Mitochondria-targeted photo-thermal/chemotherapy synergistic nano-drug delivery particle as well as preparation method and application thereof
A nano drug and chemotherapy drug technology, applied in the field of biomedicine, can solve the problems of tumor recurrence and poor curative effect, and achieve the effects of inducing cell apoptosis, uniform shape, and promoting the production of ROS
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Embodiment 1
[0038] Example 1 Preparation of nanocarrier delivery system
[0039] 1) DSPE-PEG 2000 - Synthesis of DOX (lipid-DOX)
[0040] 67.5mg DPSE-PEG 2000 - Dissolve NHS and 20mg DOX in 10mL DMSO, add 12μL triethylamine, stir at 25°C for 24h, then dialyze, add DMSO to the dialysis bag every 12h to completely dissolve the sample, repeat 6 times, then dialyze the sample for 72h, freeze the sample use after drying 1 H-NMR confirmed the structure.
[0041] 2) Synthesis of gold nanorods (AuNR)
[0042] Add 0.5mL CTAB (0.2M) solution to 0.5mL HAuCl 4 (0.5mM) solution, stir well, then add 0.06mL ice-cold NaBH dropwise 4 (0.01M) solution, until the solution turns brown, continue stirring for 2 min, and set aside. 50mL CTAB (0.2M) solution was added to a 250mL round bottom flask, and 2.8mL AgNO was added with stirring 3 (4mM) solution, 3.75mL HAuCl 4 (23mM) solution and 47.5mL of ultrapure water, stir well. 1.6 mL of ascorbic acid (0.08 M) solution was then added dropwise, during whi...
Embodiment 2
[0045] Example 2 Characterization of physical and chemical properties
[0046] 1) Transmission Electron Microscopy (TEM)
[0047] AuNR and AuNR-lipid-DOX / PTX solutions were added dropwise on a copper mesh with a carbon support film. After the solvent was evaporated, the morphology was observed under TEM and images were collected. The accelerating voltage was 120 kV. The result is as figure 1 shown. It can be seen from the results that the prepared gold nanorods have good dispersion and the length is 45-50 nm ( figure 1 A), after wrapping the lipid layer ( figure 1 B), the shape of the gold nanorods did not change.
[0048] 2) Determination of particle size and Zeta potential
[0049] The particle size and Zeta potential of AuNR and AuNR-lipid-DOX / PTX solution were measured by DLS analyzer. Disperse a small amount of sample into ultrapure water, ultrasonically disperse it uniformly, and measure it at a test temperature of 25°C. from figure 2 It can be seen from the res...
Embodiment 3
[0052] Example 3 Determination of drug loading and drug release in vitro
[0053] 1) The drug loadings of lipid-DOX and PTX in AuNR-lipid-DOX / PTX were determined by UV-vis and high performance liquid chromatography (HPLC), respectively. The prepared AuNR-lipid-DOX / PTX was destroyed and dissolved with methanol, and the AuNR was removed by centrifugation. The supernatant was measured by UV-vis and HPLC, respectively, and the content of the two drugs was calculated by the external standard method. The content of lipid-DOX was calculated by measuring the absorbance at 490 nm in UV-vis spectrum of it and the external standard. The content of PTX was calculated by measuring the peak area of PTX and the external standard in HPLC. The HPLC measurement conditions were as follows: the absorption wavelength was 227 nm; the mobile phase was methanol:acetonitrile:water=40:40:20, and the flow rate was 1 mL / min. The loading of lipid-DOX in AuNR-lipid-DOX / PTX was determined to be 20% (w / w...
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