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Targeting atherosclerotic focus cell membrane bionic drug-delivery system preparation method and product and application thereof

A technology for atherosclerosis and drug delivery system, which is applied in the field of pharmacy to achieve good in vivo and in vitro targeting effects

Inactive Publication Date: 2018-04-27
SOUTHWEST UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It has been reported in the literature that bionic nano-drug delivery systems wrapped in macrophage membranes have been used to construct bionic drug delivery systems targeting breast cancer with lung metastases, showing good potential for precise treatment of lesions, but no macrophage membrane-wrapped Nanoparticles targeting atherosclerotic lesions report

Method used

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  • Targeting atherosclerotic focus cell membrane bionic drug-delivery system preparation method and product and application thereof
  • Targeting atherosclerotic focus cell membrane bionic drug-delivery system preparation method and product and application thereof
  • Targeting atherosclerotic focus cell membrane bionic drug-delivery system preparation method and product and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1. Preparation and Characterization of Macrophage Membrane Biomimetic Nanocarriers

[0030] (1) Extraction of macrophage cell membrane:

[0031] Refer to the preparation method of Cao H et al. (Cao H, Dan Z, He X, et al. Liposomes Coated with Isolated Macrophage Membrane Can Target Lung Metastasis of Breast Cancer [J]. Acs Nano, 2016, 10:7738-7748.). Collect macrophages, add 4℃TM buffer (pH 7.4, 0.01mol L -1 Tris and 0.001mol·L -1 MgCl) was resuspended, and the concentration of the cell suspension was adjusted to 2.5×10 7 / mL, squeeze the cell membrane back and forth with a micro-homogenizer (20 times), add an appropriate amount of 1mol L -1 Sucrose was mixed with cell homogenate to make the final concentration of sucrose 0.25mol L -1 . Then proceed as follows:

[0032] 1) Centrifuge at 2000g for 10min at 4°C, and take the supernatant;

[0033] 2) Centrifuge at 3000g for 30min at 4°C, discard the supernatant;

[0034] 3) Add 0.25mol·L at 4°C -1 Resuspe...

Embodiment 2

[0039] Example 2, establishment and evaluation of human umbilical vein endothelial cell injury model

[0040] (1) Conditions for establishment of human umbilical vein endothelial cell inflammatory injury model:

[0041] Take HUVEC cells in the logarithmic growth phase, and use 1×10 per well 4 / mL density inoculated in 96-well plate, added 100 μL to each well, and adhered to the wall overnight; aspirated the medium, added serum-free medium to starve for 12 hours, and gave different concentrations of LPS (final concentrations were 0, 1.56, 3.125, 6.25, 12.5, 25 , 50, 100, 200μg·mL -1 ), three replicate wells in each group, placed in CO 2 The culture was continued in the incubator; 20 μL of MTT solution (5 g·L -1 ), 4 hours later, 150 μL of dimethyl sulfoxide was added to each well, and the absorbance (A) value was measured at a wavelength of 490 nm with a microplate reader after shaking at 37° C. for 30 minutes.

[0042] (2) α4β1 integrin and VCAM-1 protein expression detect...

Embodiment 3

[0048] Example 3. Preliminary evaluation of in vivo targeting of macrophage membrane biomimetic nanocarriers

[0049] (1) Establishment of male SPF grade ApoE in atherosclerosis model - / - Gene knockout mice and C57BL / 6J mice were fed with high-fat diet (78% basal diet+21% fat+1.25% cholesterol) for 12 weeks respectively.

[0050] (2) Evaluation of atherosclerosis model

[0051] The model mice were taken and injected intraperitoneally with 3% sodium pentobarbital (60 mg·kg -1 ). After the mouse is anesthetized, put it on an ice pack and dissect the thorax. Use a 1mL syringe to quickly collect 0.8-1mL of blood from the left ventricle, quickly transfer it to an EP tube, centrifuge at 3500g, 4°C for 15min, separate the plasma, and store at 4°C. Store in the refrigerator for later use. After fully perfused with pre-cooled PB buffer and 4% paraformaldehyde (flow rate 2mL·min -1 ), the thoracic aorta (starting from the root of the heart to the branches of the bilateral iliac art...

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Abstract

The invention relates to a targeting atherosclerotic focus cell membrane bionic drug-delivery system preparation method and a product and an application thereof. The method comprises the following steps: extracting a macrophage membrane, repeatedly extruding the macrophage membrane to prepare macrophage membrane vesicle, then mixing the material with polylactic acid-glycolic acid copolymer nanoparticles, and repeatedly extruding a filter membrane to prepare a spherical drug-delivery system. The system has an obvious core / shell structure, an average particle size is 167+ / -6.12 nm, and alpha4beta1 integrin is kept on the surface; a VCAM-1 acceptor can be effectively identified, and the system can present good in-vitro and vivo targeting, and is expected to be used for treating atherosclerosis and related diseases.

Description

technical field [0001] The invention belongs to the field of pharmacy, and relates to a preparation method for cell membrane biomimetic drug delivery targeting atherosclerotic lesions, and also relates to a product prepared by the method and its application. Background technique [0002] Atherosclerosis (AS) is a common disease that seriously endangers human health, and is the main pathological basis of ischemic cardiovascular and cerebrovascular diseases such as coronary heart disease, cerebral infarction, and peripheral vascular disease. So far, the pathogenesis of AS has not been fully elucidated, there are many theories, and among these theories, the theory of inflammation is one of the most recognized theories. This theory believes that the injury of arterial intima caused by various risk factors is the initiating link of atherosclerosis. In this process, activated vascular endothelial cells secrete some adhesion molecules, such as vascular cell adhesion molecule-1 (va...

Claims

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Application Information

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IPC IPC(8): A61K9/51A61K47/34A61K47/46A61P9/10C12N5/0786
CPCA61K9/5153A61K9/5176C12N5/0645
Inventor 李翀程立婷
Owner SOUTHWEST UNIVERSITY
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