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Rabies virus inhibition polypeptide as well as preparation method and application thereof

A technology of rabies virus and inhibitory polypeptide, which is applied in the fields of biochemistry and molecular biology, can solve the problems of lack of safe and efficient intracellular transport system, low toxicity, and no solution, so as to inhibit virus replication, strong pertinence, and inhibition Obvious effect

Inactive Publication Date: 2017-06-20
QINGDAO UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The use of siRNA technology to inhibit the replication of intracellular rabies virus is a promising new method. By destroying or silencing the expression of key viral genes, the purpose of controlling virus replication is achieved. This method is simple to operate, high in efficiency, and low in toxicity. There is a lack of safe and efficient intracellular transport system
However, its activity needs to be further improved, and as a polypeptide drug, the method of intracellular administration is still unresolved.

Method used

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  • Rabies virus inhibition polypeptide as well as preparation method and application thereof
  • Rabies virus inhibition polypeptide as well as preparation method and application thereof
  • Rabies virus inhibition polypeptide as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 optimizes the design of the rabies virus inhibitory polypeptide sequence by QSAR

[0029] (1) From published literature (Real E, Rain JC, Battaglia V, Jallet C, Perrin P, Tordo N, Chrisment P, D'Alayer J, Legrain P, Jacob Y. Antiviral drug discovery strategy using combinatorial libraries of structurally constrained peptides .JVirol.2004Jul; 78(14):7410-7.) has collected the peptide data that has been confirmed to inhibit the replication of rabies virus for modeling, a total of 29 polypeptides, and the sequences are shown in Table 1:

[0030] Table 1 Sequence information used in QSAR modeling

[0031]

[0032]

[0033] (2) The descriptor variables ( http: / / bidd2.nus.edu.sg / cgi-bin / profeat2016 / main.cgi ), four descriptor combinations of G3, G4, G5, and G9 were selected for calculation; the genetic algorithm (Genetic algorithm, GA) was used to screen the physical and chemical parameters directly related to the biological activity of the polypeptide, and...

Embodiment 2

[0040] Example 2 Activity Verification of Starting Peptides and Optimizing Peptides

[0041]The starting peptide and optimized peptide in Example 1 were synthesized by solid-phase synthesis. In order to solve the problem of poor cell permeability of polypeptide drugs, a membrane-penetrating peptide sequence RRRRRRRRR was connected to the N-terminals of the starting peptide and the optimized peptide respectively. The mass spectra of the synthesized starting peptide with membrane-penetrating peptide and the optimized fetus with membrane-penetrating peptide are shown in figure 1 , 2 , the sequences are:

[0042] SEQ ID No. 50: RRRRRRRR PPDVHTPPHALWRLHLSLRVCLVRMWIH

[0043] SEQ ID No. 51: RRRRRRRRRPPDVHTPPHALWRLHLILRVELVRMWWH.

[0044] Prepare 8 groups of BSR cells in parallel, inoculate BSR cells with 0.01 MOI of CVS, and add different concentrations of starting peptide and optimized peptide drug 1 h later (final concentrations are 0 μg / ml, 12.5 μg / ml, 25 μg / ml, 50 μg / ml, res...

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Abstract

The invention provides rabies virus inhibition polypeptide. The sequence of the rabies virus inhibition polypeptide is PPDVHTPPHALWRLHLILRVELVRMWWH, and belongs to the field of biochemistry and molecular biology. After being optimized by using QSAR (Quantitative Structure Activity Relationship), the polypeptide and cell-penetrating peptide form fusion peptide, and thus a function of inhibiting virus replication in cells can be achieved. The method is high in specificity, easy to operate and applicable to control on post-cell rabies viruses.

Description

technical field [0001] The invention belongs to the fields of biochemistry and molecular biology, and in particular relates to a rabies virus inhibitory polypeptide and its preparation method and application. Background technique [0002] Timely and effective treatment after exposure to rabies is the key to prevention and treatment, especially the combined use of inactivated rabies virus vaccine and rabies virus antibody is the only option recommended by WHO for post-exposure prophylaxis of human rabies. That is, the use of immune means to eliminate the virus before it enters the cell. Once the treatment is not timely, the virus will invade the cells, and the best time for the prevention and treatment of rabies will be lost, resulting in serious consequences. Because there is no effective method for the control of intracellular rabies virus at present. [0003] The use of siRNA technology to inhibit the replication of intracellular rabies virus is a promising new method. B...

Claims

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Application Information

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IPC IPC(8): C07K14/00C07K19/00A61K38/16A61K47/64A61P31/14G06F19/16
CPCG16B15/00C07K14/001C07K2319/10A61K38/00
Inventor 卢永忠
Owner QINGDAO UNIV OF SCI & TECH
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