Functionalized dendrimer-based SPECT-CT bimodal imaging contrast agent and preparation method thereof
A dendrimer, dual-mode imaging technology, applied in the preparation of X-ray contrast agents, radioactive carriers, etc., can solve the problems of preparation and application of SPECT/CT contrast agents that have not been found
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Embodiment 1
[0064] Such as figure 1 shown, preparation 99m Tc-Au-Ac DENPs.
[0065] (1) 20.0mg of the fifth generation polyamidoamine dendrimers (G5.NH 2 , purchased from Dendritech Company) was dissolved in 15 mL of water, and 2 mL of an aqueous solution (1.1 mg / mL) of diethylenetriaminepentaacetic acid cyclic anhydride (cDTPAA, purchased from Sigma Aldrich) was added dropwise, and the reaction was stirred at room temperature for 8 h to obtain G5- DTPA; at the same time, add 5 mL of 1-ethyl-(3-dimethylaminopropyl) carbodiimide (EDC, purchased from Sigma Aldrich) in 10 mL of mPEG-COOH in water (7.69 mg / mL) in water (7.69 mg / mL) and stirred for 2h, then the mPEG-COOH (purchased from Shanghai Yanyi Biotechnology Co., Ltd.) activated by EDC was added to the G5-DTPA solution, stirred for 1-3d, and the functionalized dendrimer (G5 -DTPA-mPEG) solution;
[0066] (2) Add 2.1mL chloroauric acid solution (30mg / mL) to the G5-DTPA-mPEG solution obtained in (1) and stir for 30min, the solution t...
Embodiment 2
[0070] Such as figure 1 shown, preparation 99m Tc-Au-Gly DENPs.
[0071] (1) 20.0mg G5.NH 2 Dissolve in 15mL water, and add 2mL cDTPAA aqueous solution (1.1mg / mL) dropwise, stir the reaction at room temperature for 8h to obtain G5-DTPA; at the same time, add 5mL EDC to 10mL mPEG-COOH aqueous solution (7.69mg / mL) Aqueous solution (7.69mg / mL) and stirred for 2h, then mPEG-COOH activated by EDC was added to the G5-DTPA solution, and stirred for 1-3d to obtain a functionalized dendrimer (G5-DTPA-mPEG) solution;
[0072] (2) Add 2.1mL chloroauric acid solution (30mg / mL) to the G5-DTPA-mPEG solution obtained in (1) and stir for 30min, the solution turns light yellow, then add 2.8mL sodium borohydride aqueous solution (10mg / mL ), the solution instantly turns dark red, and the reaction is stirred for 1-4h. Then 84 μL glycidol was added and the reaction was stirred for 24 h. Finally, the reaction product was dialyzed in phosphate buffer solution 2L×3 and distilled water 2L×3 suc...
Embodiment 3
[0076] MTT assay was used to study the cytotoxicity of the prepared Au-Ac DENPs and Au-Gly DENPs.
[0077] Collect the logarithmic phase human ovarian cancer cell line (SK-OV-3 cells), add it to a 96-well cell culture plate, add 200 μL of cell-containing RPMI1640 medium to each well to make the cell density to 8000 / well; then place in a cell culture incubator (5%CO 2 , 37°C) for 24 hours, discard the medium and add 180 μL of fresh medium, and then add 20 μL of PBS buffer containing different concentrations of Au-Ac DENPs and Au-Gly DENPs (the final gold concentrations were 0, 2.5, 5, 10, 20, 40, 80 μM) to verify the effect of the material on the growth of SK-OV-3 cells. All test groups were set with 5 wells as a parallel group; after incubation in the incubator for 24 hours, 20 μL of MTT solution (5 mg / mL) was added to each well, and after 4 hours of incubation, the culture solution in the wells was carefully sucked, and the Add 200 μL of DMSO, shake on a shaker in the dark ...
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