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A kind of liver-targeted cationic gene carrier constructed by amino-epoxy ring-opening reaction based on lactose and preparation method thereof

An epoxy ring-opening, gene carrier technology, applied in other methods of inserting foreign genetic materials, recombinant DNA technology, etc., can solve the problems of high toxicity and low effective toxicity, and achieve the effect of gene transfection and broad application prospects.

Active Publication Date: 2022-07-12
BEIJING UNIV OF CHEM TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are still many problems in the research process of using the amino-epoxy ring-opening reaction to obtain cationic gene carriers. For example, as the molecular weight increases, the corresponding transfection efficiency of this cationic carrier is higher, but the toxicity is greater. Improving the transfection efficiency as much as possible and ensuring the targeting effect while ensuring moderate toxicity have become the focus of attention; different monomers have different characteristics, some have higher transfection efficiency and less effective toxicity, how to screen for high-efficiency and high-performance The monomer is also a problem that people need to consider

Method used

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  • A kind of liver-targeted cationic gene carrier constructed by amino-epoxy ring-opening reaction based on lactose and preparation method thereof
  • A kind of liver-targeted cationic gene carrier constructed by amino-epoxy ring-opening reaction based on lactose and preparation method thereof
  • A kind of liver-targeted cationic gene carrier constructed by amino-epoxy ring-opening reaction based on lactose and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Put 513 mg of β-lactose into a 50 mL round-bottomed flask, add 5 mL of anhydrous dimethyl sulfoxide to dissolve it completely, and continuously exhaust with nitrogen for 15 min, then seal it for later use. Subsequently, 500 mg of carbonyldiimidazole was dissolved in 5 mL of anhydrous dimethyl sulfoxide, and the dissolved liquid was drawn with a syringe, and slowly poured into the previously prepared β-lactose solution. After the injection, the mixed solution was allowed to react at room temperature for 24 hours. After 24 hours of reaction, 4 g of cystamine was dissolved in 3 mL of anhydrous dimethyl sulfoxide, and it was slowly injected into the reaction solution with a syringe, and the reaction was carried out at room temperature for 24 hours. After the reaction was completed, the reaction was added dropwise to 100-200 mL of acetone for precipitation, and after repeated washing and centrifugation, a white powdery solid lactosamine was obtained after vacuum drying.

Embodiment 2

[0031]349 mg of lactose amino acid and 100 mg of triglycidyl isocyanurate were put into a 50 mL round-bottomed flask, 10 mL of dimethyl sulfoxide was added to fully dissolve, and the solution was exhausted with nitrogen gas for 15 min, and reacted at 50 °C for 24 h. After the reaction was completed, 0.2 mL of ethylenediamine was added, and the temperature was raised to 60 °C for 2 h. Finally, the reaction solution was added to 50 mL of water, and then dialyzed with a dialysis bag with a molecular weight cutoff of 3500 for 24 hours. Finally, a white flocculent polymer was obtained after freeze-drying, which was denoted as LBP. 2 Polymer (LBP 2 ) number average molecular weight (M n ) is 17500, the molecular weight distribution index (M w / M n ) is 1.85.

Embodiment 3

[0033] 438 mg of lactose amino acid and 150 mg of triglycidyl isocyanurate were placed in a 50 mL round-bottomed flask, 10 mL of dimethyl sulfoxide was added to fully dissolve, and the mixture was exhausted with nitrogen gas for 15 min, and reacted at 50 °C for 24 h. After the reaction was completed, 0.2 mL of ethylenediamine was added, and the temperature was raised to 60 °C for 2 h. Finally, the reaction solution was added to 50 mL of water, and then dialyzed with a dialysis bag with a molecular weight cut-off of 3500 for 24 hours. Finally, after freeze-drying, a white flocculent polymer LBP was obtained. 3 . Polymer (LBP 3 ) number average molecular weight (M n ) is 19600, the molecular weight distribution index (M w / M n ) is 1.91.

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Abstract

The invention discloses a preparation method for constructing a liver-targeted cationic gene carrier based on lactose through amino-epoxy ring-opening reaction. , the reaction is carried out under the protection of inert gas, the reaction temperature is 40~60 ℃, and the reaction is 24~48 hours; 2) step 1) After the reaction is completed, ethylenediamine is added, and the temperature is raised to 50~70 ℃, and the reaction is performed for 1~3 hours, Dialysis was performed after the reaction to obtain a white flocculent polymer. Based on lactose, a low-toxicity, high-efficiency, and liver-targeting cationic gene carrier is prepared by a green and mild amino-epoxy ring-opening reaction, which can not only effectively achieve gene transfection, but also achieve targeted delivery of nucleic acids. Mediated CRIPSR / Cas9 gene editing system to achieve efficient gene editing.

Description

technical field [0001] The invention belongs to the field of non-viral gene carriers, and relates to a liver-targeted cationic gene carrier constructed by amino-epoxy ring-opening reaction based on lactose and a preparation method thereof. Background technique [0002] As a tool for introducing exogenous genes into cells, the gene carrier itself should be low in toxicity and not cause immune response; secondly, it should be able to form a complex with the gene with a stable structure without causing changes in the gene structure; Finally, the carrier would ideally be able to target and degrade, so that it could target specific cells and reduce the side effects of its retention. There are two types of gene vectors widely used at present: viral vectors and non-viral vectors. Viral vectors mainly include retrovirus, adenovirus, adeno-associated virus and herpes simplex virus. Such viral vectors are generally able to easily overcome cellular barriers and immune defense mechani...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/87C08G65/26
CPCC12N15/87C08G65/2624
Inventor 徐福建祁宇俞丙然
Owner BEIJING UNIV OF CHEM TECH
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