Medicament for treating and/or preventing viral infection

A technology of drugs and viruses, applied in the field of new protein drugs and adjuvants for the treatment and/or prevention of viral infections, and drugs for the treatment and/or prevention of viral infections, which can solve the problems of weak immunogenicity and other issues

Inactive Publication Date: 2011-10-19
BEIJING ADVACCINE BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As an emerging vaccine, DNA vaccine has many advantages such as being irreplaceable by traditional vaccines. DNA vaccines are also regarded as one of the key technologies to overcome AIDS at home and abroad. However, the biggest problem restricting the development of DNA vaccines is the weak immunogenicity. It is recognized that one of the important ways to evaluate and prevent HIV virus infection is how to effectively mobilize cellular immune responses, especially to stimulate high levels of HIV-specific CD8 positive CTL cell activity and secrete high levels of γ-interferon and other effective immune responses. components

Method used

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  • Medicament for treating and/or preventing viral infection
  • Medicament for treating and/or preventing viral infection
  • Medicament for treating and/or preventing viral infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1. Cloning and expression of interleukin 17A

[0047] Since the prokaryotic expression system cannot process and cut the interleukin 17A signal peptide, when designing primers, clone and express it from the downstream of the signal peptide, so the following experiments all use IL-17A without signal peptide sequence.

[0048] The mouse IL-17A expression plasmid is a reverse transcription of RNA extracted from the mouse spleen, and the IL-17 gene is cloned from cDNA. The PCR product was recovered after electrophoresis, and ligated with pMD18-T vector to construct pMD18-T-IL-17, which was then digested with restriction enzymes BamH I and Xba I. The pMD18-T-IL-17 with the correct restriction analysis was further sequenced, and the sequencing result was aligned with the IL-17A sequence number NM_010552 on GeneBank, which was completely consistent. The human IL-17A cDNA clone was obtained by artificial synthesis using the sequence number NM_002190 on GeneBank. The artific...

Embodiment 2

[0052] Example 2. Detection of interleukin 17A biological activity

[0053] In order to prove that the expressed IL-17A is biologically active, we use IL-17 as a pro-inflammatory factor to stimulate innate immune cells to produce IL-6 and TNF-α, so we put IL-17 at different concentrations ( 50ng / ml and 100ng / ml) treat RAW264.7 cells, 48 ​​hours after stimulation, detect the secretion level of IL-6, such as image 3 As shown, the expressed IL-17A can stimulate RAW264.7 cells to secrete I1-6, and this stimulation is dose-dependent.

Embodiment 3

[0054] Example 3. Experiments of Interleukin 17A Protecting Animals Against Influenza Virus Attack

[0055] Interleukin 17A was cloned in the pET28a plasmid in the laboratory, and purified by E. coli expression. 100ug of interleukin 17A was dissolved in 10ml of 10mM pH 7.0 PBS buffer to obtain a 10ug / ml solution. Female Balb / c mice aged 6-8 weeks were divided into 3 groups, each with 15 mice, and the other group was interleukin 17A knockout Balb / c mice (IL-17KO). Each group of mice were injected with 0ul, 10ul or 50ul of the above-mentioned interleukin 17A, repeated at intervals of 6 days, a total of 2-4 times. Seven days later, each mouse was inoculated with H5N1 influenza virus with LD50=5. On the 6th day, the lungs of 3 mice from each group were extracted for RNA extraction and real-time fluorescent quantitative RT-PCR to analyze the viral load ( Figure 4 ), observe the death of the remaining animals every day ( Figure 5 ). The result is figure 1 As shown, the virus RNA co...

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Abstract

The invention provides a medicament for treating and / or preventing virus. The medicament for treating and / or preventing the virus comprises interleukin 17A and has the medicament effect generated by taking the interleukin 17A as an adjuvant together with a vaccine for enhancing the immunization effect of the vaccine, and the sequences of amino acid residues are as shown in SEQ (sequence) ID (identity) No. 2 and SEQ ID No. 3. The action mechanism of the medicament for treating and / or preventing the virus is to enable the medicament to excrete high-level gamma-interferon, Preforin and other effective immunization components by activating and improving cell immune reaction of an organism, especially by inducing high-level CD8 positive CTL (cytotoxic T lymphocyte) cell activity so as to achieve the purpose of treating and / or preventing the virus. Compared with the prior art, the medicament for treating and / or preventing the virus is not only convenient to use, low in cost and safe, but also easy to popularize.

Description

Technical field [0001] The present invention relates to a medicine for treating and / or preventing viral infection. Specifically, it relates to a novel protein drug and adjuvant for the treatment and / or prevention of viral infections. Therefore, the present invention belongs to the field of biological products. Background technique [0002] Influenza (abbreviated as influenza) is an acute respiratory infection caused by influenza virus. Influenza has brought huge disasters to human health. It has caused 4 influenza pandemics in human history, and the death toll reached tens of millions. In recent years, small-scale influenza epidemics have been introduced in some countries and regions each year, and 9% of the world's population is infected with influenza viruses each year. [0003] At present, there is no particularly effective method to prevent influenza. Influenza vaccination is considered to be the best method to prevent the occurrence and spread of influenza. Inactivated influ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/39A61K39/00A61K48/00A61P31/12
Inventor 王宾靳津俞庆龄
Owner BEIJING ADVACCINE BIOTECH
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