Viral hepatitis treatment drug interferon production technology

A viral hepatitis and therapeutic drug technology, applied in the field of biomedicine, can solve the problems of no specific drug for viral hepatitis, short half-life of interferon, insufficient targeting, etc., to reduce immunogenicity, reduce drug excretion, improve target tropism effect

Inactive Publication Date: 2015-04-22
GUANGXI UNIV
View PDF4 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] The onset of viral hepatitis infection is mostly insidious, persistent infection can lead to liver cirrhosis and primary liver cancer, and the mortality rate is very high. For patients who eventually develop liver cirrhosis, the only effective treatment is liver transplantation, but in liver transplantation In the case that surgery cannot be popularized, early treatment of chronic viral hepatitis is particularly important. At present, there is no specific drug for the treatment of viral hepatitis, and interferon, nucleoside antiviral drugs and various Chinese patent medicines are mainly used, namely Immunoenhancing drugs, but the half-life of interferon on the current market is short, and the targeting is insufficient. What is more serious is that the natural interferon drug has antigenic activity. The application number is CN200510069269.6, which discloses a hepatitis B genetic engineering targeting interferon And its production process, hepatitis B genetic engineering targeting interferon adopts the construction of human anti-HBsAg dsFv gene, which is connected with α-interferon (IFN-a) gene, and uses genetic engineering technology to express hepatitis B targeting interferon (dsFv- IFV fusion protein), the production process of hepatitis B genetic engineering targeting interferon, inoculate the prepared culture medium with the strain and then send it to the incubator for cultivation. After the completion of the cultivation, the bacteria multiply in a large number in the breeding room, and then the bacteria are collected by separation , and cleaved to separate the inclusion body protein, renatured in the folding solution, then purified, concentrated, and finally subpackaged and freeze-dried. After purification by affinity chromatography, it was proved that the fusion protein has interferon activity and reactivity with the surface antigen of hepatitis B virus , the fusion protein is of human origin, avoiding the harm caused by heterologousness, but the interferon is still antigenic, and the targeting is not enough

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] A kind of production technology of viral hepatitis therapeutic drug interferon of the present embodiment, comprises the following steps:

[0027] (1) Acquisition of the target gene: Obtain effector T cells from patients with viral hepatitis, and use restriction endonucleases to partially enzymatically digest the DNA under the conditions of temperature 37°C and pH 7.0 to obtain genes that control interferon synthesis. Target gene, spare;

[0028] (2) PCR amplification: In a microcentrifuge tube, use the 1 μg / mL target gene obtained in step (1) as a template DNA, add an appropriate amount of buffer, and a mixture of 4 kinds of dNTPs (i.e. dATP, dCTP, dGTP, dTTP) , 0.5U / 50μL DNA polymerase system, a pair of 0.1mol / L primers for synthesizing DNA, the pH of the system is 6.8, PCR amplification includes the following basic reaction steps:

[0029] ① Pre-denaturation: temperature 92°C, react for 5 minutes to dissociate the double-stranded DNA template into single strands;

...

Embodiment 2

[0042] The rest are the same as in Example 1, except that in the step (2), the DNA polymerase system is a 2.5U / 50 μL DNA polymerase system, 0.5 μmol / L synthetic DNA primers, and the system pH is 7.8,.

Embodiment 3

[0044] The rest are the same as in Example 1, except that in the step (2), the DNA polymerase system is a 2U / 50 μL DNA polymerase system, 0.3 μmol / L synthetic DNA primers, and the system pH is 7. In the step (2), the buffer is 30mmol / L phosphate buffer, 50mmol / L KCl, 5mmol / L dithiothreitol, 100μg / mL bovine serum albumin, 1.5mmol / L Mg 2+ mixed solution.

[0045] In the above three examples, the detected DNA polymerase activity was between 0.8×10 8 Above IU / mg, the effect of PCR amplification is obvious. After the chemical modification of polyethylene glycol amino group, the content and modification rate of single-chain polyethylene glycol-interferon reach 0.25mg / ml and 36.63%, respectively, which has a typical polyethylene glycol-interferon The characteristics of the alcohol-modified protein, the in vitro antigenic activity retains 15% of the natural antigenic activity, and the interferon after lyophilization has a biological activity of 1.0×10 after 6 months 8 IU / mg or mor...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses a viral hepatitis treatment drug interferon production technology which comprises the following steps: (1) acquisition of a target gene; (2) amplifying by PCR (polymerase chain reaction), to be more specific, in a microcentrifuge tube, using the 1 g / mL target gene obtained by the step (1) as template DNA, adding a right amount of a buffer solution, 4 kinds of dNTP (diethyl-nitrophenyl thiophosphate) mixtures, 0.5 2.5U / 50 muL DNA polymerase system, a pair of 0.1-0.5 mumol / L synthesized DNA primer to make the system pH 6.8-7.8; (3) gene recombination; (4) transformation; (5) separation and purification; (6) gene expression; (7) modification; and (8) freeze-drying, mass viral hepatitis treatment drug interferon can be produced in a short period of time, after modifying with polyethylene glycol, the half-life of the interferon is prolonged, hepatitis virus targeting property is improved, the antigenicity is weak, no human body immune response is caused, drug efficacy is good, production is fixed, stability is good, and the viral hepatitis treatment drug interferon production technology is suitable for popularization and application.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a production process of interferon, a medicine for treating viral hepatitis. Background technique [0002] The onset of viral hepatitis infection is mostly insidious, persistent infection can lead to liver cirrhosis and primary liver cancer, and the mortality rate is very high. For patients who eventually develop liver cirrhosis, the only effective treatment is liver transplantation, but in liver transplantation In the case that surgery cannot be popularized, early treatment of chronic viral hepatitis is particularly important. At present, there is no specific drug for the treatment of viral hepatitis, and interferon, nucleoside antiviral drugs and various Chinese patent medicines are mainly used, namely Immunoenhancing drugs, but the half-life of interferon on the current market is short, and the targeting is insufficient. What is more serious is that the natural interferon drug has a...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/555C07K1/107C12N15/70A61K38/21A61K47/48A61P1/16A61P31/14A61P31/20
CPCC07K14/555
Inventor 肖大年张政余炼
Owner GUANGXI UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap