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Preparation method of exosome capable of simultaneously loading chemical drug and nano-material

A nanomaterial and exosome technology, applied in drug combination, drug delivery, pharmaceutical formulation, etc., can solve the problems affecting imaging and anti-tumor effect, solubility, poor stability, phagocytosis of mononuclear macrophages, etc. Ease of approach, increased solubility, reduced phagocytosis and possible effects of clearance

Inactive Publication Date: 2018-06-05
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, both have some obvious disadvantages, such as: poor solubility and stability in the blood circulation system, easy to be phagocytized by mononuclear macrophages, tend to gather more in the liver, spleen and lymphoid tissue, and the concentration in the tumor site is low , affecting imaging and anti-tumor effects

Method used

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  • Preparation method of exosome capable of simultaneously loading chemical drug and nano-material
  • Preparation method of exosome capable of simultaneously loading chemical drug and nano-material
  • Preparation method of exosome capable of simultaneously loading chemical drug and nano-material

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preparation example Construction

[0031] 2) Preparation of nanometer ferric oxide:

[0032] Prepared by high-temperature pyrolysis: ① Weigh 0.7g iron acetylacetonate, 20mL benzyl ether, 2mL oleic acid and 6mL oleylamine, transfer them to a 100mL round-bottomed three-neck flask, and extend a glass dropper connected with argon into the liquid surface Below, adjust the gas flow to play a stirring role; ②Start the heating program: use 60 minutes to raise the temperature from room temperature to 220°C and keep it for 60 minutes, use 30 minutes to raise the temperature of the system to 290°C and keep it for 30 minutes; Naturally cool to room temperature, add 30mL of absolute ethanol to the system and ultrasonically disperse for 10 minutes; ④ Transfer the system to a small beaker, place it on a strong magnet for 10 minutes, use the magnetic force to obtain nano-scale ferric oxide precipitation, discard the supernatant Finally, add 30mL of ethanol again and ultrasonically disperse for 10min, then continue to place it ...

Embodiment 1

[0039] Add 100 μg of exosomes, 10 μl of 4 mg / mL nano-iron ferric oxide aqueous solution, and 5 μl of 24 mg / mL curcumin DMSO solution in 1 mL of 50 mM trehalose PBS solution; Electroporation instrument, according to the following reaction conditions: voltage 100V, capacitance 125μF, discharge time 1ms, discharge times: 1 time. After perforation, the suspension can be placed in a cell culture incubator and incubated for 1 hour, followed by ultracentrifugation at 100,000g twice for 80 minutes each time, carefully discard the supernatant, and resuspend the precipitate with PBS solution to load nano-trioxide The composite exosomes of iron and curcumin were stored at -80°C for subsequent detection and experiments;

Embodiment 2

[0041] Add 100 μg of exosomes, 10 μl of 4 mg / mL nano-iron ferric oxide aqueous solution, and 5 μl of 24 mg / mL curcumin DMSO solution in 1 mL of 50 mM trehalose PBS solution; Electroporation instrument, according to the following reaction conditions: voltage 400V, capacitance 250μF, discharge time 1ms, discharge times: 1 time. After perforation, the suspension can be placed in a cell culture incubator and incubated for 1 hour, followed by ultracentrifugation at 100,000g twice for 80 minutes each time, carefully discard the supernatant, and resuspend the precipitate with PBS solution to load nano-trioxide The composite exosomes of iron and curcumin were stored at -80°C for subsequent detection and experiments;

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Abstract

The invention discloses a preparation method of exosome capable of simultaneously loading a chemical drug and a nano-material. The preparation method comprises the following steps: 1) extracting cellsecreted exosome from cell culture supernatant by utilizing a classic differential ultra-speed centrifuging method; 2) simultaneously loading curcumin and nano ferroferric oxide in a water solution into the exosome by utilizing an electroporation method; 3) measuring OD260nm (an RNA (Ribonucleic Acid) characteristic absorption peak) to identify whether perforation is successful or not; 4) after finishing the perforation, immediately putting a mixed solution into a cell culture box and hatching for 1h, so as to facilitate exosome membrane repairing; 5) carrying out 100000g ultra-speed centrifuging to remove free curcumin and nano ferroferric oxide, so as to obtain the compound exosome capable of simultaneously loading the curcumin and the nano ferroferric oxide. The method disclosed by themethod is simple and feasible and high in success rate; in the prepared compound exosome, the loading amounts of the curcumin and the nano ferroferric oxide can meet in vivo and in vitro treatment andimaging requirements and a novel tool is provided for targeting diagnosis and treatment of various chronic diseases including tumors.

Description

technical field [0001] The invention relates to a compound exosome which has the functions of magnetic resonance imaging, magnetic hyperthermia and drug therapy. Background technique [0002] Nano-ferroferric oxide is a kind of nano-material, which has the advantages of easy synthesis, superparamagnetism, high saturation magnetization, good biocompatibility, and low toxicity, and is considered to be a good magnetic resonance imaging (MRI) contrast agent. Thermogenic material for medicament and magnetic hyperthermia. Curcumin is a plant-based polyphenolic compound with strong antitumor effects. In vitro experiments have found that curcumin has a significant inhibitory effect on the growth of various tumor cells including glioma, and its main mechanism is to inhibit tumor cell proliferation and invasion, promote tumor cell apoptosis, and resist tumor angiogenesis. It has a broad prospect in the drug treatment of glioma. However, both have some obvious disadvantages, such as...

Claims

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Application Information

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IPC IPC(8): A61K49/18A61K41/00A61K9/50A61K47/46A61K31/12A61P35/00
CPCA61K49/1896A61K9/5068A61K31/12A61K41/0052A61K2300/00
Inventor 贾刚唐秋莎安艳丽
Owner SOUTHEAST UNIV
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