Preparation method of liposome containing vancomycin, IR780 and oxygen-carrying perfluorohexane
A vancomycin and perfluorohexane technology, applied in the field of biomedicine, can solve the problems of low phototherapy efficiency, inability to directly apply, hydrophobicity limitation, etc., and achieve the effect of efficiently inhibiting the activity of anaerobic bacteria, efficient photodynamic therapy, Improve the effect of treatment
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Embodiment 1
[0058] A preparation method of liposome containing vancomycin, IR780 and perfluorohexane. In this method, vancomycin is reacted with a PEG molecule with a different functional group (DSPE-PEG2000-NHS) in advance to obtain DSPE-PEG2000-Van. When preparing liposomes, use thin film hydration method, fully dissolve sphingomyelin, DSPE-PEG2000-Van, IR780 in chloroform, fully remove the organic solvent by rotary evaporation, and then add sterile 10% sucrose solution for hydration Lipid film. Subsequently, a certain amount of PFH was added into the liposome suspension, and the PFH was ultrasonically emulsified by a probe. Finally, the liposome suspension was filled with oxygen by syringe before use. Through prescription screening, the optimal ratio of each component is: 7848.9:1:4:1461:1.744 (sphingomyelin: DSPE-PEG2000: vancomycin: perfluorohexane: IR780, mol / mol).
Embodiment 2
[0060] Synthesis of DSPE-PEG2000-Van
[0061] Accurately weigh DSPE-PEG2000-NHS and dissolve in DMF, weigh Van and dissolve in ultrapure water, according to 1:2, 1:3, 1:4, 1:5 (DSPE-PEG2000-NHS: Van, mol / mol) were mixed, and 10 μL of triethylamine was added as a catalyst to react for 18 hours.
Embodiment 3
[0063] Determination of Grafting Rate of DSPE-PEG2000-Van by HPLC
[0064] Chromatographic conditions: stationary phase: C18 column
[0065] Mobile phase: 0.025mol / L potassium dihydrogen phosphate Buffer: acetonitrile (70%-92%)
[0066] Flow rate: 1mL / min
[0067] Column temperature: 25°C
[0068] Injection volume: 30μL
[0069] Detection wavelength: 230nm
[0070] Utilize the ultrafiltration tube (3000Da) to centrifuge the different proportions of the reaction solution, and put the filtered solution and the reaction stock solution into HPLC, and compare the peak height of vancomycin.
[0071] As identified by HPLC, there was a significant difference in the grafting rate between different ratios of Van and DSPE-PEG2000-NHS ( figure 1 , image 3 , Figure 4 , Figure 5 , Figure 6 ), the 1:2 grafting ratio was 61.129%, the 1:3 grafting ratio was 83.237%, the 1:4 grafting ratio was 88.110%, and the 1:5 grafting ratio was 77.873%. Among them, the grafting rate of 1:4 is...
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