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Cationized mulberry polysaccharide nanoparticle gene vectors and manufacturing method thereof

A mulberry leaf polysaccharide, cationization technology, applied in the direction of nanotechnology, nanomedicine, nanotechnology, etc., can solve the problems of lack of biodegradable polypeptide immunogenicity, high cytotoxicity, etc., to increase biological functions, prepare The process is simple, the effect of good combination and release effect

Active Publication Date: 2014-08-06
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these cationic polymers have their own limitations, such as the lack of biodegradability of polyamines and the potential immunogenicity of polypeptides, etc.
At present, there is a synthetic material polyethyleneimine (PEI), and the test found that high molecular weight (>20000 Da) PEI transfection efficiency is very high, but the cytotoxicity is very high, and there is a problem of degradation; small molecular weight PEI (600 Da, 1300 Da, 2000 Da) have little toxicity, are easily metabolized in the body, and have almost no transfection toxicity (reference: Hongliang Huang , Hai Yu , Guping Tang , Qingqing Wang and Jun Li , Low molecular weight polyethyleneimine cross ~linked by 2~hydroxypropyl~γ~cyclodextrin coupled to peptide targeting HER2 as a gene delivery vector. Biomaterials, 2010,31(7): 1830~1838)

Method used

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  • Cationized mulberry polysaccharide nanoparticle gene vectors and manufacturing method thereof
  • Cationized mulberry polysaccharide nanoparticle gene vectors and manufacturing method thereof
  • Cationized mulberry polysaccharide nanoparticle gene vectors and manufacturing method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1. Preparation of Refined Mulberry Leaf Polysaccharide

[0033] Weigh 120g of dried mulberry leaves, crush them into powder, and pass through a 60-mesh sieve; extract according to the following process: solid-liquid ratio: 1:12, extraction temperature: 85°C, extraction time 2h / time, extraction times: 2 times; combine two The second extraction solution was centrifuged at 5000rpm for 10min, and the supernatant was taken; then the supernatant was concentrated by rotary evaporation to 1 / 6 of the original volume to obtain a concentrated solution; the concentrated solution was precipitated with 95% medical alcohol until the alcohol concentration was 75%, stand at 4°C for 12 hours; collect the precipitate, wash twice with absolute ethanol, acetone, and ether in turn; dry in vacuum to obtain the crude polysaccharide of mulberry leaves.

[0034] Weigh 3g of mulberry leaf crude polysaccharide, dissolve it completely in 40ml of double-distilled water, add 20% trichloroa...

Embodiment 2

[0037] Example 2. Preparation of Oxidized Mulberry Leaf Polysaccharide

[0038] Weigh 0.5g refined mulberry leaf polysaccharide, dissolve in 30ml double distilled water, add 0.8g KIO 4 , quickly placed in a dark room, stirred magnetically, and reacted at room temperature for 72 hours; adding 12ml ethylene glycol to the reaction solution to terminate the reaction, and continued the reaction for 30 minutes according to the aforementioned conditions; put the reaction solution into a dialysis bag, and dialyzed in double distilled water for 48 hours; Obtain oxidized mulberry leaf polysaccharide.

Embodiment 3

[0039] Example 3. Preparation of cationized mulberry leaf polysaccharide modified by ethylenediamine

[0040] Take 0.3g of oxidized mulberry leaf polysaccharide and dissolve it in 20ml of double distilled water; take 0.1ml of ethylenediamine and dissolve it in 5ml of borate buffer solution (pH=9); Add it into the mulberry leaf polysaccharide solution, and carry out magnetic stirring at the same time; after the addition, magnetically stir, and react at room temperature for 24 hours; after that, add 0.4g sodium borohydride to the reaction solution, and continue the reaction for 48 hours under the same conditions; then add to the reaction solution 0.4 g of sodium borohydride, and continue to react for 24 hours under the same conditions; put the reaction solution into a dialysis bag, and dialyze in double distilled water for 48 hours; freeze-dry the dialysate to obtain cationized mulberry polysaccharide modified with ethylenediamine.

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Abstract

A cationic mulberry leaf polysaccharide nanoparticle gene carrier, which is a gene carrier of mulberry leaf polysaccharide modified with amine compounds combined with DNA plasmid, the molecular weight distribution of mulberry leaf polysaccharide is 3~3.5×104Da, of which the mass ratio Calculated, cationized mulberry leaf polysaccharide:DNA plasmid=0.5~100:1, the particle diameter of plasmid is 50-150nm, and described amine compound is spermine, ethylenediamine or the number-average molecular weight is 600Da-2000Da poly Ethylene imine. The three cationized mulberry leaf polysaccharides of the invention all have good DNA plasmid binding effect, and become a new type of non-viral gene carrier material with biodegradability, high efficiency and low toxicity. Moreover, their preparation process is simple and the price is low. The invention discloses its preparation method.

Description

technical field [0001] The invention relates to traditional Chinese medicine polysaccharides and gene carriers, in particular to a cationized mulberry leaf polysaccharide nanoparticle gene carrier. Background technique [0002] In recent years, with the continuous development of biotechnology, gene therapy has become an effective means of clinical treatment of many major diseases such as cancer, cardiovascular and cerebrovascular diseases, and diabetes. With the deepening of gene therapy research, it has been found that the key to gene therapy is to select a suitable and effective gene carrier, that is, an appropriate introduction method, so that the target gene can be introduced in a targeted manner and expressed stably and effectively without causing toxic side effects to the body. Currently, there are two types of vectors used in gene therapy: viral vectors and non-viral vectors. Although viral vectors are widely used in the current clinical trials of gene therapy, many ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/63B82Y5/00
Inventor 徐希明邓纹纹余江南王淼曹霞
Owner JIANGSU UNIV
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