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Method for preparing chitosan nanoparticles

A technology of chitosan nanoparticles and chitosan, which is applied in the direction of medical preparations with non-active ingredients, medical preparations containing active ingredients, and pharmaceutical formulas, can solve the problems of poor clinical compliance and achieve good biological Compatibility, high drug loading, uniform distribution effect

Active Publication Date: 2010-12-08
SHANDONG SIMCERE BIO PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Poor clinical compliance due to frequent injections

Method used

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  • Method for preparing chitosan nanoparticles
  • Method for preparing chitosan nanoparticles
  • Method for preparing chitosan nanoparticles

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Weigh 500 mg of chitosan with a molecular weight of 50,000 Daltons and dissolve it in 50 mL of 1% acetic acid solution to obtain a 10 mg / mL chitosan solution. Prepare 20 mL of 5 mg / mL ammonium sulfate solution with distilled water. Slowly add ammonium sulfate solution dropwise under magnetic stirring, react for 30 minutes, sonicate the probe at 100W power for 2 minutes, centrifuge the above solution at 4 degrees Celsius and 10,000 rpm for 30 minutes to separate nanoparticles. Gained nanoparticles were redispersed in 50 mL of water to obtain chitosan nanoparticles. Use nano-ZS90 Malvern particle size analyzer to measure the particle size of nanoparticles, the average particle size is 826nm, the particle size distribution see figure 1 .

Embodiment 2

[0032] Weigh 200 mg of chitosan with a molecular weight of 100,000 Daltons and dissolve it in 20 mL of 1% acetic acid solution to obtain a 10 mg / mL chitosan solution. Prepare 20 mL of 5 mg / mL sodium sulfate solution with distilled water. Add 100mg of Endostar acetate buffer solution (pH5.5) into the chitosan solution, slowly add sodium sulfate solution dropwise under magnetic stirring, react for 10 minutes, ultrasonicate the probe for 2 minutes under 100W power, and put the above solution at 4 degrees Celsius, 15000 Centrifuge at rpm for 30 minutes to separate the nanoparticles. Gained nanoparticles were redispersed in 30 mL of water to obtain chitosan nanoparticles.

[0033] The supernatant solution after centrifugation was appropriately diluted, and the BCA method was used (the kit was from ThermoScientific, named BCA TM Protein Assay Kit) was used to measure the protein concentration, that is, the free protein concentration, and the weight percentage of recombinant human...

Embodiment 3

[0035] Weigh 200 mg of chitosan with a molecular weight of 100,000 Daltons, dissolve it in 40 mL of 1% acetic acid solution to obtain a 5 mg / mL chitosan solution, and add 200 μL of Tween 80 to dissolve it. Prepare 40 mL of 2 mg / mL sodium sulfate solution with distilled water. Take 100mg of Endostar acetate buffer solution (pH5.5) and add it to the chitosan solution containing Tween, slowly add sodium sulfate solution dropwise under magnetic stirring, react for 20 minutes, and ultrasonicate the probe for 2 minutes under 100W power, and dissolve the above solution Centrifuge at 15,000 rpm for 30 minutes at 4°C to separate nanoparticles. Gained nanoparticles were redispersed in 50 mL of water to obtain chitosan nanoparticles.

[0036] The determination of drug loading is the same as in Example 2. The particle size of the nanoparticle solution was measured with a nano-ZS90 Malvern particle size analyzer. The drug loading of the obtained nanoparticles is 20.1%, the average parti...

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Abstract

The invention relates to a method for preparing recombinant human endostatin (rh-endostatin) nanoparticles for injection. The nanoparticles comprise chitosan and the rh-endostatin. A coacervation process is adopted for preparing the nanoparticles, namely a coagulant is added into chitosan solution containing the rh-endostatin, the mixture is subjected to stirring, ultrasonic sound treatment and centrifugation, and finally the deposit is re-mixed and suspended in water to form the nanoparticles. The preparation method is simple, and the particle size of the obtained nanoparticles is 400 to 900 nanometers.

Description

technical field [0001] The invention relates to a preparation method of recombinant human endostatin chitosan nanoparticles for injection. Background technique [0002] Chitosan is the product of deacetylation of chitin, containing β-(1,4)-2-acetamido-D-glucose units and β-(1,4)-2-amino-D-glucose Units of copolymers, the latter generally exceeding 65%. According to different preparation methods, chitosan with different degrees of deacetylation and average molecular weight can be obtained. Chitosan is a cationic polymer, insoluble in common organic solvents, stable in lye, and a linear polymer electrolyte under acidic conditions. Its solution has a certain viscosity. The higher the concentration of the solution or the greater the molecular weight, the greater the viscosity. bigger. [0003] Recombinant human endostatin for injection (Endostar, Endostar) is the first new endostatin drug for lung cancer, developed by Shandong Simcere Medicin Biopharmaceutical Co., Ltd. (a ho...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/17A61K47/36A61K9/16A61P35/00
Inventor 李玲王青松刘春晖许向阳
Owner SHANDONG SIMCERE BIO PHARMA CO LTD
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