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Applications of fullerene derivatives, vaccine adjuvant, and vaccine preparation thereof

A technology of fullerene derivatives and fullerene carboxyl, which is applied in the fields of biotechnology and vaccines, can solve the problems of poor water solubility of fullerenes, achieve improved defense capabilities, good safety, and improve the level of cellular immunity and humoral immunity Effect

Active Publication Date: 2014-01-08
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore the purpose of the present invention is to provide a kind of fullerene derivative, introduces hydrophilic group on its surface, as hydroxyl (-OH), carboxyl (-COOH) etc., can solve the problem that fullerene water solubility is poor, for its Application lays the groundwork

Method used

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  • Applications of fullerene derivatives, vaccine adjuvant, and vaccine preparation thereof
  • Applications of fullerene derivatives, vaccine adjuvant, and vaccine preparation thereof
  • Applications of fullerene derivatives, vaccine adjuvant, and vaccine preparation thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Fullerenol C 60 (OH) 20 Synthesis

[0037] First, add 1 ml of 40% tetrabutylammonium hydroxide solution (TBAH) and 2 ml of sodium hydroxide solution (2.22 g / ml) to 30 ml of C 60(Sigma company) in toluene solution (1.5mg / ml), react 24h under vigorous stirring; Discard organic phase, and remove aqueous phase with vacuum rotary evaporator (RV06-ML 1-B, Germany); Wash sample with 50ml methanol, Methanol was removed by vacuum evaporation, and washed repeatedly for 3-5 times; after that, 10ml of deionized water was added to the sample, and the stirring was continued until the solution was transparent reddish-brown, then 20ml of deionized water was added, and the Sephadex G-25 chromatographic column was used to (Amersham Company, UK) purified the sample, and finally vacuum-dried to obtain fullerenol nanoparticles. Fullerenols with different numbers of hydroxyl groups can be obtained by adjusting the amount of sodium hydroxide added.

[0038] Utilize ato...

Embodiment 2

[0043] Example 2 Effects of Fullererol on the Vitality of HEK293 Cells (ATCC)

[0044] 1. Preparation of complete medium containing fullerenol:

[0045] First get the amount of fullererol obtained in Example 1, and use sterile deionized water to prepare a 2mM aqueous solution; then use complete DMEM (Dulbecco's modification of Eagle's medium) medium (containing 10% fetal bovine serum, Gibco Company) diluted fullererol aqueous solution to 500 μM, and carried out gradient dilution with complete medium to obtain a series of solutions with concentrations of 500, 100, 20, 4, 0.8, and 0.16 μM for future use.

[0046] 2. Effects of fullerenol on the viability of HEK293 cells:

[0047] First, HEK293 cells were digested with 0.25% trypsin, and then centrifuged at 1000rpm for 5min after the digestion was terminated. Single-cell suspension was prepared in DMEM medium containing 10% fetal bovine serum (FBS), and 5 × 10 4 / ml concentration was inoculated in a 96-well plate, 100 μl p...

Embodiment 3

[0049] Embodiment 3 Fullerenol is on the influence of NIH3T3 cell (ATCC) vigor

[0050] With reference to the method described in Example 2, investigate the effect of fullerenol nanoparticles on the viability of NIH3T3 cells, the results are shown in Figure 6 , similar to the results in Example 2, compared with the negative control group, the cell viability did not change significantly after the cells were treated with fullerenol nanoparticles for 48 hours, even in the highest dose group, the cell viability was still higher than 90%. Figure 7 Indicates the effect of fullerenol on the morphology of NIH3T3 cells, Figure 7 A is the normal control group, Figure 7 B represents the picture of the cells treated for 48 hours when the working concentration of fullerenol was 500 μM. It can be seen that the morphology of the cells in the test group is the same as that in the normal control group, showing a long spindle shape. All these indicate that fullerenol has good biocompati...

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Abstract

The invention provides fullerene derivatives, the fullerene derivatives are fullerol represented by a formula of C60OxHy (x is less than 50 and more than or equal to y; y is more than 10 and less than or equal to x) or fullerene carboxyl derivatives represented by a formula of C60(C(COOH)2)n (n is an integer from 1 to 4), and the diameter range of nano particles of the fullerene derivatives in a solvent is from 1 to 400 nm. The invention also provides applications of a vaccine adjuvant of the fullerene derivatives in the vaccine field; the vaccine adjuvant has a good safety, can prominently improve the vaccine immunogenicity through different immunity ways, and has a very large application prospect.

Description

[0001] This application is a divisional application of an invention patent application with an application date of December 24, 2010, an application number of 201010621120.5, and an invention title of "Application of Fullerene Derivatives and Vaccine Adjuvant and Vaccine Preparation". technical field [0002] The present invention relates to a kind of fullerene derivative, specifically, relate to fullerene alcohol particle C 60 o x h y (10<y≤x<50) and fullerene carboxyl derivatives C 60 (C(COOH) 2 ) n (n is an integer of 1-4) and the vaccine adjuvant and vaccine preparation of the fullerene derivative and its application in the field of vaccines belong to the fields of biotechnology and vaccines. Background technique [0003] Vaccines play an extremely important role in the prevention and treatment of major diseases, especially infectious diseases. In the process of vaccine research and development, safe and effective vaccines are mainly obtained through design op...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/39
Inventor 陈春英邵一鸣许利耕郝彦玲刘颖刘野
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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