Biotinylated pulullan polysaccharide nanometer grains and the preparation method

Abstract

The invention relates to a method for preparing bioepiderm pluran-polysaccharide nanometer particles, which comprises that dissolving bioepiderm pluran-polysaccharide derivant in organic solvent, adding it into water to form suspension, then dialyzing the suspension, filtering, concentrating, freezing and drying to obtain the nanometer whose average diameter is 100-300nm. And the preparation only needs some or no surface activator, with warm reaction condition and high safety. And the preparation nanometer particle has smooth and regular surface, which can combine tumor cell, as tumor target carrier material.

Description

technical field

[0001] The invention relates to the field of medicine, in particular to a biotinylated pullulan nanoparticle and a preparation method thereof. technical background

[0002] Pullulan is also translated into Chinese as pullulan, pullulan, pullulan or pullulan. It is an exopolysaccharide produced by Phyllostachys sporogenes. It is mainly composed of α-1,6-glucosidic bonds combined with maltose to form homopolysaccharides, that is, glucose is combined with α-1,4-glycosidic bonds to form maltotriose. α-1,6-glucosidic bonds are combined with other maltotriose, so that they are linked repeatedly to form high molecular polysaccharides. The ratio of α-1,4-glycosidic bond to α-1,6-glycosidic bond is 2:1, and the degree of polymerization (D.P) is 100-5000. The average molecular weight of commercial pullulan sugar is 2×10 5 , composed of about 480 maltotriose. The substance is non-toxic, non-mutagenic, colorless and tasteless, edible, has good biocompatibility, can b...

Examples

Embodiment 1

[0021] Take 2.44g of biotin, 2.06g of dicyclohexylcarbodiimide (DCC) and 611mg of 4-dimethylaminopyridine (DMAP) and add them into 50ml of anhydrous dimethyl sulfoxide (DMSO) to dissolve completely, and react for 1 hour . Subsequently, 244 mg of pullulan was added to the above reaction solution to react for 6 days, and the reaction was carried out under magnetic stirring. After the reaction was completed, the reaction solution was suction-filtered three times with a Buchner funnel to remove the precipitate DCU in the reaction solution. Put every 25ml of the filtrate into a dialysis bag, and then put the dialysis bag into 3L deionized water for dialysis for 6 days, changing the water every 30 minutes. After the dialysis, take out the white precipitate in the dialysis bag, use Buchner funnel to suction filter 3 times, collect the white solid on the funnel, wash 3 times with 200ml dichloromethane, then wash 3 times with deionized water, and dry , to obtain a white solid substan...

Embodiment 2

[0026] 10 mg of biotinylated pullulan prepared in Example 1 was completely dissolved in 2 ml of DMSO, and the 2 ml of DMSO solution was added to 4 ml of deionized water under stirring, and a nanosuspension with light blue opalescence was rapidly formed. Pour the nanosuspension into a dialysis bag (molecular weight cut off 8,000-14,000), and dialyze in 3L deionized water for 8 hours. The dialyzed nanosuspension was concentrated by ultrafiltration by centrifugation at 1200 rpm for 15 minutes using an ultrafiltration tube with a molecular weight cut off of 30,000. 5% mannitol was added to the ultrafiltrate and freeze-dried to obtain biotinylated pullulan nanoparticles in the form of white powder. Take a certain amount of white powder and redisperse it in a certain volume of deionized water, take the dispersed nano suspension and put it into the sample cell of the laser particle size analyzer for particle size analysis. The measured average particle diameter is 123.6±0.7 nm, and ...

