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SiRNA (small interfering Ribose Nucleic Acid) capable of preventing and controlling influenza, and pharmaceutical composition and medical application thereof

A composition and influenza technology, applied in the field of medicine, can solve problems such as easy generation of drug resistance, and achieve the effect of reducing adsorption and infection

Inactive Publication Date: 2014-03-26
GUANGZHOU INST OF RESPIRATORY DISEASE +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the above two types of compound drugs have various side effects and limitations, such as easy to produce drug resistance, which has also become an important issue in clinical treatment and new drug development.

Method used

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  • SiRNA (small interfering Ribose Nucleic Acid) capable of preventing and controlling influenza, and pharmaceutical composition and medical application thereof
  • SiRNA (small interfering Ribose Nucleic Acid) capable of preventing and controlling influenza, and pharmaceutical composition and medical application thereof
  • SiRNA (small interfering Ribose Nucleic Acid) capable of preventing and controlling influenza, and pharmaceutical composition and medical application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1: Screening of siRNA targeting ST6GAL1 and ST3GAL4 genes

[0050] Find ST6GAL1 and ST3GAL4 gene sequence from GenBank database, and determine suitable target sequence by bioinformatics technology; Design siRNA for the target sequence finding out, these siRNA sequences are as follows:

[0051] Sequence name: siST6GAL1_001

[0052] siRNA sense strand: 5'CAGCCAACUUCCAACAAGAdTdT 3'(SEQ ID NO.1)

[0053] siRNA antisense strand: 3' dTdT GUCGGUUGAAGGUUGUUCU 5' (SEQ ID NO.2)

[0054] Sequence name: siST6GAL1_002

[0055] siRNA sense strand: 5' GUACCAGAAUCCGGAUUAU dTdT 3' (SEQ ID NO.3)

[0056] siRNA antisense strand: 3' dTdT CAUGGUCUUAGGCCUAAUA 5' (SEQ ID NO.4)

[0057] Sequence name: siST3GAL4_001

[0058] siRNA sense strand: 5' GGUCAUCAGAUUGAACAAU dTdT 3' (SEQ ID NO.5)

[0059] siRNA antisense strand: 3' dTdT CCAGUAGUCUAACUUGUUA 5' (SEQ ID NO.6)

[0060] Sequence name: siST3GAL4_002

[0061] siRNA sense strand: 5' GCAGACCAUUCACUACUAU dTdT 3' (SEQ ID NO.7)

...

Embodiment 2

[0063] Embodiment 2. The inhibition efficiency of the siRNA to ST6GAL1 and ST3GAL4 gene expression

[0064] These synthesized and modified siRNA molecules were transfected into representative airway epithelial cell lines A549, HBE and HEp-2 cells by liposome transfection method. and liposomes, and then mix the two together, let stand at room temperature for 20 minutes, and add the liposomes to the cells to complete the transfection after the siRNA is encapsulated. After 48 hours, real-time quantitative RT-PCR and western blot were used to detect the expression of ST6GAL1 and ST3GAL4 genes at the protein level to evaluate the silencing effects of different siRNAs ( Figure 1A and Figure 1B , Figure 2A and Figure 2B ).

[0065] The results showed that the relative expression levels of ST6GAL1 and ST3GAL4 in respiratory epithelial cells transfected with targeting siRNA were significantly reduced, and the inhibition efficiency of the siRNA on the target gene was above 80%. ...

Embodiment 3

[0075] Example 3. The inhibitory effect of the siRNA on the expression of sialic acid receptor molecules on the cell surface

[0076]The method of introducing the siRNA into respiratory epithelial cells was the same as in Example 2. After 56 hours, the inhibitory effect of siRNA on cell surface sialic acid receptor molecules was detected by immunofluorescence and flow cytometry. like Figure 3A , Figure 3B , Figure 3C , Figure 3D , Figure 3E , Figure 3F ,and Figure 4A , Figure 4B As shown, the results show that siRNA has a significant inhibitory effect on the expression of sialic acid receptor molecules on the cell surface, and the inhibition efficiency is above 80%, as follows:

[0077] Figure 3A It is the effect of immunofluorescence detection of the inhibitory effects of non-transfected targeting siRNA and transfected targeting ST6GAL1--siRNA on the sialic acid receptor molecule SAa2, 6GAL on the surface of HBE cells respectively. Figure 3A The left column...

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Abstract

The invention discloses a siRNA (small interfering Ribose Nucleic Acid) of targeting genes ST6GAL1 and ST3GAL4 capable of preventing and controlling influenza. The siRNA comprises at least one of siST6GAL1_001, siST6GAL1_002, siST3GAL4_001, and siST3GAL4_002. The invention also provides a pharmaceutical composition of which the siRNA serves as an effective ingredient. The invention also provides an application of the siRNA in preparation of the pharmaceutical composition, a medicine and a prevention preparation for preventing and controlling influenza virus. By adopting the pharmaceutical composition, medicine and prevention preparation, the expression of a human influenza A virus sialic acid receptor and an avian influenza A virus sialic acid receptor on the surface of epithelial cell of human respiratory tract can be inhibited, thus the adsorption and infection of human and avian influenza A virus can be reduced effectively; in addition, the effective rate of the siRNA reaches more than 80% in the respiratory tract epithelial cell line.

Description

technical field [0001] The invention relates to the technical field of medicine, in particular to a siRNA capable of preventing and treating influenza, a pharmaceutical composition and a medical application thereof. Background technique [0002] Influenza virus is called for short influenza virus, is a kind of RNA virus that causes human and animal to suffer from influenza; Influenza virus can be divided into four genera according to nucleoprotein (NP) and matrix protein (M) wherein: ), Beta (B), C (C) and Togotovirus-like genera. Influenza A widely exists in humans and other animals, and it has a wide range of infections and appears in an epidemic form with the most serious harm; Influenza B exists only in humans and often causes local outbreaks of influenza; Influenza C exists in humans and pigs , rarely popular. [0003] Influenza viruses enter the host body mainly by infecting human respiratory epithelial cells, causing various systemic symptoms and even serious compli...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113A61K48/00A61P31/16
Inventor 钟南山吴东杨子峰周荣关文达王玉涛李海涛秦笙李润峰招惠珊伍时冠
Owner GUANGZHOU INST OF RESPIRATORY DISEASE
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