A drug-loaded delivery nano-platform and its preparation method and application
A drug-loaded and platform-based technology, applied in the fields of gene therapy and chemotherapy nanomaterials and their preparation and application, to achieve the effects of inhibiting cell proliferation, good dispersion and biocompatibility, and easy synthesis and purification
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Embodiment 1
[0085] (1) Weigh 4.22mg Ad-COOH, 41.62mg EDC·HCl, and 21.98mg NHS respectively, and dissolve them in 5mL of DMSO solution, then add EDC·HCl and NHS solution dropwise to the Ad-COOH solution, at room temperature Stir for 3h. Then weigh 100mg G3.NH 2 Dissolved in 5 mL of DMSO solution, the resulting activated Ad-COOH solution was added dropwise to G3.NH 2 solution, continue to react for 3 days, transfer the obtained product to a dialysis bag with a molecular weight cut-off of 1000, dialyze in distilled water for three days (4L×3), and then freeze-dry to obtain dry Ad-G3 at last,- Store at 20°C for later use.
[0086] (2) Weigh 43.64 mg β-CD and 62.34 mg CDI respectively, dissolve them in 5 mL of DMSO solution, then add the CDI solution dropwise into the β-CD solution, and stir at room temperature for 6 hours. Weigh 40mg G5.NH 2 Dissolved in 5 mL of DMSO solution, the resulting activated β-CD solution was added dropwise to G5.NH 2 solution, continue to react for 60h, transfe...
Embodiment 2
[0092] Carry out NMR characterization to embodiment 1 step (1), step (2), Ad-G3 prepared by step (3), G5-CD, G5-CD / Ad-G3, 1 H NMR characterization results are as attached to the instructions figure 2 As shown in a, there is a proton peak at the chemical shift of 1.48-1.9ppm, which is a characteristic group proton peak in the molecular structure of Ad, according to G3.NH 2 The integral area ratio between each G3.NH can be calculated 2 1.1 Ad molecules were linked to it. Instructions attached figure 2 As shown in b, there are proton peaks of CD at chemical shifts of 3.5-4.1 and 5.0ppm, and methylene proton peaks of G5 at chemical shifts of 2.2-3.4ppm. According to the integrated area, it can be calculated that 7.55 CDs are connected to the surface of G5 molecular. See attached figure 2c: According to the integrated area corresponding to the proton peaks of CD and Ad, it can be finally calculated that 4.2 Ad-G3 molecules are connected to the surface of G5-CD. 2D NOESY is us...
Embodiment 3
[0094] The G5-CD / Ad-G3 / miRNA 21i complex prepared in step (4) of Example 1 was subjected to a gel retardation experiment. Prepare 8 wells of agarose gel (1.0% w / v) containing ethidium bromide (1 mg / mL), and place at room temperature until the agarose gel solidifies. The amount of siRNA was 1 μg / well, G5-CD / Ad-G3 / miRNA 21i complexes were prepared according to different N / P ratios of 0, 0.125, 0.25, 0.5, 1, 2 and 5, incubated for 30 min, and naked miRNA 21i was used as control. Then the corresponding G5-CD / Ad-G3 / miRNA 21i complexes were added to the wells of the agarose gel respectively, with a voltage of 80V and a time of 30min. The migration of miRNA 21i in the gel was analyzed using a gel imager. The result is attached to the manual Figure 4 shown. The results showed that when the N / P ratio was greater than or equal to 1, G5-CD / Ad-G3 / miRNA 21i could completely compress miRNA 21i and prevent miRNA 21i from electromigration.
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