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TMC/genipin/pDNA nanometer carrier and preparation method and application thereof

A nanocarrier and genipin technology, which is applied in the field of TMC/genipin/pDNA nanocarriers and their preparation, can solve the problems of not being able to ensure that genes are not degraded, no targeted immune response, etc., and achieve low cost, decentralized Good performance and stability

Inactive Publication Date: 2017-02-15
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 body, have no targeting, and will cause the body's immune response. At present, cationic polymers As a non-viral carrier, the vector has been paid attention to by people, and the safety hazard of the potential viral vector is eliminated

Method used

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  • TMC/genipin/pDNA nanometer carrier and preparation method and application thereof
  • TMC/genipin/pDNA nanometer carrier and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] 1. Preparation of iodide trimethyl chitosan

[0044] Weigh 2g of ground chitosan (DD=80%-95%), 4.8gNaI, 11ml15%NaOH solution, 11.5mlCH3I dissolved in 80ml1-methyl-2-pyrrolidone in a water bath at 60°C for 1h. Condensation is required during the reaction CH3I was precipitated by ethanol solution and separated by centrifugation. The product obtained above was washed twice with ether and filtered to remove ethanol.

[0045] 2. Dissolve the product in 1 in 80ml of 1-methyl-2-pyrrolidone solution in a 60°C water bath to remove excess ether adsorbed on trimethyl chitosan iodide. Measure 4.8gNaI, 11ml15%NaOH solution, and 7mlCH3I into 80ml1-methyl-2-pyrrolidone solution in turn, and bathe in water at 60°C for 0.5h. Then add 2mlCH3I and 0.6gNaOH and continue stirring for 1h.

[0046] 3. Preparation of deiodinated trimethyl chitosan

[0047] Dissolve the iodinated trimethyl chitosan product prepared in 2 in 10% NaCl solution, carry out deiodination, then precipitate with etha...

Embodiment 2

[0055] 1. Preparation of iodide trimethyl chitosan

[0056] Weigh 2g of ground chitosan (DD=80%-95%), dissolve 4.8gNaI, 11ml15%NaOH solution, 11.5mlCH3I in 80ml1-methyl-2-pyrrolidone in a water bath at 40°C for 1h. Condensation is required during the reaction CH3I was precipitated by ethanol solution and separated by centrifugation. The product obtained above was washed twice with ether and filtered to remove ethanol.

[0057] 2. Dissolve the product in 1 in 80ml of 1-methyl-2-pyrrolidone solution in a 40°C water bath to remove excess ether adsorbed on iodized trimethyl chitosan. Measure 4.8gNaI, 11ml15%NaOH solution, 7mlCH3I into 80ml1-methyl-2-pyrrolidone solution successively, and bathe in 40℃ water for 0.5h. Then add 2mlCH3I and 0.6gNaOH and continue to stir for 1h.

[0058] 3. Preparation of deiodinated trimethyl chitosan

[0059] Dissolve the iodinated trimethyl chitosan product prepared in 2 in 10% NaCl solution, carry out deiodination, then precipitate with ethanol,...

Embodiment 3

[0067] 1. Preparation of iodide trimethyl chitosan

[0068] Weigh 1g of ground chitosan (DD=80%-95%), dissolve 2.4gNaI, 5.5ml15%NaOH solution, 5.75mlCH3I in 40ml1-methyl-2-pyrrolidone in a water bath at 60°C for 1h. During the reaction process, Condensed CH3I was precipitated by ethanol solution and separated by centrifugation. The product obtained above was washed twice with ether and filtered to remove ethanol.

[0069] 2. Dissolve the product in 1 in 80ml of 1-methyl-2-pyrrolidone solution in a 60°C water bath to remove excess ether adsorbed on trimethyl chitosan iodide. Measure 2.4gNaI, 5.5ml15%NaOH solution, 3.5mlCH3I into 40ml1-methyl-2-pyrrolidone solution in sequence, and bathe in water at 60°C for 0.5h. Then add 1mlCH3I and 0.3gNaOH and continue stirring for 1h;

[0070] 3. Preparation of deiodinated trimethyl chitosan

[0071] Dissolve the iodinated trimethyl chitosan product prepared in 2 in 10% NaCl solution, carry out deiodination, then precipitate with ethanol...

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Abstract

The invention discloses a TMC / genipin / pDNA nanometer carrier which is prepared from a TMC / genipin nanometer sphere for wrapping the pDNA. The particle diameter of the TMC / genipin / pDNA nanometer carrier is 100nm to 500nm. The preparation method comprises the steps: the monomer chitosan is synthesized into trimethyl chitosan, the nanometer sphere is formed through a cross-linking agent genipin, and then the DNA is carried into the nanometer sphere. The preparation method is simple in technology and low in cost; the prepared nanometer particles are good in stability, good in dispersion in water and controllable in particle diameter; the prepared pDNA nanometer carrier is smaller in effects on of the size and the electric charge quantity in PBS and oil phase; the prepared nanometer particles are better in matter stability than dual monomers, can change the charge distribution of the whole matter and are more suitable for gene therapy and antibody production.

Description

technical field [0001] The invention relates to the field of gene therapy and antibody production, in particular to TMC / genipin / pDNA 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 advantage...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K39/00A61K47/69B82Y5/00
CPCA61K48/0008A61K39/00A61K2039/53A61K2039/55555B82Y5/00
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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