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Immunogene therapeutic drug for chronic hepatitis B and preparation method for immunogene therapeutic drug

A chronic hepatitis B, gene therapy technology, applied in gene therapy, drug combination, pharmaceutical formula, etc., can solve the problems of long treatment course, inability to suppress drug-resistant virus, and low sustained response rate, so as to enhance immune response and realize The effect of effective continuous control, increased safety and high efficiency

Inactive Publication Date: 2015-09-30
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

Nucleoside analogs (such as lamivudine, adefovir, entecavir, etc.) can inhibit the replication of hepatitis B virus by inhibiting the activity of DNA reverse transcriptase, although they can effectively reduce the number of viral DNA copies in the serum of patients, and even reach the level of detection. However, it cannot inhibit the production of drug-resistant virus, and nucleoside analogs can rarely achieve the purpose of long-term immune control of HBV infection by effectively eliminating the residual virus in infected liver cells, and it is difficult to achieve the purpose of hepatitis B surface antigen (HBsAg) and HBeAg serum turned negative or completely cleared, therefore, long-term use of nucleoside analogs is prone to drug resistance, and rebound is prone to occur after drug withdrawal
Interferon (interferon, IFN) has direct antiviral and immunomodulatory effects, and can effectively improve the body's immunity, but it can only be administered by injection, and has the disadvantages of low sustained response rate, many side effects, long treatment course, and high price.

Method used

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  • Immunogene therapeutic drug for chronic hepatitis B and preparation method for immunogene therapeutic drug
  • Immunogene therapeutic drug for chronic hepatitis B and preparation method for immunogene therapeutic drug
  • Immunogene therapeutic drug for chronic hepatitis B and preparation method for immunogene therapeutic drug

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Example 1. Construction and expression of antibody targeting interferon (dsFvα) and interleukin-12 fusion expression vector pVAX-HBVE

[0063] 1. Experimental materials and instruments

[0064] 1. Bacterial strains and cell lines

[0065] E.coli.trans5α competent cells were purchased from Quanshijin Company; HEK293T cells were purchased from Beijing Union Medical College Cell Bank.

[0066] 2. Enzymes and reagents

[0067] DNA restriction endonucleases were purchased from New England Biolabs (NEB) biological company; plasmid recovery kit and gel recovery kit were purchased from Beijing Kangwei Century Company; fetal bovine serum for experiment was purchased from Hyclone Company; MEM medium was purchased from Hyclone Company; PCR enzymes and dNTPs were purchased from TaKaRa Company; human IFN-αELISA detection kits and human IL-12ELISA detection kits were purchased from Beijing Xinbosheng Co., Ltd.; kanamycin and ampicillin were purchased from Biotech Biotech Co., Ltd....

Embodiment 2

[0147] Example 2. Construction, expression and preliminary functional verification of replicable vector pSVK-HBVE

[0148] 1. Experimental materials and instruments

[0149] 1. Bacterial strains, cell lines and experimental animals

[0150] E.coli.DH5α competent cells were purchased from Beijing Quanshijin Company; HEK293T cells were purchased from Beijing Union Medical College Cell Bank. Female transgenic mice (6-8 weeks old, with 1.3 copies of HBV whole genome transferred), weighing about 20 g, were purchased from Guangzhou Air Force General Hospital.

[0151] 2. Enzymes and reagents

[0152] DNA restriction endonucleases were purchased from New England Biolabs (NEB) biological company; plasmid recovery kit and gel recovery kit were purchased from Beijing Kangwei Century Company; fetal bovine serum for experiment was purchased from Hyclone Company; MEM medium was purchased from Hyclone Company; enzymes and dNTPs used in PCR were purchased from TaKaRa Company; human IFN-αE...

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Abstract

The invention relates to an immunogene therapeutic drug for chronic hepatitis B and a preparation method for the immunogene therapeutic drug. The active component of the drug is a replication-competent recombinant vector pSVK-dSFValpha-IRES-hIL-12 (for short, pSVK-HBVE), which is constructed by taking a pSVK carrier as a starting carrier and carries a fusion gene dSFValpha-IRES-hIL12. An antibody targeting interferon alpha and human IL-12 are co-expressed in the replication-competent recombinant vector pSVK to construct a humanized HBsAg dsFv antibody, human interferon-alpha and interleukin-12 fusion expression vector pSVK-dSFValpha-IRES-hIL-12, the fusion expression vector is efficiently expressed in an eukaryotic cell and in vitro, the combined application of human IL-12 and human IFN-alpha has an important synergistic effect during the tumor control process, and a safe and efficient immunogene therapeutic drug is provided for gene therapy of chronic hepatitis B.

Description

technical field [0001] The invention belongs to the gene therapy drug in the field of biopharmaceuticals, in particular to an immune gene therapy drug for chronic hepatitis B and a preparation method thereof. Background technique [0002] Chronic hepatitis B is a serious infectious disease caused by hepatitis B virus (HBV). There are currently 2 billion people infected with hepatitis B virus in the world, about 400 million of whom are chronically infected with hepatitis B virus. The persistent infection of HBV will cause some patients to deteriorate into liver cirrhosis or even hepatocellular carcinoma, and die of liver-related diseases caused by HBV every year. The number reached one million people. [0003] At present, antiviral therapy is the most effective clinical treatment for chronic hepatitis B. Commonly used antiviral drugs for hepatitis B include interferon and nucleoside analogs. Nucleoside analogs and interferon inhibit the replication and synthesis of hepatitis...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61P1/16A61P31/20C12N15/85C12N15/66
Inventor 于继云阎瑾琦王宇朱晓明武帅王籽橙杜芝燕
Owner INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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