Immuno liposome therapeutic hepatitis B vaccine and its preparing method

Abstract

The therapeutic hepatitis B vaccine is liposome containing hepatitis B surface antigen with or without immunoregulation molecule. It may be used to treat chronic hepatitis B caused by HIV infection and to prevent HIV infection. The technological process of inverse phase evaporation and freeze drying to prepare the said vaccine is also provided.

Description

technical field

[0001] The invention relates to a hepatitis B vaccine, in particular to an immune liposome therapeutic hepatitis B vaccine, and simultaneously the invention also relates to a preparation method of the vaccine. Background technique

[0002] Hepatitis B is one of the most prevalent infectious diseases in the world, which seriously threatens human health. Chronic hepatitis B patients and HBV virus carriers are the main sources of infection. At present, people mainly take preventive means to hepatitis B virus, that is, vaccination of hepatitis B vaccine for prevention. Preventive vaccines are mainly used on organisms that have never been infected, so viral proteins of natural structure can be directly used as vaccine antigens. But the hepatitis B virus vaccine that has been developed and applied can play a good preventive effect on HBV infection, but it is difficult to treat and eliminate the virus for the body infected with HBV. The main reason is that the bo...

Examples

Embodiment 1

[0063] The specific embodiment embodiment 1 freeze-drying hydration method prepares immune liposome therapeutic hepatitis B vaccine

[0064] Take soybean lecithin, cholesterol, phospholipid A, etc. (the molar ratio is 1:1:1), dissolve them in ethanol solution; remove the organic solvent by rotary evaporation, and form a phospholipid film; add an appropriate amount of phosphate buffer (0.1M, pH7. 4), at 45°C, ultrasonic for 5 minutes to form a small monolayer liposome suspension; mix the liposome suspension with the HBsAg S+pre-S1+pre-S2 solution at a molar ratio of 3:1, then add the frozen Protective agent mannitol, 7°C / min, quickly frozen to -30°C, freeze-dried; cobalt 60 irradiation sterilization; then add sterile water, shake with a shaker for 5 minutes, rehydrate to form a large monolayer or few layers The liposome suspension is the therapeutic hepatitis B vaccine of immune liposome. Embodiment 2 freeze-drying hydration method prepares immune liposome therapeutic hepatiti...

Embodiment 2

[0065] Take soybean lecithin and cholesterol (1:1 molar ratio), dissolve them in ethanol solution; remove the organic solvent by rotary evaporation, and form a phospholipid film; For 5 minutes, a small monolayer liposome suspension was formed; the liposome suspension was mixed with the HBsAg S+pre-S1+pre-S2 solution, the molar ratio was 3:1, then the cryoprotectant mannitol was added, and the temperature was 7°C / minute, quickly frozen to -30 ° C, and freeze-dried; cobalt 60 irradiation sterilization; then add sterile water, shake with a shaker for 5 minutes, and rehydrate to form a large monolayer or few-layer liposome suspension. It is immune liposome therapeutic hepatitis B vaccine. Embodiment 3 reverse phase evaporation method prepares immune liposome therapeutic hepatitis B vaccine

Embodiment 3

[0066] Take soybean lecithin, cholesterol, phospholipid A, etc. (the molar ratio is 1:1:1), dissolve in the mixed solution of chloroform and isopropanol; add the aqueous solution of hepatitis B surface antigen S+pre-S1+pre-S2 into the above-mentioned mixed solution, The molar ratio of the mixed solution to the antigen solution is 2:1, and treated in a bath ultrasonic apparatus at 45°C for 5 minutes to form a stable water-in-oil emulsion, and then evaporate under pressure at 45°C to remove the organic solvent to obtain a thick For the thick jelly, add an appropriate amount of phosphate buffer (0.1M, pH 7.4) to dissolve the jelly, continue rotary evaporation at 45°C, and then evaporate under reduced pressure for 15 minutes to remove a small amount of organic solvent. After standing for 30 minutes, the resulting liposomes were put into a dialysis bag, and with phosphate buffer (0.1M, pH7.4), dialyzed at room temperature for a total time of 24 hours to remove free unwrapped liposom...

