Preparation for enhancing hypoxic tumor photodynamic therapy and preparation method and application of preparation
A photodynamic therapy and tumor technology, applied in the field of medicine, can solve the problems of oxygen consumption and insufficient supply, achieve low cost, good stability, and enhance the effect of photodynamic therapy
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Embodiment 1
[0040] 1. Synthesis method of ROS-sensitive material dextrin-modified phenyl borate (Dex-PBA)
[0041] Step 1, Synthesis of imidazole carbamate (CDI-PBA): Dissolve 3.51 g of 4-(hydroxymethyl)phenylboronic acid pinacol ester (PBA) in 22 mL of anhydrous dichloromethane, and then add 4.86 g of N , N-carbonyldiimidazole (CDI), after stirring at 25°C for 30 minutes, the solvent was removed by distillation under reduced pressure to obtain a crude product. The product was then dissolved in 100 mL of ethyl acetate and washed three times with 7.5 mL each. After drying over anhydrous magnesium sulfate and filtering, the final product was obtained.
[0042] The molar ratio of PBA to CDI is 1:1-3, preferably 1:2.
[0043] Step 2, synthesis of dextrin-modified phenyl borate (Dex-PBA): 197.02 mg dextrin, 298.6 mg 4-dimethylaminopyridine (DMAP) and 729.2 mg CDI-PBA obtained in step 1 were dissolved in 3.5 mL in anhydrous dimethyl sulfoxide. After the mixture was stirred and reacted overn...
Embodiment 2
[0063] Production of singlet oxygen in solutions of nanoparticles in different formulation groups
[0064] Using DPBF as a singlet oxygen detection probe, the ethanol solution containing DPBF (12 μL, 5 mM) was added to the aqueous solutions of PBS, DCe6, DPCe6 and DPCe6@ATO (6 mL, 5 μM Ce6 concentration), respectively. Then a 650 nm laser (15 mW cm -2 ), the absorption spectrum of the aqueous solution at 407nm was detected every 5s with a UV-Vis spectrophotometer.
[0065] Figure 9 It is a diagram of the generation of singlet oxygen in different nanoparticle solutions. It can be seen that the 407nm absorption of DPBF in the DPCe6 and DPCe6@ATO solutions of the preparation group was significantly reduced under light, indicating that a large amount of singlet oxygen was generated.
Embodiment 3
[0067] In vitro cytotoxicity assay of DPCe6@ATO
[0068] Step 1, human non-small cell lung cancer cell A549 cells were inoculated in a 96-well plate at a cell density of 2500 cells / well, using 1640 medium containing 10% fetal bovine serum and 1% penicillin / streptomycin under normoxic conditions (21%O 2 ) under cultivation. After 12 h, the cells were divided into two groups, half of which were under hypoxic conditions (1% O 2 ), and the rest of the cells were cultured under normoxic conditions (21% O 2 )to cultivate. When the cell density increased to 60%-70%, the PBS solution containing three kinds of nanoparticles with different concentrations of Ce6 and ATO was co-incubated with the cells for 4 hours in the dark.
[0069] Step 2, replace the spent medium with fresh culture medium, set up the light group and the dark group, and irradiate the cells with a 650 nm laser at a power density of 6.75 mW / cm2 for 5 minutes in the light group. Repeat the same operation after 4h. ...
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