Hydrogen peroxide-responsive liposomal nanoprobe and preparation method and application thereof

A technology of hydrogen peroxide and nano-probes, which is applied in the field of biomedicine to achieve high-sensitivity detection of inflammation and tumors, long blood circulation time, and good dispersibility

Active Publication Date: 2017-05-17
JE & NA BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the detection of hydrogen peroxide in vivo using photoacoustic imaging is rarely reported.

Method used

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  • Hydrogen peroxide-responsive liposomal nanoprobe and preparation method and application thereof
  • Hydrogen peroxide-responsive liposomal nanoprobe and preparation method and application thereof
  • Hydrogen peroxide-responsive liposomal nanoprobe and preparation method and application thereof

Examples

Experimental program
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preparation example Construction

[0043] The preparation method of the liposome nanoprobe of hydrogen peroxide response provided by the present invention comprises the following steps:

[0044] Dipalmitoylphosphatidylcholine (DPPC), cholesterol, distearoylphosphatidylethanolamine-polyethylene glycol (DSPE-PEG) and 2,2-azino-bis(3-ethyl-benzothiazole- 6-sulfonic acid) diammonium salt (ABTS) is dissolved in methanol with a molar ratio of 6:4:0.5:12, and then forms a lipid film by rotary evaporation;

[0045] Then 0.2mg / ml of horseradish peroxidase (HRP) phosphate buffered saline (PBS) was added thereto, and the mixed solution was frozen, then thawed at 65°C;

[0046] Filter the solution after five times of freezing and thawing on a 200-nm filter membrane for ~50 times, and then purify it through an S-300 gel column to remove free 2,2-azino-bis(3-ethyl-benzo Thiazole-6-sulfonic acid) diammonium salt (ABTS) and horseradish peroxidase (HRP).

[0047] The hydrogen peroxide-responsive liposome nanoprobe provided by...

Embodiment 1

[0049] Example 1: Preparation of hydrogen peroxide-responsive liposome nanoprobes (Lipo@HRP&ABTS):

[0050] Based on liposome, horseradish peroxidase (HRP) is loaded in its hydrophilic cavity, and the hydrophobic layer is used to load 2,2-azino-bis(3-ethyl-benzothiazole) -6-sulfonic acid) diammonium salt (ABTS), forming Lipo@HRP&ABTS. Specifically include the following steps:

[0051] Dipalmitoylphosphatidylcholine (DPPC), cholesterol, distearoylphosphatidylethanolamine-polyethylene glycol (DSPE-PEG) and 2,2-azino-bis(3-ethyl-benzothiazole- 6-sulfonic acid) diammonium salt (ABTS) was dissolved in methanol with a molar ratio of 6:4:0.5:12, and then a lipid film was formed by rotary evaporation; then 0.2 mg / ml of horseradish peroxidase was added thereto (HRP) Phosphate buffered saline solution (PBS) and treat the mixture at 65°C; filter the solution after five times of freezing and thawing on a 200nm filter membrane for about 50 times, and then pass through the S-300 gel colum...

Embodiment 2

[0053] Example 2: Specificity of the hydrogen peroxide response of Lipo@HRP & ABTS

[0054] In order to study the specificity of the hydrogen peroxide response of Lipo@HRP&ABTS, this example incubated Lipo@HRP&ABTS or Lipo@ABTS with different reactive oxygen species, including hydrogen peroxide (H 2 o 2 ), tert-butyl hydroperoxide (TBHP), hypochlorite ( - OCl), hydroxyl radical ( OH), tert-butyl radical ( OtBu), nitric oxide radical (NO ), peroxide radical (O 2 - ).

[0055] attached image 3 It is the absorption at 800nm ​​measured after the above Lipo@HRP&ABTS was incubated with different reactive oxygen species for half an hour. From image 3 From the absorption data, it can be seen that Lipo@ABTS does not respond to almost all reactive oxygen species. The absorption of Lipo@HRP&ABTS at 800nm ​​was significantly enhanced after incubation with hydrogen peroxide, while the response to other types of reactive oxygen species was not very strong. Therefore, Lipo@HRP&ABTS...

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Abstract

The invention relates to the technical field of biomedicine, in particular to a hydrogen peroxide-responsive liposomal nanoprobe and a preparation method and application thereof. The liposomal nanoprobe comprises a liposome, HRP and ABTS, wherein a hydrophilic cavity of the liposome is loaded with HRP, and a hydrophobic layer of the liposome is loaded with ABTS. The hydrogen peroxide-responsive liposomal nanoprobe prepared by the invention has very good hydrogen peroxide responsiveness, and can realize the detection of nano hydrogen peroxide. Compared with the traditional method for detecting hydrogen peroxide by fluorescence or bioluminescence, the photoacoustic imaging detection can significantly improve the depth of penetration of tissues to achieve the hydrogen peroxide imaging of deep tissues; the photoacoustic imaging of the liposome nanoprobe can monitor the occurrence and development of inflammation in real time, and the liposome nanoprobe can react with hydrogen peroxide in a tumor microenvironment to achieve the photoacoustic imaging of tumors.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a hydrogen peroxide-responsive liposome nanoprobe and its preparation method and application. Background technique [0002] Hydrogen peroxide plays an important role in regulating various physiological processes including cell growth, immune response and aging. Abnormal secretion of hydrogen peroxide is often closely related to many diseases, such as: cancer, diabetes, inflammation, cardiovascular disease and neurological disease. Therefore, the development of accurate and sensitive hydrogen peroxide detection probes will have very important clinical value. It can not only be used for disease diagnosis, but also help doctors understand the mechanism and development of diseases well. However, since the concentration of hydrogen peroxide in physiological environment is quite low (∼<50 μM), it is still a very challenging task to develop a technique that can be used for clini...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/22A61K41/00A61P35/00
CPCA61K41/0052A61K49/227
Inventor 刘庄陈倩
Owner JE & NA BIOTECH CO LTD
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