A kind of bionic exosome and its preparation and application

An exosome and biomimetic technology, applied in the field of biomimetic exosomes, can solve problems such as undiscovered tumor treatment, and achieve the effect of enhancing the infiltration of immune cells into tumors, enhancing infiltration, and improving anti-tumor efficacy.

Active Publication Date: 2022-04-22
THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are no reports on this type of biomimetic exosomes, let alone the use of such biomimetic exosomes in the treatment of tumors

Method used

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  • A kind of bionic exosome and its preparation and application
  • A kind of bionic exosome and its preparation and application
  • A kind of bionic exosome and its preparation and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] This example is used to illustrate the preparation of liposomes.

[0061] Liposomes were prepared by the thin film dispersion method. 20mg lipid material DPPC, cholesterol and DSPE-PEG 2000 (mass ratio 80:15:5) was dissolved in 2 mL of chloroform:methanol (3:1 V / V), and the solvent was evaporated by a rotary evaporator to form a lipid film.

[0062] Add 2mL PBS to the lipid film for hydration to form a lipid suspension.

[0063] The lipid suspension was passed through a cellulose acetate membrane with a pore size of 100 nm through a liposome extruder at 25°C for 20 times, and the obtained homogeneous liposome solution was stored at 4°C for use.

[0064] The particle size distribution of liposomes ( figure 1 A), zeta potential ( figure 2 ) was characterized by a nanometer particle size analyzer, and the morphology ( image 3 A) Characterized by transmission electron microscopy.

[0065] In the particle size distribution diagram, the average particle size of liposo...

Embodiment 2

[0067] This example is used to illustrate the preparation of biomimetic exosomes.

[0068] 4T1 breast tumor cell membrane protein was extracted by cell membrane protein and cytoplasmic protein extraction kit, and the concentration of cell membrane protein was determined by BCA method.

[0069] In Example 1, tumor cell membrane protein was added while PBS was added to the lipid film for hydration, the mass ratio of membrane protein to lipid material was 1:100, and the mixture was fully mixed to form a suspension.

[0070] Add NE to the membrane protein-lipid suspension, the mass ratio of NE to lipid material is 1:40, incubate at 25°C for 30min, then pass through a cellulose acetate membrane with a pore size of 100nm through a liposome extruder for 20 times , and the obtained homogeneous biomimetic exosome solution was stored at 4°C for future use.

[0071] Particle size distribution of biomimetic exosomes ( figure 1 B), Zeta potential ( figure 2 ) was characterized by a nan...

Embodiment 3

[0074] This example is used to illustrate the protein characterization of membrane protein chimeric liposomes in the biomimetic exosomes of the present invention.

[0075] The tumor cell membrane proteins on the biomimetic exosomes prepared in Example 2 of the present invention were determined by Coomassie brilliant blue staining. Configure 10% SDS polyacrylamide gel, load 10 μg of 4T1 breast tumor cell total protein, 4T1 breast tumor cell membrane protein, 4T1 breast tumor cell cytoplasmic protein and biomimetic exosomes, and infiltrate with Coomassie brilliant blue staining solution after electrophoresis Soak and stain for 2 hours, then decolorize with decolorizing solution. Take pictures to observe the protein bands between groups (results see Figure 4 ). The results showed that the protein of the biomimetic exosome was very similar to the membrane protein of 4T1 breast tumor cells, but significantly different from the cytoplasmic protein of 4T1 breast tumor cells, indic...

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Abstract

The invention discloses a bionic exosome and its preparation and application. In the present invention, artificial lipid materials are chimerized with tumor cell membrane proteins to prepare nano-liposomes with a structure particle size similar to that of exosomes, and NE is bound to the surface of nano-liposomes through electrostatic adsorption to serve as bionic exosomes. The biomimetic exosomes have stable lipid membrane structure, tumor targeting and tumor matrix degradation functions. The biomimetic exosomes can enhance the penetration of anticancer drugs into the tumor and the infiltration of immune cells into the tumor after targeting and degrading the tumor matrix, greatly improving the antitumor efficacy, and have great transformational value and application in combined chemotherapy / immune antitumor prospect.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and specifically relates to a bionic exosome targeting to degrade tumor matrix and its preparation and application. Background technique [0002] Tumor is a major disease that seriously endangers human health. Desmoplastic tumors have a dense tumor stroma mainly composed of abundant tumor extracellular matrix and tumor-associated fibroblasts, which encloses relatively few tumor cells, and blood vessels are buried in the tumor stroma, not directly adjacent to tumor cells , not only makes it difficult for drugs to penetrate into the tumor, but also hinders the infiltration of immune cells in the tumor site. Using nanotechnology to break through the tumor matrix can promote the entry of anticancer drugs into the tumor, and at the same time promote the infiltration of immune cells in the tumor, which is of great significance for tumor treatment. [0003] Neutrophils are important immune cells i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K9/127A61K47/24A61K47/42A61K38/48A61P35/00
CPCA61K9/1271A61K47/24A61K47/42A61K38/486C12Y304/21037A61P35/00
Inventor 向大雄李泳江吴军勇胡雄彬
Owner THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV
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