Quaternary ammonium type cationic starch gene controlled release vector material and preparation method and application thereof

A technology of cationic starch and controlled release carrier, applied in the field of natural polymer materials in medicine, can solve the problems of degradation, unsolved gene transfection efficiency, inability to escape the recognition and clearance of the reticuloendothelial system, etc., and achieves good biocompatibility Effect

Active Publication Date: 2012-04-11
SOUTH CHINA UNIV OF TECH
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  • Abstract
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Problems solved by technology

However, since the DNA is adsorbed on the surface of the nanosphere, which is similar in form to the general plasmid vector, it cannot escape the recognition and removal of the reticuloendothelial system during the transfection process.
Therefore, there are disadvantages such as low DNA transfection rate and easy degradation by intracellular restriction endonucleases and capture by the body's immune system.
Liu Jun et al. (Liu Jun, Liu Xuanming, Xiao Suyao, etc. Gene transduction based on starch nanoparticles as carriers under ultrasound. Chemical Journal of Chinese Universities. 2005, 26: 634-637) Based on ultrasound energy, cell walls, cell membranes and even nuclear membranes can be instantly Channel, through which exogenous DNA can enter cells and express, established a method for ultrasound-mediated starch nanoparticles to improve the efficiency of exogenous gene transfection, but it did not substantially solve the problem of starch carrier itself on gene transfection. inefficiency problem

Method used

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  • Quaternary ammonium type cationic starch gene controlled release vector material and preparation method and application thereof
  • Quaternary ammonium type cationic starch gene controlled release vector material and preparation method and application thereof
  • Quaternary ammonium type cationic starch gene controlled release vector material and preparation method and application thereof

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Experimental program
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Effect test

Embodiment 1

[0022] The water content is 4.7%, the weight average molecular mass (M w ) is 100g (dry basis) of starch of 13000Da in a kneader and the mass percent of stirring and mixing in an ice-water bath is 18.5% NaOH (18.5g), 20.3% water (20.3g) and 87.0% 3-chloro- The mixed system of dihydroxypropyltrimethylammonium chloride (87.0g) was dry-mixed and reacted for 48 hours at a temperature of 25°C and a rotating speed of 100 rpm; After filtering, wash with absolute ethanol until AgNO 3 Check for Cl - Dry at 60° C., crush and pass through an 80-mesh sieve to obtain a quaternary ammonium cationic starch gene controlled-release carrier material with a degree of substitution (DS) of 0.43 (Table 1). Other conditions are constant, change the weight-average molecular weight of starch and the consumption (table 1) of NaOH and 3-chloro-dihydroxypropyltrimethyl ammonium chloride, the quaternary ammonium type cationic starch gene controlled release carrier material of gained different degree of ...

Embodiment 2

[0027] The water content is 13.5%, the weight average molecular mass (M w) is 100g (dry basis) of starch 100g (dry base) of 64000Da in a kneader and the mass percent of stirring and mixing in an ice-water bath is 18.5% KOH (18.5g), 11.5% water (11.5g) and 87.0% 3-chloro- Dihydroxypropyltrimethylammonium chloride (87.0g) mixed system, at a temperature of 30°C and a rotation speed of 40 rpm, dry mixed reaction for 48 hours; after the reaction was complete, the sample was precipitated with absolute ethanol, and suction filtered After that, wash with absolute ethanol until AgNO 3 Check for Cl - Dry at 60° C., crush and pass through an 80-mesh sieve to obtain a quaternary ammonium cationic starch gene controlled-release carrier material with a degree of substitution (DS) of 0.19 (Table 2). Other conditions are constant, change the weight-average molecular weight of starch and the consumption (table 9) of NaOH and 3-chloro-dihydroxypropyltrimethyl ammonium chloride, the quaternary...

Embodiment 3

[0031] The water content is 4.7%, the weight average molecular mass (M w ) is 100g (dry basis) of starch of 17000Da in a kneader and the mass percent of stirring and mixing in an ice-water bath is 18.5% NaOH (18.5g), 20.3% water (20.3g) and 87.0% 3-chloro- Dihydroxypropyltrimethylammonium chloride (87.0g) mixed system, at a temperature of 30°C and a rotation speed of 75 rpm, dry mixed reaction for 48 hours; after the reaction was complete, the sample was precipitated with absolute ethanol, and suction filtered After that, wash with absolute ethanol until AgNO 3 Check for Cl - Dry at 60° C., crush and pass through an 80-mesh sieve to obtain a quaternary ammonium cationic starch gene controlled-release carrier material with a degree of substitution (DS) of 0.35 (Table 3). Other conditions are constant, change the weight-average molecular weight of starch and the consumption (table 3) of NaOH and 3-chloro-dihydroxypropyl trimethyl ammonium chloride, the quaternary ammonium type...

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Abstract

The invention discloses a quaternary ammonium type cationic starch gene controlled release vector material, and a preparation method and an application thereof; the preparation method particularly comprises the following steps: allowing starch which has a water content of 4.7-13.5% and a weight-average molecular weight of 13000-64000 Da to mix and react with a mixed system of solid caustic soda, water, and a quaternary ammonium type cationic etherifying agent which are stirred and well mixed through an ice water bath in a kneader by a dry method under a condition with a temperature of 25-30 DEG C and a rotating speed of 40-100 r/min for 12-48 hours; after the reaction is completed, washing, drying, and crushing to obtain the quaternary ammonium type cationic starch gene controlled releasevector material. An application of the vector material prepared by the invention as a non-viral vector material in gene therapy realizes the controlled release of gene drugs, improves the drug bioavailability, reduces the toxic and side effect of the drugs and the vector.

Description

technical field [0001] The invention relates to the field of research and development of natural polymer materials in the field of medicine, especially the application of starch carrier materials in the field of controlled release of genes. Background technique [0002] In the process of gene therapy, gene drugs must successfully enter the nuclei of recipient cells at the lesion site to participate in regulation or expression, in order to reflect the efficacy of gene therapy. However, genetic medicines are mostly nucleic acid substances such as DNA or RNA. The exposed nucleic acid substances are easily degraded by nucleases in the external environment and the human body. Negatively charged, the same as the charge of the cell membrane, resulting in homosexual repulsion, so it is difficult to pass through the outer membrane of the recipient cell and enter the nucleus. Therefore, the delivery of gene drugs requires a certain delivery system. The gene drug delivery systems curr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08B31/12A61K48/00
Inventor 李晓玺杜云翔黄晓仪陈玲李琳李冰苏健裕
Owner SOUTH CHINA UNIV OF TECH
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