Anti-virus agent

An antiviral and reagent technology, applied in antiviral agents, viruses, viral peptides, etc., can solve problems such as unproven effects and unproven drugs

Inactive Publication Date: 2004-09-08
INTEGRAL TECHNOLOGY INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it has not been proven that these drugs alone are very effective, nor has the combination of the two drugs been shown to produce the desired effect (Samuel et al., Viology 130(1983), 474-484)

Method used

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Use Transfectin transformation method (frequency 2500-1200 clones / 10 5 Cell) The DNA plasmid (pCI-New / AVA) was introduced into HepG2 line cells. Transformed cells were selected in RPMI 1640 medium containing 10% calf serum and geneticin (0.5 mg / ml). Geneticin resistant clone (G418 R ) Used to determine HCV-specific antigen, RNAAVA.

[0053] After 24 to 72 hours of transformation, the G418 R The cells identify HCV-specific antigens and RNA as well as diphtheria toxin. The results showed that after 72 hours or longer (up to 14 days), no diphtheria toxin caused G418 R Morphological characteristics of cell death. In 14 days, in G418 R SRNA was found in the cell culture medium HCV HCV-like particles. Studies have shown that the expression of non-structural protein (HepG2NS line) of human liver cancer cells HepG2 R The medium on which the cells have been grown is highly sensitive and dissolves to a titer as high as 1:1600.

Embodiment 2

[0055] AVA is used as antiviral RNA. Amplification was performed by using thermal polymerase Pwo II and plasmid pCI-new / AVAPCR. T7 phage RNA polymerase is used to complete the transcription of the amplicon. The RNA obtained by electroporation was inserted into HepG2NS cells. After 24 hours of electroporation, diphtheria toxin can be detected in the cells. The initial diphtheria toxin is detected 48 to 52 hours after electroporation, and it can be detected for 120 hours.

[0056] Within 6 hours, HCV-specific antigens and SRNA can be detected in the cells HCV , And diphtheria toxin. ELISA can also detect toxins. Obvious signs of cell destruction were detected 28 hours after electroporation. Within 14 days, SRNA was detected in the perforated cell culture medium of the HepG2NS line HCV HCV is similar to particles. The non-tracted HepG2NS line is highly sensitive to the medium in which the perforated cells have been grown, dissolving to a titer as high as 1:100.

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Abstract

The present invention pertains to an anti-virus agent acting against single stranded RNA viruses. In particular, the present invention relates to an anti-virus agent containing a construct comprising a nucleotide sequence derived from the single stranded virus and capable to direct the synthesis of the (-) strand and a gene encoding a toxin operably linked to a regulon, wherein the gene encoding the toxin is oriented antisense in the construct. The present invention also pertains to the use of said anti-virus agent for preparing a medicament for treating viral diseases.

Description

Technical field [0001] The present invention relates to an antiviral agent (AVA) having anti-single-stranded RNA(+) virus activity. In particular, the antiviral agent of the present invention has a structure containing a nucleotide sequence derived from a single-stranded virus and capable of directing the synthesis of the (-) strand, and a gene encoding a toxin operably linked to a regulator , Wherein the gene encoding the toxin is antisense oriented in the structure. The present invention also relates to the use of said antiviral agent to prepare drugs for treating viral diseases. Background technique [0002] Single-stranded RNA (+) viruses are (+)-stranded viruses that are amplified in host cells but have no DNA stage. This virus uses the enzymes encoded by the virus itself, usually referred to as RNA-dependent RNA polymerase (RdRp), to replicate their RNA. In most cases, the RdRp is a complex of so-called non-structural (NS) proteins, which does not form part of the virion. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/09A61K31/7088A61K35/76A61K38/00A61K38/16A61K47/64A61K48/00A61P31/12A61P31/14C07K14/18
CPCA61K38/164C07K14/005A61K48/00A61K47/48261C12N2770/24222A61K47/6415A61P31/12A61P31/14Y02A50/30
Inventor 马瑞娜·N.·巴布什维林
Owner INTEGRAL TECHNOLOGY INC
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