Charge inversion type polypeptide composite nano drug as well as preparation method and application thereof
A charge inversion, nano-drug technology, applied in the field of biomedicine, can solve problems such as multi-drug resistance, and achieve the effect of enhancing internalization, mild photothermal therapy, and reversing tumor drug resistance
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Embodiment 1
[0037] Preparation of charge reversal polypeptide compound nano medicine
[0038] 1. The preparation method of polyethylene glycol-b-poly(L-lysine)-b-poly(L-cysteine) is as follows figure 2 Shown.
[0039] Step 1: Refer to the existing literature to obtain ε-benzyloxycarbonyl-L-lysine anhydride and S-o-nitrobenzyl-L-cysteine. In the glove box, take 100 mg of amino polyethylene glycol, Dissolve in 3 mL of anhydrous N,N-dimethylformamide, then add 73.5 mg of ε-benzyloxycarbonyl-L-lysine anhydride, react at room temperature for 24 h, then add S-o-nitrobenzyl -L-cysteine 259.5 mg, after continuing the reaction for 48 h, the reaction solution was settled in 24 mL of anhydrous ether, centrifuged again, repeated 3 times, and vacuum dried for 24 h to obtain a white solid. The yield is 80.2~85.5%.
[0040] Step 2: Take 50 mg of the white solid obtained in Step 1 and dissolve it in 100 mL of a mixed solvent of N,N-dimethylformamide / acetonitrile (volume ratio 4: 1) and place it in an ultra...
Embodiment 2
[0052] Effect of charge-reversal polypeptide compound nanomedicine on breast cancer cells
[0053] The charge-reversal polypeptide composite nanomedicine and doxorubicin prepared in Example 1 were prepared with cell culture medium to prepare the concentration of doxorubicin to be 0.5, 1, 2, 4, 8, 16, 32, 64 µg, respectively. / mL, and then cultured with MCF-7 cells (breast cancer) and MCF-7 / ADR cells (adriamycin-resistant breast cancer) for 48 h. In addition, a separate charge-reversal polypeptide composite nano-drug is required Group, after culturing for 4 h, irradiate it with near-infrared laser for 5 min (808nm, 1.6W / cm 2 ), continue to cultivate for 48 h. Using MTT method for cell viability test, the results are as follows Image 6 with Figure 7 Shown. Image 6 with Figure 7 In the figure, the abscissa DOX refers to the concentration of doxorubicin. In the figure, pH 7.4 and pH 6.8 refer to the experimental group of charge-reversal polypeptide composite nanomedicine at pH 7....
Embodiment 3
[0056] Experiment on the effect of charge-reversal polypeptide compound nano-medicine on MCF-7 / ADR tumor growth
[0057] The mice vaccinated with MCF-7 / ADR tumor-bearing mice were divided into seven groups: normal saline, normal saline + NIR, doxorubicin (5 mg / kg), charge reversal polypeptide composite nano-drug (2 mg / mL) ), charge reversal polypeptide compound nano medicine + NIR (2mg / mL). One injection on day 0, and 12 hours after injection, the saline + NIR and the charge-reversal polypeptide composite nano-drug + NIR were illuminated for 5 min (808 nm, 1.6 W / cm2), and the mice were exposed every other day Weigh and measure the tumor volume, the results are as follows Figure 8 Shown.
[0058] Figure 8 In the figure, the abscissa represents the number of days the mice received the experiment, the ordinate in A panel represents the tumor volume of the mouse, and the ordinate in panel B represents the weight of the mouse. In the figure, PBS refers to the normal saline injection...
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