Biodegradable crosslinked polyethylenimine and its uses

A technology of polyethyleneimine and biodegradation, applied in the biological field, can solve problems such as cell membrane damage and achieve the effect of reducing cytotoxicity

Inactive Publication Date: 2007-05-30
NANJING WISEGEN BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Zhong Z's research found that adding hydrophobic segments to the polymer can strengthen its hydrophobic interaction with the cell membrane, th...

Method used

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  • Biodegradable crosslinked polyethylenimine and its uses
  • Biodegradable crosslinked polyethylenimine and its uses
  • Biodegradable crosslinked polyethylenimine and its uses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Synthesis and properties of embodiment 1 cross-linked PEI

[0037] 1 gram of PEI (Mw: 2000 Daltons) was dissolved in 3ml of freshly distilled dichloromethane, and then an appropriate proportion of diglycidyl adipate and hexanediol diacrylate was added, and after a proper reaction time at 40°C, the solution Gradually turn yellow. The solution was then transferred to a Spectra / PorMwco 10,000 membrane and dialyzed against double distilled water for 4 days at 4°C. It was then lyophilized to remove water to obtain a light-colored solid (CLPEI). Store at -70°C. Various cross-linked PEI polymers (Table 1) can be obtained by using different cross-linking agents and PEI combinations. The reaction combination is shown in Formula 1, and the obtained polymer structure is shown in Formula 2. Product carries out HNMR (varian 300MHz, D 2 (0) analysis, it was found that there was no double bond hydrogen (5.5-6.0ppm CH in the raw material) in the product 2 =CH-) absorption peak, bu...

Embodiment 2

[0047] Embodiment 2 Lactobionic acid and folic acid modification of cross-linked PEI

[0048] The cross-linked PEI is connected with lactobionic acid by amide bond through EDC (1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide) activated amino group. 200 mg of the cross-linked PEI synthesized in Example 1 was dissolved in 6 ml of 10 mM TEMED / HCl buffer. Then 180 mg of EDC was added and stirred at 25°C for 24 hours. Then add an appropriate amount of lactobionic acid and stir for 72 hours at 25°C. Finally, the obtained product was dialyzed with double distilled water for 4 days, lyophilized to remove water, and galactosyl-modified cross-linked PEI (Gal-PEI) was obtained.

[0049] 10 mg of folic acid and an appropriate amount of DCC were dissolved in freshly distilled DMSO and reacted at low temperature for a period of time, and then 200 mg of the PEI synthesized in Example 1 was added. After reacting for 12 hours under stirring, dialyze with double distilled water for 4 days at ...

Embodiment 3

[0050] Example 3 Preparation and Characterization of Crosslinked PEI and pEGFP-Cl Complex

[0051] Preparation of complex

[0052] The biodegradable cross-linked PEI or PEI 25kDa synthesized in Example 1 were respectively dissolved in PBS (140mM NaCl, 2.7mM KCl, 10mM NaCl 2 HPO 4 , 1.8mM KH 2 PO 4 , pH 7.4), made into a 1mg / ml stock solution. ddH 2 O diluted to 1 mg / ml stock solution. According to a certain PEI / DNA (mass ratio), prepare a complex of cross-linked PEI or PEI 25kDa PEI and DNA, for example, prepare a complex of cross-linked PEI / DNA=2:1, the specific process is as follows: Take 1 μl of plasmid DNA solution (1mg / ml) was dissolved in 50μl PBS, mixed gently, then 2μl cross-linked PEI was dissolved in 50μl PBS, mixed gently, then the two were mixed, oscillated for 10 seconds, and stood at room temperature for 10-15 Minutes, ready for experiments characterizing its performance and cell transfection.

[0053] Electrophoretic retardation experiment

[0054] Prep...

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Abstract

The invention discloses a decomposable crosslinking polyethylene imine and making method and application under physiological condition, which is characterized by the following: adopting one or more composition of glycidyl ester of each kind diacid, acroleic acid/methacrylic acid glycidyl ester or polyol or polyatomic condensed alcohol/polybasic ester of methyl acroleic acid as crosslinking agent; crosslinking with linear or branched polyethylene imine to form non-virus gene infection-transmitting carrier for each cell and biological tissue.

Description

technical field [0001] The present invention belongs to the field of biotechnology, in particular, relates to a kind of delivery carrier and implementation method of biologically active substances that can be used in various cells in vitro and in vivo tissues or organs. RNA, and expression plasmids encoding therapeutic genes are transferred into cells or tissues and organs. The invention also relates to the preparation and use of this delivery carrier. Another object of the present invention is to provide a gene delivery composition and method wherein the gene carrier is biodegradable and biocompatible. technical background [0002] Gene therapy refers to the transfer of exogenous genes into cells, through the restoration or increase of gene expression to correct the disorder of human gene structure or function, prevent the development of disease, kill diseased cells, or inhibit the genetic material of foreign pathogens Copy, so as to achieve the purpose of curing diseases...

Claims

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Application Information

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IPC IPC(8): C08F126/02C08J3/24C08K5/10A61K47/48A61K48/00C12N15/87
CPCA61K47/48346A61K47/48176A61K47/48692B82Y5/00A61K47/48192C08G73/0206C12N15/87A61K47/58A61K47/59A61K47/66A61K47/6883
Inventor 董伟成立珍
Owner NANJING WISEGEN BIOTECH
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