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Human serum albumin complex hydrophobically modified pullulan nanoparticles and preparation method thereof

A technology of human serum albumin and pullulan polysaccharide, which can be used in drug combinations, steroids, inactive components of polymer compounds, etc., can solve problems such as no literature reports, and achieve stable blood drug concentration and delayed release. Effect

Inactive Publication Date: 2011-08-10
INST OF BIOMEDICAL ENG CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the research on hydrophobically modified pullulan nanoparticles is mainly to use it as a drug carrier to explore its drug release and targeting, and further to prepare human serum albumin compounded by compounding human serum albumin and nanoparticles. Cholesterol hydrophobically modified pullulan nanoparticles to improve its drug release and pharmacodynamic properties, so far no literature reports

Method used

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  • Human serum albumin complex hydrophobically modified pullulan nanoparticles and preparation method thereof
  • Human serum albumin complex hydrophobically modified pullulan nanoparticles and preparation method thereof
  • Human serum albumin complex hydrophobically modified pullulan nanoparticles and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1: Synthesis of cholesterol hydrophobically modified pullulan and calculation of degree of substitution

[0034]Dissolve 2.5g (6.5mmol) of cholesterol modification and 2.0g (20mmol) of succinic anhydride in 20mL of anhydrous pyridine, react at room temperature for 48h, stop the reaction, drop the reaction solution into glacial hydrochloric acid solution with pH=1~2, and precipitate White flocculent precipitate. Refrigerate for 16 hours, filter with suction, and collect the precipitate. The precipitate was washed with distilled water until pH>5, recrystallized in ethyl acetate / ethanol, and dried at 80°C to obtain 2.0 g of pure white needle-like succinylcholesterol (CHS).

[0035] Take 0.5g of a pullulan sample and dissolve it in 15ml of dehydrated dimethyl sulfoxide for subsequent use; take succinylcholesterol (CHS), 4-lutidine (DMAP / CHS=1, mmol / m mol) , (EDC / CHS=1.2, mmol / mmol), dissolved in 10ml DMSO, stirred at room temperature, and activated for 1h, the act...

Embodiment 2

[0038] Example 2: Preparation method and characterization of cholesterol hydrophobically modified pullulan nanoparticles

[0039] Take 20 mg of each CHP polymer and add 1 mL of DMSO solvent to dissolve. The solution was transferred into a dialysis bag, put into 3L distilled water, changed the water every 8 hours, dialyzed for 48 hours, and dialyzed the DMSO clean. After the dialysis, the volume was constant, 100W power ultrasonic for 10s, and filtered with a 0.45μm microporous membrane. figure 2 It shows that under dynamic light scattering, the CHP nanoparticles present a unimodal distribution with a particle size of about 100nm. Samples were prepared on a copper grid, stained with 2% phosphopentanoic acid, and the morphology of the nanoparticles was spherical in TEM observation. The particle size and Zeta potential of the nanoparticles were measured by dynamic light scattering, and the Zeta potential of the CHP nanoparticles was about -3mV.

Embodiment 3

[0040] Example 3: Determination of changes in the fluorescence intensity of nanoparticles and proteins complexed and changes in the secondary structure of proteins

[0041] CHP nanoparticles were mixed with human serum albumin, and HSA nanoparticles were set as a blank control group with the same protein concentration. A fluorescence spectrophotometer was used to measure the fluorescence spectra of the two, and the excitation wavenumber was set to 275nm, and the emission wavenumber was set to 287-345nm. The mixed solution was inserted into a 2ml EP tube, and then the mixed solution was placed in a shaker at 37° C. with a rotational speed of 20 rpm for 10 h. The samples were taken out every 2 hours, and the fluorescence intensity of all the samples was recorded in the above-mentioned manner, and the maximum fluorescence intensity was taken to determine the time when the protein and the nanometer complexed completely. The circular dichroism spectrophotometer measures the circul...

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Abstract

The invention relates to human serum albumin complex hydrophobically modified pullulan nanoparticles and a preparation method thereof. Hydrophobic group cholesterol, pullulan polysaccharides, succinic anhydride and human serum albumin are taken as raw materials for synthesis; the specific synthesis process is as follows: enabling the cholesterol and the succinic anhydride in the mass ratio of (1-2.5): 2 to react to firstly synthesize succinic anhydride cholesterol, and enabling the succinic anhydride cholesterol to react with the pullulan polysaccharides to synthesize cholesterol hydrophobically modified pullulan polysaccharides in the presence of a catalyst, wherein the mass ratio of cholesterol succinate (CHS) to sugar moiety of the pullulan polysaccharides is (1-4): 20; and performing further self-assembly on the hydrophobically modified pullulan polysaccharides to form a solution of the nanoparticles, then incubating the nanoparticles and the human serum albumin in a shaker for 9 hours, and finally getting the human serum albumin complex hydrophobically modified pullulan nanoparticles. The nanoparticles after complexation of the human serum albumin and the hydrophobically modified pullulan polysaccharide nanoparticles are shaped like spheres, and can maintain slow release effect of the medicament-loaded nanoparticles in a long lasting manner and stabilize plasma concentration.

Description

Technical field: [0001] The invention relates to a hydrophobically modified pullulan nanoparticle compounded with human serum albumin which can be used as a drug carrier and a preparation method. Specifically, pullulan is hydrophobically modified to generate a polymer, which self-assembles into nanospheres with a shell-core structure in aqueous solution, and further obtains a complex hydrophobic modification by compounding human serum albumin (Human Serum Albumin, HSA). Sexual pullulan nanoparticles. Background technique: [0002] As a drug carrier, the preparation of self-aggregating nanoparticles based on natural polymer materials has attracted more and more attention. Polysaccharides are relatively abundant natural polymers in nature. Many polysaccharides also have anti-tumor and immunomodulatory activities. Therefore, the modification of polysaccharides and their derivatives to form amphiphilic graft copolymers has a wide range of applications in the fields of medicine ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K47/42A61K9/16C07J9/00C08B37/00A61P35/00
Inventor 张其清陶晓军李学敏王建华
Owner INST OF BIOMEDICAL ENG CHINESE ACAD OF MEDICAL SCI
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