min to remove dead cells; take the supernatant after centrifugation and place it in a 15mL centrifuge tube, and centrifuge at 20000g for 30min to remove cell debris and microvesicles; take the supernatant and filter it through a filter of 0.22μm filter to further remove impurities; use PBS buffer to balance the centrifuge tube, and centrifuge a...

Embodiment 3

[0063] A preparation method of the exosome-nucleic acid aptamer liposome composite drug delivery system of the present invention, comprising the following steps:

[0064] (1) Culture of exosome cells

[0065] Hep2 cells were incubated at 37°C, 5% CO 2 cultured at 75cm 2 In the culture flask, the serum concentration is 10%. When the cell confluence reaches 50%-60%, replace it with exosome-free serum and cultivate for 2-3 days. Collect the cell culture medium and remove it by centrifugation at 300g for 10min at 4°C. cell;

[0066] (2) Extraction of exosomes

[0067] Centrifuge the cell culture medium at 2000g (centrifugal force) for 10min to remove dead cells; take the supernatant after centrifugation and place it in a 15mL centrifuge tube, and centrifuge at 20000g for 30min to remove cell debris and microvesicles; take the supernatant and filter it through a filter of 0.22μm filter to further remove impurities; use PBS buffer to balance the centrifuge tube, and centrifuge a...

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Abstract

The invention discloses an exosome-nucleic acid aptamer liposome composite drug carrier system which comprises an anti-tumor medicine, wherein the surface of the anti-tumor medicine is wrapped by nano-liposomes, a nucleic acid aptamer is connected with the surfaces of the nano-liposomes through chemical bonds to form a nucleic acid aptamer liposome on which the anti-tumor medicine is carried; thenucleic acid aptamer liposome on which the anti-tumor medicine is carried is compounded with exosome. The invention further discloses a preparation method and an application of the composite drug carrier system. The composite drug carrier system is good in targeting property, good in immune evasion property, free of cytotoxicity and not liable to be degraded by macrophage, can be applied to anti-tumor medicine chemotherapy of cancer patients, is capable of greatly improving the treatment effect of anti-tumor medicines, reducing the situations of chemotherapy effect degradation and disease reoccurrence caused by drug resistance, and alleviating the bottle neck problems of conventional chemotherapy of cancer patients, and has great application values and application prospects in intensifyingcancer treatment effects.

Description

technical field [0001] The present invention relates to the technical field of cancer targeted drug delivery system, in particular to an exosome-nucleic acid aptamer liposome composite drug delivery system and its preparation method and application. Background technique [0002] In recent years, the development and research of nano-drug carriers has received extensive attention. Nano-drug carriers have many advantages, such as: nanoparticles are easy to prepare, modified and processed with good controllability, and can protect drugs, genes or functional molecules from degradation by the body or some enzymes, etc. Some nanoparticles, while acting as "carriers", also have the function of immune adjuvants. [0003] Liposome is an artificial vesicle structure. When the liposome is in water, the hydrophilic head of the phospholipid molecule is inserted into the water, and the hydrophobic tail of the liposome extends to the air. After stirring, a spherical liposome with bilayer l...

Claims

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Application Information

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IPC IPC(8): A61K45/00A61K9/127A61K47/54A61K47/46A61P35/00
Inventor 张翼袁孟颖王珊马领然胡欣贾国萍江仁庭竺敏
Owner CENT SOUTH UNIV
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