A kind of ATP responsive nano micelles, preparation method and application
A technology of nanomicelle and nanomicelle solution, applied in the field of bioengineering, can solve the problems of non-viral gene carrier targeting and low loading efficiency, achieve high drug loading and encapsulation rate, enhance stability and smooth surface Effect
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Embodiment 1
[0048] Embodiment 1 Preparation of polylysine-phenylboronic acid and dopamine-hyaluronic acid
[0049] (1) Preparation of polylysine-phenylboronic acid:
[0050]
[0051] According to the above polymer synthesis route, the specific description is as follows:
[0052] Add 4-(bromomethyl)phenylboronic acid and polylysine into anhydrous dimethyl sulfoxide, stir and react at 80°C for 24 hours, the 4-(bromomethyl)phenylboronic acid and polylysine The molar ratio of lysine is 8:1-10:1 respectively, and finally the product is dialyzed and freeze-dried to complete the preparation of polylysine-phenylboronic acid. H NMR spectrum such as image 3 showed that polylysine-phenylboronic acid (PLL-PBA) was successfully synthesized. Note: PLL-polylysine, PBA-phenylboronic acid.
[0053] (2) Preparation of dopamine-hyaluronic acid:
[0054]
[0055] The synthetic route of dopamine-hyaluronic acid (HA-Cat), wherein HA is hyaluronic acid, Dopamine is dopamine, and EDC is 1-(3-dimethyl...
Embodiment 2
[0058] Example 2 Preparation of HA / Bcl-2 siRNA-PLL-PBA-p-gp siRNA-DOX nanomicelles
[0059] Put 5 mg of PLL-PBA and 2.5 mg of Bcl-2 siRNA and p-gp siRNA into 10 mg of DOX aqueous solution to form a Bcl-2 siRNA-PLL-PBA-p-gp siRNA-DOX nanocomplex loaded with genes and drugs, Then put HA into the above-mentioned nano-micelle solution and stir to form HA / Bcl-2 siRNA-PLL-PBA-p-gp siRNA-DOX nano-micelle. The results of the nanomicelles by transmission electron microscopy are as follows: Figure 4 As shown, the prepared nanoparticles are circular and have good dispersibility, and the particle size is mainly concentrated between 100-200nm, especially the particle size of 150nm. Note: Dox is doxorubicin hydrochloride, nm-nanometer.
Embodiment 3
[0060] Example 3 Cytotoxicity of HA / PLL-PBA
[0061] MCF-7 cells were plated in 96-well plates, the density of cells per well was 1×105, and cultured overnight at 37°C in a cell culture incubator with a volume fraction of CO2 of 5%. Nanomicelles of different concentrations were added to the 96-well plate (the cell group with only the culture medium was the negative control group), and 5 replicate wells were set for each experimental condition. After 24 hours of culture, 20 μL of CCK8 (5 mg / mL) solution was added, and the culture was continued for 2 hours. After that, the culture solution was aspirated, tested at 450 nm in a microplate reader, and the cell survival rate was calculated. Among them, CCK8 is a rapid and highly sensitive kit for cell viability and cytotoxicity detection.
[0062] The experimental results are shown in 5. Compared with the control group, the OD value of the cells incubated with nanoparticles within 24 hours did not change significantly, indicating t...
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