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Lipid peroxide generator and preparation method and application of lipidosome

A technology of peroxide and generator, which is applied in the direction of liposome delivery, medical preparations containing active ingredients, and pharmaceutical formulas, etc. Oxidation, limited Fenton reaction efficiency and other problems, to achieve the effect of overcoming the low efficiency of Fenton reaction, simple preparation method and high safety

Inactive Publication Date: 2019-10-22
CHINA PHARM UNIV
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  • Abstract
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  • Application Information

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Problems solved by technology

[0005] However, although the pH of the tumor microenvironment is weakly acidic, it is difficult to meet the optimal reaction conditions (pH = 3-4) required for the Fenton reaction; and the intracellular H 2 o 2 The content is limited, which greatly limits the efficiency of the Fenton reaction, and usually requires the addition of a large amount of iron and the introduction of exogenous H 2 o 2 or stimulate endogenous H 2 o 2 The generation of is used to enhance the Fenton reaction
Based on the iron ion-dependent lipid peroxide is the main executor of ferroptosis, and the general ferroptosis delivery system cannot effectively realize the lipid peroxidation triggered by the Fenton reaction, therefore, the development of a method that can overcome the Fenton reaction A ferroptosis delivery system that is limited by low efficiency and can generate a large amount of lipid peroxides is of great significance for improving the treatment of tumor ferroptosis

Method used

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  • Lipid peroxide generator and preparation method and application of lipidosome

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Embodiment 1

[0059] Detection of Fe using o-phenanthroline reagent 2+ , first add a series of concentrations of GSH and o-phenanthroline detection reagents to the FAC solution, vortex and shake at 37 o C was reacted on a constant temperature shaker for 24 h, and the absorbance at 510 nm was detected using a microplate reader. figure 1 For the purpose of adding different concentrations of GSH to FAC in this example, the amount of ferrous iron produced, the results verified that GSH can reduce ferric iron to ferrous iron. The results showed that: with the increase of GSH concentration, the absorbance of the mixture at 510 nm gradually increased, and when GSH was greater than 1 mM, the absorbance of the mixture was the same as that of the positive control FeSO 4 The same, thus indicating that FAC can be effectively reduced to Fe in tumor cells with high concentration of GSH 2+ , and then two valence states of iron ions exist simultaneously to form an iron redox pair.

Embodiment 2

[0061] First add Fe to Lip and Lip@FAC 2+ , at 37 o C reacted overnight in a constant temperature shaker, and the blank liposome Lip solution was added to Fe 2+ The front and back are clear and transparent, and the appearance has no obvious change. At the same time, the appearance of the Lip@FAC solution has no obvious change after overnight. When adding Fe to the Lip@FAC solution 2+ , a yellow precipitate appeared in the solution after reacting overnight. Then GSH was added to Lip and Lip@FAC respectively, and the Lip solution did not change significantly before and after adding GSH, but a large amount of yellow precipitate appeared in Lip@FAC before and after adding GSH; figure 2 shown. Perform mass spectrometry analysis on the above substances, such as image 3 As shown, the results show that Lip added Fe 2+ , before and after GSH and Lip@FAC itself, the molecular weight did not change significantly, and the molecular ion peak of 804 appeared, but the molecular ion p...

Embodiment 3

[0063] In this experiment, the membrane extrusion method was used, and the ethanol solution of soybean lecithin was added dropwise to the FAC physiological saline solution under the vortex state, and a dialysis bag with a molecular weight cut-off of 14 KDa was used for dialysis in normal saline overnight to remove ethanol and Unencapsulated FAC; on this basis, a liposome extruder is further used to prepare liposomes with a smaller particle size and uniformity.

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Abstract

The invention discloses a lipid peroxide generator and a preparation method and application of a lipidosome. The lipid peroxide generator comprises lipid and an iron source. The preparation method ofthe lipidosome comprises the steps that the iron source is wrapped by the lipid to form the lipidosome. The lipidosome is taken into and enters tumor cells through caveolin-mediated endocytosis, ironions are released in an acid lysosome environment, an iron oxidation-reduction pair formed in the cytoplasm can catalyze lipid peroxidation of unsaturated lipid, accumulation of the lipid peroxide inthe cells is caused, finally, iron apoptosis of the tumor cells is caused, and the aim of treating the tumors is achieved. Through in vitro representation, it is explained that the lipid peroxide generator can generate the lipid peroxide under triggering of the iron oxidation-reduction pair; through anti-tumor activity evaluation on the cell level and animal cell, it is proved that the lipid peroxide generator can generate a large amount of lipid peroxide in the tumor cells, and iron apoptosis-mediated tumor treatment is effectively achieved.

Description

technical field [0001] The invention relates to a lipid peroxide generator and its preparation method and application, belonging to the technical field of liposomes. Background technique [0002] Cancer is a disease that seriously threatens human health. According to the "Global Cancer Report 2018", there were 18.1 million new cancer cases and 9.6 million deaths worldwide, further increasing the global cancer burden. Currently, the commonly used clinical treatment methods are surgery, radiotherapy, and chemotherapy. Among them, chemotherapy mostly kills tumor cells through apoptosis, necrosis, and autophagy. However, most chemotherapeutic drugs have defects such as low solubility, poor stability, strong toxic and side effects, and multi-drug resistance after long-term use. Therefore, discovering and utilizing a new cell death mode is a new way to realize cancer therapy. [0003] In 2012, Stockwell et al. defined the process of programmed cell death caused by the accumulat...

Claims

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Application Information

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IPC IPC(8): A61K9/127A61K33/26A61K45/06A61P35/00
CPCA61K9/127A61K33/26A61K45/06A61P35/00A61K2300/00
Inventor 姜虎林何玉静刘笑影万星
Owner CHINA PHARM UNIV
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