Application of a pH-sensitive liposome producing NO
A liposome and sensitive technology, applied in the application field of biomedical materials, can solve the problems of normal cytotoxicity and side effects of tumor cells, poor selectivity of tumor tissue, multidrug resistance, etc., to overcome multidrug resistance of tumors, benefit Ingestion, the effect of enhancing the curative effect
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Embodiment 1
[0020] Embodiment 1: A kind of preparation method of the pH-sensitive liposome that produces NO
[0021] (1) Chol 2.01 mg, HSPC 47.03 mg, DSPE-PEG 2000 13.75 mg, DOTAP 7.0 mg and paclitaxel 4.05 mg were dissolved in 30 mL chloroform solution;
[0022] (2) Use a rotary evaporator to rotate at a speed of 100 r / min for 4 h at room temperature to obtain a uniform film-like material;
[0023] (3) After elution with 6.78 mL of PBS buffer solution containing 2.2 mM DETA NONOate at pH = 8.0, ultrasonic treatment was performed for 10 min at a power of 300 W by an ultrasonic cell disruptor to form cells loaded with NO donors and Paclitaxel liposomes;
[0024] (4) HSPC, DOTAP and DSPE-PEG 2000 The ratio of the sum of the mass of PEG-PLL-DMA to the mass of PEG-PLL-DMA is 1:6. Add PEG-PLL-DMA and stir at room temperature for 1 h to obtain pH-sensitive liposomes that produce NO.
Embodiment 2
[0025] Embodiment 2: A kind of preparation method of the pH-sensitive liposome that produces NO
[0026] (1) Chol 2.01 mg, HSPC 47.03 mg, DSPE-PEG 2000 13.75 mg, DOTAP 15.05 mg and paclitaxel 4.05 mg were dissolved in 30 mL chloroform solution;
[0027] (2) Use a rotary evaporator to rotate at a speed of 100 r / min for 4 h at room temperature to obtain a uniform film-like material;
[0028] (3) After elution with 7.58 mL of PBS buffer solution containing 1.375 mM DETA NONOate at pH=8.0, ultrasonic treatment was performed for 10 min at a power of 300 W using an ultrasonic cell disruptor to form packets loaded with NO donors and Paclitaxel liposomes;
[0029] (4) HSPC, DOTAP and DSPE-PEG 2000 The ratio of the sum of the mass of PEG-PLL-DMA to the mass of PEG-PLL-DMA is 1:30. Add PEG-PLL-DMA and stir at room temperature for 1 h to obtain pH-sensitive liposomes that produce NO.
Embodiment 3
[0030] Example 3: Cell uptake experiment of pH-sensitive liposomes producing NO
[0031] A549 / T human lung cancer drug-resistant strain cells in the logarithmic growth phase were digested with 0.25% trypsin, centrifuged (1000rpm, 5 min), and diluted with RPMI 1640 medium containing 10% FBS to a concentration of 1×10 5 / mL of single cell suspension at 2×10 5 The density of cells / well was seeded in a 6-well culture plate, 2 mL per well, cultured at 37°C for 24 h, the original medium was discarded, washed with PBS, and 2 mL of medium containing different liposomes at pH 7.4 and 6.5 were added respectively (PEG-PLL-DMA modified liposomes containing DETANONOate, PEG-PLL-SA modified liposomes containing DETA NONOate, PEG-PLL-DMA modified liposomes without DETA NONOate), at the same time, the uptake experiment Coumarin-6 was used instead of paclitaxel, and the concentration of coumarin-6 was 1 μg / mL. After culturing for 1 h, the medium was discarded, washed twice with PBS, digested ...
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