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Nano cell membrane drug-loaded vesicle, preparation method and application thereof

A cell membrane and vesicle technology is applied in the field of nano-drug-loading vesicles prepared based on cell membranes and the preparation thereof, which can solve the problems of limited yield and unsuitability for large-scale extraction, and achieves avoiding damage, good biocompatibility and immunity. Escape ability, the effect of enhancing the effect of drug treatment

Inactive Publication Date: 2020-08-07
NANKAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The nanovesicle does not affect the activity of the encapsulation components, and can avoid the attack of the body's immune system. The outer shell component is the cell membrane, but its yield is very limited, which is not suitable for large-scale extraction

Method used

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  • Nano cell membrane drug-loaded vesicle, preparation method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] 1) Acquisition of cell membrane: Take 10 normal cultured human bone marrow mesenchymal stem cells 7 Put each in a centrifuge tube, centrifuge at 1000rpm for 5min, discard the supernatant to obtain spare cells; prepare cell lysate: 0.01M Tris, 0.001M MgCl 2 , 2mMPMSF, 0.35M sucrose, DNase and RNaseA with a final concentration of 10 μg / mL were dissolved in distilled water, and the final pH=7.4; 10 7 Add 1mL cell lysate to each spare cell, homogenize at 22000rpm for 2min, then centrifuge at 4000rpm at 4°C for 10min, collect the supernatant, then centrifuge at 10000rpm at 4°C for 20min, discard the supernatant, and wash the obtained pellet twice with cell lysate times, the resulting precipitate is the purified cell membrane;

[0015] 2) Preparation of cell membrane nanovesicles: 10 7 After the cell membrane of each cell was resuspended with 0.5mL PBS, it was sonicated for 25s with a sonicator (Sonics VCX150) at a power of 25%, and then mixed with an equal volume of the dr...

Embodiment 2

[0017] 1) Acquisition of cell membrane: After human umbilical cord mesenchymal stem cells were subcultured and spread for 6 hours, the induction medium (20ng / mL INFγ, 10ng / mL TNFα) was replaced for 48 hours to induce the cell membrane to obtain the targeting function, and the functionalized cells were dispersed with PBS , 10 7 Dispense each cell / tube into a centrifuge tube, centrifuge at 1000rpm for 5min, discard the supernatant to obtain spare cells; prepare cell lysate: 0.01M Tris, 0.001M MgCl 2 , 2mM PMSF, 0.35M sucrose, DNase and RNaseA with a final concentration of 10 μg / mL were dissolved in distilled water, and the final pH=7.4; 10 7 Add 1mL cell lysate to each spare cell, homogenize at 22000rpm for 2min, then centrifuge at 4000rpm at 4°C for 10min, collect the supernatant, then centrifuge at 10000rpm at 4°C for 20min, discard the supernatant, and wash the obtained pellet twice with cell lysate times, the resulting precipitate is the purified cell membrane;

[0018] 2)...

Embodiment 3

[0020] 1) Acquisition of cell membrane: Take 10 normal cultured human adipose-derived mesenchymal stem cells 7 Put each in a centrifuge tube, centrifuge at 1000rpm for 5min, discard the supernatant to obtain spare cells; prepare cell lysate: 0.01M Tris, 0.001M MgCl 2 , 2mMPMSF, 0.35M sucrose, DNase and RNaseA with a final concentration of 10 μg / mL were dissolved in distilled water, and the final pH=7.4; 10 7 Add 1mL cell lysate to each spare cell, homogenize at 22000rpm for 2min, then centrifuge at 4000rpm at 4°C for 10min, collect the supernatant, then centrifuge at 10000rpm at 4°C for 20min, discard the supernatant, and wash the obtained pellet twice with cell lysate times, the resulting precipitate is the purified cell membrane;

[0021] 2) Preparation of cell membrane nanovesicles: 10 7 After the cell membrane of each cell was resuspended with 0.5mL PBS, it was sonicated with a sonicator (Sonics VCX150) at a power of 35% for 25s, then mixed with an equal volume of dexame...

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Abstract

The invention discloses a nano cell membrane drug-loaded vesicle, a preparation method and application thereof. The preparation method is characterized in that through a physical extrusion technology,drug coating is carried out in the process of forming nanoscale cell membrane vesicles, no influence is generated on the activity of a coated drug, the in-vivo circulation time of the drug can be prolonged, the drug targeting ability is improved, and directional slow release of the drug is realized. The preparation method comprises the steps of cell membrane acquisition and cell membrane nano-vesicle preparation. Specific cell delivery of the drug targeting specific part can be realized, so that the drug fully plays a therapeutic role, and liver and kidney injury caused by one-time massive drug aggregation in the liver or kidney is prevented. Dynamic changes in a drug body can be tracked through probe wrapping.

Description

technical field [0001] The field of the present invention relates to nanocarriers and drug encapsulation, specifically to nanometer drug-loaded vesicles prepared based on cell membranes and their preparation methods and applications. [0002] technical background [0003] Biomimetic nano-drug carriers can not only release drugs slowly to prolong the timeliness of drugs in vivo, but also assist drugs with low solubility to prepare injectable dosage forms. Most of the existing nanocarriers are artificially synthesized, and the system contains non-biological substances, which cannot completely simulate the natural components of the body, and are easily recognized and cleared by the immune system when they enter the body. Cellular exosomes are small vesicles of hundreds of nanometers formed by cell membrane-wrapped proteins, DNA, mRNA, microRNA and other biologically active molecules. It can efficiently and directionally transport biologically active factors to target cells to p...

Claims

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Application Information

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IPC IPC(8): A61K9/51A61K47/46A61K38/18
CPCA61K9/5176A61K38/1866
Inventor 齐春晓刘祥胜王淑芳孔德领
Owner NANKAI UNIV
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