Anti-tumor composition based on nano-silver material and application of anti-tumor composition
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, achieve the effect of enhancing curative effect, reducing infection risk and improving biological safety
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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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