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Erythrocyte membrane-encapsulated rapamycin nanoparticle and preparation method and application thereof

A technology of rapamycin and red blood cell membrane, which is applied in the field of rapamycin nanoparticles and its preparation, which can solve the problems of lack, targeting of toxic and side effects of drugs, and unclear effectiveness and safety of chronic diseases in vivo.

Pending Publication Date: 2020-05-22
CHONGQING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

To this end, researchers have tried many different types of RAP drug delivery systems. For example, drug-eluting stents constructed by coating RAP on the surface of metal stents are currently widely used in coronary artery surgical implants, which can increase their Bioavailability, but long-term treatment with other drugs is required after stent implantation, so effective drug delivery is still a problem to be faced
In addition, some researchers have prepared a variety of RAP drug-loaded particles by physically encapsulating or chemically bonding drugs into liposomes, polymer nanoparticles, and polyme

Method used

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  • Erythrocyte membrane-encapsulated rapamycin nanoparticle and preparation method and application thereof
  • Erythrocyte membrane-encapsulated rapamycin nanoparticle and preparation method and application thereof
  • Erythrocyte membrane-encapsulated rapamycin nanoparticle and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1: Construction and property characterization of erythrocyte membrane-coated nanomedicine

[0039] ①Preparation of nano-erythrocyte capsule

[0040] The red blood cells were collected from the whole blood through multiple centrifugation, and the nano-red blood cell capsule was prepared by further freeze-thaw cycle technology: a) Red blood cell separation: fresh whole blood was anticoagulated with heparin and ethylenediaminetetraacetic acid (EDTA), and kept at 4°C Centrifuge under the conditions (4000 rpm, 5min), wash with PBS repeatedly 3 times, and collect red blood cells. Red blood cell membranes are prepared by multiple "freeze-thaw cycles". b) Preparation of nano-erythrocyte capsules by freeze-thaw cycles: the erythrocyte suspension was first frozen at -80°C, and then thawed at room temperature. Subsequently, the nano-red blood cell capsule was obtained by centrifugation (14000rpm, 20min) at 4°C, and the PBS solution was washed repeatedly for 3 times. . ...

Embodiment 2

[0056] Example 2: In vitro evaluation of erythrocyte membrane-coated nanomedicine

[0057] ①Characterization of surface membrane proteins of RBC / RAP@PLGA nanoparticles

[0058] In order to confirm that the surface of RBC / RAP@PLGA nanoparticles maintains the original functional membrane proteins, the erythrocyte envelope, nano-erythrocyte envelope, and RBC / RAP@PLGA surface proteins were determined by SDS-PAGE. SDS sample buffer (Invitrogen) was prepared first. They were then run in 4-12% Bis-Tris 10-well mini gels on a BIO-RAD electrophoresis system. The samples were run at 75V for 0.5h and then at 140V for 1h to achieve sufficient protein separation. Finally, polyacrylamide gels were stained with SimplyBlue (Invitrogen) to visualize protein bands.

[0059] ② Protein adsorption

[0060] Serum was used to characterize the protein adsorption properties of nanoparticles. RAP@PLGA and RBC / RAP@PLGA were incubated with serum solutions in PBS (0.01M), respectively, to keep the na...

Embodiment 3

[0077] Example 3: In vivo evaluation of erythrocyte membrane-coated nanomedicine

[0078] ①ApoE - / - mouse AS model

[0079] All animal care and experimental protocols were reviewed and approved. Male C57BL / 6 mice (25-30 g, 8 weeks) were purchased from Peking University Health Science Center (Beijing, China) male apolipoprotein-E deficient (ApoE - / - ) mice. ApoE fed a high-fat diet (HFD, normal diet with 0.5% cholesterol and 5% fat) - / - Mouse model AS.

[0080] ②ApoE - / - mouse AS treatment

[0081] 20 ApoE - / - Mice were randomly divided into 4 groups (5 mice in each group) and fed with HFD for 10 weeks. Mice were treated with RAP@PLGA, RBC / RAP@PLGA, and equivalent RAP (0.7 mg / kg) via the tail vein every three days for 30 days. Inject 5% sucrose as the model control group. During the treatment, the body weight changes of the mice were weighed simultaneously.

[0082] ③ Quantitative analysis of atherosclerotic plaque

[0083] At the end of treatment, ApoE - / - Mice we...

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Abstract

The invention discloses a erythrocyte membrane-encapsulated rapamycin nanoparticles. The nanoparticles comprise a rapamycin nano-drug core, and the nano-drug core takes PLGA as a carrier and is encapsulated in an erythrocyte membrane. The nanoparticles disclosed by the invention cooperate with an efficient drug solubilizing effect and an autologous RBC nano-surface immune escape effect of an artificial nano-drug carrier; long-acting blood circulation can be shown, targeted atherosclerosis drug delivery and local controllable drug functionalization are achieved; plaque development is remarkablyimproved; and the effect of safely and efficiently treating atherosclerosis is achieved.

Description

technical field [0001] The invention belongs to the technical field of medicines, and in particular relates to rapamycin nanoparticles encapsulated by red blood cell membranes, a preparation method and an application thereof. Background technique [0002] According to the World Health Statistics Report released by the World Health Organization (WHO) in 2017, non-communicable diseases accounted for 70% of the total death toll in the world, among which cardiovascular disease (Cardiovascular disease, CVD) still ranks first in the death of residents, with a high mortality rate. 45%. The prevalence and mortality of cardiovascular diseases in China are still on the rise, and it is estimated that there are 290 million people with cardiovascular diseases. Atherosclerosis (AS) is the pathological basis of many CVD diseases, such as coronary artery disease, carotid artery disease and peripheral vascular disease, and it is a major problem that human beings urgently need to overcome. ...

Claims

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Application Information

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IPC IPC(8): A61K9/50A61K31/436A61K47/34A61K47/46A61P9/10
CPCA61K9/5068A61K31/436A61K47/34A61P9/10
Inventor 王贵学王溢吴伟
Owner CHONGQING UNIV
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