Choroidal neovascularization targeting nanoparticle coated with macrophage membrane and preparation method of choroidal neovascularization targeting nanoparticle

A technology targeting nanoparticles and neovascularization, which is applied in the field of choroidal neovascularization targeting nanoparticles and its preparation, can solve the problems that there is no intravenous nano-preparation for the treatment of retinal diseases, poor drug compliance of patients, etc., and achieve high drug loading efficiency , strong drug-loading ability, and efficient treatment of diseases

Pending Publication Date: 2021-09-24
赵晨
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the delivery of small molecule drugs is limited to intravitreal injection, but under this mode of administration, the patient's medication compliance is poor
There are no reports of intravenous administration of nanoformulations for retinal diseases

Method used

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  • Choroidal neovascularization targeting nanoparticle coated with macrophage membrane and preparation method of choroidal neovascularization targeting nanoparticle
  • Choroidal neovascularization targeting nanoparticle coated with macrophage membrane and preparation method of choroidal neovascularization targeting nanoparticle
  • Choroidal neovascularization targeting nanoparticle coated with macrophage membrane and preparation method of choroidal neovascularization targeting nanoparticle

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Experimental program
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Effect test

Embodiment 1

[0046] (1) Extraction of macrophages: 25 g of C57 male mice were intraperitoneally injected with 1 mL of 4% thioglycolate broth. After 3 days, the mice were sacrificed, the whole body was immersed in 75% ethanol for disinfection, and 5 mL of medium was injected twice to extract peritoneal macrophages. Cells were extracted at 37°C. 5%CO 2 Cultivate to 80%-90% under certain conditions;

[0047] (2) Collect the obtained cells into a centrifuge tube, centrifuge 3 times at 500g×5min, freeze and thaw repeatedly 3 times with liquid nitrogen to lyse the cells, and then repeatedly grind 20 times with a Dawns homogenizer to further break the cells;

[0048] Collect the cell lysate in a centrifuge tube, centrifuge at 3200g×5min at 4°C to remove large cell debris;

[0049] Collect the supernatant, centrifuge at 20000g×25min at 4°C to remove organelles;

[0050] Collect the supernatant, centrifuge at 100000g×60min, 4°C, discard the supernatant, and the resulting white precipitate is the...

Embodiment 2

[0059] (1) Extraction of macrophages: SD rats were injected intraperitoneally with 1 mL of 4% thioglycollate broth, sacrificed 3 days later, immersed in 75% ethanol for disinfection, and injected 5 mL of medium twice to extract peritoneal macrophages Cells were extracted at 37°C. 5%CO 2 Cultivate to 80%-90% under certain conditions;

[0060] (2) Collect the obtained cells into a centrifuge tube, centrifuge 3 times at 500g×5min, freeze and thaw repeatedly 3 times with liquid nitrogen to lyse the cells, and then repeatedly grind 20 times with a Dawns homogenizer to further break the cells;

[0061] Collect the cell lysate in a centrifuge tube, centrifuge at 3200g×5min at 4°C to remove large cell debris;

[0062] Collect the supernatant, centrifuge at 20000g×25min at 4°C to remove organelles;

[0063] Collect the supernatant, centrifuge at 100000g×60min, 4°C, discard the supernatant, and the resulting white precipitate is the cell membrane; resuspend the cell membrane pellet i...

Embodiment 3

[0069] Preparation of membrane-coated nanoparticle preparations of rapamycin-loaded macrophages labeled with cell membrane orange-red fluorescent probe DiI.

[0070] (1) Obtain macrophages directly from the mouse mononuclear macrophage leukemia cell line Raw264.7 cells, at 37°C, 5% CO 2Cultivate to 80%-90% under certain conditions;

[0071] (2) Scrape the macrophages obtained in (1), collect them in a centrifuge tube, centrifuge 3 times at 500g×5min, freeze and thaw them 3 times with liquid nitrogen to lyse the cells, and grind them repeatedly with a Dawns homogenizer 20 times to further break the cells; collect the cell lysate in a centrifuge tube, centrifuge at 3200g×5min, 4°C to remove large cell debris;

[0072] Collect the supernatant, centrifuge at 20000g×25min at 4°C to remove organelles;

[0073] Collect the supernatant, centrifuge at 100000g×60min, 4°C, discard the supernatant, and the resulting white precipitate is the cell membrane; resuspend the cell membrane pel...

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Abstract

The invention discloses a choroidal neovascularization targeting nanoparticle coated with a macrophage membrane and a preparation method of the choroidal neovascularization targeting nanoparticle. A polylactic acid-glycolic acid copolymer entraps rapamycin to prepare rapamycin-PLGA nanoparticles, the rapamycin-PLGA nanoparticles are used as an inner core, and a macrophage membrane is used as a capsid to coat the surface of the rapamycin-PLGA nanoparticles; wherein the mass ratio of the rapamycin to the polylactic acid-glycolic acid copolymer is 1: (9-20), and the mass ratio of the protein in the macrophage membrane to the polylactic acid-glycolic acid copolymer is 1: (1-1.5). The prepared nanoparticles are good in biocompatibility, the preparation method is simple, and the particle size distribution of the nanoparticles is uniform. The key lesion of age-related macular degeneration, namely choroidal neovascularization, is actively targeted by using biomolecules on the surface of a macrophage membrane, so that the active recruitment of a drug-containing carrier to the neovascularization is effectively improved, and the effect of protecting retinal pigment epithelium under an inflammatory condition is also achieved while the neovascularization is inhibited.

Description

technical field [0001] The invention belongs to the technical field of pharmaceutical preparations, and in particular relates to choroidal neovascularization targeting nanoparticles based on macrophage membrane-coated loaded rapamycin and a preparation method thereof. Background technique [0002] With the development of economy and society and the deepening of population aging process, blinding eye diseases have become one of the diseases that mainly affect human health and quality of life. Among them, age-related macular degeneration (AMD) is the leading cause of visual impairment in the elderly worldwide. At present, about 200 million people in the world are threatened with blindness caused by age-related macular degeneration. In my country, with the acceleration of population aging, the prevalence of AMD increases year by year, and the prevalence of AMD among people over 65 years old is about 13.65%. The pathogenesis of AMD is diverse and complex, and it is still incon...

Claims

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Application Information

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IPC IPC(8): A61K9/52A61K31/436A61K47/46A61K47/34A61P27/02
CPCA61K9/5176A61K9/5153A61K9/5192A61K9/0019A61K31/436A61P27/02
Inventor 赵晨夏韦艺蒋晨李潮楚永超
Owner 赵晨
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