LPEIS/DNA/HA nano-carrier and preparation method and application thereof

A nano-carrier and nano-sphere technology, which is applied in the direction of pharmaceutical formulations, medical preparations of non-active ingredients, gene therapy, etc., can solve problems such as the inability to ensure that genes are not degraded, and there is no targeted immune response, achieving low cost, Controllable size and good dispersion

Inactive Publication Date: 2017-03-29
HUBEI UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Some common methods such as gene gun method, electroporation method, microinjection and other methods, but the current disadvantages of these methods are that they cannot ensure that the gene will not be degraded when entering the bod

Method used

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  • LPEIS/DNA/HA nano-carrier and preparation method and application thereof
  • LPEIS/DNA/HA nano-carrier and preparation method and application thereof
  • LPEIS/DNA/HA nano-carrier and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] 1. Sulfonation

[0058] 1.48gN,N-bis(2-hydroxyethyl)ethylenediamine (10mmol) was dissolved in 40ml of dichloromethane, 1.2217g of 4-dimethylaminopyridine (10mmol) was added, 9.9g of trimethylaminopyridine (92mmol ) and 8.3886 g p-toluenesulfonyl chloride (44 mmol). The above mixed solution was stirred at 0°C for 1d. The dichloromethane was evaporated using a rotary evaporator, and the obtained substance was redissolved in 100ml of chloroform, extracted three times with 0.1M hydrochloric acid, and the organic phase was extracted with MgSO 4 Dry and volatilize the organic phase to obtain a white or yellowish powder.

[0059] 2. Desulfonation

[0060] Completely dissolve the powder obtained in 1 in 20ml of N,N-methyleneformamide solution, add excess ethanolamine dropwise, mix and stir the mixture at 40-60°C for 1-2d, and redissolve the obtained substance after volatilizing the organic phase In dimethyl sulfoxide, the organic phase is volatilized to remove residual ethan...

Embodiment 2

[0076] 1. Sulfonation

[0077] 1.48gN,N-bis(2-hydroxyethyl)ethylenediamine (10mmol) was dissolved in 40ml of dichloromethane, 1.2217g of 4-dimethylaminopyridine (10mmol) was added, 9.9g of trimethylaminopyridine (92mmol ) and 8.3886 g p-toluenesulfonyl chloride (44 mmol). The above mixed solution was stirred at 0°C for 1d. The dichloromethane was evaporated using a rotary evaporator, and the obtained substance was redissolved in 100ml of chloroform, extracted three times with 0.1M hydrochloric acid, and the organic phase was extracted with MgSO 4 Dry and volatilize the organic phase to obtain a white or yellowish powder.

[0078] 2. Desulfonation

[0079] Completely dissolve the powder obtained in 1 in 20ml of N,N-methyleneformamide solution, add excess ethanolamine dropwise, mix and stir the mixture at 40-60°C for 1-2d, and redissolve the obtained substance after volatilizing the organic phase In dimethyl sulfoxide, the organic phase is volatilized to remove residual ethan...

Embodiment 3

[0095] 1. Sulfonation

[0096] Dissolve 0.74g N,N-bis(2-hydroxyethyl)ethylenediamine (5mmol) in 20ml of dichloromethane, add 0.6109g of 4-dimethylaminopyridine (5mmol), 4.95g trimethylaminopyridine (46mmol ) and 4.1943 g p-toluenesulfonyl chloride (22 mmol). The above mixed solution was stirred at 0°C for 1d. The dichloromethane was evaporated using a rotary evaporator, and the obtained substance was redissolved in 50ml of chloroform, extracted three times with 0.1M hydrochloric acid, and the organic phase was extracted with MgSO 4 Dry and volatilize the organic phase to obtain a white or yellowish powder.

[0097] 2. Desulfonation

[0098] Completely dissolve the powder obtained in 1 in 20ml of N,N-methyleneformamide solution, add excess ethanolamine dropwise, mix and stir the mixture at 40-60°C for 1-2d, and redissolve the obtained substance after volatilizing the organic phase In dimethyl sulfoxide, the organic phase is volatilized to remove residual ethanolamine to obta...

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Abstract

The invention discloses an LPEIS/pDNA/HA nano-carrier and a preparation method and application thereof, and belongs to the field of gene therapy and DNA vaccine antibody expression. The LPEIS/pDNA/HA nano-carrier takes polyethyleneimine as a body, LPEIS is synthesized through a polymerization reaction, and under the electrostatic interaction, the LPEIS is combined with negatively charged DNA and HA to form nanospheres; and the particle size of the LPEIS/pDNA/HA nano-carrier is 100 nm-400 nm. According to the method, the process is simple, and cost is low; the prepared product is good in stability, good in dispersity in warter and controllable in particle size; and the procedure that the nano-carrier enters the human body for gene therapy and antibody expression is simple, the preparation procedure is simple, the effects that the PEI/pDNA/HA nano-composite carries positive charges, is small in particle size and enters the human body easily are guaranteed, the nano-carrier can give expression to green fluorescent protein after entering a cell nucleus, an immune response of an organism is stimulated, and the treatment purpose is realized.

Description

technical field [0001] The invention relates to the field of gene therapy and antibody production, in particular to LPEIS / DNA / HA nanometer carrier and its preparation method and application. Background technique [0002] With the prosperity and progress of social economy, people pay more and more attention to health problems. Among the top 10 deadly diseases, the top ones are heart disease, malignant tumors, cerebrovascular diseases, and diabetes, which seriously affect people's lives, and these diseases are still on the rise, and the mortality rate is also getting higher and higher, tending to be younger. These diseases often cause many complications. Gene therapy has brought dawn to people. Since 1990, gene therapy has been realized in the field of biomedicine and clinical gene intervention for human diseases, and its potential in treating diseases has been recognized. There are many methods for treating diseases compared with traditional ones. Incomparable advantages. ...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K9/51A61K47/36A61K47/32
CPCA61K9/5138A61K9/5161A61K48/0025A61K48/0041
Inventor 孙宏浩王甜甜廖天作祝红达刘明星郭惠玲孙红梅
Owner HUBEI UNIV OF TECH
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