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A cucumber exosome-like vesicle containing cucurbitacin b as an anticancer drug

A technology of exosomes and cucurbitacin, which is applied in the direction of medical preparations containing active ingredients, antineoplastic drugs, drug combinations, etc., can solve the problems of expensive DNA, poor stability in the body, and limited applications, so as to increase drug resistance , low cost and easy operation

Active Publication Date: 2022-07-19
FUJIAN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] 2. However, there are still some problems in the practical application of animal exosome acquisition and isolation
However, DNA is expensive, complex in design, poor in vivo stability, and easily digested by enzymes, which to some extent limits the application of DNA self-assembly technology in drug carriers.

Method used

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  • A cucumber exosome-like vesicle containing cucurbitacin b as an anticancer drug
  • A cucumber exosome-like vesicle containing cucurbitacin b as an anticancer drug
  • A cucumber exosome-like vesicle containing cucurbitacin b as an anticancer drug

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Embodiment 1 of the present invention provides a method for isolating exosome-like vesicles from cucumber, the preparation steps of which are as follows:

[0063] (1) Squeeze cucumber juice through a juicer and filter through gauze to obtain a filtrate;

[0064] (2) centrifuging the filtrate and removing the precipitate to obtain a supernatant;

[0065] (3) After ultracentrifuging the supernatant, resuspending the precipitate with an appropriate amount of PBS to obtain cucumber exosome-like vesicles.

[0066] Juicing in the step (1) requires pre-cooling PBS or PBS ice cubes to be mixed with cucumber.

[0067] The centrifugation in the step (2) is carried out at 5000 g for 10 min and at 10000 g for 35 min.

[0068] The ultracentrifugation in the step (3) was carried out at 30,000 g for 30 min.

[0069] The characterization results of the obtained cucumber exosome-like vesicles are as follows: figure 1 shown, (A) dynamic light scattering detection of size distribution...

Embodiment 2

[0071] The process of analyzing the active ingredients in the cucumber exosome-like vesicles prepared in Example 1 by high performance liquid chromatography is as follows:

[0072] (1) Centrifuge the cucumber exosome-like vesicles prepared in Example 1, discard the supernatant PBS, and obtain a cucumber exosome-like vesicle precipitate;

[0073] (2) Resuspend the cucumber exosome-like vesicles with an appropriate amount of methanol to obtain a cucumber exosome-like vesicle suspension, and ultrasonically;

[0074] (3) centrifuging the sonicated suspension, the cucumber exosome-like vesicles are precipitated for use, and the supernatant is collected;

[0075] (4) Repeat steps (2) and (3) twice, and combine the supernatants collected three times.

[0076] In the steps (2) and (4), the ratio of the total amount of methanol to the total protein of cucumber exosome-like vesicles is 1 mL: 6 mg (the total protein of cucumber exosome-like vesicles was determined by using the BCA prote...

Embodiment 3

[0082] The cytotoxic effects of cucurbitacin B standard substance and cucumber exosome-like vesicles on cancer cells were verified by MTT method. The specific operation methods are as follows:

[0083] (1) The cancer cell suspension was evenly added to the 96-well plate, and the culture medium was aspirated and discarded after the cells adhered to the wall;

[0084] (2) Administration: The cucurbitacin B standard product and the cucumber exosome-like vesicles prepared in Example 1 were respectively added to different wells of the above 96-well plate, and were aspirated and discarded after reacting for 24 hours;

[0085] (3) MTT solution was added to the above 96-well plate, and the solution was aspirated and discarded after 4 hours of reaction;

[0086] (4) Add dimethyl sulfoxide (DMSO) to the above-mentioned 96-well plate, measure the absorbance with a microplate reader after shaking, and calculate the cell viability.

[0087] In the step (1), the concentration of the cell s...

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Abstract

The invention discloses a cucumber exosome-like vesicle containing cucurbitacin B and can be used as an anticancer drug. Cucurbitacin B can be used to inhibit the proliferation of various cancer cells, but its extraction method is complicated and time-consuming. The method of isolating cucumber exosome-like vesicles to obtain cucurbitacin B has the advantages of simple and rapid operation, and the lipid bilayer of exosome-like vesicles, as the natural protective film of cucurbitacin B, can be used for a long time. maintain its stability. Moreover, the presence of the lipid bilayer also increased the uptake rate of cucurbitacin B by cells to a certain extent through the principle of similar compatibility. When acting on adenocarcinoma human alveolar basal epithelial cells (A549 cells), the maximum cell activity inhibition rate of cucumber exosome-like vesicles is twice that of cucurbitacin B standard, and the dosage is lower, which can reduce the toxicity to normal cells It has good anticancer potential and industrialization prospects, and is of great significance in clinical tumor treatment applications.

Description

technical field [0001] The invention relates to the technical field of separation and extraction of plant exosome-like vesicles and anticancer drug research, and more particularly, to the extraction of cucumber exosome-like vesicles containing active pharmaceutical ingredients, and the exosome-like vesicles as Feasibility study of anticancer drugs. Background technique [0002] 1. Exosomes are nano-scale vesicle structures actively secreted by cells. In recent years, they have been very popular as drug carriers because they can participate in various biological processes such as intercellular communication. Moreover, the lipid bilayer on the outside of the exosome can effectively protect the active ingredients inside and ensure that they are not decomposed before reaching the target site. In addition, the presence of lipid bilayers provides modification sites for a variety of functional molecules. [0003] 2. However, the acquisition and isolation of animal exosomes still ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K36/42A61K9/127A61K31/575A61P35/00
CPCA61K36/42A61K9/127A61K31/575A61P35/00
Inventor 陈敬华陈婷婷夏垚坤曾鲁鹏马丙香王华英施婉华
Owner FUJIAN MEDICAL UNIV
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