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Cucumber exosome-like vesicles containing cucurbitacin B and capable of being used as anti-cancer drugs

A technology of exosomes and cucurbitacin, which is applied to medical preparations containing active ingredients, antineoplastic drugs, drug combinations, etc., can solve the problems of expensive DNA, poor stability in the body, and easy enzymatic hydrolysis, etc., to inhibit proliferation , low cost and easy operation

Active Publication Date: 2021-03-12
FUJIAN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology allows extractive bodies like cocoonfishes (Crocuss sp.) or sea urchin be separated into their own components that are protected against chemotherapy agents used on other types of tumors such as lung carcinoma. These extravesicular particles contain both hydrophilic and lipophillic compounds that help keep them stable during storage at room temperature. Additionally, this new type of material also helps prevent certain forms of cancer called suckers from growing inside these materials due to its ability to stimulate immune responses. Overall, it provides technical benefits over existing methods of isolate pure chemical substances from crops.

Problems solved by technology

The technical problem addressed in this patents relates to finding ways to produce stable and effective substances (plant extracts) containing these types of lipids called cocursolides or lignanols while avoiding the drawbacks associated with current methods including complicated steps involved at different levels of purification and concentration stages during production process.

Method used

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  • Cucumber exosome-like vesicles containing cucurbitacin B and capable of being used as anti-cancer drugs
  • Cucumber exosome-like vesicles containing cucurbitacin B and capable of being used as anti-cancer drugs
  • Cucumber exosome-like vesicles containing cucurbitacin B and capable of being used as anti-cancer drugs

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 the cucumber juice through a juicer and filter it through gauze to obtain the filtrate;

[0064] (2) centrifuging the filtrate and obtaining a supernatant by removing the precipitate;

[0065] (3) After performing ultracentrifugation on the supernatant, an appropriate amount of PBS was used to resuspend the pellet to obtain cucumber exosome-like vesicles.

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

[0067] The centrifugal separation in the step (2) was 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 30000 g for 30 min.

[0069] The characterization results of the obtained cucumber exosome-like vesicles are as follows: figure 1 Shown, (A) The particle size d...

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 the cucumber exosome-like vesicle precipitate;

[0073] (2) Use an appropriate amount of methanol to resuspend the cucumber exosome-like vesicle pellet to obtain a cucumber exosome-like vesicle suspension, and sonicate;

[0074] (3) Centrifuge the ultrasonic suspension, precipitate the cucumber exosome-like vesicles for use, and collect the supernatant;

[0075] (4) Repeat steps (2) and (3) twice, and combine the supernatants collected 3 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 using the BCA protein quantificati...

Embodiment 3

[0082] The cytotoxic effect of cucurbitacin B standard and cucumber exosome-like vesicles on cancer cells was verified by MTT method. The specific operation method is as follows:

[0083] (1) Evenly add the cancer cell suspension into the 96-well plate, and discard the culture medium after the cells adhere to the wall;

[0084] (2) Administration: Add the standard cucurbitacin B and the cucumber exosome-like vesicles prepared in Example 1 into different wells of the above-mentioned 96-well plate, and discard after reacting for 24 hours;

[0085] (3) Add MTT solution to the above-mentioned 96-well plate, react for 4 hours, and discard by suction;

[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 survival rate.

[0087] In the step (1), the concentration of the cell suspension is 100,000 cells / mL, and the volume of the cell suspension in each well is 100 μL.

[...

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Abstract

The invention discloses cucumber exosome-like vesicles containing cucurbitacin B and capable of being used as anti-cancer drugs. Cucurbitacin B can be used for inhibiting proliferation of various cancer cells, but an extraction method of cucurbitacin B is complex and time-consuming. The method of separating the cucumber exosome-like vesicles to obtain cucurbitacin B has the advantages of easinessand quickness in operation and the like, and the lipid bilayer of the exosome-like vesicles is used as a natural protective film of cucurbitacin B, so that the stability of cucurbitacin B can be maintained for a long time. Moreover, through the principle of similarity and intermiscibility, the intake rate of cells to cucurbitacin B is also improved to a certain extent due to the existence of the lipid bilayer. When the cucumber exosome-like vesicles act on human alveolar basal epithelial cells (A549 cells) of adenocarcinoma, the maximum cell activity inhibition rate of the cucumber exosome-like vesicles is twice that of a cucurbitacin B standard substance, the dosage is lower, the toxicity to normal cells and the drug resistance to cancer cells can be reduced, and the cucumber exosome-likevesicles have good anti-cancer potential and industrialization prospect, and have an important significance in the clinical tumor treatment application.

Description

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Claims

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Application Information

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Owner FUJIAN MEDICAL UNIV
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