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Blood-brain barrier penetrating drug delivery system as well as preparation method and application thereof

A blood-brain barrier permeation and delivery system technology, applied in the field of blood-brain barrier drug delivery system and its preparation, can solve the problems of cytotoxicity and difficult clinical transformation, and achieve high-efficiency permeation of the blood-brain barrier and excellent anti-glioma effect. Effect

Pending Publication Date: 2022-06-21
SHENZHEN INST OF ADVANCED TECH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the effectiveness, safety, large-scale preparation feasibility and stability of the carrier materials that constitute the aforementioned drug delivery system still need to be further improved and clarified, and most of the carrier materials are in the laboratory research stage, and clinical transformation is difficult
On the other hand, if immune cells are used as drug delivery carriers, chemotherapy drugs are often toxic to cells, and immune cells play multifaceted roles in the tumor microenvironment, which makes it difficult to directly use immune cells as drug carriers. challenge

Method used

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  • Blood-brain barrier penetrating drug delivery system as well as preparation method and application thereof
  • Blood-brain barrier penetrating drug delivery system as well as preparation method and application thereof
  • Blood-brain barrier penetrating drug delivery system as well as preparation method and application thereof

Examples

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Effect test

Embodiment 1

[0059] In this example, a neutrophil exosome is extracted, and the method is as follows:

[0060] (1) Take 25g Kunming mice, kill them by neck dislocation after anesthesia, put them in 75% ethanol for sterilization for 5min, and move them into an ultra-clean workbench;

[0061] (2) Carefully separate the mouse leg skin with sterilized surgical equipment, cut out the entire thigh, put it in pre-cooled PBS, peel off the mouse leg muscle, take out the femur and tibia, and put it into the femur and tibia containing RPMI1640 the petri dish;

[0062] (3) Rinse out the bone marrow with a fresh amount of RPMI1640, filter the cells with a 70 μm filter, centrifuge at 450 × g for 10 min, and resuspend the pellet in PBS solution;

[0063] (4) Use erythrocyte lysate to lyse erythrocytes, centrifuge at 450 × g for 5 min, and resuspend the pellet in PBS solution to obtain a single-cell suspension;

[0064] (5) Prepare percoll stock solution with 10×PBS and percoll cell separation solution ...

Embodiment 2

[0070] The present embodiment prepares a blood-brain barrier drug delivery system, and the method is as follows:

[0071] (1) Use BCA kit to measure neutrophil exosome protein concentration, take 0.4 mg / mL neutrophil exosomes extracted according to the method of Example 1 (exosomes diluted with 1×PBS, 0.22 μm Use after filtration with a sterile filter), add an equivalent concentration of 0.4 mg / mL DOX solution (DOX is prepared with 1×PBS containing 5% DMSO);

[0072] (2) Put the mixture of neutrophil exosomes and DOX in a centrifuge tube, and perform sonication in an ultrasonic cell disruptor; ultrasonic conditions: 20% amplitude, 6 cycles, 4s on, 2s off, each Cool the samples on ice for 2 min between cycles;

[0073] (3) After the treatment, the samples were incubated at 37°C for 1 h to ensure the recovery of the plasma membrane of NEs-Exos;

[0074] (4) After incubation at 37°C, the samples were centrifuged at 100,000 × g for 70 min, washed with PBS, and centrifuged again ...

Embodiment 3

[0076] Observation of transmission electron microscopy on the neutrophil exosomes and blood-brain barrier drug delivery systems prepared in Example 1 and Example 2: combining neutrophil exosomes or blood-brain barrier drug delivery systems with etc. The amount of 4% paraformaldehyde was mixed and fixed on a sample-loaded copper grid (200 mesh, coated with a formaldehyde metal film). After 20min washing with PBS, the copper mesh was placed on a drop of 1% glutaraldehyde for 5min. The copper mesh was then washed in ultrapure water. 2min each time, a total of 8 times. Images were captured using a JEM 1200EX TEM, respectively as figure 1 and figure 2 As shown in the figure, it can be seen from the figure that the neutrophil exosomes are cup-shaped or saucer-shaped ( figure 1 ), while the particle size of neutrophil exosomes loaded with DOX increased, but the morphology was also consistent with the exosome morphology ( figure 2 ).

[0077] Western blot analysis was performe...

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Abstract

The invention relates to a blood-brain barrier penetrating drug delivery system as well as a preparation method and application thereof. The blood-brain barrier penetrating drug delivery system comprises a neutrophil exosome and a drug entrapped in the neutrophil exosome, the medicine is a medicine for treating central nervous system diseases. The invention constructs a neutrophil exosome drug-loading delivery system with good biocompatibility, and the drug-loading system has biological characteristics similar to that of neutrophil, can efficiently penetrate through a blood-brain barrier, effectively respond to inflammation, and target an inflammation site, so that the drug-loading system has a good application prospect. The compound is a drug carrier with great application prospect for treating glioma and other central nervous system diseases. The invention provides a new strategy with a transformation application prospect for non-invasive inflammatory microenvironment targeted drug delivery of glioma and other central nervous system diseases.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a blood-brain barrier drug delivery system and a preparation method and application thereof. Background technique [0002] Gliomas are the most common primary malignant tumors of the central nervous system, with high invasiveness and poor prognosis. Traditional tumor resection is greatly limited due to its infiltration and aggressiveness. At the same time, the blood-brain barrier system composed of brain capillary endothelial cells, pericytes, astrocytes, etc. through tight junctions prevents toxic substances from invading the brain and also prevents most drugs from entering the brain, making glial The therapeutic drugs for cancer and many central system diseases (such as Parkinson's disease and Alzheimer's disease, etc.) are limited in application. Therefore, designing safe carriers that can assist drugs to efficiently penetrate the blood-brain barrier and effe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K47/46A61K31/704A61P35/00A61P9/10A61P25/16A61P25/28C12N5/0787
CPCA61K47/46A61K31/704A61P35/00A61P9/10A61P25/16A61P25/28C12N5/0642C12N2509/00
Inventor 唐为王亚洲王君李光林
Owner SHENZHEN INST OF ADVANCED TECH CHINESE ACAD OF SCI
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