Bionic nanometer erythrocyte genophore and preparation method and application thereof

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 effects of easy-to-obtain raw materials, good biocompatibility, and simple material components

Active Publication Date: 2018-09-21
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 ...

Method used

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  • Bionic nanometer erythrocyte genophore and preparation method and application thereof
  • Bionic nanometer erythrocyte genophore and preparation method and application thereof
  • Bionic nanometer erythrocyte genophore and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[0075] Weigh 50 mg of bovine serum albumin (BSA) and dissolve it 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) and dissolve it in 2 mL of water with pH=4.75, add it dropwise to solution A to obtain solution B . Weigh 2.17 mg DIPEA (N'N-diisopropylethylenediamine) and dissolve it in 2 mL of water with pH=4.75, add it dropwise to solution B, and react at 25°C for 2 hours to obtain solution C. Weigh 70 mg of EDC and dissolve it in 2 mL of water with pH=4.75, and add it dropwise into solution C to obtain solution D. Weighed 3.6 mg PEI600 and dissolved it in 2 mL of water with pH=4.75, added it dropwise to solution D under 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 unreacted E...

Embodiment 2

[0077] High-resolution mass spectrometry characterization of embodiment 2 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. Such as figure 1As shown, the maximum peak value of BSA is 66820Da; the maximum peak value of BSA grafted only with N'N-diisopropylethylenediamine is 68802Da; The largest peak is 72402Da. figure 1 The results proved that the cationic bovine serum albumin (cBSA) was successfully synthesized in this step reaction.

Embodiment 3

[0079] Example 3 Preparation of cBSA-siRNA nanocomplex

[0080] Weigh 10 mg of cBSA and dissolve it in 10 mL of ultrapure water, and prepare siRNA (VEGF-siRNA, purchased from Suzhou Gemma Gene Co., Ltd.), whose sequence is 5'-GGAUCAAACCUCACCAAAGTTCUUUGGUGAGGUUUGAUCCTT-3' (SEQ ID NO.1) into 1 mg / mL of stock solution. The cBSA-siRNA nanocomplexes were prepared under the environment of pH 5 or 7.4 respectively. According to a series of mass ratios set, the cBSA solution was quickly added to the siRNA stock solution, and incubated at 25°C for 30 minutes to obtain a series of Nanocomposites with different surface charge properties in a neutral environment, wherein the mass ratios of cBSA to siRNA are 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 bionic nano red blood cell gene carrier and its preparation method and application. 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 the gene 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 ability, and easy preparation and modification. Toxicity and low transfection efficiency. At present, more non-viral gene carriers have been reported, including cationic liposomes, chitosan, polyethyleneimine (PEI), polylysine (PLL), polyamine dendrimers (PAMAM) and the like. These cationic ...

Claims

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Application Information

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IPC IPC(8): A61K48/00
CPCA61K48/0025
Inventor 戴箭纪鑫王艳明薛巍
Owner JINAN UNIVERSITY
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