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Method for preparing recombinant human endostatin chitosan nanoparticles for injection

A technology of chitosan nanoparticles and vascular endothelium, applied in the direction of non-active ingredient medical preparations, medical preparations containing active ingredients, drug combinations, etc., can solve the problems of short half-life and poor clinical compliance, and achieve High drug loading, good biocompatibility, and uniform distribution

Active Publication Date: 2011-01-26
SHANDONG SIMCERE BIO PHARMA CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, as an exogenous protein, recombinant human endostatin is easily recognized by the immune system in the body and then degraded. Therefore, the half-life in the body is very short. In clinical use, patients need to inject once a day for 14 consecutive days as a course of treatment. After one week, proceed to the next course of treatment
Poor clinical compliance due to frequent injections

Method used

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  • Method for preparing recombinant human endostatin chitosan nanoparticles for injection
  • Method for preparing recombinant human endostatin chitosan nanoparticles for injection
  • Method for preparing recombinant human endostatin chitosan nanoparticles for injection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Weigh 500 mg of chitosan with a molecular weight of 100,000 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 sodium polyphosphate solution with distilled water. Add acetate buffer (pH 5.5) containing 200 mg of Endostar into the chitosan solution, slowly add sodium polyphosphate solution dropwise under magnetic stirring, and react for 30 minutes to obtain recombinant human endostatin nanoparticles. The above nanoparticle solution was centrifuged at 4°C and 10,000 rpm for 30 minutes to separate the nanoparticles. The obtained nanoparticles were redispersed in 50 mL of water, 1 g of trehalose was added, and freeze-dried to obtain recombinant human endostatin nanoparticles with uniform particle size distribution.

[0037] 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...

Embodiment 2

[0039]Weigh 500 mg of chitosan with a molecular weight of 100,000 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 sodium polyphosphate solution with distilled water. Take 200mg of Endostar acetate buffer solution (pH5.5) and add it to the chitosan solution, slowly add sodium polyphosphate solution dropwise under magnetic stirring, react for 5 minutes, and ultrasonicate the probe at 100W for 2 minutes to obtain recombinant human blood vessels Endostatin nanoparticles. The above nanoparticle solution was centrifuged at 4°C and 10,000 rpm for 30 minutes to separate the nanoparticles. The obtained nanoparticles were redispersed in 50 mL of water, 1 g of trehalose was added, and freeze-dried to obtain recombinant human endostatin nanoparticles with uniform particle size distribution.

[0040] The determination of drug loading is the same as in Example 1. The freeze-dried nanoparticles were redissolved i...

Embodiment 3

[0042] Weighing 250 mg of chitosan with a molecular weight of 100,000 Daltons was dissolved in 50 mL of 1% acetic acid solution to obtain a 5 mg / mL chitosan solution. Prepare 15 mL of 5 mg / mL sodium polyphosphate solution with distilled water. Add 200 mg of Endostar phosphate buffer solution (pH5.0) into the chitosan solution, slowly add sodium polyphosphate solution dropwise under propeller stirring (300 rpm), react for 10 minutes, and ultrasonically probe for 2 minutes under 100W power, that is Recombinant human endostatin nanoparticles were obtained. The nanoparticle solution was centrifuged at 4000 rpm for 10 minutes with an ultrafiltration membrane with a molecular weight cut-off of 50,000 Daltons to separate the nanoparticles. The obtained nanoparticle concentrate was dispersed in 30 mL of water, 1.5 g of trehalose was added, and freeze-dried to obtain recombinant human endostatin nanoparticles with uniform particle size distribution.

[0043] The ultrafiltrate after c...

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Abstract

The invention relates to a method for preparing recombinant human endostatin chitosan nanoparticles for injection, in particular to the preparation of the recombinant human endostatin chitosan nanoparticles by taking biodegradable chitosan as a carrier and sodium tripolyphosphate as an ion crosslinking agent and performing an ion crosslinking reaction on chitosan solution of recombinant human endostatin and the solution of sodium tripolyphosphate under a continuously stirring condition.

Description

technical field [0001] The invention relates to a preparation method of recombinant human endostatin chitosan nanoparticle for injection, in particular to a preparation method using biodegradable chitosan as a matrix. The particle size range of the nanoparticles prepared by the method is controllable, the drug loading capacity is high, and the nanoparticles are safe, non-toxic and easy to operate. Background technique [0002] Recombinant human endostatin for injection (Endostar, Endostar) was developed by Shandong Simcere Maidejin Biopharmaceutical Co., Ltd. (a holding subsidiary of Simcere Pharmaceuticals), and obtained the new drug certificate issued by the State Food and Drug Administration in 2005. Obtained the production approval in 2006, and is currently the world's first recombinant human endostatin approved by the national food and drug regulatory agency for marketing. [0003] Recombinant human endostatin is produced by modifying the nucleotide coding sequence of ...

Claims

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Application Information

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IPC IPC(8): A61K9/14A61K38/17A61K47/36A61P35/00
Inventor 李玲王青松刘春晖许向阳
Owner SHANDONG SIMCERE BIO PHARMA CO LTD
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