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Biologically orthogonal targeted cytomembrane biomimetic nanoparticle and preparation method and application thereof

A bio-orthogonal, biomimetic nanotechnology, applied in the direction of luminescence/biological dyeing preparations, preparations for in vivo experiments, medical preparations of non-active ingredients, etc., can solve the problem of insufficient targeting recognition effect of heterologous cell membrane biomimetic nanoparticles, Biological safety issues and other issues, to achieve the effect of enhancing the effect of photothermal therapy, good biological activity, and improving the ability of target recognition and uptake

Pending Publication Date: 2019-08-09
SHENZHEN INST OF ADVANCED TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the antigen mutation and immune escape of tumor cells, the above-mentioned biomimetic nanoparticles with heterologous cell membranes have insufficient target recognition effect or potential biological safety problems.

Method used

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  • Biologically orthogonal targeted cytomembrane biomimetic nanoparticle and preparation method and application thereof
  • Biologically orthogonal targeted cytomembrane biomimetic nanoparticle and preparation method and application thereof
  • Biologically orthogonal targeted cytomembrane biomimetic nanoparticle and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0059] Example 1: Synthesis of Lipid / Amino Acid / Saccharide Derivatives Modified with Chemical Reporter Groups

[0060]Synthesis of lipid derivatives: mainly refer to the method disclosed in Analytical Chemistry 2013 85(10), 5263-5270. The specific preparation method is as follows: dissolve a certain amount of 1,2-dibromoethane and sodium azide in DMF, react at 80°C for 20 hours, add ice water and sodium chloride solution after the reaction, and extract to obtain the intermediate 2' - Ethyl bromide azide. The obtained 2'-bromoethane azide was dissolved in tetrahydrofuran, dimethylmethanolamine was added under argon protection, the reaction was carried out at 0 °C for 6 h, and the target product azidoethylcholine AE-Cho was obtained by precipitation with ether. Synthetic routes for choline derivatives are shown below. Among them, acetylation of choline analogs can modify the azide group into the cell membrane system through cellular lipid metabolism.

[0061]

[0062] Sy...

example 2

[0068] Example 2: Preparation of metabolically labeled T cells with azide functional groups

[0069] The obtained human T cells (1 × 10 7 A) cultured in AIM-V medium containing interleukin-2 (IL-2) and 2% fetal bovine serum (FBS), adding CD3 / CD28 antibody magnetic beads to stimulate T cell proliferation and activation, and co-cultured for 72h After adding azide-modified carbohydrate derivatives Ac 4 GalNAz (50 μM), after co-incubating for 48 h, the cells were washed twice with PBS to obtain azide (-N 3 ) functional group-labeled activated T cells (N 3 -T cell), and use flow and confocal imaging for analysis and identification, such as figure 2 shown, Ac 4 The surface of GalNAz sugar-modified T cells showed green Fluor 488 fluorescence signal, while the surface of T cells without sugar modification was almost undetectable; similarly, flow cytometry analysis showed that Ac 4 GalNAz sugar-modified T cells detected more fluorescent signals, consistent with the results of c...

example 3

[0070] Example 3: Bioorthogonally targeted T cell membrane biomimetic nanoparticles (N 3 -TINPs) synthesis

[0071] Take 2×10 respectively 7 N 3 -T cells and T cells were centrifuged at 1000 rpm for 4 min, hypotonic lysis buffer (containing protease inhibitors) was added to the obtained cell pellet, placed on ice for lysis, and then the cells were further disrupted by ultrasonication. In order to obtain purified cell membranes, we use differential centrifugation: first, at 4°C, centrifuge at 3200 × g for 5 min and then take the supernatant; Take the supernatant. The supernatant was finally centrifuged at 50,000-200,000 × g for 50 min (4°C), and the precipitate was collected to obtain purified N 3 -T cell membrane and T cell membrane. respectively N 3 -T cell membrane, T cell membrane was dissolved and then added 90 μg of distearoyl phosphatidyl ethanolamine-polyethylene glycol (DSPE-PEG 2000 ), extruded about 10 times through a 220nm polycarbonate film to obtain N 3 ...

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Abstract

The invention provides a biologically orthogonal targeted cytomembrane biomimetic nanoparticle. The biological orthogonally targeted cytomembrane biomimetic nanoparticle is characterized by comprisinga nano core composed of a high polymer and an active ingredient and cytomembrane wrapping the nano core; a basal metabolite modified with biologically orthogonal functional groups is embedded in thecytomembrane. The basal metabolite with the biologically orthogonal functional groups is added into the basal metabolite modified with the biologically orthogonal functional groups during cell culture, and through metabolism, the basal metabolite modified with the biologically orthogonal functional groups is embedded in the cytomembrane.

Description

technical field [0001] The invention provides a preparation technology of cell membrane biomimetic nanoparticles (BINPs) with bioorthogonal targeting and tumor photothermal therapy effect, the core of which is polyglycolide lactide wrapped with indocyanine green (ICG) Lactide copolymer (Poly(lactic-co-glycolic acid), PLGA), the surface is covered with a chemical reporter group (-N 3 / -BCN / -Tz / -alkynyl, etc.) functionalized cell membrane, and has good optical imaging and tumor photothermal therapy effect. The reporter group on the surface of the cell membrane is covalently bound to the paired group on the surface of the tumor cell, which effectively improves the target recognition and uptake of the biomimetic nanoparticles on the immune cell membrane by tumor cells in vivo, and enhances the photothermal therapy effect of BINPs on tumor cells. And it has the biological activity function of natural cell membrane and good biocompatibility. Background technique [0002] With th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/51A61K47/46A61K47/34A61K47/26A61K47/18A61K47/28A61K41/00A61P35/00A61K49/00
CPCA61K9/5123A61K9/5153A61K9/5176A61K9/5192A61K41/0052A61K49/0034A61K49/005A61K49/0054A61K49/0093A61P35/00
Inventor 蔡林涛潘宏郑明彬韩雨彤李文军陈泽罗英梅
Owner SHENZHEN INST OF ADVANCED TECH
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