Embodiment 3

[0028] Take 10 mg of biotinylated pullulan and completely dissolve it in 2 ml of DMSO, and add the 2 ml of DMSO solution dropwise to the water phase mixed with 5 ml of deionized water and 5 ml of ethanol under low-speed magnetic stirring, and quickly form a light blue opalescent Nanosuspension. Pour the nanosuspension into a dialysis bag (molecular weight cut off 8,000-14,000), and dialyze in 3L deionized water for 8 hours. The dialyzed nanosuspension was concentrated by ultrafiltration by centrifugation at 1200 rpm for 15 minutes using an ultrafiltration tube with a molecular weight cut off of 30,000. 5% mannitol was added to the ultrafiltrate and freeze-dried to obtain biotinylated pullulan nanoparticles in the form of white powder. The average particle size is 170.7±0.5 nanometers, and the dispersion coefficient is 0.065.

technical field

[0001] The invention relates to the field of medicine, in particular to a biotinylated pullulan nanoparticle and a preparation method thereof. technical background

[0002] Pullulan is also translated into Chinese as pullulan, pullulan, pullulan or pullulan. It is an exopolysaccharide produced by Phyllostachys sporogenes. It is mainly composed of α-1,6-glucosidic bonds combined with maltose to form homopolysaccharides, that is, glucose is combined with α-1,4-glycosidic bonds to form maltotriose. α-1,6-glucosidic bonds are combined with other maltotriose, so that they are linked repeatedly to form high molecular polysaccharides. The ratio of α-1,4-glycosidic bond to α-1,6-glycosidic bond is 2:1, and the degree of polymerization (D.P) is 100-5000. The average molecular weight of commercial pullulan sugar is 2×10 5 , composed of about 480 maltotriose. The substance is non-toxic, non-mutagenic, colorless and tasteless, edible, has good biocompatibility, can b...

Examples

Embodiment 1

[0021] Take 2.44g of biotin, 2.06g of dicyclohexylcarbodiimide (DCC) and 611mg of 4-dimethylaminopyridine (DMAP) and add them into 50ml of anhydrous dimethyl sulfoxide (DMSO) to dissolve completely, and react for 1 hour . Subsequently, 244 mg of pullulan was added to the above reaction solution to react for 6 days, and the reaction was carried out under magnetic stirring. After the reaction was completed, the reaction solution was suction-filtered three times with a Buchner funnel to remove the precipitate DCU in the reaction solution. Put every 25ml of the filtrate into a dialysis bag, and then put the dialysis bag into 3L deionized water for dialysis for 6 days, changing the water every 30 minutes. After the dialysis, take out the white precipitate in the dialysis bag, use Buchner funnel to suction filter 3 times, collect the white solid on the funnel, wash 3 times with 200ml dichloromethane, then wash 3 times with deionized water, and dry , to obtain a white solid substan...

Embodiment 2

[0026] 10 mg of biotinylated pullulan prepared in Example 1 was completely dissolved in 2 ml of DMSO, and the 2 ml of DMSO solution was added to 4 ml of deionized water under stirring, and a nanosuspension with light blue opalescence was rapidly formed. Pour the nanosuspension into a dialysis bag (molecular weight cut off 8,000-14,000), and dialyze in 3L deionized water for 8 hours. The dialyzed nanosuspension was concentrated by ultrafiltration by centrifugation at 1200 rpm for 15 minutes using an ultrafiltration tube with a molecular weight cut off of 30,000. 5% mannitol was added to the ultrafiltrate and freeze-dried to obtain biotinylated pullulan nanoparticles in the form of white powder. Take a certain amount of white powder and redisperse it in a certain volume of deionized water, take the dispersed nano suspension and put it into the sample cell of the laser particle size analyzer for particle size analysis. The measured average particle diameter is 123.6±0.7 nm, and ...

Embodiment 3

[0028] Take 10 mg of biotinylated pullulan and completely dissolve it in 2 ml of DMSO, and add the 2 ml of DMSO solution dropwise to the water phase mixed with 5 ml of deionized water and 5 ml of ethanol under low-speed magnetic stirring, and quickly form a light blue opalescent Nanosuspension. Pour the nanosuspension into a dialysis bag (molecular weight cut off 8,000-14,000), and dialyze in 3L deionized water for 8 hours. The dialyzed nanosuspension was concentrated by ultrafiltration by centrifugation at 1200 rpm for 15 minutes using an ultrafiltration tube with a molecular weight cut off of 30,000. 5% mannitol was added to the ultrafiltrate and freeze-dried to obtain biotinylated pullulan nanoparticles in the form of white powder. The average particle size is 170.7±0.5 nanometers, and the dispersion coefficient is 0.065.

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