Application of exosome in preparation of anti-infective drugs

An exosome, anti-infection technology, applied in the field of biomedicine, can solve the problems of difficult treatment, treatment failure drug resistance, drug resistance and so on

Inactive Publication Date: 2020-05-26
CHONGQING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Infections caused by intracellular pathogens continue to attract clinical attention, because the infections caused by them are often caused by the low concentration of most antibacterial drugs in the cells and cannot be inhibited, making the treatment difficult, and the treatment of intracellular bacterial infections is compli...

Method used

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  • Application of exosome in preparation of anti-infective drugs
  • Application of exosome in preparation of anti-infective drugs
  • Application of exosome in preparation of anti-infective drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: Culture of RAW264.7 cells and extraction and identification of exosomes

[0024] 1. Experimental materials

[0025] DMEM high-glucose medium was purchased from Hyclone Company; fetal bovine serum was purchased from Gibco Company; mouse macrophage RAW264.7 was purchased from Shanghai Cell Bank of Chinese Academy of Sciences; Exosome Rapid Extraction Reagent (ExoQuick) was purchased from SBI Company.

[0026] 2. Isolation and Characterization of Exosomes

[0027] RAW264.7 cells were cultured with 10% volume fraction of exosome-free fetal bovine serum DMEM medium + mass fraction of 1% penicillin-streptomycin complete medium. The cell culture fluid was collected, and exosomes were extracted according to the instructions of the ExoQuick kit. The specific steps are as follows: transfer the culture supernatant to a centrifuge tube, and centrifuge to remove cell debris. Take the supernatant and transfer it to a new centrifuge tube, add the reagent according to the...

Embodiment 2

[0031] Example 2: Encapsulation of Linezolid (LZD) by co-incubation

[0032] Quantitative standard curve determination of LZD: Accurately weigh LZD powder and dissolve it in DMSO to make a stock solution of 10 mg / mL, and store it at -80°C after aliquoting. Precisely draw an appropriate amount of LZD stock solution, dilute with acetonitrile to make standard solutions with final concentrations of 0.5, 1, 2, 4, 10, 20, 40 μg / mL, and inject 50 μL. Chromatographic conditions: Chromatographic column: Extend-C18, 250mm×4.6mm, 5μm, Agilent; mobile phase: acetonitrile: water = 20:80; flow rate: 1 mL / min; column temperature: 30° C.; detection wavelength: 251 nm. After obtaining the corresponding experimental results, linear regression is performed with the concentration of the analyte as the ordinate and the peak area of ​​the analyte as the abscissa, and the linear regression equation is obtained: Y=187.45x+10.227, R 2 =1, the linear relationship is good, and its linear range is 0...

Embodiment 3

[0035] Example 3: Application of linezolid-loaded exosomes (ExoLZD) to treat MRSA infection in RAW264.7 cells

[0036] RAW264.7 intracellular MRSA infection model construction: RAW264.7 cells were completely cultured with DMEM with a volume fraction of 10% fetal bovine serum at 37°C, 5% CO2 cultured in an incubator. Bacterial MRSA was cultured to the logarithmic phase, centrifuged at 4000rpm / min for 10min, discarded the supernatant, resuspended with the above-mentioned complete medium, and cultured again at 37°C for 30min. Replace the original macrophage culture medium with this bacteria-containing medium. After 2 hours of infection, wash the extracellular bacteria with sterile PBS buffer, and check whether the extracellular bacteria are eliminated by the live bacteria agar plate method.

[0037] Observation of ExoLZD treatment of MRSA infection in RAW264.7 cells by plate colony counting method: 2 hours after MRSA infected RAW264.7 cells, culture medium containing LZD, ExoLZD,...

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PUM

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Abstract

The invention relates to a new application of exosome secreted by cells, and relates to a preparation method of the exosome and an application of the exosome to treatment of infectious related diseases after being wrapped with antibacterial drugs. Compared with the prior art, the exosome is used as an antibacterial drug carrier, so that the biological effect of the antibacterial drug can be improved, and the effect of the antibacterial drug in the aspect of treating refractory infectious related diseases is enhanced.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to a new application of exosomes secreted by cells, including its preparation method and its application in the treatment of infectious-related diseases after wrapping drugs. Background technique [0002] Exosomes are a type of extracellular vesicles secreted by cells with a diameter of 30-150 nm. Exosomes can be secreted by almost all types of cells, and can be continuously secreted under physiological and pathological conditions of tissue cells, and exist in various body fluids such as saliva, plasma, and milk. Exosomes have potential biological applications in cell communication. They can transport biologically active molecules back and forth between cells, deliver miRNAs and proteins to other cells, and act as a bridge for inter-cell communication. Based on these characteristics, exosomes have unique advantages as natural endogenous carriers, such as low immunogenicity, high stability i...

Claims

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Application Information

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IPC IPC(8): A61K9/51A61K47/46A61K31/5377A61K38/14A61P31/00A61P31/04A61P31/10A61P31/12
CPCA61K9/5176A61K31/5377A61K38/14A61P31/00A61P31/04A61P31/10A61P31/12Y02A50/30
Inventor 贺耘杨晓洪石功名
Owner CHONGQING UNIV
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