Antigen covalently bound chitosan nanoparticle-based nasal immune carrier

A technology of chitosan nanoparticles and covalent bonding, applied in the field of medicine, can solve the problems of unsatisfactory immune effect, incomplete release, low encapsulation rate, etc., and achieve nasal cavity immune effect, not easy to fall off, and simple preparation method Effect

Active Publication Date: 2015-01-14
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the chitosan nanoparticle is used as the main way of mucosal immune adjuvant to encapsulate the antigen inside the nanoparticle, but this method has obvious disadvantages: (1) the encapsulation efficiency is low; (3) The antigens that can contact APC and be recognized and presented by APC are less, making the immune effect unsatisfactory

Method used

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  • Antigen covalently bound chitosan nanoparticle-based nasal immune carrier
  • Antigen covalently bound chitosan nanoparticle-based nasal immune carrier
  • Antigen covalently bound chitosan nanoparticle-based nasal immune carrier

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Weigh 0.1 g of chitosan, dissolve it in 50 mL of 1% acetic acid, add 3-maleimido propionate hydroxysuccinimide solution dropwise under stirring, and react for 1 h. After the reaction, the reaction solution was dialyzed for 24 h and freeze-dried to obtain maleimide-chitosan (Mal-CS).

[0057] Weighed an appropriate amount of Mal-CS and dissolved it in sodium acetate buffer (pH5.5), added an equal volume of sodium tripolyphosphate solution, and stirred at room temperature for 30 min to prepare a blank nanoparticle suspension. The suspension was mixed with thiolated ovalbumin (OVA) and reacted at room temperature for 10 min to obtain chitosan nanoparticles covalently bound to OVA.

[0058]

Embodiment 2

[0060] Weigh 2.5 g of chitosan, add to 250 mL of 1% acetic acid to dissolve. Add 0.10~0.14 g ammonium persulfate, fill with N 2 Protect, heat and stir, then add 1.2~2.0 g methacryloyloxyethyltrimethylammonium chloride (TMAEMC), and react for 2~5 h. The reaction solution was dialyzed for 24 h and freeze-dried to obtain quaternized chitosan (TMC).

[0061] Weigh 0.5 g of TMC, dissolve it in 50 mL of sodium acetate buffer solution, add 3-maleimidopropionic acid hydroxysuccinimide ester solution dropwise under stirring, and react for 1 h. After the reaction, the reaction solution was dialyzed for 24 h and freeze-dried to obtain maleimide-quaternized chitosan (Mal-TMC).

[0062] The prepared TMC, Mal-TMC were dissolved in deuterated acetic acid / heavy water, 1 H NMR identification of its structure, the results are as follows figure 1 shown; TMC showed a strong (-N + (CH 3 ) 3 ) peak, confirming that TMAEMC has been attached to chitosan. Mal-TMC has an obvious peak (maleimide...

Embodiment 3

[0065] Weighed an appropriate amount of Mal-TMC and dissolved it in HEPES buffer (pH 7.0), added an equal volume of sodium tripolyphosphate solution, and stirred at room temperature for 30 min to prepare a blank nanoparticle suspension. The suspension was mixed with thiolated ovalbumin (OVA), and reacted at room temperature for 10 min to obtain TMC nanoparticles covalently linked by OVA.

[0066]

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Abstract

The invention belongs to the technical field of medicines, and relates to an antigen covalently bound chitosan nanoparticle-based nasal immune carrier and a preparation method thereof. The carrier is composed of a nanoparticle and an antigen, the antigen is covalently bound to the surface of the nanoparticle, and the nanoparticle is prepared by adopting a raw material chitosan and its derivatives; and the antigen is selected from from infectious diseases or tumors related proteins or polypeptides. The carrier can significantly improve the neck lymph node targeting of people nasally dosed with the antigen proteins or polypeptides, enhances the mucosal and systemic immune response, has the advantages of simple preparation, stable process, high antigen utilization rate, difficult antigen shedding, benefiting of the improvement of the cervical lymph node transportation of the antigens, good nasal immune effect and no nasal mucosa toxicity, and is suitable for preventing respiratory infectious diseases.

Description

technical field [0001] The invention belongs to the technical field of medicine, and relates to a nasal immune carrier based on antigen covalently binding chitosan nanoparticles and a preparation method thereof. Background technique [0002] In recent years, respiratory infectious diseases caused by bacteria, fungi, viruses, etc. have occurred frequently, such as the SARS outbreak in 2003, the H5N1 avian influenza outbreak in 2005, the H1N1 influenza pandemic in 2009, and the recent H7N9 new avian influenza, etc. , a serious threat to human health; therefore, how to prevent the spread of respiratory infectious diseases is still a difficult task for medical workers. At present, there is still a lack of effective large-scale air disinfection methods, so the focus of its prevention lies in the vaccination of corresponding vaccines. [0003] Respiratory tract infectious diseases are mainly caused by pathogens entering or infecting the respiratory mucosa, and the nasal mucosa is...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/385A61K39/39A61K47/48
CPCY02A50/30
Inventor 张奇志刘青锋郑晓瑶邵夏炎蒋新国
Owner FUDAN UNIV
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