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Lipidosome nucleic acid vaccine adjuvant as well as preparation method and application thereof

A vaccine adjuvant and body nucleic acid technology, applied in the directions of liposome delivery, antiviral agents, pharmaceutical formulations, etc., can solve the problems of low effective delivery rate, weak immune response, no immune response, etc., and achieves simplified preparation process, Improves immune response and improves presentation efficiency

Inactive Publication Date: 2015-12-02
INST OF PROCESS ENG CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Although cationic liposomes can be complexed by the electrostatic interaction between positively charged cationic lipids and negatively charged nucleic acid molecules, they can effectively carry nucleic acid molecules; yet, there are two difficulties in it: (1) nucleic acid vaccines need to be placed in the nucleus However, the cationic liposome nucleic acid system cannot effectively escape from the inclusion body after entering the cell, so it cannot enter the nucleus, and eventually leads to weak or no immune response; (2) the cationic liposome nucleic acid system lacks Targeting, cannot be enriched at the target release site, so the effective delivery rate is low and the immune response is weak

Method used

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  • Lipidosome nucleic acid vaccine adjuvant as well as preparation method and application thereof
  • Lipidosome nucleic acid vaccine adjuvant as well as preparation method and application thereof
  • Lipidosome nucleic acid vaccine adjuvant as well as preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0053] The preparation of liposome nucleic acid vaccine adjuvant comprises the steps:

[0054] Weigh cationic liposome DOTAP55.84mg, cholesterol 30.932mg, DSPE-PCB2061.46mg and DSPE-PCB20-mannose 26.34mg in a 100mL round bottom flask, add chloroform to fully dissolve, at 45°C, speed 122rpm / min The chloroform was removed by rotary evaporation under reduced pressure to form a thin film, and dried overnight in a vacuum oven to remove residual chloroform. Then, 10 mL of sterile phosphate buffered saline (1×PBS, pH=7.4) was added to the flask, and the flask was sonicated at 37° C. for 30 min to obtain a light yellow translucent emulsion. Add the emulsion into a high-pressure homogenizer, and under the condition of a pressure of 100 MPa, homogenize under high pressure for 3 times to prepare the liposome nucleic acid vaccine adjuvant.

Embodiment 2

[0056] Weigh cationic liposome DDAB55.84mg, 1,2-oleoylphosphatidylethanolamine 30.932mg, DG-PCB2071.556mg and DG-PCB20-folate 107.326mg in a 100mL round bottom flask, add chloroform to fully dissolve, and Under the conditions of 55° C. and 152 rpm / min, the organic solvent was evaporated under reduced pressure to form a thin film, and dried overnight in a vacuum oven to remove residual chloroform. Then, 10 mL of sterile phosphate buffered saline (1×PBS, pH=7.4) was added to the flask, and the flask was sonicated at 37° C. for 60 min to obtain a light yellow translucent emulsion. Add the emulsion into a high-pressure homogenizer, and under the condition of a pressure of 100 MPa, homogenize under high pressure for 3 times to prepare a novel liposome nucleic acid vaccine adjuvant.

Embodiment 3

[0058] Weigh cationic liposome DOGS55.84mg, distearoylphosphatidylethanolamine 30.932mg, DC-PCB50107.318mg and DC-PCB50-RGD160.977mg in a 100mL round-bottomed flask, add chloroform to fully dissolve, and place at 60℃ , under the condition of rotating speed 182rpm / min, the organic solvent was removed by rotary evaporation under reduced pressure to form a thin film, and dried overnight in a vacuum oven to remove residual chloroform. Then, 10 mL of sterile phosphate buffered saline (1×PBS, pH=7.4) was added to the flask, and the flask was sonicated at 37° C. for 1 h to obtain a light yellow translucent emulsion. Add the emulsion into a high-pressure homogenizer, and under the condition of a pressure of 100 MPa, homogenize under high pressure for 3 times to prepare a novel liposome nucleic acid vaccine adjuvant.

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Abstract

The invention relates to a lipidosome nucleic acid vaccine adjuvant as well as a preparation method and application thereof. The lipidosome nucleic acid vaccine adjuvant is composed of cationic lipid, auxiliary lipid, polycarboxylic abromine neutral lipid and targeting molecule modified polycarboxylic abromine lipid molecules. The targeting molecules facilitate the system to target the immune cells; the cationic lipid supports the nucleic acid vaccine through electrostatic action; polycarboxylic abromine neutral lipid facilitates the escape of the nucleic acid vaccine from cell endosomes, so that the nucleic acid vaccine can enter cell nucleus for expression, and the nucleic acid is improved in presentation efficiency, generates an adjuvant effect, and greatly improves immune response. According to the invention, the problems of vaccination infection caused by inactivation failure during conventional attenuation and inactivation of the vaccine and the strict requirement for biosafety during the production process are solved; in addition, the lipidosome nucleic acid vaccine adjuvant provided by the invention is simplified in preparation technology and good in biosafety, and can be developed and applied to the prevention and immunity treatment of human serious diseases, such as AIDS and tumors.

Description

technical field [0001] The invention relates to the field of nucleic acid vaccines, in particular to a liposome nucleic acid vaccine adjuvant, a preparation method thereof and an application in the field of prevention and immunotherapy vaccines. Background technique [0002] At present, the occurrence and prevalence of infectious diseases (such as AIDS, SARS, etc.) or certain chronic diseases (such as tumors) seriously threaten human health and social stability. Vaccines (a type of biologically active product) are currently the only weapon that humans can control an infectious disease. Vaccination not only protects the body from the invasion of infectious disease pathogens, but also limits the spread of pathogenic microorganisms in the population. [0003] Traditional attenuated vaccines or inactivated vaccines made from attenuated or inactivated pathogenic microorganisms that can stimulate the body to produce specific antibodies / cellular immunity against pathogenic microorg...

Claims

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Application Information

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IPC IPC(8): A61K9/127A61K39/39A61K48/00A61P31/18A61P35/00
Inventor 张欣阳俊乔晨萌
Owner INST OF PROCESS ENG CHINESE ACAD OF SCI
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