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A bionic nano-erythrocyte gene carrier and its preparation method and application

A gene carrier and biomimetic nanotechnology, which is applied in the field of biomedical engineering materials, can solve the problems of lack of gene carriers, etc., and achieve the effect of easy-to-obtain raw materials, simple material components, and good biocompatibility

Active Publication Date: 2020-07-03
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, tumor gene therapy (such as siRNA therapy) lacks a safe and efficient gene carrier. Combining cell bionics with the concept of charge reversal, the construction of red blood cell bionic nano-gene carrier and its application have not been reported.

Method used

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  • A bionic nano-erythrocyte gene carrier and its preparation method and application
  • A bionic nano-erythrocyte gene carrier and its preparation method and application
  • A bionic nano-erythrocyte gene carrier and its preparation method and application

Examples

Experimental program
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Effect test

Embodiment 1

[0074] Example 1 Preparation of charge-reversed cationized bovine serum albumin (cBSA)

[0075] 50 mg of bovine serum albumin (BSA) was weighed and dissolved in 5 mL of water with pH=4.75 to obtain A solution. Weigh 3.6 mg of EDC (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride), dissolve it in 2 mL of pH=4.75 water, and add it dropwise to solution A to obtain solution B . 2.17 mg of DIPEA (N'N-diisopropylethylenediamine) was weighed and dissolved in 2 mL of water with pH=4.75, added dropwise to solution B, and reacted at 25°C for 2 hours to obtain solution C. Weigh 70 mg of EDC, dissolve it in 2 mL of water with pH=4.75, and add it dropwise to solution C to obtain solution D. 3.6 mg of PEI600 was weighed and dissolved in 2 mL of water with pH=4.75, added dropwise to solution D in an ice-water bath, and reacted at 25° C. for 2 hours to obtain solution E. The reaction was stopped by adding acetate buffer. The solution was ultrafiltered four times, and the unreac...

Embodiment 2

[0077] Example 2 High-resolution mass spectrometry characterization of cationized bovine serum albumin (cBSA)

[0078] The cationized bovine serum albumin grafted with different cationic units obtained in Example 1 was dissolved in ultrapure water (1 mg / mL) for high-resolution mass spectrometry characterization. like figure 1As shown, the maximum peak of BSA is 66820 Da; the maximum peak of BSA grafted with N'N-diisopropylethylenediamine only is 68802 Da; the maximum peak of cationized bovine serum albumin (cBSA) (with both DIPEA and PEI600 grafted) The maximum peak is 72402Da. figure 1 The results showed that the cationized bovine serum albumin (cBSA) was successfully synthesized in this step.

Embodiment 3

[0079] Example 3 Preparation of cBSA-siRNA nanocomplexes

[0080] Weigh 10 mg of cBSA and dissolve it in 10 mL of ultrapure water. siRNA (VEGF-siRNA, purchased from Suzhou Gene Gene Co., Ltd.), whose sequence is 5'-GGAUCAAACCUCACCAAAGTTCUUUGGUGAGGUUUGAUCCTT-3' (SEQ ID NO.1), is configured to 1 mg / mL of stock solution. The cBSA-siRNA nanocomplexes were prepared at pH 5 or 7.4, respectively. According to a set mass ratio, the cBSA solution was quickly added to the siRNA stock solution, and incubated at 25°C for 30 minutes to obtain a series of The nanocomposites with different surface charge properties in a neutral environment, wherein the mass ratio of cBSA to siRNA were 0.5, 2, 5, and 8, respectively.

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Abstract

The invention provides a bionic nanometer erythrocyte genophore and a preparation method and application thereof. The bionic nanometer erythrocyte genophore comprises an erythrocyte membrane and a kernel which is wrapped by the erythrocyte membrane and can achieve charge reversal. The invention further the preparation method of the bionic nanometer erythrocyte genophore. The method comprises the steps that the genophore which can achieve charge reversal is synthesized through an amide reaction; then, the kernel of the genophore which can achieve charge reversal is wrapped by the erythrocyte membrane through an extrusion method. According to the bionic nanometer erythrocyte genophore, a biological membrane wrapping treatment gene is applied to disease treatment for the first time, the electronegativity of the kernel can be guaranteed, the gene is successfully wrapped by the erythrocyte membrane, long circulation of gene medicine in vivo is achieved, and the charge reversal can be achieved under the focus microenvironment, and accordingly nucleic acid medicine is released. Meanwhile, the genophore is free of cytotoxicity, non-toxic metabolism in vivo can be completed, the brand-new,safe and efficient genophore is provided for gene treatment, and the genophore has wide application prospects.

Description

technical field [0001] The invention belongs to the field of biomedical engineering materials, in particular to a biomimetic nano-red blood cell gene carrier and a preparation method and application thereof. Background technique [0002] Gene therapy has been extensively studied as a potential treatment for genetic diseases, malignancies, viral infections, and neurological diseases [1]. In gene therapy, whether the gene carrier can effectively guide and control the endocytosis and expression of genes by the lesion cells is the key to clinical application. Compared with viral gene carriers, non-viral gene carriers have relatively high biocompatibility, low immunogenicity, strong gene-carrying capacity, and easy preparation and modification. toxicity and low transfection efficiency. The most reported non-viral gene carriers include cationic liposomes, chitosan, polyethyleneimine (PEI), polylysine (PLL), polyamine dendrimers (PAMAM) and so on. These cationic gene carriers ar...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K48/00
CPCA61K48/0025
Inventor 戴箭纪鑫王艳明薛巍
Owner JINAN UNIVERSITY
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