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Replicon DNA (Deoxyribonucleic Acid) vaccine for treating chronic hepatitis B

A chronic hepatitis B and DNA vaccine technology, applied in the direction of medical preparations containing active ingredients, antibody medical ingredients, pharmaceutical formulations, etc., can solve problems such as unsuitable hepatitis B therapeutic DNA vaccines, and achieve development potential and application value. The effect of improving efficiency and enhancing immune activation ability

Inactive Publication Date: 2013-08-14
INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, both the modified pSCA1 and the original vector pSFV1 are ampicillin-resistant vectors, and the antibiotics commonly used in clinical practice are still ampicillin drugs, so ampicillin-resistant vectors will have certain risks in clinical application and are not suitable for use. To obtain a therapeutic DNA vaccine for hepatitis B

Method used

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  • Replicon DNA (Deoxyribonucleic Acid) vaccine for treating chronic hepatitis B
  • Replicon DNA (Deoxyribonucleic Acid) vaccine for treating chronic hepatitis B
  • Replicon DNA (Deoxyribonucleic Acid) vaccine for treating chronic hepatitis B

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1, Construction of recombinant replicon DNA vaccine vector pSVK-HBVA and conventional DNA vaccine vector pVAX-HBVA

[0056] 1. Construction of conventional DNA vaccine plasmid pVAX-HBVA

[0057] Taking pVAX1-IRES-GM / B7 (see literature: Liu Ning, Yan Jinqi, Ran Duoliang, etc. Construction and expression of pVAX1-IRES-GM / B7 immunopotentiation vector for tumor gene vaccines. Journal of the Academy of Military Medical Sciences, 2008, 32( 3):249-52) was used as the starting vector to construct a conventional DNA vaccine vector pVAX-HBVA carrying the fusion gene FL-CS, human IgG Fc gene, GPI gene, GM-CSF gene and B7.1 gene from upstream to downstream sequentially ( see Figure 9 The physical map of the recombinant plasmid pVAX-HBVA), the specific construction method includes the following steps:

[0058] 1. Design and acquisition of HBV complex antigen CS gene

[0059] Search for related sequences of HBV C, S, PreS and other genes on the NCBI website (Genebank numb...

Embodiment 2

[0098] Embodiment 2, the immunological function detection of pSVK-HBVA immune BALB / c mice

[0099] Referring to Example 1, in the process of constructing the new replicon DNA vaccine pSVK-HBVA, a conventional DNA vaccine pVAX1-HBVA based on the common plasmid vector pVAX1 was also constructed. Three dose groups of high (100 μg), medium (10 μg) and low (1 μg) of the two vaccines were set up to immunize healthy Balb / c mice, and the humoral and cellular immune responses of the mice after immunization were detected.

[0100] Immune grouping: 6-8 weeks old Balb / c female mice were randomly divided into 10 groups: ① blank group, ② normal saline group, ③ pVAX1 empty vector group, ④ pSVK empty vector group, ⑤ pVAX-HBVA1 μg, ⑥ pVAX-HBVA 10 μg , ⑦pVAX-HBVA100μg, ⑧pSVK-HBVA1μg, ⑨pSVK-HBVA10μg, ⑩pSVK-HBVA100μg, 10 animals / group.

[0101] Immunization scheme: The leg muscles of the mice were immunized according to the dosage of each group, and the vaccine was delivered by in vivo electropo...

Embodiment 3

[0112] Example 3, Cellular Immunology Detection of pSVK-HBVA Immunized HBV Tg Transgenic Mice

[0113] Example 2 shows that the new replicon DNA vaccine pSVK-HBVA can activate strong humoral and cellular immune responses in healthy Balb / c mice at a low dose of 1 μg. This example further explores the effect of the new replicon DNA vaccine pSVK-HBVA on transgenic The ability of HBV Tg (Transgenic, Tg) transgenic mice with full-length hepatitis B gene to activate immune response in vivo. HBV Tg transgenic mice (purchased from 458 Hospital) are similar to patients with chronic hepatitis B. Transgenic mice carry hepatitis B genes for a long time and are highly immune-tolerant to hepatitis B antigens. The ability to activate cellular immune responses in this model is important for the treatment of chronic hepatitis B important basis. The immunization grouping and immunization scheme formulation of the transgenic mice were the same as the immunization procedures of the normal Balb / c...

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Abstract

The invention discloses a replicon DNA (Deoxyribonucleic Acid) vaccine for treating chronic hepatitis B. The replicon DNA vaccine is a recombinant replicon DNA vaccine vector plasmid carrying a composite hepatitis B antigen gene. The composite hepatitis B antigen gene is named as HBV-CS, the nucleotide sequence of the composite hepatitis B antigen gene is shown as the sequence 1 in a sequence table; a vector of the replicon DNA vaccine is named as pSVK. The recombinant replicon DNA vaccine vector plasmid which sequentially carries a fusion gene FL-CS, a human IgGFc-segment gene, a GPI gene and a GM-CSF gene and a B7.1 gene from upstream to downstream and is constructed by using the pSVK as a starting gene is named as pSVK-HBVA, and the nucleotide sequence of the recombinant replicon DNA vaccine vector plasmid is shown as the sequence 7 in the sequence table.

Description

technical field [0001] The invention belongs to a therapeutic DNA vaccine in the field of biopharmaceuticals, in particular to a replicon DNA vaccine for treating chronic hepatitis B. Background technique [0002] Chronic infection with hepatitis B virus (HBV) is the leading cause of liver disease worldwide. At present, the main drugs for clinical treatment of chronic hepatitis B are antiviral drugs such as α-interferon, lamivudine, and adefovir. Once the drug is stopped, it will immediately face the dilemma of disease recurrence and viral rebound (Margarita Pardo, Javier Bartolome, Vicente Carreno. Current Therapy of Chronic Hepatitis B. Archives of Medical Research, 2007, 38:661-677.). Compared with various existing therapeutic drugs, therapeutic vaccines have unique advantages and effects in reversing hepatitis B immune tolerance, activating immune responses, and clearing the virus, and are considered to be a potential effective strategy for controlling chronic hepatitis...

Claims

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Application Information

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IPC IPC(8): A61K39/29A61P31/20
Inventor 于继云阎瑾琦王宇张巍徐元基贾锐张亮肖毅朱晓明
Owner INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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