Polyethyleneimine (PEI) derivative taking amphipathic chitosan as cross linker and preparation method and application thereof

A technology of polyethylenimine and chitosan, applied in other methods of inserting foreign genetic materials, recombinant DNA technology, etc., to achieve the effects of increasing stability, reducing cytotoxicity, and low toxicity

Active Publication Date: 2012-06-27
SHANGHAI OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But about a kind of polyethylenimine derivatives with amphiphilic chitosan as cross-linking agent and its preparation method and application have not yet been reported

Method used

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  • Polyethyleneimine (PEI) derivative taking amphipathic chitosan as cross linker and preparation method and application thereof
  • Polyethyleneimine (PEI) derivative taking amphipathic chitosan as cross linker and preparation method and application thereof
  • Polyethyleneimine (PEI) derivative taking amphipathic chitosan as cross linker and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Preparation of Polyethyleneimine Derivatives Using Amphiphilic Chitosan as a Crosslinking Agent

[0026] Add 6g of chitosan and 17.0ml of n-octanal into 125 mL of methanol, stir at 30°C for 12h, then add KBH in batches 4 3 g, stirred overnight, filtered, washed the filter cake repeatedly with water and hot methanol, and dried under vacuum at 50°C. Take 2 g of the above product, put it in a three-necked flask, add 30 mL of N-methylpyrrolidone, 5 g of KI, 10 mL of 15% NaOH aqueous solution and 10.4 mL of CH 3 1. Heat up to 60°C and react for 1 h under full stirring, cool to room temperature, centrifuge for 30 min (1000 r / min), collect the lower layer solid and dissolve it in distilled water, dialyze for 5 days, filter out the insoluble matter and freeze-dry the filtrate to obtain pale Yellow amphipathic chitosan.

[0027] Weigh 0.1 mmol of amphiphilic chitosan after water removal, add 1 mmol of triphosgene, dissolve in a mixed solution of anhydrous toluene ...

Embodiment 2

[0028] Example 2 Preparation of Polyethyleneimine Derivatives Using Amphiphilic Chitosan as Crosslinking Agent

[0029] Add 6g of chitosan and 17.0ml of n-decyl aldehyde into 125 mL of methanol, stir at 30°C for 12h, then add KBH in batches 43 g, stirred overnight, filtered, washed the filter cake repeatedly with water and hot methanol, and dried under vacuum at 50°C. Take 2 g of the above product, put it in a three-necked flask, add 30 mL of N-methylpyrrolidone, 5 g of KI, 10 mL of 15% NaOH aqueous solution and 10.4 mL of CH 3 1. Heat up to 60°C and react for 1 h under full stirring, cool to room temperature, centrifuge for 30 min (1000 r / min), collect the lower layer solid and dissolve it in distilled water, dialyze for 5 days, filter out the insoluble matter and freeze-dry the filtrate to obtain pale Yellow amphipathic chitosan.

[0030] Weigh 0.1 mmol of amphiphilic chitosan after water removal, add 0.1 mmol of triphosgene, dissolve in a mixed solution of anhydrous tol...

Embodiment 3

[0031] Example 3 Preparation of Polyethyleneimine Derivatives Using Amphiphilic Chitosan as Crosslinking Agent

[0032] Add 6g of chitosan and 17.0ml of lauric aldehyde into 125 mL of methanol, stir at 30°C for 12h, then add KBH in batches 4 3 g, stirred overnight, filtered, washed the filter cake repeatedly with water and hot methanol, and dried under vacuum at 50°C. Take 2 g of the above product, put it in a three-necked flask, add 30 mL of N-methylpyrrolidone, 5 g of KI, 10 mL of 15% NaOH aqueous solution and 10.4 mL of CH 3 1. Heat up to 60°C and react for 1 h under full stirring, cool to room temperature, centrifuge for 30 min (1000 r / min), collect the lower layer solid and dissolve it in distilled water, dialyze for 5 days, filter out the insoluble matter and freeze-dry the filtrate to obtain pale Yellow amphipathic chitosan.

[0033] Weigh 0.1 mmol of amphiphilic chitosan after water removal, add 0.5 mmol of triphosgene, dissolve in a mixed solution of anhydrous to...

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Abstract

The invention relates to a polyethyleneimine (PEI) derivative taking an amphipathic chitosan as a cross linker. The molar ratio of the amphipathic chitosan to PEI in the derivative is 1:(1-20), the molecular weight range of the PEI in the raw materials for the preparation of the derivative is 600-70,000, and the molecular weight range of the chitosan in the raw materials for the preparation of the derivative is 1,000-50,000. The invention further provides a preparation method and application of the derivative. The PEI derivative has the advantages that: the PEI derivative is connected with low-molecular-weight PEI through a chemical cross-linking method by using the advantages of the chitosan, and the cytotoxicity arising from high-molecular-weight PEI is greatly lowered under the premise that a certain transfection efficiency is ensured; and particles can be prevented from aggregating in a blood circulation fluid environment due to the introduction of a hydrophilic surface by the chitosan, the stability is improved, the size of a complex formed by deoxyribonucleic acid (DNA) and the PEI derivative connected with the low-molecular-weight PEI can reach nanoscale, and thus, the PEI derivative is suitable for the in-vivo transmission of gene drug.

Description

technical field [0001] The invention relates to a gene carrier material, in particular to a polyethyleneimine derivative using amphiphilic chitosan as a crosslinking agent, a preparation method and application thereof. Background technique [0002] Gene therapy is a new treatment method based on genetic engineering technology and molecular genetics in recent years. There are three important links in gene therapy, namely, target gene, transgenic carrier and target cell. Gene transfer system is the core technology of gene therapy. The biggest problem at this stage is that the ideal gene carrier has not yet been found. Currently applied vectors include two categories: viral vectors and non-viral vectors. Viral vectors have high transfection efficiency but have problems such as low carrying capacity and potential safety threats. Therefore, non-viral vectors have developed rapidly in recent years, especially cationic polymers. [0003] Polyethyleneimine (PEI) is the most widel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08G73/04C08G81/00C08B37/08C12N15/87
Inventor 刘克海朱青吴文惠
Owner SHANGHAI OCEAN UNIV
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