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Vaccine for treating chronic hepatitis B and preparation method and application thereof

A chronic hepatitis B and vaccine technology, applied in the field of vaccines for the treatment of chronic hepatitis B and their preparation, can solve the problems of inability to induce antibody responses, complicated preparation methods, and clinical data with no therapeutic effect, etc.

Pending Publication Date: 2019-06-14
成都凡益康生物医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The drawback of these techniques is that they generally do not induce good antibody responses
The hepatitis B nucleic acid vaccine of the invention can be used to prepare preparations for the prevention and treatment of hepatitis B vaccine, but the patent uses genetic engineering methods, the preparation method is complicated and there is no clinical data on the relevant therapeutic effect
At the same time, this technology does not induce antibody responses. Many literature reports suggest that therapeutic hepatitis B vaccines need to induce both antibody responses and cellular immune responses

Method used

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  • Vaccine for treating chronic hepatitis B and preparation method and application thereof
  • Vaccine for treating chronic hepatitis B and preparation method and application thereof
  • Vaccine for treating chronic hepatitis B and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 2

[0136] Example 2 Preparation of the vaccine containing TLR4 agonist and QS-21 liposome complex adjuvant Preparation of various ingredients:

[0137] Monophosphoryl Lipid A, Monophosphoryl Lipid A (MPLA) is purchased from the market, and the concentration after being dissolved in 100% alcohol is 1.0 mg per milliliter. QS-21 is purchased from the market, and the concentration of its aqueous solution is 1.0 mg per milliliter.

[0138] Weigh 0.0489g of cholesterol, measure 4.89ml of chloroform in a fume hood, add it to the EP tube containing cholesterol, pipette to dissolve, and mix well.

[0139] Weigh 0.0676g of dioleoyl lecithin 1,2-Dioleoyl-sn-glycero-3-phosphocholine (DOPC), absorb 1.352ml of chloroform in a fume hood, add to DOPC, mix with a pipette, and make a mark 4 ℃ and protected from light.

[0140] Antigen preparation: HBsAg is a virus-like particle expressed by yeast, its purity meets the standards for vaccine production, and it is diluted with normal saline to a con...

Embodiment 3

[0146] Example 3 Immunogenicity of therapeutic hepatitis B vaccine formulated with TLR4 agonist and QS-21 liposome complex adjuvant

[0147] Eight mice per group were treated with C57 (about 6-7 weeks). According to the method of Example 1, 3 days before the vaccine injection, the hepatitis B virus plasmid (paav-HBV1.2) was injected into the tail vein under high pressure to infect C57 mice. On day 1, day 14 and day 28, each mouse was subcutaneously injected with 0.3 ml of the vaccine three times. See Example 2 for the preparation method of the vaccine. Set up a control group without infection, a control group with infection but no immunity, and a control group with only antigen immunization (without adjuvant) after infection.

[0148] On the 10th day after the second immunization (that is, the 24th day of the experiment), blood was collected from the vein, the serum was separated, and the antibody titer of HBsAg was detected by ELISA (results in figure 1 ). 100% of mice imm...

Embodiment 4

[0151] Example 4 The therapeutic effect of the therapeutic hepatitis B vaccine formulated with TLR4 agonist and QS-21 liposome complex adjuvant in the hepatitis B mouse model

[0152] Eight mice per group were treated with C57 (about 6-7 weeks). According to the method of Example 1, three days before the vaccine injection, the hepatitis B virus plasmid (paav-HBV1.2) was injected into the tail vein under high pressure to infect C57 mice. Each mouse was subcutaneously injected with 0.3 ml of the vaccine on day 1, day 14 and day 28, respectively. See Example 2 for the preparation method of the vaccine. A control group without infection, a control group infected but not immunized, and a control group immunized with antigen without adjuvant after infection are set.

[0153] Blood samples were collected before the first immunization and on days 3, 10, 17, 24 and 31 after immunization. The virus components of HBsAg and HBeAg in serum were detected by ELISA method.

[0154] There w...

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Abstract

The invention relates to a vaccine for treating chronic hepatitis B and a preparation method and application thereof. The vaccine is prepared from a hepatitis B virus antigen and a compound adjuvant containing a molecular adjuvant. The vaccine can not only induce strong antibody response, but also induce strong cellular immune response. A carrier of the compound adjuvant used can deliver the antigen to a lymphatic system very effectively. The molecular adjuvant can activate T cell subsets and effectively increase the level and the persistence of an antibody and cellular immune response. The antibody against the hepatitis B virus antigen produced after immunization can eliminate the virus and the antigen in the blood circulation. The vaccine can quickly reduce or even eliminate the antigenof the virus in the blood, eliminate the hepatitis B virus in the liver cells, and prevent the patient from carrying the hepatitis B virus.

Description

technical field [0001] The invention belongs to the technical field of biological and chemical pharmacy, and relates to a vaccine for treating chronic hepatitis B and its preparation method and application. Background technique [0002] Hepatitis B is chronic hepatitis caused by HBV hepatitis B virus (HBV). According to the statistics of the World Health Organization, about 2 billion people in the world have been infected with HBV, of which 350 million people are chronic HBV infected people, and about 1 million people die every year from liver failure, cirrhosis and primary hepatocytes caused by HBV infection. cancer. my country is a high prevalence area of ​​HBV infection, and the positive rate of HBsAg in the general population is 9%. The HBsAg positive rates of the vaccinated and non-vaccinated groups were 4.51% and 9.51%, respectively. There are about 120 million HBV carriers in my country, accounting for one-third of the world's total. Chronic hepatitis B (HBV) infe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/12A61K39/39A61P31/20A61P35/00
Inventor 何勇刚刘晓宇
Owner 成都凡益康生物医药科技有限公司
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