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Oligoarginine covalently modified chitosan, and preparation method, screening and application thereof

A technology of oligoarginine and chitosan, which is applied in the field of biomedical materials, can solve the undiscovered effect of 2-7 oligoarginine on chitosan nucleic acid delivery efficiency, low transfection efficiency, and limitation of chitosan Sugar application and other issues, to achieve good anti-tumor effect, simple preparation method, and improved cell uptake efficiency

Inactive Publication Date: 2016-01-06
INST OF MATERIA MEDICA CHINESE ACAD OF MEDICAL SCI
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

[0004] The application of chitosan to deliver siRNA was first reported in 2006, (see KatasH, AlparHO, Development and characterization of nanoparticles for siRNA delivery [J], JControlRelease, 2006, 115 (2): 216-225.) But lower transfection efficiency limits chitosan in Applications in the field of siRNA delivery
The present invention has been searched by patent and literature, and no report has been found to explore the influence of 2-7 oligoarginine on the delivery efficiency of chitosan nucleic acid

Method used

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  • Oligoarginine covalently modified chitosan, and preparation method, screening and application thereof
  • Oligoarginine covalently modified chitosan, and preparation method, screening and application thereof
  • Oligoarginine covalently modified chitosan, and preparation method, screening and application thereof

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Embodiment 1

[0055] Embodiment 1: the preparation of the Rn modified chitosan whose degree of substitution is 10%

[0056] (1) Weigh a certain 5×10 -4 Mole of Rn, put in a 25ml round bottom flask, add 10ml of acetic acid / sodium acetate buffer solution with pH 5.5, add EDC and NHS at a molar ratio of 1:1.5, stir and activate at room temperature for 4 hours;

[0057] (2) Add chitosan to the above mixture at a molar ratio of Rn: CS (Mw: 11.3kDa) at a ratio of 1:5, stir and dissolve at room temperature, and continue to react for 20 hours;

[0058] (3) After the reaction, the reaction liquid was transferred to a dialysis bag of 3500-12000 ℃, dialyzed with deionized water for 48 hours, and the product after dialysis was freeze-dried for 36 hours to prepare oligoarginine-modified chitosan.

[0059] The polymer synthesis route is attached figure 1 Shown, the degree of substitution of the product by 1 HNMR calculation, the results are shown in Table 1 (the degree of substitution of oligoarginine...

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Abstract

The invention provides a novel carrier CS-g-Rn a series of oligoarginine (Arginine, called R for short, R2-R7) covalently modified chitosan, and an in-vivo and in-vitro application research for preparing nanoparticles through loading DNA and functional siRNA by using the highly effective carrier. The oligoarginine is covalently combined to the free amino group of a chitosan molecule through a one-step synthesis method, the prepared new carrier maintains the safety of chitosan, and the new carrier and nucleic acid with the negatively charged surface undergo complex coacervation to prepare the nanoparticles, so the nucleic acid delivery efficiency is effectively improved. The nanoparticles prepared through loading of the functional siRNA on CS-g-Rn have good treatment effects in vivo and in vitro, and have good application prospect.

Description

technical field [0001] The invention belongs to the field of biomedical materials and relates to a polymer material, in particular to a series of chitosan derivatives covalently modified by oligoarginine, its preparation method, screening and application. Background technique [0002] Gene therapy (GeneTherapy), as a new treatment method, has become the research frontier in the field of medicine. Among them, nucleic acid therapy provides a new treatment method for many intractable diseases including cancer. However, the lack of safe and efficient nucleic acid delivery vectors has always been a bottleneck that limits its application. At present, nucleic acid delivery vectors are mainly divided into two categories: viral vectors and non-viral vectors. Viral vectors have high transfection efficiency, but there are safety issues such as immune response, cytopathic changes, teratogenicity, and mutagenicity in application. Based on the above reasons, non-viral vectors are ideal ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08B37/08C12N15/87C12N15/85
Inventor 高钟镐黄伟杨飞飞柳珊金明姬
Owner INST OF MATERIA MEDICA CHINESE ACAD OF MEDICAL SCI
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