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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

Active Publication Date: 2021-07-23
THE SECOND HOSPITAL AFFILIATED TO WENZHOU MEDICAL COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, photosensitizers such as IR780 encounter the following problems during use: (1) limited by their own hydrophobicity, they cannot be directly applied in vivo
(2) It is easily metabolized by the liver and kidney in the body, and the biological half-life is short
(3) When light is applied, the local tissue cannot provide enough oxygen, so the efficiency of phototherapy is low
[0006] Patent application number CN202010907422.2 discloses a preparation method of a liposome composite material using light-controlled release of tungsten sulfide quantum dots and vancomycin, wherein distearoylphosphatidylethanolamine-polyethylene glycol- Hydroxyl (DSPE-PEG-OH) and cholesterol cooperate with dipalmitoylphosphatidylcholine (DPPC) to obtain a lipid film, but under this scheme, cholesterol is added to reduce the toxin neutralization effect of phospholipids, and the distearoyl phospholipids used Ethanolamine-polyethylene glycol-hydroxyl (DSPE-PEG-OH) is a hydroxyl group, its properties are relatively stable, and the reaction effect is poor

Method used

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  • Preparation method of liposome containing vancomycin, IR780 and oxygen-carrying perfluorohexane
  • Preparation method of liposome containing vancomycin, IR780 and oxygen-carrying perfluorohexane
  • Preparation method of liposome containing vancomycin, IR780 and oxygen-carrying perfluorohexane

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Effect test

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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Abstract

The invention discloses a preparation method of a liposome containing vancomycin, IR780 and oxygen-carrying perfluorohexane. The preparation method comprises the following steps: step 1, carrying out a reaction on vancomycin (Van) and a PEG (Polyethylene Glycol) molecule (DSPE-PEG2000-NHS) to obtain DSPE-PEG2000-Van; step 2, preparing a liposome suspension from sphingomyelin, DSPE-PEG2000-Van and IR780 by adopting a thin film hydration method; step 3, adding perfluorohexane (PFH) and carrying out ultrasonic emulsification on the PFH; and step 4, injecting oxygen to obtain a finished product. According to the invention, the Van reaction efficiency can be improved, the toxin neutralization effect is improved, the photodynamic therapy efficiency is high, and the treatment effect on anaerobic bacteria is better.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a preparation method of liposomes containing vancomycin, IR780 and perfluorohexane. Background technique [0002] In recent years, nanotechnology has been increasingly used in the diagnosis and treatment of diseases, playing an important role in the fields of tumors or bacterial infections. Methicillin-resistant Staphylococcus aureus (MRSA) is a common type of "super bacteria" in clinical practice, which shows heterogeneous drug resistance and broad-spectrum drug resistance, and currently there is no specific drug for clinical use For the treatment of infections caused by MRSA. [0003] Vancomycin (Vancomycin, Van) is clinically used as a narrow-spectrum antibiotic, which is only effective against Gram-positive bacteria. MRSA is particularly sensitive to Van, so Van is mainly used clinically for serious infections caused by MRSA, such as pneumonia, endocarditis and sepsis. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/14A61K41/00A61K33/00A61K9/127A61K47/24A61K47/06A61P31/04A61P29/00
CPCA61K38/14A61K41/0057A61K33/00A61K9/127A61K47/24A61K47/06A61P31/04A61P29/00A61K2300/00
Inventor 陈一杰诸葛德力赵应征陈梦纯张铭瑶
Owner THE SECOND HOSPITAL AFFILIATED TO WENZHOU MEDICAL COLLEGE
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