A kind of antitumor composition and application thereof based on nano silver material
A nano-silver, anti-tumor technology, applied in the field of biomedicine, can solve the problems of severe infection and high safety risk of patients
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Embodiment 1
[0037] An anti-tumor composition comprising attenuated Salmonella and sialic acid-modified nano-silver. Among them, sialic acid modified silver nanoparticles (SP-AgNPs), including sialic acid (SA) and lipoic acid polyethylene glycol (LA-PEG-NH 2 ) through amide reaction to form sialic acid-modified polyethylene glycol (LA-PEG-SA), and then modify this polymer on the surface of silver nanoparticles (AgNPs) prepared by sodium citrate reduction method through Ag-S bonds.
[0038] The sialic acid-modified nano silver (SP-AgNPs) of the present embodiment is prepared by the following method:
[0039] (1) Dissolve 85.9mg SA, 63.9mg EDC·HCl, 38.4mg NHS in 5.6mL DMSO, stir at room temperature for 2h to activate the carboxyl group of SA. 200mg LA-PEG-NH 2 Dissolved in 2mL DMSO, added to the above reaction solution, and continued to react for 24h. After the reaction, the reaction solution was transferred to a dialysis bag (MWCO=3500), and dialyzed in ultrapure water for 48 hours. The...
experiment example
[0043] (1) Proton NMR spectrum: SA, LA-PEG-NH 2 1. LA-PEG-SA was dissolved in deuterated DMSO, with tetramethylsilane (TMS) as internal standard, and analyzed by proton nuclear magnetic resonance spectrometer. figure 1 SA, LA-PEG-NH 2 , the hydrogen spectrum of LA-PEG-SA, it can be seen from the figure: LA-PEG-SA has SA and LA-PEG-NH 2 All the characteristic peaks, prove the successful synthesis of LA-PEG-SA.
[0044] (2) Morphology: observe the morphology of SP-AgNPs, the detection method of morphology: drop the sample on a 400-mesh copper grid covered with carbon film, place it in a desiccator, and place it under a transmission electron microscope Titan G2-F20 after it dries naturally Observed. figure 2 It is a transmission electron microscope image of SP-AgNPs, and it can be seen from the figure that the SP-AgNPs particles of the present invention are spherical in shape, relatively uniform in distribution, and have a particle size of about 15nm.
[0045] (3) Potential:...
Embodiment 2
[0049] Investigate the targeting effect of the SP-AgNPs of Example 1 on neutrophils and B16F10 cells, the specific steps are:
[0050] (1) Preparation of fluorescein isothiocyanate (FITC)-labeled PEG-AgNPs and SP-AgNPs. The specific steps are:
[0051] AgNPs were obtained according to the method in Example 1 and redispersed in water. Add 20 mL of LA-PEG-SA (or LA-PEG-NH 2 ) aqueous solution and 2mL of LA-PEG-FITC aqueous solution with a concentration of 10mg / mL, stirred at room temperature in the dark for 24h, and centrifuged at a high speed (20000rpm, 30min) to obtain FITC-labeled PEG-AgNPs or SP-AgNPs (FITC-PEG-AgNPs, FITC -SP-AgNPs).
[0052] (2) Take the peripheral blood of the mouse, use the mouse peripheral blood neutrophil separation liquid kit to separate the neutrophils therein, and dilute it to 2×10 with an appropriate amount of 1640 complete medium containing 10% FBS 4 cells / mL of cell suspension, 0.5 mL per well was inoculated in a 24-well plate, and a total of...
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