Trimethyl chitosan-graft-polyethylene glycol/nucleic acid brain-targeting micellar and preparation method thereof

A technology of trimethyl chitosan and polyethylene glycol, which can be used in gene therapy, pharmaceutical formulations, non-effective ingredients of polymer compounds, etc., and can solve problems such as easy degradation and unstable transfection efficiency

Inactive Publication Date: 2013-07-03
SHENYANG PHARMA UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] The purpose of the present invention is to prepare a brain-targeted micelle of trimethyl chitosan-graft-polyethylene glycol-brain-targeted functional peptide / nucleic acid mediated by acetylcholine receptors, so as to realize the passage of nucleic acid drugs Active targeting of the blood-brain barrier and the brain, and solving its clinical application defects such as easy degradation, instability and low transfection efficiency in vivo

Method used

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  • Trimethyl chitosan-graft-polyethylene glycol/nucleic acid brain-targeting micellar and preparation method thereof
  • Trimethyl chitosan-graft-polyethylene glycol/nucleic acid brain-targeting micellar and preparation method thereof
  • Trimethyl chitosan-graft-polyethylene glycol/nucleic acid brain-targeting micellar and preparation method thereof

Examples

Experimental program
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Effect test

example 1

[0045] The purification of example 1 chitosan

[0046] Add 13.8g of chitosan into 1120mL of 1% acetic acid aqueous solution, stir magnetically for 40 minutes to dissolve it, filter it with suction, add ammonia water dropwise to adjust the pH value of the filtrate to 7.0 to precipitate chitosan, and filter it with a 0.45 μm filter membrane. The chitosan product on the filter membrane was washed 3 times with double distilled water, and freeze-dried.

example 2

[0047] The preparation of example 2 N-trimethyl chitosan

[0048] sample 1

[0049] Weigh 0.5 g of purified chitosan (molecular weight 50 kD, degree of deacetylation 95%) into a 50 mL three-necked flask, add 20 mL of N-methylpyrrolidone, and stir and swell at room temperature for 12 hours. Add 1.2 g of sodium iodide to 2.8 mL with a mass concentration of 0.2 g·mL -1 sodium hydroxide solution and 3 mL of methyl iodide, and stirred in a water bath at 60 °C for 45 minutes to complete the first step of methylation. Add 2.8 mL of sodium hydroxide solution and 1.25 mL of methyl iodide to react for 45 minutes to complete the second step of methylation. The reaction solution was added to 200 mL of absolute ethanol and stirred for 1 hour to precipitate trimethyl chitosan. The precipitate was centrifuged (5000 rpm, 15 minutes) and washed three times with ethanol, three times with ether, and then dried in vacuum. After drying, dissolve the product in 20 mL of 50 g L -1 I ions were ex...

example 3 3

[0056] The preparation of example 3 trimethyl chitosan-graft-polyethylene glycol

[0057] sample 1

[0058] 10 mg of trimethyl chitosan and 15 mg of maleimide-polyethylene glycol-succinimide (MAL-PEG-SCM) of sample 1 in Example 2 were dissolved in 2 mL of distilled water, and magnetically stirred at room temperature for 6 hours , ultrafiltration and centrifugation to remove unreacted PEG modifiers, the obtained concentrated solution was diluted with distilled water and then ultrafiltration and centrifugation, repeated 3 times, and finally the ultrafiltrate was freeze-dried.

[0059] Get the trimethyl chitosan (TMC) of example 2 sample 1, the synthetic trimethyl chitosan-graft-polyethylene glycol (TMC-PEG-MAL) and maleimide-polyethylene glycol - Succinimide (MAL-PEG-SCM) 5 mg each, in D 2 O is the solvent 1 H-NMR (300 MHz) characterization, the results entered Figure 4 . Figure 4 Compared with TMC in TMC-PEG-MAL, the extra peak with a chemical shift of 3.713 is -CH in po...

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Abstract

The invention belongs to the technical field of medicine, and relates to an acetylcholine-receptor-mediated trimethyl chitosan-graft-polyethylene glycol / nucleic acid brain-targeting micellar and a preparation method thereof. According to the invention, trimethyl chitosan-graft-polyethylene glycol-brain-targeting functional peptide is obtained through the steps that: chitosan is sequentially subjected to quaternary ammonium modification such that trimethyl chitosan is obtained; the amino group of trimethyl chitosan is subjected to a reaction with an active group of two-end-activated polyethylene glycol, such that trimethyl chitosan-graft-polyethylene glycol is obtained; and another active group of polyethylene glycol is subjected to a reaction with brain-targeting functional peptide RVG. According to the invention, ion composite micellar is adopted as a carrier, and brain-targeting functional peptide RVG is adopted as a targeting molecule, such that nucleic acid medicine can actively target and position through blood-brain barrier and brain. The ion composite micellar provided by the invention is formed through static composition of positively charged trimethyl chitosan-graft-polyethylene glycol-brain-targeting functional peptide and negatively charged nucleic acid. With the invention, nucleic acid medicine defects such as easy in-vivo degradation, poor stability, and low transfection efficiency are solved.

Description

technical field [0001] The invention belongs to the field of medical gene therapy, and relates to a preparation method and technology of a novel high-efficiency non-viral vector, in particular to a trimethyl chitosan-graft-polyethylene glycol-brain targeting function mediated by an acetylcholine receptor Peptide / nucleic acid brain targeting micelles and methods for their preparation. Background technique [0002] The number of people suffering from neurodegenerative and other brain diseases is increasing year by year in the world, but there are still few effective drugs for treating such diseases. Nucleic acid drugs have become the forefront of medical research due to their high specificity, rapid action, safety, and high efficiency. [0003] However, there are insurmountable problems in the application of nucleic acid drugs: first, there are a large number of nucleases in the body, which can hydrolyze the phosphodiester bond of nucleic acid, making the drug degraded and in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K47/42A61K48/00A61K9/10A61P25/28
Inventor 姜同英王思玲高亦鲲
Owner SHENYANG PHARMA UNIVERSITY
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