technical field

[0001] The invention relates to a hepatitis B vaccine, in particular to an immune liposome therapeutic hepatitis B vaccine, and simultaneously the invention also relates to a preparation method of the vaccine. Background technique

[0002] Hepatitis B is one of the most prevalent infectious diseases in the world, which seriously threatens human health. Chronic hepatitis B patients and HBV virus carriers are the main sources of infection. At present, people mainly take preventive means to hepatitis B virus, that is, vaccination of hepatitis B vaccine for prevention. Preventive vaccines are mainly used on organisms that have never been infected, so viral proteins of natural structure can be directly used as vaccine antigens. But the hepatitis B virus vaccine that has been developed and applied can play a good preventive effect on HBV infection, but it is difficult to treat and eliminate the virus for the body infected with HBV. The main reason is that the bo...

Examples

Embodiment 1

[0063] The specific embodiment embodiment 1 freeze-drying hydration method prepares immune liposome therapeutic hepatitis B vaccine

[0064] Take soybean lecithin, cholesterol, phospholipid A, etc. (the molar ratio is 1:1:1), dissolve them in ethanol solution; remove the organic solvent by rotary evaporation, and form a phospholipid film; add an appropriate amount of phosphate buffer (0.1M, pH7. 4), at 45°C, ultrasonic for 5 minutes to form a small monolayer liposome suspension; mix the liposome suspension with the HBsAg S+pre-S1+pre-S2 solution at a molar ratio of 3:1, then add the frozen Protective agent mannitol, 7°C / min, quickly frozen to -30°C, freeze-dried; cobalt 60 irradiation sterilization; then add sterile water, shake with a shaker for 5 minutes, rehydrate to form a large monolayer or few layers The liposome suspension is the therapeutic hepatitis B vaccine of immune liposome. Embodiment 2 freeze-drying hydration method prepares immune liposome therapeutic hepatiti...

Embodiment 2

[0065] Take soybean lecithin and cholesterol (1:1 molar ratio), dissolve them in ethanol solution; remove the organic solvent by rotary evaporation, and form a phospholipid film; For 5 minutes, a small monolayer liposome suspension was formed; the liposome suspension was mixed with the HBsAg S+pre-S1+pre-S2 solution, the molar ratio was 3:1, then the cryoprotectant mannitol was added, and the temperature was 7°C / minute, quickly frozen to -30 ° C, and freeze-dried; cobalt 60 irradiation sterilization; then add sterile water, shake with a shaker for 5 minutes, and rehydrate to form a large monolayer or few-layer liposome suspension. It is immune liposome therapeutic hepatitis B vaccine. Embodiment 3 reverse phase evaporation method prepares immune liposome therapeutic hepatitis B vaccine

Embodiment 3

[0066] Take soybean lecithin, cholesterol, phospholipid A, etc. (the molar ratio is 1:1:1), dissolve in the mixed solution of chloroform and isopropanol; add the aqueous solution of hepatitis B surface antigen S+pre-S1+pre-S2 into the above-mentioned mixed solution, The molar ratio of the mixed solution to the antigen solution is 2:1, and treated in a bath ultrasonic apparatus at 45°C for 5 minutes to form a stable water-in-oil emulsion, and then evaporate under pressure at 45°C to remove the organic solvent to obtain a thick For the thick jelly, add an appropriate amount of phosphate buffer (0.1M, pH 7.4) to dissolve the jelly, continue rotary evaporation at 45°C, and then evaporate under reduced pressure for 15 minutes to remove a small amount of organic solvent. After standing for 30 minutes, the resulting liposomes were put into a dialysis bag, and with phosphate buffer (0.1M, pH7.4), dialyzed at room temperature for a total time of 24 hours to remove free unwrapped liposom...